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Abd Rahman F, Azwa FN. Comparative Dental Pulp Stem Cells (DPSCs) and Periodontal Ligament Stem Cells (PDLSCs): Difference in effect of aspirin on osteoblast potential of PDLSCs and DPSCs. Tissue Cell 2025; 94:102776. [PMID: 40022908 DOI: 10.1016/j.tice.2025.102776] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/14/2024] [Revised: 01/27/2025] [Accepted: 02/01/2025] [Indexed: 03/04/2025]
Abstract
Periodontal Ligament Stem Cells (PDLSCs) and Dental Pulp Stem Cells (DPSCs) are mesenchymal stem cells with the ability to self-renew and differentiate into three lineages. One significant advantage of dental stem cells, such as PDLSCs and DPSCs, is their ease of harvest compared to other types of mesenchymal stem cells (MSCs). While MSCs are highly valued in bone tissue engineering, MSCs sourced from dental tissues, such as PDLSCs and DPSCs, offer promising options for periodontal regeneration because they are more easily accessible and can be collected through minimally invasive methods. Currently, PDLSCs and DPSCs exhibit a strong ability to undergo osteogenic differentiation when stimulated by factors such as growth factors, chemicals, and paracrine signaling. It has been shown that aspirin (ASA) can enhance the osteoblastic potential of PDLSCs and DPSCs, although the exact mechanism remains unclear. This article examines the origin and features of mesenchymal stem cells, the bone regeneration potential of DPSCs and PDLSCs, the factors that enhance their osteogenic differentiation, and a comparison of PDLSCs and DPSCs regarding their proliferation and differentiation abilities. Additionally, we will examine the effects of aspirin on PDLSCs and DPSCs. In conclusion, PDLSCs show a greater effect on osteoblast differentiation.
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Affiliation(s)
- Fazliny Abd Rahman
- School of Dentistry (SoD), Management & Science University (MSU), University Drive, Off Persiaran Olahraga, 40100 Shah Alam, Selangor.
| | - Fatin Nur Azwa
- Faculty of Dentistry, Oral Cancer Research Centre (ORCC), University of Malaya (UM), Wilayah Persekutuan, Kuala Lumpur 50603, Malaysia
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Mutlu Özçınar B, Özükoç C, Türkmen E, Çakır R. Dental pulp stem cells (DPSCs), stem cells from human exfoliated deciduous teeth (SHEDSCs), and periodontal ligament stem cells (PDLSCs) isolation, characterization and the effectiveness of allantoin as bioactive molecule for dental regeneration. J Dent 2025; 154:105604. [PMID: 39904472 DOI: 10.1016/j.jdent.2025.105604] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/28/2024] [Revised: 01/20/2025] [Accepted: 01/30/2025] [Indexed: 02/06/2025] Open
Abstract
INTRODUCTION Dental stem cells are valuable tools in regenerative medicine due to their pluripotency and self-renewal properties. This study aimed to investigate the effects of allantoin (Al) on Dental pulp stem cells (DPSCs), stem cells from human exfoliated deciduous teeth (SHEDSCs), and periodontal ligament stem cells (PDLSCs) regarding cytotoxicity, proliferation, wound healing, and osteogenic differentiation. METHODS Human dental stem cells were isolated from three dental tissues using the explant culture method and cultured in DMEM-F12 medium supplemented with 15 % fetal bovine serum (FBS) and antibiotics. The cytotoxicity and proliferation of allantoin were assessed using the XTT cell viability assay at concentrations ranging from 0.25 to 5 mg/mL. Wound healing was evaluated through a scratch assay at 1 mg/mL, and osteogenic differentiation was assessed using Alizarin Red S staining at 0.5 mg/mL and 1 mg/mL. RESULTS Al exhibited no cytotoxic effects across the tested concentrations. It enhanced cell proliferation, particularly in SHEDSCs at 5 mg/mL. DPSCs also showed significant improvement in wound healing in the scratch assay. At 1 mg/mL, Al inhibited osteogenic differentiation in DPSCs and PDLSCs, as indicated by reduced mineralization. CONCLUSION Al shows potential as a non-cytotoxic agent for enhancing the proliferation of dental stem cells, especially SHEDSCs. However, its limited effect on wound healing of SHEDSCs and PDLSCs and inhibition of osteogenic differentiation at higher concentrations suggest that further optimization is required for its application in bone regeneration. STATEMENT OF CLINICAL RELEVANCE Evaluation of the effects of plant-based therapeutic compounds on various types of dental stem cells may have the potential to increase the success of stem cell-based therapies in clinical applications in regenerative dentistry.
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Affiliation(s)
- Betül Mutlu Özçınar
- Department of Bioengineering, Faculty of Chemical and Metallurgical Engineering, Yıldız Technical University, İstanbul, Turkey.
| | - Can Özükoç
- Department of Pediatric Dentistry, Faculty of Dentistry, Istanbul Medipol University, Istanbul, Turkey
| | - Emrah Türkmen
- Department of Periodontology, Faculty of Dentistry, Istanbul Medipol University, Istanbul, Turkey
| | - Rabia Çakır
- Department of Bioengineering, Faculty of Chemical and Metallurgical Engineering, Yıldız Technical University, İstanbul, Turkey
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Lu H, Shi F, Wang B, Zheng Y, Lu J, Zeng B, Zhao W. Molecular Biological Comparison of Pulp Stem Cells from Supernumerary Teeth, Permanent Teeth, and Deciduous Teeth for Endodontic Regeneration. Int J Mol Sci 2025; 26:1933. [PMID: 40076559 PMCID: PMC11901064 DOI: 10.3390/ijms26051933] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/17/2025] [Revised: 02/17/2025] [Accepted: 02/22/2025] [Indexed: 03/14/2025] Open
Abstract
Supernumerary tooth-derived pulp stem cells (SNTSCs) hold promise for endodontic regeneration, yet little is known about the similarities and diversities of SNTSCs relative to other dental-derived mesenchymal stem cells. Herein, we compare the biological characteristics of SNTSCs with dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED). Cell proliferation, migration, and odontogenic differentiation potential, as well as viability and aging-related phenotype after long-term storage, were evaluated. Additionally, gene expressions during induced odontogenic differentiation were profiled by transcriptome sequencing. Our findings indicated that the SNTSCs outperformed the DPSCs but were inferior to the SHED in cell proliferation. The SNTSCs exhibited comparable migratory capacity to the SHED and surpassed the DPSCs. Of particular interest, the odontogenic differentiation potential followed the pattern of SHED > SNTSCs > DPSCs. After two years of storage, the SNTSCs showed weakness in resistance to apoptosis induced by lipopolysaccharide, whereas difference between the SNTSCs and SHED in stemness and senescence was not obvious. Transcriptome analysis revealed that upregulated genes in the SNTSCs were particularly enriched in inflammatory signaling pathways compared to both the DPSCs and SHED. Collectively, SNTSCs share many satisfactory features in proliferation and differentiation with SHED, which may serve as a promising alternative cell source for endodontic regeneration.
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Affiliation(s)
- Hui Lu
- Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055, China; (H.L.); (F.S.)
- Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
| | - Fangyang Shi
- Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055, China; (H.L.); (F.S.)
- Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
| | - Boqun Wang
- Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055, China; (H.L.); (F.S.)
- Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
| | - Yexin Zheng
- Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055, China; (H.L.); (F.S.)
- Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
| | - Jiaxuan Lu
- Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055, China; (H.L.); (F.S.)
- Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
| | - Binghui Zeng
- Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055, China; (H.L.); (F.S.)
- Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
| | - Wei Zhao
- Hospital of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou 510055, China; (H.L.); (F.S.)
- Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China
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Wang Z, Ren L, Li Z, Qiu Q, Wang H, Huang X, Ma D. Impact of Different Cell Types on the Osteogenic Differentiation Process of Mesenchymal Stem Cells. Stem Cells Int 2025; 2025:5551222. [PMID: 39980864 PMCID: PMC11842143 DOI: 10.1155/sci/5551222] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/27/2024] [Revised: 10/15/2024] [Accepted: 01/17/2025] [Indexed: 02/22/2025] Open
Abstract
The skeleton is an important organ in the human body. Bone defects caused by trauma, inflammation, tumors, and other reasons can impact the quality of life of patients. Although the skeleton has a certain ability to repair itself, the current most effective method is still autologous bone transplantation due to factors such as blood supply and defect size. Modern medicine is attempting to overcome these limitations through cell therapy, with mesenchymal stem cells (MSCs) playing a crucial role. MSCs can be extracted from different tissues, and their differentiation potential varies depending on the source. Various cells and cell secretions can influence this process. This article, based on previous research, reviews the effects of macrophages, endothelial cells (ECs), nerve cells, periodontal cells, and even some bacteria on MSC osteogenic differentiation, aiming to provide a reference for multicell coculture strategies related to osteogenesis.
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Affiliation(s)
- Zixin Wang
- School of Stomatology, Lanzhou University, Lanzhou, China
| | - Lina Ren
- School of Stomatology, Lanzhou University, Lanzhou, China
| | - Zhengtao Li
- School of Stomatology, Lanzhou University, Lanzhou, China
| | - Qingyuan Qiu
- School of Stomatology, Lanzhou University, Lanzhou, China
| | - Haonan Wang
- School of Stomatology, Lanzhou University, Lanzhou, China
| | - Xin Huang
- School of Stomatology, Lanzhou University, Lanzhou, China
| | - Dongyang Ma
- School of Stomatology, Lanzhou University, Lanzhou, China
- Department of Oral and Maxillofacial Surgery, The 940th Hospital of Joint Logistics Support Force of PLA, Lanzhou, China
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Gallo MC, Elias A, Reynolds J, Ball JR, Lieberman JR. Regional Gene Therapy for Bone Tissue Engineering: A Current Concepts Review. Bioengineering (Basel) 2025; 12:120. [PMID: 40001640 PMCID: PMC11852166 DOI: 10.3390/bioengineering12020120] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/24/2024] [Revised: 01/20/2025] [Accepted: 01/24/2025] [Indexed: 02/27/2025] Open
Abstract
The management of segmental bone defects presents a complex reconstruction challenge for orthopedic surgeons. Current treatment options are limited by efficacy across the spectrum of injury, morbidity, and cost. Regional gene therapy is a promising tissue engineering strategy for bone repair, as it allows for local implantation of nucleic acids or genetically modified cells to direct specific protein expression. In cell-based gene therapy approaches, a variety of different cell types have been described including mesenchymal stem cells (MSCs) derived from multiple sources-bone marrow, adipose, skeletal muscle, and umbilical cord tissue, among others. MSCs, in particular, have been well studied, as they serve as a source of osteoprogenitor cells in addition to providing a vehicle for transgene delivery. Furthermore, MSCs possess immunomodulatory properties, which may support the development of an allogeneic "off-the-shelf" gene therapy product. Identifying an optimal cell type is paramount to the successful clinical translation of cell-based gene therapy approaches. Here, we review current strategies for the management of segmental bone loss in orthopedic surgery, including bone grafting, bone graft substitutes, and operative techniques. We also highlight regional gene therapy as a tissue engineering strategy for bone repair, with a focus on cell types and cell sources suitable for this application.
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Affiliation(s)
- Matthew C. Gallo
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
| | - Aura Elias
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
| | - Julius Reynolds
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
| | - Jacob R. Ball
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
| | - Jay R. Lieberman
- Department of Orthopaedic Surgery, Keck School of Medicine of the University of Southern California, Los Angeles, CA 90033, USA; (M.C.G.); (A.E.); (J.R.); (J.R.B.)
- Alfred E. Mann Department of Biomedical Engineering, Viterbi School of Engineering, University of Southern California, Los Angeles, CA 90089, USA
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Kadkhoda Z, Motie P, Rad MR, Mohaghegh S, Kouhestani F, Motamedian SR. Comparison of Periodontal Ligament Stem Cells with Mesenchymal Stem Cells from Other Sources: A Scoping Systematic Review of In vitro and In vivo Studies. Curr Stem Cell Res Ther 2024; 19:497-522. [PMID: 36397622 DOI: 10.2174/1574888x17666220429123319] [Citation(s) in RCA: 2] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/08/2021] [Revised: 12/31/2021] [Accepted: 03/11/2022] [Indexed: 11/22/2022]
Abstract
OBJECTIVE The application of stem cells in regenerative medicine depends on their biological properties. This scoping review aimed to compare the features of periodontal ligament stem cells (PDLSSCs) with stem cells derived from other sources. DESIGN An electronic search in PubMed/Medline, Embase, Scopus, Google Scholar and Science Direct was conducted to identify in vitro and in vivo studies limited to English language. RESULTS Overall, 65 articles were included. Most comparisons were made between bone marrow stem cells (BMSCs) and PDLSCs. BMSCs were found to have lower proliferation and higher osteogenesis potential in vitro and in vivo than PDLSCs; on the contrary, dental follicle stem cells and umbilical cord mesenchymal stem cells (UCMSCs) had a higher proliferative ability and lower osteogenesis than PDLSCs. Moreover, UCMSCs exhibited a higher apoptotic rate, hTERT expression, and relative telomerase length. The immunomodulatory function of adipose-derived stem cells and BMSCs was comparable to PDLSCs. Gingival mesenchymal stem cells showed less sensitivity to long-term culture. Both pure and mixed gingival cells had lower osteogenic ability compared to PDLSCs. Comparison of dental pulp stem cells (DPSCs) with PDLSCs regarding proliferation rate, osteo/adipogenesis, and immunomodulatory properties was contradictory; however, in vivo bone formation of DPSCs seemed to be lower than PDLSCs. CONCLUSION In light of the performed comparative studies, PDLSCs showed comparable results to stem cells derived from other sources; however, further in vivo studies are needed to determine the actual pros and cons of stem cells in comparison to each other.
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Affiliation(s)
- Zeinab Kadkhoda
- Department of Periodontology, School of Dentistry, Shahid Beheshti University of Medical Science, Tehran, Iran
| | - Parisa Motie
- Student Research Committee, School of Dentistry, Shahid Beheshti University of Medical Science, Tehran, Iran
| | - Maryam Rezaei Rad
- Dental Research Center, Research Institute of Dental Sciences, School of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Sadra Mohaghegh
- Student Research Committee, School of Dentistry, Shahid Beheshti University of Medical Science, Tehran, Iran
| | - Farnaz Kouhestani
- Department of Periodontics, School of Dentistry, Bushehr University of Medical Sciences, Tehran, Iran
| | - Saeed Reza Motamedian
- Dentofacial Deformities Research Center, Research Institute of Dental Sciences, Department of Orthodontics, School of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Alkharobi H. Exploring Various Transfection Approaches and Their Applications in Studying the Regenerative Potential of Dental Pulp Stem Cells. Curr Issues Mol Biol 2023; 45:10026-10040. [PMID: 38132472 PMCID: PMC10742526 DOI: 10.3390/cimb45120626] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/23/2023] [Revised: 12/05/2023] [Accepted: 12/08/2023] [Indexed: 12/23/2023] Open
Abstract
Transfection is a contemporary approach for introducing foreign genetic material into target cells. The effective transport of genetic materials into cells is mostly influenced by (a) the characteristics of the genetic material (quantity and quality), (b) the transfection procedure (incubation time, ratio of the reagents to the introduced genetic material, and components of cell culture), and (c) targeted cells for transfection (cell origin and cell type). This review summarizes the findings of different studies focusing on various transfection approaches and their applications to explore the regenerative potential of dental pulp stem cells (DPSCs). Several databases, including Scopus, Google Scholar, and PubMed, were searched to obtain the literature for the current review. Different keywords were used as key terms in the search. Approximately 200 articles were retained after removing duplicates from different databases. Articles published in English that discussed different transfection approaches were included. Several sources were excluded because they did not meet the inclusion criteria. Approximately 70 relevant published sources were included in the final stage to achieve the study objectives. This review demonstrated that no single transfection system is applicable to all cases and the various cell types with no side effects. Further studies are needed to focus on optimizing process parameters, decreasing the toxicity and side effects of available transfection techniques, and increasing their efficiencies. Moreover, this review sheds light on the impact of using different valuable transfection approaches to investigate the regenerative potential of DPSCs.
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Affiliation(s)
- Hanaa Alkharobi
- Department of Oral Biology, College of Dentistry, King Abdul-Aziz University, Jeddah 21589, Saudi Arabia
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8
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Zhang J, Ye F, Ye A, He B. Lysyl oxidase inhibits BMP9-induced osteoblastic differentiation through reducing Wnt/β-catenin via HIF-1a repression in 3T3-L1 cells. J Orthop Surg Res 2023; 18:911. [PMID: 38031108 PMCID: PMC10688138 DOI: 10.1186/s13018-023-04251-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/07/2023] [Accepted: 09/28/2023] [Indexed: 12/01/2023] Open
Abstract
BACKGROUND Bone morphogenetic protein 9 (BMP9) is a promising growth factor in bone tissue engineering, while the detailed molecular mechanism underlying BMP9-oriented osteogenesis remains unclear. In this study, we investigated the effect of lysyl oxidase (Lox) on the BMP9 osteogenic potential via in vivo and in vitro experiments, as well as the underlying mechanism. METHODS PCR assay, western blot analysis, histochemical staining, and immunofluorescence assay were used to quantify the osteogenic markers level, as well as the possible mechanism. The mouse ectopic osteogenesis assay was used to assess the impact of Lox on BMP9-induced bone formation. RESULTS Our findings suggested that Lox was obviously upregulated by BMP9 in 3T3-L1 cells. BMP9-induced Runx2, OPN, and mineralization were all enhanced by Lox inhibition or knockdown, while Lox overexpression reduced their expression. Additionally, the BMP9-induced adipogenic makers were repressed by Lox inhibition. Inhibition of Lox resulted in an increase in c-Myc mRNA and β-catenin protein levels. However, the increase in BMP9-induced osteoblastic biomarkers caused by Lox inhibition was obviously reduced when β-catenin knockdown. BMP9 upregulated HIF-1α expression, which was further enhanced by Lox inhibition or knockdown, but reversed by Lox overexpression. Lox knockdown or HIF-1α overexpression increased BMP9-induced bone formation, although the enhancement caused by Lox knockdown was largely diminished when HIF-1α was knocked down. Lox inhibition increased β-catenin levels and decreased SOST levels, which were almost reversed by HIF-1α knockdown. CONCLUSION Lox may reduce the BMP9 osteoblastic potential by inhibiting Wnt/β-catenin signaling via repressing the expression HIF-1α partially.
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Affiliation(s)
- Jie Zhang
- Department of Pharmacology, School of Pharmacy, Chongqing Medical University, No. 1 Yixueyuan Road, Yuzhong, Chongqing, 400016, People's Republic of China
- Key Laboratory of Biochemistry and Molecular Pharmacology of Chongqing, Chongqing Medical University, Chongqing, 400016, People's Republic of China
| | - FangLin Ye
- Department of Pharmacology, School of Pharmacy, Chongqing Medical University, No. 1 Yixueyuan Road, Yuzhong, Chongqing, 400016, People's Republic of China
- Key Laboratory of Biochemistry and Molecular Pharmacology of Chongqing, Chongqing Medical University, Chongqing, 400016, People's Republic of China
| | - AiHua Ye
- Department of Pharmacology, School of Pharmacy, Chongqing Medical University, No. 1 Yixueyuan Road, Yuzhong, Chongqing, 400016, People's Republic of China
- Key Laboratory of Biochemistry and Molecular Pharmacology of Chongqing, Chongqing Medical University, Chongqing, 400016, People's Republic of China
| | - BaiCheng He
- Department of Pharmacology, School of Pharmacy, Chongqing Medical University, No. 1 Yixueyuan Road, Yuzhong, Chongqing, 400016, People's Republic of China.
- Key Laboratory of Biochemistry and Molecular Pharmacology of Chongqing, Chongqing Medical University, Chongqing, 400016, People's Republic of China.
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Bae KB, Choi Y, Lee BN, Chang HS, Hwang IN, Oh WM, Hwang YC. A comparison of osteogenic effect of newly manufactured calcium silicate-based sealers in vitro. Dent Mater J 2023; 42:860-867. [PMID: 37914232 DOI: 10.4012/dmj.2023-048] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/03/2023]
Abstract
This study aimed to assess the effect of different calcium silicate-based root canal sealers (CSRS) on osteogenic effect in human periodontal ligament cells (hPDLCs). hPDLCs were cultured in a medium containing extract of 5 types of CSRS. The specimens were assessed by the cell cytotoxicity test, alkaline phosphatase staining, alizarin red S staining, quantitative real-time PCR, Western blot analysis, and enzyme-linked immunosorbent assay. The diluted concentrations of extracted solutions had no significant effect on the viability of hPDLCs. There was a statistically significant difference in the mRNA expression level of bone sialoprotein (BSP), osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2) among some groups. The protein expressions of BSP, OCN, and RUNX2 were significantly higher in some groups compared to the control group. The CSRS did not interfere with the osteogenic differentiation of hPDLCs, compared to the control group. CSRS are shown to have biocompatibility and osteogenic differentiation effect on hPDLCs.
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Affiliation(s)
- Kkot-Byeol Bae
- Department of Conservative Dentistry, School of Dentistry, Dental Science Research Institute, Chonnam National University
| | - Yoorina Choi
- Department of Conservative Dentistry, College of Dentistry, Wonkwang University
| | - Bin-Na Lee
- Department of Conservative Dentistry, School of Dentistry, Dental Science Research Institute, Chonnam National University
| | - Hoon-Sang Chang
- Department of Conservative Dentistry, School of Dentistry, Dental Science Research Institute, Chonnam National University
| | - In-Nam Hwang
- Department of Conservative Dentistry, School of Dentistry, Dental Science Research Institute, Chonnam National University
| | - Won-Mann Oh
- Department of Conservative Dentistry, School of Dentistry, Dental Science Research Institute, Chonnam National University
| | - Yun-Chan Hwang
- Department of Conservative Dentistry, School of Dentistry, Dental Science Research Institute, Chonnam National University
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Saber SM, Gomaa SM, Elashiry MM, El-Banna A, Schäfer E. Comparative biological properties of resin-free and resin-based calcium silicate-based endodontic repair materials on human periodontal ligament stem cells. Clin Oral Investig 2023; 27:6757-6768. [PMID: 37796335 PMCID: PMC10630253 DOI: 10.1007/s00784-023-05288-5] [Citation(s) in RCA: 12] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2023] [Accepted: 09/27/2023] [Indexed: 10/06/2023]
Abstract
OBJECTIVES To investigate the effect of three different calcium silicate-based materials (CSBM) on the biological behavior of human periodontal ligament stem cells (hPDLSCs). METHODS Eluates of Biodentine, NeoPutty and TheraCal PT prepared at 1:1, 1:2, and 1:4 ratios were extracted under sterile conditions. The cytotoxicity of the extracts to the hPDLSCs was assessed using the MTT assay. Scratch wound healing assay was utilized for assessing cell migration. Scanning electron microscopy was used to detect cell attachment and morphology. Calcium ion release was measured using inductively coupled plasma-optical emission spectrometry; the pH-value was evaluated with a pH-meter. ANOVA with post hoc Tukey test was used for statistical analysis. RESULTS Cell viability was significantly higher for Biodentine and NeoPutty at day 1 with all dilutions (p < 0.05), while at day 3 and day 7 with dilutions 1:2 and 1:4; all materials showed similar behavior (p > 0.05). Biodentine had the highest percentage of cell migration into the scratched area at day 1 for all dilutions (p < 0.05). Stem cells were attached favorably on Biodentine and NeoPutty with evident spreading, and intercellular communications; however, this was not shown for TheraCal PT. Biodentine showed the highest pH values and calcium ion release (p < 0.05). CONCLUSIONS The resin-free CSBM showed better performance and favorable biological effects on hPDLSCs and were therefore considered promising for usage as endodontic repair materials. CLINICAL SIGNIFICANCE Proper selection of materials with favorable impact on the host stem cells is crucial to ensure outcome in different clinical scenarios.
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Affiliation(s)
- Shehabeldin M Saber
- Department of Endodontics, Faculty of Dentistry, The British University in Egypt (BUE), Cairo, Egypt
- Dental Science Research Group, Health Research Centre of Excellence, The British University in Egypt (BUE), Cairo, Egypt
- Department of Endodontics, Faculty of Dentistry, Ain Shams University, Egypt, Cairo, Egypt
| | - Shaimaa M Gomaa
- Dental Science Research Group, Health Research Centre of Excellence, The British University in Egypt (BUE), Cairo, Egypt
| | - Mohamed M Elashiry
- Department of Endodontics, Faculty of Dentistry, Ain Shams University, Egypt, Cairo, Egypt
- Department of Endodontics, Dental College of Georgia, Augusta University, Augusta, Georgia, USA
| | - Ahmed El-Banna
- Department of Biomaterials, Faculty of Dentistry, Ain Shams University, Cairo, Egypt
| | - Edgar Schäfer
- Central Interdisciplinary Ambulance in the School of Dentistry, University of Münster, Münster, Germany.
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11
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Roi A, Roi C, Negruțiu ML, Rusu LC, Riviș M. Mesenchymal Stem Cells Derived from Human Periapical Cysts and Their Implications in Regenerative Medicine. Biomedicines 2023; 11:2436. [PMID: 37760877 PMCID: PMC10525783 DOI: 10.3390/biomedicines11092436] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/03/2023] [Revised: 08/27/2023] [Accepted: 08/28/2023] [Indexed: 09/29/2023] Open
Abstract
Mesenchymal stem cells currently play an important role in the tissue engineering field in developing new regenerative approaches. The oral cavity is a rich source of mesenchymal stem cells, and introducing the use of dental stem cells, characterized by a multilineage differentiation potential, immunomodulatory activity and repair capacity, offers a good perspective for clinical dentistry. Human periapical cyst mesenchymal stem cells (hPCy-MSCs) represent a new category of dental stem cells, being collected from pathological tissue and exhibiting MSCs-like properties. As studies have described, these new identified cells possess the same characteristics as those described in MSCs, exhibiting plasticity, a high proliferation rate and the potential to differentiate into osteogenic, adipogenic and neural lineages. Reusing the biological tissue that is considered pathologic offers a new perspective for the development of further clinical applications. The identification and characterization of MSCs in the human periapical cysts allows for a better understanding of the molecular interactions, the potential healing capacity and the mechanisms of inducing the local osteogenic process, integrated in the microenvironment. Although their involvement in regenerative medicine research is recent, they exhibit important properties that refer them for the development of clinical applications in dentistry.
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Affiliation(s)
- Alexandra Roi
- Department of Oral Pathology, “Victor Babeș” University of Medicine and Pharmacy, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania; (A.R.); (L.C.R.)
- Multidisciplinary Center for Research, Evaluation, Diagnosis and Therapies in Oral Medicine, “Victor Babeș” University of Medicine and Pharmacy, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania;
| | - Ciprian Roi
- Multidisciplinary Center for Research, Evaluation, Diagnosis and Therapies in Oral Medicine, “Victor Babeș” University of Medicine and Pharmacy, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania;
- Department of Anesthesiology and Oral Surgery, “Victor Babeș” University of Medicine and Pharmacy, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania
| | - Meda Lavinia Negruțiu
- Department of Prostheses Technology and Dental Materials, Faculty of Dental Medicine, “Victor Babes” University of Medicine and Pharmacy Timisoara, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania;
- Research Center in Dental Medicine Using Conventional and Alternative Technologies, “Victor Babes” University of Medicine and Pharmacy Timisoara, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania
| | - Laura Cristina Rusu
- Department of Oral Pathology, “Victor Babeș” University of Medicine and Pharmacy, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania; (A.R.); (L.C.R.)
- Multidisciplinary Center for Research, Evaluation, Diagnosis and Therapies in Oral Medicine, “Victor Babeș” University of Medicine and Pharmacy, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania;
| | - Mircea Riviș
- Multidisciplinary Center for Research, Evaluation, Diagnosis and Therapies in Oral Medicine, “Victor Babeș” University of Medicine and Pharmacy, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania;
- Department of Anesthesiology and Oral Surgery, “Victor Babeș” University of Medicine and Pharmacy, Eftimie Murgu Sq. No. 2, 300041 Timișoara, Romania
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12
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Zhang Y, Chen X, Yang X, Huang L, Qiu X. Mesenchymal Stem Cell-Derived from Dental Tissues-Related lncRNAs: A New Regulator in Osteogenic Differentiation. J Tissue Eng Regen Med 2023; 2023:4622584. [PMID: 40226409 PMCID: PMC11919082 DOI: 10.1155/2023/4622584] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/13/2023] [Revised: 06/12/2023] [Accepted: 06/22/2023] [Indexed: 04/15/2025]
Abstract
Odontogenic stem cells are mesenchymal stem cells (MSCs) with multipotential differentiation potential from different dental tissues. Their osteogenic differentiation is of great significance in bone tissue engineering. In recent years, it has been found that long noncoding RNAs (lncRNAs) participate in regulating the osteoblastic differentiation of stem cells at the epigenetic level, transcriptional level, and posttranscriptional level. We reviewed the existing lncRNA related to the osteogenic differentiation of odontogenic stem cells and emphasized the critical mechanism of lncRNA in the osteogenic differentiation of odontogenic stem cells. These findings are expected to be an important target for promoting osteoblastic differentiation of odontogenic stem cells in bone regeneration therapy with lncRNA.
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Affiliation(s)
- Yinchun Zhang
- Department of Endodontics, Stomatological Hospital, School of Stomatology, Southern Medical University, Guangdong 510280, China
| | - Xuan Chen
- Department of Endodontics, Stomatological Hospital, School of Stomatology, Southern Medical University, Guangdong 510280, China
| | - XiaoXia Yang
- Department of Endodontics, Stomatological Hospital, School of Stomatology, Southern Medical University, Guangdong 510280, China
| | - Lei Huang
- Department of Endodontics, Stomatological Hospital, School of Stomatology, Southern Medical University, Guangdong 510280, China
| | - Xiaoling Qiu
- Department of Endodontics, Stomatological Hospital, School of Stomatology, Southern Medical University, Guangdong 510280, China
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13
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Alves L, Machado V, Botelho J, Mendes JJ, Cabral JMS, da Silva CL, Carvalho MS. Enhanced Proliferative and Osteogenic Potential of Periodontal Ligament Stromal Cells. Biomedicines 2023; 11:biomedicines11051352. [PMID: 37239023 DOI: 10.3390/biomedicines11051352] [Citation(s) in RCA: 5] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/08/2023] [Revised: 04/24/2023] [Accepted: 04/27/2023] [Indexed: 05/28/2023] Open
Abstract
Cell-based therapies using periodontal ligament stromal cells (PDLSC) for periodontal regeneration may represent an alternative source for mesenchymal stromal cells (MSC) to MSC derived from bone marrow (MSC(M)) and adipose tissue (MSC(AT)). We aimed to characterize the osteogenic/periodontal potential of PDLSC in comparison to MSC(M) and MSC(AT). PDLSC were obtained from surgically extracted healthy human third molars, while MSC(M) and MSC(AT) were obtained from a previously established cell bank. Flow cytometry, immunocytochemistry, and cell proliferation analyses provided cellular characteristics from each group. Cells from the three groups presented MSC-like morphology, MSC-related marker expression, and multilineage differentiation capacity (adipogenic, chondrogenic, and osteogenic). In this study, PDLSC expressed osteopontin, osteocalcin, and asporin, while MSC(M) and MSC(AT) did not. Of note, only PDLSC expressed CD146, a marker previously applied to identify PDLSC, and presented higher proliferative potential compared to MSC(M) and MSC(AT). Upon osteogenic induction, PDLSC exhibited higher calcium content and enhanced upregulation of osteogenic/periodontal genes compared to MSC(M) and MSC(AT), such as Runx2, Col1A1 and CEMP-1. However, the alkaline phosphatase activity of PDLSC did not increase. Our findings suggest that PDLSC might be a promising cell source for periodontal regeneration, presenting enhanced proliferative and osteogenic potential compared to MSC(M) and MSC(AT).
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Affiliation(s)
- Laura Alves
- Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
- Associate Laboratory i4HB-Institute for Health and Bioeconomy, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
| | - Vanessa Machado
- Clinical Research Unit, Egas Moniz Center for Interdisciplinary Research, Egas Moniz School of Health and Science, 2829-511 Almada, Portugal
- Evidence-Based Hub, Egas Moniz Center for Interdisciplinary Research, Egas Moniz School of Health and Science, 2829-511 Almada, Portugal
| | - João Botelho
- Clinical Research Unit, Egas Moniz Center for Interdisciplinary Research, Egas Moniz School of Health and Science, 2829-511 Almada, Portugal
- Evidence-Based Hub, Egas Moniz Center for Interdisciplinary Research, Egas Moniz School of Health and Science, 2829-511 Almada, Portugal
| | - José João Mendes
- Clinical Research Unit, Egas Moniz Center for Interdisciplinary Research, Egas Moniz School of Health and Science, 2829-511 Almada, Portugal
- Evidence-Based Hub, Egas Moniz Center for Interdisciplinary Research, Egas Moniz School of Health and Science, 2829-511 Almada, Portugal
| | - Joaquim M S Cabral
- Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
- Associate Laboratory i4HB-Institute for Health and Bioeconomy, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
| | - Cláudia L da Silva
- Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
- Associate Laboratory i4HB-Institute for Health and Bioeconomy, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
| | - Marta S Carvalho
- Department of Bioengineering and iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
- Associate Laboratory i4HB-Institute for Health and Bioeconomy, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
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14
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Carvalho S, Santos JI, Moreira L, Gonçalves M, David H, Matos L, Encarnação M, Alves S, Coutinho MF. Neurological Disease Modeling Using Pluripotent and Multipotent Stem Cells: A Key Step towards Understanding and Treating Mucopolysaccharidoses. Biomedicines 2023; 11:biomedicines11041234. [PMID: 37189853 DOI: 10.3390/biomedicines11041234] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/08/2023] [Revised: 04/18/2023] [Accepted: 04/19/2023] [Indexed: 05/17/2023] Open
Abstract
Despite extensive research, the links between the accumulation of glycosaminoglycans (GAGs) and the clinical features seen in patients suffering from various forms of mucopolysaccharidoses (MPSs) have yet to be further elucidated. This is particularly true for the neuropathology of these disorders; the neurological symptoms are currently incurable, even in the cases where a disease-specific therapeutic approach does exist. One of the best ways to get insights on the molecular mechanisms driving that pathogenesis is the analysis of patient-derived cells. Yet, not every patient-derived cell recapitulates relevant disease features. For the neuronopathic forms of MPSs, for example, this is particularly evident because of the obvious inability to access live neurons. This scenario changed significantly with the advent of induced pluripotent stem cell (iPSC) technologies. From then on, a series of differentiation protocols to generate neurons from iPSC was developed and extensively used for disease modeling. Currently, human iPSC and iPSC-derived cell models have been generated for several MPSs and numerous lessons were learnt from their analysis. Here we review most of those studies, not only listing the currently available MPS iPSC lines and their derived models, but also summarizing how they were generated and the major information different groups have gathered from their analyses. Finally, and taking into account that iPSC generation is a laborious/expensive protocol that holds significant limitations, we also hypothesize on a tempting alternative to establish MPS patient-derived neuronal cells in a much more expedite way, by taking advantage of the existence of a population of multipotent stem cells in human dental pulp to establish mixed neuronal and glial cultures.
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Affiliation(s)
- Sofia Carvalho
- Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal
- Center for the Study of Animal Science-Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal
- Associate Laboratory for Animal and Veterinary Sciences, AL4AnimalS, Faculdade de Medicina Veterinária Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal
- Faculty of Pharmacy, University of Coimbra, Polo das Ciências da Saúde, Azinhaga de SantaComba, 3000-548 Coimbra, Portugal
| | - Juliana Inês Santos
- Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal
- Center for the Study of Animal Science-Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal
- Associate Laboratory for Animal and Veterinary Sciences, AL4AnimalS, Faculdade de Medicina Veterinária Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal
- Biology Department, Faculty of Sciences, University of Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal
| | - Luciana Moreira
- Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal
- Center for the Study of Animal Science-Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal
- Associate Laboratory for Animal and Veterinary Sciences, AL4AnimalS, Faculdade de Medicina Veterinária Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal
| | - Mariana Gonçalves
- Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal
- Center for the Study of Animal Science-Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal
- Associate Laboratory for Animal and Veterinary Sciences, AL4AnimalS, Faculdade de Medicina Veterinária Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal
- Centre for the Research and Technology of Agro-Environmental and Biological Sciences, CITAB, Inov4Agro, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal
| | - Hugo David
- Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal
- Center for the Study of Animal Science-Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal
- Associate Laboratory for Animal and Veterinary Sciences, AL4AnimalS, Faculdade de Medicina Veterinária Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal
- Biology Department, Faculty of Sciences, University of Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal
| | - Liliana Matos
- Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal
- Center for the Study of Animal Science-Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal
- Associate Laboratory for Animal and Veterinary Sciences, AL4AnimalS, Faculdade de Medicina Veterinária Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal
| | - Marisa Encarnação
- Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal
- Center for the Study of Animal Science-Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal
- Associate Laboratory for Animal and Veterinary Sciences, AL4AnimalS, Faculdade de Medicina Veterinária Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal
| | - Sandra Alves
- Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal
- Center for the Study of Animal Science-Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal
- Associate Laboratory for Animal and Veterinary Sciences, AL4AnimalS, Faculdade de Medicina Veterinária Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal
| | - Maria Francisca Coutinho
- Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal
- Center for the Study of Animal Science-Instituto de Ciências, Tecnologias e Agroambiente da Universidade do Porto, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal
- Associate Laboratory for Animal and Veterinary Sciences, AL4AnimalS, Faculdade de Medicina Veterinária Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal
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15
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Zhao X, Xie Z, Rao N, Zhang S, Zhang Y. Effect of dermatopontin on osteogenic differentiation of periodontal ligament stem cells. Gene 2023; 858:147185. [PMID: 36632910 DOI: 10.1016/j.gene.2023.147185] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/13/2022] [Revised: 01/03/2023] [Accepted: 01/05/2023] [Indexed: 01/11/2023]
Abstract
Human periodontal ligament stem cells (hPDLSCs) are promising seed cells for oral bone tissue engineering. Dermatopontin (DPT) is a small-molecule protein recognized as a non-collagenous component of the extracellular matrix and is associated with a variety of biological processes. In this study, we first determined that DPT was elevated during the osteogenic differentiation of hPDLSCs. HPDLSCs interfering with DPT expression were established by lentiviral infection. It was found that the proliferation and osteogenic differentiation ability of hPDLSCs were inhibited after interfering DPT with lentivirus. Exogenous recombinant DPT treatment could not alter the proliferation of hPDLSCs. Coincidentally, exogenous DPT can only enhance the osteogenic differentiation of hPDLSCs in the control lentivirus group, but had no significant effect on the DPT interference group. This study expands the understanding of DPT function and implicates DPT as an important target for enhancing osteogenic differentiation of hPDLSCs.
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Affiliation(s)
- Xuechun Zhao
- Department of Oral Implantology, School and Hospital of Stomatology, Kunming Medical University, Kunming, PR China; Yunnan Key Laboratory of Stomatology, Kunming, PR China
| | - Zhigang Xie
- Department of Oral Implantology, School and Hospital of Stomatology, Kunming Medical University, Kunming, PR China; Yunnan Key Laboratory of Stomatology, Kunming, PR China
| | - Nanquan Rao
- Department of Oral Implantology, School and Hospital of Stomatology, Kunming Medical University, Kunming, PR China; Yunnan Key Laboratory of Stomatology, Kunming, PR China
| | - Shu Zhang
- Department of Oral Implantology, School and Hospital of Stomatology, Kunming Medical University, Kunming, PR China; Yunnan Key Laboratory of Stomatology, Kunming, PR China
| | - Yunpeng Zhang
- Department of Oral Implantology, School and Hospital of Stomatology, Kunming Medical University, Kunming, PR China; Yunnan Key Laboratory of Stomatology, Kunming, PR China.
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16
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Kong H, Liu P, Li H, Zeng X, Xu P, Yao X, Liu S, Cheng CK, Xu J. Mesenchymal Stem Cell-Derived Extracellular Vesicles: The Novel Therapeutic Option for Regenerative Dentistry. Stem Cell Rev Rep 2023; 19:46-58. [PMID: 35132538 DOI: 10.1007/s12015-022-10342-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 01/23/2022] [Indexed: 01/29/2023]
Abstract
Dental mesenchymal stem cells (MSCs) are characterized by unlimited self-renewal ability and high multidirectional differentiation potential. Since dental MSCs can be easily isolated and exhibit a high capability to differentiate into odontogenic cells, they are considered as attractive therapeutic agents in regenerative dentistry. Recently, MSC-derived extracellular vesicles (MSC-EVs) have attracted widespread attention as carriers for cell-free therapy due to their potential functions. Many studies have shown that MSC-EVs can mediate microenvironment at tissue damage site, and coordinate the regeneration process. Additionally, MSC-EVs can mediate intercellular communication, thus affecting the phenotypes and functions of recipient cells. In this review, we mainly summarized the types of MSCs that could be potentially applied in regenerative dentistry, the possible molecular cargos of MSC-EVs, and the major effects of MSC-EVs on the therapeutic induction of osteogenic differentiation.
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Affiliation(s)
- Haiying Kong
- Department of Dentistry, Longgang E.N.T. Hospital & Shenzhen Key Laboratory of E.N.T, Institute of E.N.T, Shenzhen, Guangdong, China
| | - Peiqi Liu
- Department of Dentistry, Longgang E.N.T. Hospital & Shenzhen Key Laboratory of E.N.T, Institute of E.N.T, Shenzhen, Guangdong, China.,Second School of Clinical Medicine, Guangdong Medical University, Dongguan, Guangdong, China
| | - Hongwen Li
- Department of Dentistry, Longgang E.N.T. Hospital & Shenzhen Key Laboratory of E.N.T, Institute of E.N.T, Shenzhen, Guangdong, China.,Shenzhen Longgang Institute of Stomatology, Shenzhen, Guangdong, China
| | - Xiantao Zeng
- Department of Dentistry, Longgang E.N.T. Hospital & Shenzhen Key Laboratory of E.N.T, Institute of E.N.T, Shenzhen, Guangdong, China
| | - Peiwu Xu
- Department of Dentistry, Longgang E.N.T. Hospital & Shenzhen Key Laboratory of E.N.T, Institute of E.N.T, Shenzhen, Guangdong, China
| | - Xinhui Yao
- Department of Dentistry, Longgang E.N.T. Hospital & Shenzhen Key Laboratory of E.N.T, Institute of E.N.T, Shenzhen, Guangdong, China
| | - Senqing Liu
- Department of Dentistry, Longgang E.N.T. Hospital & Shenzhen Key Laboratory of E.N.T, Institute of E.N.T, Shenzhen, Guangdong, China
| | - Chak Kwong Cheng
- School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong, China.
| | - Jian Xu
- Department of Dentistry, Longgang E.N.T. Hospital & Shenzhen Key Laboratory of E.N.T, Institute of E.N.T, Shenzhen, Guangdong, China. .,Shenzhen Longgang Institute of Stomatology, Shenzhen, Guangdong, China.
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17
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Sugiaman VK, Djuanda R, Pranata N, Naliani S, Demolsky WL. Tissue Engineering with Stem Cell from Human Exfoliated Deciduous Teeth (SHED) and Collagen Matrix, Regulated by Growth Factor in Regenerating the Dental Pulp. Polymers (Basel) 2022; 14:polym14183712. [PMID: 36145860 PMCID: PMC9503223 DOI: 10.3390/polym14183712] [Citation(s) in RCA: 7] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/07/2022] [Revised: 08/25/2022] [Accepted: 09/02/2022] [Indexed: 11/16/2022] Open
Abstract
Maintaining dental pulp vitality and preventing tooth loss are two challenges in endodontic treatment. A tooth lacking a viable pulp loses its defense mechanism and regenerative ability, making it more vulnerable to severe damage and eventually necessitating extraction. The tissue engineering approach has drawn attention as an alternative therapy as it can regenerate dentin-pulp complex structures and functions. Stem cells or progenitor cells, extracellular matrix, and signaling molecules are triad components of this approach. Stem cells from human exfoliated deciduous teeth (SHED) are a promising, noninvasive source of stem cells for tissue regeneration. Not only can SHEDs regenerate dentin-pulp tissues (comprised of fibroblasts, odontoblasts, endothelial cells, and nerve cells), but SHEDs also possess immunomodulatory and immunosuppressive properties. The collagen matrix is a material of choice to provide structural and microenvironmental support for SHED-to-dentin pulp tissue differentiation. Growth factors regulate cell proliferation, migration, and differentiation into specific phenotypes via signal-transduction pathways. This review provides current concepts and applications of the tissue engineering approach, especially SHEDs, in endodontic treatment.
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Affiliation(s)
- Vinna K Sugiaman
- Department of Oral Biology, Faculty of Dentistry, Maranatha Christian University, Bandung 40164, Indonesia
| | - Rudy Djuanda
- Department of Conservative Dentistry and Endodontic, Faculty of Dentistry, Maranatha Christian University, Bandung 40164, Indonesia
| | - Natallia Pranata
- Department of Oral Biology, Faculty of Dentistry, Maranatha Christian University, Bandung 40164, Indonesia
| | - Silvia Naliani
- Department of Prosthodontics, Faculty of Dentistry, Maranatha Christian University, Bandung 40164, Indonesia
| | - Wayan L Demolsky
- Department of Oral Biology, Faculty of Dentistry, Maranatha Christian University, Bandung 40164, Indonesia
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18
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Zhong X, Wang H. circSKIL promotes osteoblastic differentiation of periodontal ligament cells by sponging miR-532-5p to activate Notch signaling. J Periodontal Res 2022; 57:1148-1158. [PMID: 36063416 DOI: 10.1111/jre.13052] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/21/2022] [Revised: 08/08/2022] [Accepted: 08/24/2022] [Indexed: 12/09/2022]
Abstract
BACKGROUND AND OBJECTIVE Periodontal ligament cells (PDLCs) possess the capacity to differentiate into a variety of cell types to benefit periodontal regeneration. In this study, we examined the circSKIL/miR-532-5p/Notch1 axis in controlling the osteoblastic differentiation of PDLCs. METHODS Primary human PDLCs (hPDLCs) were isolated and induced to differentiate into osteoblasts. Osteogenic responses were assessed for the expressions of osteoblast-related marker proteins (including alkaline phosphatase (ALP), osteocalcin (OCN), bone morphogenetic protein-2 (BMP2), and runt-related transcription factor 2 (RUNX2) by RT-PCR. The formation of mineralized nodules was examined by Alizarin Red S (ARS) staining and ALP activity. Expressions of circSKIL, miR-532-5p, and Notch1 were measured by RT-PCR and western blotting, and their regulations by combining bioinformatic analysis and luciferase reporter assay. Notch signaling was assessed for the expressions of hairy and enhancer of split-1 (HES1) and Notch intracellular domain (NICD). RESULTS During osteoblastic differentiation of hPDLCs, circSKIL, and Notch1 were up-regulated, while miR-532-5p down-regulated. By sponging miR-532-5p, circSKIL activated Notch signaling, increasing levels of Notch1, HES1, and NICD. Functionally, knocking down circSKIL or overexpressing miR-532-5p inhibited osteoblastic differentiation of PDLCs, down-regulating ALP, OCN, BMP2, and RUNX2, and reducing ARS staining or ALP activity. The impacts of circSKIL knockdown were rescued by miR-532-5p inhibitor or overexpressing Notch1, while those caused by up-regulating miR-532-5p were reversed by overexpressing Notch1. CONCLUSION By targeting miR-532-5p and up-regulating Notch1, circSKIL critically controls osteoblastic differentiation of hPDLCs. Therefore, modulating this axis may maximize the differentiation of PDLCs into osteoblasts and benefit periodontal regeneration.
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Affiliation(s)
- Xiaohuan Zhong
- Center of Stomatology, Xiangya Hospital, Central South University, Changsha, China
| | - Huixin Wang
- Center of Stomatology, Xiangya Hospital, Central South University, Changsha, China
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19
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Different Sources of Mesenchymal Stem Cells for Tissue Regeneration: A Guide to Identifying the Most Favorable One in Orthopedics and Dentistry Applications. Int J Mol Sci 2022; 23:ijms23116356. [PMID: 35683035 PMCID: PMC9181542 DOI: 10.3390/ijms23116356] [Citation(s) in RCA: 59] [Impact Index Per Article: 19.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/08/2022] [Revised: 05/31/2022] [Accepted: 06/03/2022] [Indexed: 12/04/2022] Open
Abstract
The success of regenerative medicine in various clinical applications depends on the appropriate selection of the source of mesenchymal stem cells (MSCs). Indeed, the source conditions, the quality and quantity of MSCs, have an influence on the growth factors, cytokines, extracellular vesicles, and secrete bioactive factors of the regenerative milieu, thus influencing the clinical result. Thus, optimal source selection should harmonize this complex setting and ensure a well-personalized and effective treatment. Mesenchymal stem cells (MSCs) can be obtained from several sources, including bone marrow and adipose tissue, already used in orthopedic regenerative applications. In this sense, for bone, dental, and oral injuries, MSCs could provide an innovative and effective therapy. The present review aims to compare the properties (proliferation, migration, clonogenicity, angiogenic capacity, differentiation potential, and secretome) of MSCs derived from bone marrow, adipose tissue, and dental tissue to enable clinicians to select the best source of MSCs for their clinical application in bone and oral tissue regeneration to delineate new translational perspectives. A review of the literature was conducted using the search engines Web of Science, Pubmed, Scopus, and Google Scholar. An analysis of different publications showed that all sources compared (bone marrow mesenchymal stem cells (BM-MSCs), adipose tissue mesenchymal stem cells (AT-MSCs), and dental tissue mesenchymal stem cells (DT-MSCs)) are good options to promote proper migration and angiogenesis, and they turn out to be useful for gingival, dental pulp, bone, and periodontal regeneration. In particular, DT-MSCs have better proliferation rates and AT and G-MSC sources showed higher clonogenicity. MSCs from bone marrow, widely used in orthopedic regenerative medicine, are preferable for their differentiation ability. Considering all the properties among sources, BM-MSCs, AT-MSCs, and DT-MSCs present as potential candidates for oral and dental regeneration.
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20
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Faruangsaeng T, Thaweesapphitak S, Khamwachirapitak C, Porntaveetus T, Shotelersuk V. Comparative transcriptome profiles of human dental pulp stem cells from maxillary and mandibular teeth. Sci Rep 2022; 12:8860. [PMID: 35614192 PMCID: PMC9133121 DOI: 10.1038/s41598-022-12867-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/30/2022] [Accepted: 05/11/2022] [Indexed: 11/09/2022] Open
Abstract
The molecular control of tooth development is different between the maxilla and mandible, contributing to different tooth shapes and locations; however, whether this difference occurs in human permanent teeth is unknown. The aim of this study was to investigate and compare the transcriptome profiles of permanent maxillary and mandibular posterior teeth. Ten participants who had a pair of opposing premolars or molars extracted were recruited. The RNA obtained from cultured dental pulp stem cells underwent RNA-sequencing and qRT-PCR. The transcriptome profiles of two opposing premolar pairs and two molar pairs demonstrated that the upper premolars, lower premolars, upper molars, and lower molars expressed the same top-ranked genes, comprising FN1, COL1A1, COL1A2, ACTB, and EEFIA1, which are involved in extracellular matrix organization, immune system, signal transduction, hemostasis, and vesicle-mediated transport. Comparative transcriptome analyses of each/combined tooth pairs demonstrated that PITX1 was the only gene with different expression levels between upper and lower posterior teeth. PITX1 exhibited a 64-fold and 116-fold higher expression level in lower teeth compared with their upper premolars and molars, respectively. These differences were confirmed by qRT-PCR. Taken together, this study, for the first time, reveals that PITX1 is expressed significantly higher in mandibular posterior teeth compared with maxillary posterior teeth. The difference is more evident in the molars compared with premolars and consistent with its expression pattern in mouse developing teeth. We demonstrate that differences in lower versus upper teeth gene expression during odontogenesis occur in permanent teeth and suggest that these differences should be considered in molecular studies of dental pulp stem cells. Our findings pave the way to develop a more precise treatment in regenerative dentistry such as gene-based therapies for dentin/pulp regeneration and regeneration of different tooth types.
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Affiliation(s)
- Thira Faruangsaeng
- International Graduate Program in Geriatric Dentistry and Special Patients Care, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.,Center of Excellence in Genomics and Precision Dentistry, Department of Physiology, Faculty of Dentistry, Chulalongkorn University, Bangkok, 10330, Thailand
| | - Sermporn Thaweesapphitak
- Center of Excellence in Genomics and Precision Dentistry, Department of Physiology, Faculty of Dentistry, Chulalongkorn University, Bangkok, 10330, Thailand
| | - Chompak Khamwachirapitak
- Center of Excellence in Genomics and Precision Dentistry, Department of Physiology, Faculty of Dentistry, Chulalongkorn University, Bangkok, 10330, Thailand
| | - Thantrira Porntaveetus
- International Graduate Program in Geriatric Dentistry and Special Patients Care, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand. .,Center of Excellence in Genomics and Precision Dentistry, Department of Physiology, Faculty of Dentistry, Chulalongkorn University, Bangkok, 10330, Thailand.
| | - Vorasuk Shotelersuk
- Center of Excellence for Medical Genomics, Medical Genomics Cluster, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.,Excellence Center for Genomics and Precision Medicine, King Chulalongkorn Memorial Hospital, The Thai Red Cross Society, Bangkok, Thailand
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21
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Guo R, Yu J. Multipotency and Immunomodulatory Benefits of Stem Cells From Human Exfoliated Deciduous Teeth. FRONTIERS IN DENTAL MEDICINE 2022. [DOI: 10.3389/fdmed.2022.805875] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022] Open
Abstract
Stem cells derived from human exfoliated deciduous teeth (SHEDs) are considered a promising cell population for cell-based or cell-free therapy and tissue engineering because of their proliferative, multipotency and immunomodulator. Based on recent studies, we find that SHEDs show the superior ability of nerve regeneration in addition to the potential of osteogenesis, odontogenesis owing to their derivation from the neural crest. Besides, much evidence suggests that SHEDs have a paracrine effect and can function as immunomodulatory regents attributing to their capability of secreting cytokines and extracellular vesicles. Here, we review the characteristic of SHEDs, their multipotency to regenerate damaged tissues, specifically concentrating on bones or nerves, following the paracrine activity or immunomodulatory benefits of their potential for clinical application in regenerative medicine.
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22
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Okić-Đorđević I, Obradović H, Kukolj T, Petrović A, Mojsilović S, Bugarski D, Jauković A. Dental mesenchymal stromal/stem cells in different microenvironments— implications in regenerative therapy. World J Stem Cells 2021; 13:1863-1880. [PMID: 35069987 PMCID: PMC8727232 DOI: 10.4252/wjsc.v13.i12.1863] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/17/2021] [Revised: 06/15/2021] [Accepted: 11/25/2021] [Indexed: 02/06/2023] Open
Abstract
Current research data reveal microenvironment as a significant modifier of physical functions, pathologic changes, as well as the therapeutic effects of stem cells. When comparing regeneration potential of various stem cell types used for cytotherapy and tissue engineering, mesenchymal stem cells (MSCs) are currently the most attractive cell source for bone and tooth regeneration due to their differentiation and immunomodulatory potential and lack of ethical issues associated with their use. The microenvironment of donors and recipients selected in cytotherapy plays a crucial role in regenerative potential of transplanted MSCs, indicating interactions of cells with their microenvironment indispensable in MSC-mediated bone and dental regeneration. Since a variety of MSC populations have been procured from different parts of the tooth and tooth-supporting tissues, MSCs of dental origin and their achievements in capacity to reconstitute various dental tissues have gained attention of many research groups over the years. This review discusses recent advances in comparative analyses of dental MSC regeneration potential with regards to their tissue origin and specific microenvironmental conditions, giving additional insight into the current clinical application of these cells.
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Affiliation(s)
- Ivana Okić-Đorđević
- Laboratory for Experimental Hematology and Stem Cells, Institute for Medical Research, University of Belgrade, Belgrade 11129, Serbia
| | - Hristina Obradović
- Laboratory for Experimental Hematology and Stem Cells, Institute for Medical Research, University of Belgrade, Belgrade 11129, Serbia
| | - Tamara Kukolj
- Laboratory for Experimental Hematology and Stem Cells, Institute for Medical Research, University of Belgrade, Belgrade 11129, Serbia
| | - Anđelija Petrović
- Laboratory for Experimental Hematology and Stem Cells, Institute for Medical Research, University of Belgrade, Belgrade 11129, Serbia
| | - Slavko Mojsilović
- Laboratory for Experimental Hematology and Stem Cells, Institute for Medical Research, University of Belgrade, Belgrade 11129, Serbia
| | - Diana Bugarski
- Laboratory for Experimental Hematology and Stem Cells, Institute for Medical Research, University of Belgrade, Belgrade 11129, Serbia
| | - Aleksandra Jauković
- Laboratory for Experimental Hematology and Stem Cells, Institute for Medical Research, University of Belgrade, Belgrade 11129, Serbia
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23
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Földes A, Reider H, Varga A, Nagy KS, Perczel-Kovach K, Kis-Petik K, DenBesten P, Ballagi A, Varga G. Culturing and Scaling up Stem Cells of Dental Pulp Origin Using Microcarriers. Polymers (Basel) 2021; 13:3951. [PMID: 34833250 PMCID: PMC8622966 DOI: 10.3390/polym13223951] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/30/2021] [Revised: 11/08/2021] [Accepted: 11/09/2021] [Indexed: 02/07/2023] Open
Abstract
Ectomesenchymal stem cells derived from the dental pulp are of neural crest origin, and as such are promising sources for cell therapy and tissue engineering. For safe upscaling of these cells, microcarrier-based culturing under dynamic conditions is a promising technology. We tested the suitability of two microcarriers, non-porous Cytodex 1 and porous Cytopore 2, for culturing well characterized dental pulp stem cells (DPSCs) using a shake flask system. Human DPSCs were cultured on these microcarriers in 96-well plates, and further expanded in shake flasks for upscaling experiments. Cell viability was measured using the alamarBlue assay, while cell morphology was observed by conventional and two-photon microscopies. Glucose consumption of cells was detected by the glucose oxidase/Clark-electrode method. DPSCs adhered to and grew well on both microcarrier surfaces and were also found in the pores of the Cytopore 2. Cells grown in tissue culture plates (static, non-shaking conditions) yielded 7 × 105 cells/well. In shake flasks, static preincubation promoted cell adhesion to the microcarriers. Under dynamic culture conditions (shaking) 3 × 107 cells were obtained in shake flasks. The DPSCs exhausted their glucose supply from the medium by day seven even with partial batch-feeding. In conclusion, both non-porous and porous microcarriers are suitable for upscaling ectomesenchymal DPSCs under dynamic culture conditions.
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Affiliation(s)
- Anna Földes
- Department of Oral Biology, Semmelweis University, H-1089 Budapest, Hungary; (A.F.); (H.R.); (A.V.); (K.S.N.); (K.P.-K.)
| | - Hajnalka Reider
- Department of Oral Biology, Semmelweis University, H-1089 Budapest, Hungary; (A.F.); (H.R.); (A.V.); (K.S.N.); (K.P.-K.)
- Department of Applied Biotechnology and Food Science, University of Technology and Economics, H-1089 Budapest, Hungary;
| | - Anita Varga
- Department of Oral Biology, Semmelweis University, H-1089 Budapest, Hungary; (A.F.); (H.R.); (A.V.); (K.S.N.); (K.P.-K.)
- Department of Applied Biotechnology and Food Science, University of Technology and Economics, H-1089 Budapest, Hungary;
| | - Krisztina S. Nagy
- Department of Oral Biology, Semmelweis University, H-1089 Budapest, Hungary; (A.F.); (H.R.); (A.V.); (K.S.N.); (K.P.-K.)
- Institute of Biophysics and Radiation Biology, Semmelweis University, H-1089 Budapest, Hungary;
| | - Katalin Perczel-Kovach
- Department of Oral Biology, Semmelweis University, H-1089 Budapest, Hungary; (A.F.); (H.R.); (A.V.); (K.S.N.); (K.P.-K.)
- Department of Community Dentistry, Semmelweis University, H-1089 Budapest, Hungary
| | - Katalin Kis-Petik
- Institute of Biophysics and Radiation Biology, Semmelweis University, H-1089 Budapest, Hungary;
| | - Pamela DenBesten
- Department of Orofacial Science, University of California, San Francisco, CA 94143, USA;
| | - András Ballagi
- Department of Applied Biotechnology and Food Science, University of Technology and Economics, H-1089 Budapest, Hungary;
- Gedeon Richter Plc, H-1089 Budapest, Hungary
| | - Gábor Varga
- Department of Oral Biology, Semmelweis University, H-1089 Budapest, Hungary; (A.F.); (H.R.); (A.V.); (K.S.N.); (K.P.-K.)
- Centre for Translational Medicine, Semmelweis University, H-1089 Budapest, Hungary
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24
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Winning L, El Karim IA, Linden GJ, Irwin CR, Killough SA, Lundy FT. Differential regulation of NPY and SP receptor expression in STRO-1+ve PDLSCs by inflammatory cytokines. J Periodontal Res 2021; 57:186-194. [PMID: 34773642 DOI: 10.1111/jre.12952] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/09/2021] [Revised: 09/30/2021] [Accepted: 10/30/2021] [Indexed: 12/17/2022]
Abstract
OBJECTIVES The aims of this study were to investigate neuropeptide receptor expression regulation on STRO-1 +ve periodontal ligament stem cells (PDLSCs) in response to inflammatory cytokines and to investigate a potential osteogenic effect of neuropeptides. BACKGROUND Nerve fibres innervating the periodontal tissues in humans contain several neuropeptides including neuropeptide Y and substance P. The role of neuropeptide receptors on PDLSCs, including their response to the local inflammatory environment of periodontitis, is currently unknown. METHODS A homogenous population of STRO-1 +ve PDLSCs was prepared by immunomagnetic separation of cells obtained by the tissue out-growth method from healthy premolar teeth from a single donor. Regulation of gene expression of the neuropeptide Y Y1 receptor and substance P receptor tachykinin receptor 1 was investigated. A potential osteogenic effect of neuropeptide Y and substance P was also investigated by measuring alkaline phosphatase (ALP) activity, Alizarin red staining and quantifying osteogenic gene expression. RESULTS Treatment of STRO-1 +ve PDLSCs with tumour necrosis factor-alpha or interleukin 1-beta up-regulated the expression of the neuropeptide Y's Y1 receptor, but down-regulated substance P's receptor. Significantly increased ALP activity was observed in STRO-1 +ve PDLSCs treated with neuropeptide Y but not substance P. Further studies showed that neuropeptide Y had a modest osteogenic effect on cells at both a functional level and a gene level. CONCLUSIONS Expression of the neuropeptide Y Y1 receptor gene on STRO-1 +ve PDLSCs was sensitive to local inflammatory cytokines. Treatment of cells with neuropeptide Y was found to produce a modest enhanced osteogenic effect.
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Affiliation(s)
- Lewis Winning
- The Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, Northern Ireland.,Dublin Dental University Hospital, Trinity College Dublin, Dublin, Ireland
| | - Ikhlas A El Karim
- The Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, Northern Ireland
| | - Gerard J Linden
- Centre for Dentistry, School of Medicine Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, Northern Ireland
| | - Christopher R Irwin
- Centre for Dentistry, School of Medicine Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, Northern Ireland
| | - Simon A Killough
- Centre for Dentistry, School of Medicine Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, Northern Ireland
| | - Fionnuala T Lundy
- The Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, Northern Ireland
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25
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Afami ME, El Karim I, About I, Krasnodembskaya AD, Laverty G, Lundy FT. Multicomponent Peptide Hydrogels as an Innovative Platform for Cell-Based Tissue Engineering in the Dental Pulp. Pharmaceutics 2021; 13:1575. [PMID: 34683868 PMCID: PMC8539061 DOI: 10.3390/pharmaceutics13101575] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/14/2021] [Revised: 09/20/2021] [Accepted: 09/22/2021] [Indexed: 11/17/2022] Open
Abstract
In light of the increasing levels of antibiotic resistance, nanomaterials and novel biologics are urgently required to manage bacterial infections. To date, commercially available self-assembling peptide hydrogels have not been studied extensively for their ability to inhibit micro-organisms relevant to tissue engineering sites such as dental root canals. In this work, we assess the biocompatibility of dental pulp stem/stromal cells with commercially available multicomponent peptide hydrogels. We also determine the effects of dental pulp stem/stromal cell (DPSC) culture in hydrogels on growth factor/cytokine expression. Furthermore, to investigate novel aspects of self-assembling peptide hydrogels, we determine their antimicrobial activity against the oral pathogens Staphylococcus aureus, Enterococcus faecalis, and Fusobacterium nucleatum. We show that self-assembling peptide hydrogels and hydrogels functionalized with the adhesion motif Arg-Gly-Asp (RGD) are biocompatible with DPSCs, and that cells grown in 3D hydrogel cultures produce a discrete secretome compared with 2D-cultured cells. Furthermore, we show that soluble peptides and assembled hydrogels have antimicrobial effects against oral pathogens. Given their antibacterial activity against oral pathogens, biocompatibility with dental pulp stem/stromal cells and enhancement of an angiogenic secretome, multicomponent peptide hydrogels hold promise for translational use.
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Affiliation(s)
- Marina E. Afami
- Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen’s University Belfast, 97 Lisburn Road, Belfast BT9 7BL, UK; (M.E.A.); (I.E.K.); (A.D.K.)
| | - Ikhlas El Karim
- Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen’s University Belfast, 97 Lisburn Road, Belfast BT9 7BL, UK; (M.E.A.); (I.E.K.); (A.D.K.)
| | - Imad About
- Aix Marseille Univ, CNRS, ISM, Inst Movement Sci, 13385 Marseille, France;
| | - Anna D. Krasnodembskaya
- Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen’s University Belfast, 97 Lisburn Road, Belfast BT9 7BL, UK; (M.E.A.); (I.E.K.); (A.D.K.)
| | - Garry Laverty
- School of Pharmacy, Queen’s University Belfast, 97 Lisburn Road, Belfast BT9 7BL, UK;
| | - Fionnuala T. Lundy
- Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen’s University Belfast, 97 Lisburn Road, Belfast BT9 7BL, UK; (M.E.A.); (I.E.K.); (A.D.K.)
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26
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Bordini EAF, Ferreira JA, Dubey N, Ribeiro JS, de Souza Costa CA, Soares DG, Bottino MC. Injectable Multifunctional Drug Delivery System for Hard Tissue Regeneration under Inflammatory Microenvironments. ACS APPLIED BIO MATERIALS 2021; 4:6993-7006. [PMID: 35006932 DOI: 10.1021/acsabm.1c00620] [Citation(s) in RCA: 17] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/27/2022]
Abstract
Engineering multifunctional hydrogel systems capable of amplifying the regenerative capacity of endogenous progenitor cells via localized presentation of therapeutics under tissue inflammation is central to the translation of effective strategies for hard tissue regeneration. Here, we loaded dexamethasone (DEX), a pleotropic drug with anti-inflammatory and mineralizing abilities, into aluminosilicate clay nanotubes (halloysite clay nanotubes (HNTs)) to engineer an injectable multifunctional drug delivery system based on photo-cross-linkable gelatin methacryloyl (GelMA) hydrogel. In detail, a series of hydrogels based on GelMA formulations containing distinct amounts of DEX-loaded nanotubes was analyzed for physicochemical and mechanical properties and kinetics of DEX release as well as compatibility with mesenchymal stem cells from human exfoliated deciduous teeth (SHEDs). The anti-inflammatory response and mineralization potential of the engineered hydrogels were determined in vitro and in vivo. DEX conjugation with HNTs was confirmed by FTIR analysis. The incorporation of DEX-loaded nanotubes enhanced the mechanical strength of GelMA with no effect on its degradation and swelling ratio. Scanning electron microscopy (SEM) images demonstrated the porous architecture of GelMA, which was not significantly altered by DEX-loaded nanotubes' (HNTs/DEX) incorporation. All GelMA formulations showed cytocompatibility with SHEDs (p < 0.05) regardless of the presence of HNTs or HNTs/DEX. However, the highest osteogenic cell differentiation was noticed with the addition of HNT/DEX 10% in GelMA formulations (p < 0.01). The controlled release of DEX over 7 days restored the expression of alkaline phosphatase and mineralization (p < 0.0001) in lipopolysaccharide (LPS)-stimulated SHEDs in vitro. Importantly, in vivo data revealed that DEX-loaded nanotube-modified GelMA (5.0% HNT/DEX 10%) led to enhanced bone formation after 6 weeks (p < 0.0001) compared to DEX-free formulations with a minimum localized inflammatory response after 7 days. Altogether, our findings show that the engineered DEX-loaded nanotube-modified hydrogel may possess great potential to trigger in situ mineralized tissue regeneration under inflammatory conditions.
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Affiliation(s)
- Ester A F Bordini
- Department of Cariology, Restorative Sciences, and Endodontics, School of Dentistry, University of Michigan, 1011 N. University Ave., Ann Arbor, Michigan 48109, United States
| | - Jessica A Ferreira
- Department of Cariology, Restorative Sciences, and Endodontics, School of Dentistry, University of Michigan, 1011 N. University Ave., Ann Arbor, Michigan 48109, United States
| | - Nileshkumar Dubey
- Department of Cariology, Restorative Sciences, and Endodontics, School of Dentistry, University of Michigan, 1011 N. University Ave., Ann Arbor, Michigan 48109, United States
| | - Juliana S Ribeiro
- Department of Cariology, Restorative Sciences, and Endodontics, School of Dentistry, University of Michigan, 1011 N. University Ave., Ann Arbor, Michigan 48109, United States
| | - Carlos A de Souza Costa
- Department of Physiology and Pathology, Araraquara School of Dentistry, Universidade Estadual Paulista (UNESP), 1680 Humaitá Street, Araraquara, Sao Paulo 14801-903, Brazil
| | - Diana G Soares
- Department of Operative Dentistry, Endodontics and Dental Materials, Bauru School of Dentistry, Sao Paulo University (USP), Al. Dr. Octavio Pinheiro Brizola, 9-75, Bauru, Sao Paulo 17012-901, Brazil
| | - Marco C Bottino
- Department of Cariology, Restorative Sciences, and Endodontics, School of Dentistry, University of Michigan, 1011 N. University Ave., Ann Arbor, Michigan 48109, United States.,Department of Biomedical Engineering, College of Engineering, University of Michigan, Carl A. Gerstacker Building, 2200 Bonisteel Blvd., Ann Arbor, Michigan 48109, United States
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27
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Sanz JL, Guerrero-Gironés J, Pecci-Lloret MP, Pecci-Lloret MR, Melo M. Biological interactions between calcium silicate-based endodontic biomaterials and periodontal ligament stem cells: A systematic review of in vitro studies. Int Endod J 2021; 54:2025-2043. [PMID: 34338339 DOI: 10.1111/iej.13600] [Citation(s) in RCA: 42] [Impact Index Per Article: 10.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/15/2021] [Revised: 07/16/2021] [Accepted: 07/29/2021] [Indexed: 02/07/2023]
Abstract
BACKGROUND Most recently, the biological interactions, that is cytocompatibility, cell differentiation and mineralization potential, between calcium silicate-based biomaterials and periodontal ligament stem cells (PDLSCs) have been studied at an in vitro level, in order to predict their clinical behaviour during endodontic procedures involving direct contact with periodontal tissues, namely root canal treatment, endodontic surgery and regenerative endodontic treatment. OBJECTIVE The aim of the present systematic review was to present a qualitative synthesis of available in vitro studies assessing the biological interaction of PDLSCs and calcium silicate-based biomaterials. METHODOLOGY The present review followed PRISMA 2020 guidelines. An advanced database search was performed in Medline, Scopus, Embase, Web of Science and SciELO on 1 July 2020 and last updated on 22 April 2021. Studies assessing the biological interactions of PDLSCs with calcium silicate-based sealers (CSSs) and/or cements (CSCs) at an in vitro level were considered for inclusion. The evaluation of the 'biological interaction' was defined as any assay or test on the cytotoxicity, cytocompatibility, cell plasticity or differentiation potential, and bioactive properties of PDLSCs cultured in CSC or CSS-conditioned media. Quality (risk of bias) was assessed using a modified CONSORT checklist for in vitro studies of dental materials. RESULTS A total of 20 studies were included for the qualitative synthesis. CSCs and CSSs, as a group of endodontic materials, exhibit adequate cytocompatibility and favour the osteo/cementogenic differentiation and mineralization potential of PDLSCs, as evidenced from the in vitro studies included in the present systematic review. DISCUSSION The influence of the compositional differences, inclusion of additives, sample preparation, and varying conditions and manipulations on the biological properties of calcium silicate-based materials remain a subject for future research. CONCLUSIONS Within the limitations of the in vitro nature of the included studies, this work supports the potential use of calcium silicate-based endodontic materials in stem cell therapy and biologically based regenerative endodontic procedures. REGISTRATION OSF Registries; https://doi.org/10.17605/OSF.IO/SQ9UY.
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Affiliation(s)
- José Luis Sanz
- Departament d'Estomatologia, Facultat de Medicina I Odontologia, Universitat de València, Valencia, Spain
| | - Julia Guerrero-Gironés
- Department of Dermatology, Stomatology, Radiology and Physical Medicine, Faculty of Medicine, Morales Meseguer Hospital, University of Murcia, Murcia, Spain
| | - María P Pecci-Lloret
- Department of Dermatology, Stomatology, Radiology and Physical Medicine, Faculty of Medicine, Morales Meseguer Hospital, University of Murcia, Murcia, Spain
| | - Miguel R Pecci-Lloret
- Department of Dermatology, Stomatology, Radiology and Physical Medicine, Faculty of Medicine, Morales Meseguer Hospital, University of Murcia, Murcia, Spain
| | - María Melo
- Departament d'Estomatologia, Facultat de Medicina I Odontologia, Universitat de València, Valencia, Spain
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28
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Mercado-Rubio MD, Pérez-Argueta E, Zepeda-Pedreguera A, Aguilar-Ayala FJ, Peñaloza-Cuevas R, Kú-González A, Rojas-Herrera RA, Rodas-Junco BA, Nic-Can GI. Similar Features, Different Behaviors: A Comparative In VitroStudy of the Adipogenic Potential of Stem Cells from Human Follicle, Dental Pulp, and Periodontal Ligament. J Pers Med 2021; 11:jpm11080738. [PMID: 34442382 PMCID: PMC8401480 DOI: 10.3390/jpm11080738] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/07/2021] [Revised: 07/24/2021] [Accepted: 07/24/2021] [Indexed: 12/21/2022] Open
Abstract
Dental tissue-derived mesenchymal stem cells (DT-MSCs) are a promising resource for tissue regeneration due to their multilineage potential. Despite accumulating data regarding the biology and differentiation potential of DT-MSCs, few studies have investigated their adipogenic capacity. In this study, we have investigated the mesenchymal features of dental pulp stem cells (DPSCs), as well as the in vitro effects of different adipogenic media on these cells, and compared them to those of periodontal ligament stem cells (PLSCs) and dental follicle stem cells (DFSCs). DFSC, PLSCs, and DPSCs exhibit similar morphology and proliferation capacity, but they differ in their self-renewal ability and expression of stemness markers (e.g OCT4 and c-MYC). Interestingly, DFSCs and PLSCs exhibited more lipid accumulation than DPSCs when induced to adipogenic differentiation. In addition, the mRNA levels of adipogenic markers (PPAR, LPL, and ADIPOQ) were significantly higher in DFSCs and PLSCs than in DPSCs, which could be related to the differences in the adipogenic commitment in those cells. These findings reveal that the adipogenic capacity differ among DT-MSCs, features that might be advantageous to increasing our understanding about the developmental origins and regulation of adipogenic commitment.
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Affiliation(s)
- Melissa D. Mercado-Rubio
- Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Periférico Norte Kilómetro 33.5, Tablaje Catastral 13615, Chuburná de Hidalgo Inn, Mérida 97203, Yucatán, Mexico; (M.D.M.-R.); (E.P.-A.); (A.Z.-P.); (R.A.R.-H.)
| | - Erick Pérez-Argueta
- Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Periférico Norte Kilómetro 33.5, Tablaje Catastral 13615, Chuburná de Hidalgo Inn, Mérida 97203, Yucatán, Mexico; (M.D.M.-R.); (E.P.-A.); (A.Z.-P.); (R.A.R.-H.)
| | - Alejandro Zepeda-Pedreguera
- Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Periférico Norte Kilómetro 33.5, Tablaje Catastral 13615, Chuburná de Hidalgo Inn, Mérida 97203, Yucatán, Mexico; (M.D.M.-R.); (E.P.-A.); (A.Z.-P.); (R.A.R.-H.)
| | - Fernando J. Aguilar-Ayala
- Laboratorio Translacional de Células Troncales-Facultad de Odontología, Universidad Autónoma de Yucatán, Calle 61-A X Av. Itzaes Costado Sur “Parque de la Paz”, Col. Centro, Mérida 97000, Yucatán, Mexico; (F.J.A.-A.); (R.P.-C.)
| | - Ricardo Peñaloza-Cuevas
- Laboratorio Translacional de Células Troncales-Facultad de Odontología, Universidad Autónoma de Yucatán, Calle 61-A X Av. Itzaes Costado Sur “Parque de la Paz”, Col. Centro, Mérida 97000, Yucatán, Mexico; (F.J.A.-A.); (R.P.-C.)
| | - Angela Kú-González
- Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán, Calle 43 No. 130, Col. Chuburná de Hidalgo, Mérida 97200, Yucatán, Mexico;
| | - Rafael A. Rojas-Herrera
- Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Periférico Norte Kilómetro 33.5, Tablaje Catastral 13615, Chuburná de Hidalgo Inn, Mérida 97203, Yucatán, Mexico; (M.D.M.-R.); (E.P.-A.); (A.Z.-P.); (R.A.R.-H.)
| | - Beatriz A. Rodas-Junco
- Laboratorio Translacional de Células Troncales-Facultad de Odontología, Universidad Autónoma de Yucatán, Calle 61-A X Av. Itzaes Costado Sur “Parque de la Paz”, Col. Centro, Mérida 97000, Yucatán, Mexico; (F.J.A.-A.); (R.P.-C.)
- CONACYT-Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Periférico Norte Kilómetro 33.5, Tablaje Catastral 13615, Chuburná de Hidalgo Inn, Mérida 97203, Yucatán, Mexico
- Correspondence: (B.A.R.-J.); or (G.I.N.-C.)
| | - Geovanny I. Nic-Can
- Laboratorio Translacional de Células Troncales-Facultad de Odontología, Universidad Autónoma de Yucatán, Calle 61-A X Av. Itzaes Costado Sur “Parque de la Paz”, Col. Centro, Mérida 97000, Yucatán, Mexico; (F.J.A.-A.); (R.P.-C.)
- CONACYT-Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Periférico Norte Kilómetro 33.5, Tablaje Catastral 13615, Chuburná de Hidalgo Inn, Mérida 97203, Yucatán, Mexico
- Correspondence: (B.A.R.-J.); or (G.I.N.-C.)
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PDLCs and EPCs Co-Cultured on Ta Discs: A Golden Fleece for "Compromised" Osseointegration. Int J Mol Sci 2021; 22:ijms22094486. [PMID: 33925774 PMCID: PMC8123461 DOI: 10.3390/ijms22094486] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/17/2021] [Revised: 04/09/2021] [Accepted: 04/19/2021] [Indexed: 12/04/2022] Open
Abstract
Material research in tissue engineering forms a vital link between basic cell research and animal research. Periodontal ligament cells (PDLCs, P) from the tooth have an osteogenic effect, whereas endothelial progenitor cells (EPCs, E) assist in neovascularization. In the present study, the interaction of PDLCs and EPCs with Tantalum (Ta, I) discs, either alone (IP or IE group) or in 1:1 (IPE) ratio was explored. Additionally, surface analysis of Ta discs with respect to different types and cycles of sterilization and disinfection procedures was evaluated. It was observed that Ta discs could be used for a maximum of three times, after which the changes in properties of Ta discs were detrimental to cell growth, irrespective of the type of the protocol. Cell-Disc’s analysis revealed that cell proliferation in the IE group at day 6 and day 10 was significantly higher (p < 0.05) than other groups. A cell viability assay revealed increased live cells in the IPE group than in the IP or IE group. Similarly, adhesion and penetration of cells in the IPE group were not only higher, but also had an increased thickness of cellular extensions. RT-PCR analysis revealed that on day 8, both osteogenic (ALP, RUNX-2, and BSP) and angiogenic genes (VEGFR-2, CD31) increased significantly in the IPE group as compared to the IP or IE groups (p < 0.05). In conclusion, Ta discs promoted cellular proliferation and increased osteogenic and angiogenic activity by augmenting RUNX-2 and VEGFR-2 activity.
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Zhao D, Wang X, Tie C, Cheng B, Yang S, Sun Z, Yin M, Li X, Yin M. Bio-functional strontium-containing photocrosslinked alginate hydrogels for promoting the osteogenic behaviors. MATERIALS SCIENCE & ENGINEERING. C, MATERIALS FOR BIOLOGICAL APPLICATIONS 2021; 126:112130. [PMID: 34082947 DOI: 10.1016/j.msec.2021.112130] [Citation(s) in RCA: 16] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/06/2021] [Revised: 04/05/2021] [Accepted: 04/20/2021] [Indexed: 12/23/2022]
Abstract
In recent years, photocrosslinked alginate hydrogel has been widely studied in bone tissue engineering, owing to its numerous advantages. However, there are still some shortcomings like insufficient mechanical strength and lack of bone induction. To compensate for these deficiencies, in this work, a novel doped strontium (Sr) photocrosslinked methacrylated alginate (Sr-PMA) hydrogel was developed. Photocrosslinked alginate hydrogel fabricated via crosslinking methacrylate-modified alginate under ultraviolet (UV) light was placed into strontium solutions to prepare Sr-PMA gel by chelating reaction. The chemical structures, swelling behaviors, degradation profiles, elastic moduli, Sr2+ ion release and surface morphology of the Sr-PMA hydrogel were characterized, and we found that physical properties of the gels can be tailored by varying concentration of Sr2+ ions. And MC3T3-E1 cell viability, proliferation and mineralization outside the hydrogel were also investigated. Further research on cell survival, multiplication, osteogenic differentiation of the cells encapsulated in Sr-PMA hydrogels were explored. In vitro studies of biological properties revealed that incorporation of Sr2+ into photocrosslinked alginate gels significantly improved osteogenic differentiation capabilities and mineralization via stimulating expression of osteogenesis related genes and proteins of the cells compared to strontium-free photocrosslinked alginate gels. The research demonstrates that the innovative Sr-PMA hydrogels possessing adjustable physical performances, excellent biocompatibility and osteogenic differentiation capabilities could be potentially applied to bone tissue engineering and regenerative medicine. Meanwhile, it also provides a reference for the modification of biological properties of biomaterials.
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Affiliation(s)
- Delu Zhao
- Hubei Tumor Biological Behavior Key Laboratory, Center of stomatology, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China; Department of Prosthodontics, Hefei Stomatological Clinic Hospital, Anhui Medical University, & Hefei Stomatological Hospital, Hefei 230001, Anhui Province, China
| | - Xin Wang
- Hubei Tumor Biological Behavior Key Laboratory, Center of stomatology, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China
| | - Chaorong Tie
- Hubei Tumor Biological Behavior Key Laboratory, Center of stomatology, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China
| | - Bo Cheng
- Hubei Tumor Biological Behavior Key Laboratory, Center of stomatology, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China
| | - Sisi Yang
- Hubei Tumor Biological Behavior Key Laboratory, Center of stomatology, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China
| | - Zhen Sun
- Hubei Tumor Biological Behavior Key Laboratory, Center of stomatology, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China
| | - Miaomiao Yin
- Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Sauvage Center for Molecular Sciences, College of Chemistry and Molecular Science, Wuhan University, Wuhan 430072, Hubei Province, China
| | - Xiaobao Li
- Department of Stomatology, Affiliated Wuhan Children's Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, Hubei Province, China
| | - Miao Yin
- Hubei Tumor Biological Behavior Key Laboratory, Center of stomatology, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China.
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Assessment of a PCL-3D Printing-Dental Pulp Stem Cells Triplet for Bone Engineering: An In Vitro Study. Polymers (Basel) 2021; 13:polym13071154. [PMID: 33916576 PMCID: PMC8038447 DOI: 10.3390/polym13071154] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/07/2021] [Revised: 03/29/2021] [Accepted: 04/01/2021] [Indexed: 12/18/2022] Open
Abstract
The search of suitable combinations of stem cells, biomaterials and scaffolds manufacturing methods have become a major focus of research for bone engineering. The aim of this study was to test the potential of dental pulp stem cells to attach, proliferate, mineralize and differentiate on 3D printed polycaprolactone (PCL) scaffolds. A 100% pure Mw: 84,500 ± 1000 PCL was selected. 5 × 10 × 5 mm3 parallelepiped scaffolds were designed as a wood-pilled structure composed of 20 layers of 250 μm in height, in a non-alternate order ([0,0,0,90,90,90°]). 3D printing was made at 170 °C. Swine dental pulp stem cells (DPSCs) were extracted from lower lateral incisors of swine and cultivated until the cells reached 80% confluence. The third passage was used for seeding on the scaffolds. Phenotype of cells was determined by flow Cytometry. Live and dead, Alamar blue™, von Kossa and alizarin red staining assays were performed. Scaffolds with 290 + 30 μm strand diameter, 938 ± 80 μm pores in the axial direction and 689 ± 13 μm pores in the lateral direction were manufactured. Together, cell viability tests, von Kossa and Alizarin red staining indicate the ability of the printed scaffolds to support DPSCs attachment, proliferation and enable differentiation followed by mineralization. The selected material-processing technique-cell line (PCL-3D printing-DPSCs) triplet can be though to be used for further modelling and preclinical experiments in bone engineering studies.
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Al Natour B, Lundy FT, Moynah PN, About I, Jeanneau C, Irwin CR, Domberoski Y, El Karim IA. Odontoblast cell death induces NLRP3 inflammasome-dependent sterile inflammation and regulates dental pulp cell migration, proliferation and differentiation. Int Endod J 2021; 54:941-950. [PMID: 33503274 DOI: 10.1111/iej.13483] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/29/2020] [Accepted: 01/25/2021] [Indexed: 12/13/2022]
Abstract
AIM To investigate the ability of dead odontoblasts to initiate NLRP3 inflammasome-dependent sterile inflammation and to explore the effect on dental pulp cell (DPCs) migration, proliferation and odontogenic differentiation. METHODS Odontoblast-like cells were subjected to freezing-thawing cycles to produce odontoblast necrotic cell lysate (ONCL). DPCs were treated with ONCL to assess proliferation and migration. THP-1 differentiated macrophages stimulated with ONCL and live cell imaging and western blotting were used to assess NLRP3 inflammasome activation. Cytokines were measured with multiplex arrays and ELISA. qPCR, alkaline phosphatase and Alizarin red assays were used to assess odontogenic differentiation of DPCs. Data were analysed using the t-test or anova followed by a Bonferroni post hoc test with the level of significance set at P ≤ 0.05. RESULTS ONCL induced migration and proliferation of DPCs. Treatment of THP-1 macrophages with ONCL resulted in the release of the inflammatory cytokines IL-1β, IL-6, IL-8, TNFα, IFN-γ, CCL2 and angiogenic growth factors, angiogenin and angiopoietin. This inflammatory response was associated with activation of NFκB, p38MAPK and NLRP3 inflammasome. To confirm that ONCL induced inflammatory response is NLRP3 inflammasome-dependent, treatment with a caspase-1 inhibitor and a specific NLRP3 inhibitor significantly reduced IL-1β release in THP-1 macrophages (P = 0.01 and 0.001). Inflammasome activation product, IL-1β, induced odontogenic differentiation of DPCS as evident by the increase in odontogenic genes expression DMP-1, RUNX-2, DSPP and SPP, alkaline phosphatase activity and mineralization. CONCLUSION Dead odontoblasts induced NLRP3 inflammasome-dependent sterile inflammation and activated the migration, proliferation and differentiation of DPCs.
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Affiliation(s)
- B Al Natour
- School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK.,Department of Oral Medicine and Oral Surgery, Faculty of Dentistry, Jordan University of Science and Technology, Irbid, Jordan
| | - F T Lundy
- School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK
| | - P N Moynah
- School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK.,Department of Biology, The Kathleen Lonsdale Institute for Human Health Research, National University of Ireland Maynooth, Maynooth, Ireland
| | - I About
- UMR 7287 CNRS, Faculté d'Odontologie, Université d'Aix-Marseille, Marseille, France
| | - C Jeanneau
- UMR 7287 CNRS, Faculté d'Odontologie, Université d'Aix-Marseille, Marseille, France
| | - C R Irwin
- School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK
| | - Y Domberoski
- School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK
| | - I A El Karim
- School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK
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Berbéri A, Fayyad-Kazan M, Ayoub S, Bou Assaf R, Sabbagh J, Ghassibe-Sabbagh M, Badran B. Osteogenic potential of dental and oral derived stem cells in bone tissue engineering among animal models: An update. Tissue Cell 2021; 71:101515. [PMID: 33657504 DOI: 10.1016/j.tice.2021.101515] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/21/2020] [Revised: 02/21/2021] [Accepted: 02/21/2021] [Indexed: 12/20/2022]
Abstract
Small bone defects can heal spontaneously through the bone modeling process due to their physiological environmental conditions. The bone modeling cycle preserves the reliability of the skeleton through the well-adjusted activities of its fundamental cell. Stem cells are a source of pluripotent cells with a capacity to differentiate into any tissue in the existence of a suitable medium. The concept of bone engineering is based on stem cells that can differentiate into bone cells. Mesenchymal stromal cells have been evaluated in bone tissue engineering due to their capacity to differentiate in osteoblasts. They can be isolated from bone marrow and from several adults oral and dental tissues such as permanent or deciduous teeth dental pulp, periodontal ligament, apical dental papilla, dental follicle precursor cells usually isolated from the follicle surrounding the third molar, gingival tissue, periosteum-derived cells, dental alveolar socket, and maxillary sinus Schneiderian membrane-derived cells. Therefore, a suitable animal model is a crucial step, as preclinical trials, to study the outcomes of mesenchymal cells on the healing of bone defects. We will discuss, through this paper, the use of mesenchymal stem cells obtained from several oral tissues mixed with different types of scaffolds tested in different animal models for bone tissue engineering. We will explore and link the comparisons between human and animal models and emphasized the factors that we need to take into consideration when choosing animals. The pig is considered as the animal of choice when testing large size and multiple defects for bone tissue engineering.
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Affiliation(s)
- Antoine Berbéri
- Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Lebanese University, Beirut, Lebanon.
| | - Mohammad Fayyad-Kazan
- Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, Beirut, Lebanon; Laboratory of Cancer Biology and Molecular Immunology, Faculty of Sciences-I, Lebanese University, Hadath- Beirut, Lebanon.
| | - Sara Ayoub
- Department of Prosthodontics, Faculty of Dentistry, Lebanese University, Beirut, Lebanon.
| | - Rita Bou Assaf
- Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Lebanese University, Beirut, Lebanon.
| | - Joseph Sabbagh
- Department of Restorative Dentistry and Endodontics, Faculty of Dental Medicine, Lebanese University, Beirut, Lebanon.
| | - Michella Ghassibe-Sabbagh
- Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, Beirut, Lebanon.
| | - Bassam Badran
- Laboratory of Cancer Biology and Molecular Immunology, Faculty of Sciences-I, Lebanese University, Hadath- Beirut, Lebanon.
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Zhang W, Jia L, Zhao B, Xiong Y, Wang YN, Liang J, Xu X. Quercetin reverses TNF‑α induced osteogenic damage to human periodontal ligament stem cells by suppressing the NF‑κB/NLRP3 inflammasome pathway. Int J Mol Med 2021; 47:39. [PMID: 33537804 PMCID: PMC7891819 DOI: 10.3892/ijmm.2021.4872] [Citation(s) in RCA: 21] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/16/2020] [Accepted: 01/21/2021] [Indexed: 11/17/2022] Open
Abstract
Quercetin (Quer) is a typical antioxidant flavonoid from plants that is involved in bone metabolism, as well as in the progression of inflammatory diseases. Elevated levels of tumor necrosis factor-α (TNF-α), a typical pro-inflammatory cytokine, can affect osteogenesis. In the present study, TNF-α was used to establish an in vitro model of periodontitis. The effects of Quer on, as well as its potential role in the osteogenic response of human periodontal ligament stem cells (hPDLSCs) under TNF-α-induced inflammatory conditions and the underlying mechanisms were then investigated. Within the appropriate concentration range, Quer did not exhibit any cytotoxicity. More importantly, Quer significantly attenuated the TNF-α induced the suppression of osteogenesis-related genes and proteins, alkaline phosphatase (ALP) activity and mineralized matrix in the hPDLSCs. These findings were associated with the fact that Quer inhibited the activation of the NF-κB signaling pathway, as well as the expression of NLRP3 inflammation-associated proteins in the inflammatory microenvironment. Moreover, the silencing of NLRP3 by small interfering RNA (siRNA) was found to protect the hPDLSCs against TNF-α-induced osteogenic damage, which was in accordance with the effects of Quer. On the whole, the present study demonstrates that Quer reduces the impaired osteogenesis of hPDLSCs under TNF-α-induced inflammatory conditions by inhibiting the NF-κB/NLRP3 inflammasome pathway. Thus, Quer may prove to be a potential remedy against periodontal bone defects.
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Affiliation(s)
- Wenjing Zhang
- School of Stomatology, Shandong University, Jinan, Shandong 250012, P.R. China
| | - Linglu Jia
- School of Stomatology, Shandong University, Jinan, Shandong 250012, P.R. China
| | - Bin Zhao
- School of Stomatology, Shandong University, Jinan, Shandong 250012, P.R. China
| | - Yixuan Xiong
- School of Stomatology, Shandong University, Jinan, Shandong 250012, P.R. China
| | - Ya-Nan Wang
- School of Stomatology, Shandong University, Jinan, Shandong 250012, P.R. China
| | - Jin Liang
- School of Stomatology, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan, Shandong 250117, P.R. China
| | - Xin Xu
- School of Stomatology, Shandong University, Jinan, Shandong 250012, P.R. China
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Sordi MB, Curtarelli RB, da Silva IT, Fongaro G, Benfatti CAM, de Souza Magini R, Cabral da Cruz AC. Effect of dexamethasone as osteogenic supplementation in in vitro osteogenic differentiation of stem cells from human exfoliated deciduous teeth. JOURNAL OF MATERIALS SCIENCE. MATERIALS IN MEDICINE 2021; 32:1. [PMID: 33469820 PMCID: PMC7815568 DOI: 10.1007/s10856-020-06475-6] [Citation(s) in RCA: 23] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/20/2020] [Accepted: 12/10/2020] [Indexed: 05/05/2023]
Abstract
In in vitro culture systems, dexamethasone (DEX) has been applied with ascorbic acid (ASC) and β-glycerophosphate (βGLY) as culture media supplementation to induce osteogenic differentiation of mesenchymal stem cells. However, there are some inconsistencies regarding the role of DEX as osteogenic media supplementation. Therefore, this study verified the influence of DEX culture media supplementation on the osteogenic differentiation, especially the capacity to mineralize the extracellular matrix of stem cells from human exfoliated deciduous teeth (SHED). Five groups were established: G1-SHED + Dulbecco's Modified Eagles' Medium (DMEM) + fetal bovine serum (FBS); G2-SHED + DMEM + FBS + DEX; G3-SHED + DMEM + FBS + ASC + βGLY; G4-SHED + DMEM + FBS + ASC + βGLY + DEX; G5-MC3T3-E1 + α Minimal Essential Medium (MEM) + FBS + ASC + βGLY. DNA content, alkaline phosphatase (ALP) activity, free calcium quantification in the extracellular medium, and extracellular matrix mineralization quantification through staining with von Kossa, alizarin red, and tetracycline were performed on days 7 and 21. Osteogenic media supplemented with ASC and β-GLY demonstrated similar effects on SHED in the presence or absence of DEX for DNA content (day 21) and capacity to mineralize the extracellular matrix according to alizarin red and tetracycline quantifications (day 21). In addition, the presence of DEX in the osteogenic medium promoted less ALP activity (day 7) and extracellular matrix mineralization according to the von Kossa assay (day 21), and more free calcium quantification at extracellular medium (day 21). In summary, the presence of DEX in the osteogenic media supplementation did not interfere with SHED commitment into mineral matrix depositor cells. We suggest that DEX may be omitted from culture media supplementation for SHED osteogenic differentiation in vitro studies.
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Affiliation(s)
- Mariane Beatriz Sordi
- Center for Research on Dental Implants, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil
- Laboratory of Applied Virology, Federal University of Santa Catarina, Henrique da Silva Fontes Avenue, Florianópolis, 88040-900, Brazil
| | - Raissa Borges Curtarelli
- Center for Research on Dental Implants, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil
- Laboratory of Applied Virology, Federal University of Santa Catarina, Henrique da Silva Fontes Avenue, Florianópolis, 88040-900, Brazil
| | - Izabella Thaís da Silva
- Laboratory of Applied Virology, Federal University of Santa Catarina, Henrique da Silva Fontes Avenue, Florianópolis, 88040-900, Brazil
- Department of Pharmaceutics Science, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil
| | - Gislaine Fongaro
- Laboratory of Applied Virology, Federal University of Santa Catarina, Henrique da Silva Fontes Avenue, Florianópolis, 88040-900, Brazil
- Department of Microbiology, Immunology, and Parasitology, Federal University of Santa Catarina, Henrique da Silva Fontes Avenue, Florianópolis, 88040-900, Brazil
| | - Cesar Augusto Magalhães Benfatti
- Center for Research on Dental Implants, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil
- Department of Dentistry, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil
| | - Ricardo de Souza Magini
- Center for Research on Dental Implants, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil
- Department of Dentistry, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil
| | - Ariadne Cristiane Cabral da Cruz
- Center for Research on Dental Implants, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil.
- Laboratory of Applied Virology, Federal University of Santa Catarina, Henrique da Silva Fontes Avenue, Florianópolis, 88040-900, Brazil.
- Department of Dentistry, Federal University of Santa Catarina, Delfino Conti Street, Florianópolis, 88040-900, Brazil.
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Ercal P, Pekozer GG. A Current Overview of Scaffold-Based Bone Regeneration Strategies with Dental Stem Cells. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2020; 1288:61-85. [PMID: 32185698 DOI: 10.1007/5584_2020_505] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
Bone defects due to trauma or diseases still pose a clinical challenge to be resolved in the current tissue engineering approaches. As an alternative to traditional methods to restore bone defects, such as autografts, bone tissue engineering aims to achieve new bone formation via novel biomaterials used in combination with multipotent stem cells and bioactive molecules. Mesenchymal stem cells (MSCs) can be successfully isolated from various dental tissues at different stages of development including dental pulp, apical papilla, dental follicle, tooth germ, deciduous teeth, periodontal ligament and gingiva. A wide range of biomaterials including polymers, ceramics and composites have been investigated for their potential as an ideal bone scaffold material. This article reviews the properties and the manufacturing methods of biomaterials used in bone tissue engineering, and provides an overview of bone tissue regeneration approaches of scaffold and dental stem cell combinations as well as their limitations.
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Affiliation(s)
- Pınar Ercal
- Faculty of Dentistry, Department of Oral Surgery, Altinbas University, Istanbul, Turkey.
| | - Gorke Gurel Pekozer
- Faculty of Electrical and Electronics Engineering, Department of Biomedical Engineering, Yıldız Technical University, Istanbul, Turkey
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Schlesinger PH, Braddock DT, Larrouture QC, Ray EC, Riazanski V, Nelson DJ, Tourkova IL, Blair HC. Phylogeny and chemistry of biological mineral transport. Bone 2020; 141:115621. [PMID: 32858255 PMCID: PMC7771281 DOI: 10.1016/j.bone.2020.115621] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/26/2020] [Revised: 08/24/2020] [Accepted: 08/24/2020] [Indexed: 02/08/2023]
Abstract
Three physiologically mineralizing tissues - teeth, cartilage and bone - have critical common elements and important evolutionary relationships. Phylogenetically the most ancient densely mineralized tissue is teeth. In jawless fishes without skeletons, tooth formation included epithelial transport of phosphates, a process echoed later in bone physiology. Cartilage and mineralized cartilage are skeletal elements separate from bone, but with metabolic features common to bone. Cartilage mineralization is coordinated with high expression of tissue nonspecific alkaline phosphatase and PHOSPHO1 to harvest available phosphate esters and support mineralization of collagen secreted locally. Mineralization in true bone results from stochastic nucleation of hydroxyapatite crystals within the cross-linked collagen fibrils. Mineral accumulation in dense collagen is, at least in major part, mediated by amorphous aggregates - often called Posner clusters - of calcium and phosphate that are small enough to diffuse into collagen fibrils. Mineral accumulation in membrane vesicles is widely suggested, but does not correlate with a definitive stage of mineralization. Conversely mineral deposition at non-physiologic sites where calcium and phosphate are adequate has been shown to be regulated in large part by pyrophosphate. All of these elements are present in vertebrate bone metabolism. A key biological element of bone formation is an epithelial-like cellular organization which allows control of phosphate, calcium and pH during mineralization.
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Affiliation(s)
- Paul H Schlesinger
- Dept of Cell Biology, Washington University, Saint Louis, MO, United States of America
| | - Demetrios T Braddock
- Dept. of Pathology, Yale New Haven Hospital, 310 Cedar Street, New Haven, CT, United States of America
| | - Quitterie C Larrouture
- Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, Botnar Research Centre, Windmill Road, Oxford OX3 7LD, UK
| | - Evan C Ray
- Renal Electrolyte Division, Department of Medicine, University of Pittsburgh, Pittsburgh, PA, United States of America
| | - Vladimir Riazanski
- Dept of Neurobiology, Pharmacology & Physiology, University of Chicago, Chicago, IL, United States of America
| | - Deborah J Nelson
- Dept of Neurobiology, Pharmacology & Physiology, University of Chicago, Chicago, IL, United States of America
| | - Irina L Tourkova
- Veteran's Affairs Medical Center, Pittsburgh PA and Department of Pathology, University of Pittsburgh, Pittsburgh, PA, United States of America
| | - Harry C Blair
- Veteran's Affairs Medical Center, Pittsburgh PA and Department of Pathology, University of Pittsburgh, Pittsburgh, PA, United States of America.
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Sanz JL, Forner L, Llena C, Guerrero-Gironés J, Melo M, Rengo S, Spagnuolo G, Rodríguez-Lozano FJ. Cytocompatibility and Bioactive Properties of Hydraulic Calcium Silicate-Based Cements (HCSCs) on Stem Cells from Human Exfoliated Deciduous Teeth (SHEDs): A Systematic Review of In Vitro Studies. J Clin Med 2020; 9:jcm9123872. [PMID: 33260782 PMCID: PMC7761433 DOI: 10.3390/jcm9123872] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/03/2020] [Revised: 11/20/2020] [Accepted: 11/25/2020] [Indexed: 02/06/2023] Open
Abstract
The implementation of hydraulic calcium silicate-based endodontic cements (HCSCs) in biologically based endodontic procedures for the primary dentition has been recently investigated, focusing on the biological response of stem cells from human exfoliated deciduous teeth (SHEDs) towards them. The present systematic review aimed to present a qualitative synthesis of the available literature consisting of in vitro assays, which assessed the cytocompatibility and bioactive properties of HCSCs in direct contact with SHEDs. Following the PRISMA statement, an electronic database search was carried out in Medline, Scopus, Embase, Web of Science, and SciELO on March 31st and updated on November 16th, 2020. In vitro studies evaluating the biological response of SHEDs to the treatment with HCSCs were eligible. Within the term biological response, assays assessing the cytocompatibility (i.e., cell viability, migration, proliferation), cell plasticity or differentiation (i.e., osteo/odontogenic marker expression), and bioactivity or biomineralization (i.e., mineralized nodule formation) were included. A total of seven studies were included after the selection process. The study sample comprised an extensive range of cell viability, migration, proliferation, adhesion, and bioactivity assays regarding the biological response of SHEDs towards five different commercially available HCSCs (MTA, ProRoot MTA, Biodentine, iRoot BP Plus, and Theracal LC). Biodentine, MTA, and iRoot BP Plus showed significant positive results in cytocompatibility and bioactivity assays when cultured with SHEDs. The results from in vitro assays assessing the cytocompatibility and bioactivity of the HCSCs MTA, Biodentine, and iRoot BP Plus towards SHEDs support their use in vital pulp treatment for the primary dentition.
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Affiliation(s)
- José Luis Sanz
- Departament d’Estomatologia, Facultat de Medicina I Odontologia, Universitat de València, 46010 Valencia, Spain; (J.L.S.); (C.L.); (M.M.)
| | - Leopoldo Forner
- Departament d’Estomatologia, Facultat de Medicina I Odontologia, Universitat de València, 46010 Valencia, Spain; (J.L.S.); (C.L.); (M.M.)
- Correspondence: ; Tel.: +34-963864175
| | - Carmen Llena
- Departament d’Estomatologia, Facultat de Medicina I Odontologia, Universitat de València, 46010 Valencia, Spain; (J.L.S.); (C.L.); (M.M.)
| | - Julia Guerrero-Gironés
- Cellular Therapy and Hematopoietic Transplant Research Group, Biomedical Research Institute, Virgen de la Arrixaca Clinical University Hospital, IMIB-Arrixaca, University of Murcia, 30120 Murcia, Spain; (J.G.-G.); (F.J.R.-L.)
- Department of Dermatology, Stomatology, Radiology and Physical Medicine, Morales Meseguer Hospital, Faculty of Medicine, University of Murcia, 30100 Murcia, Spain
| | - María Melo
- Departament d’Estomatologia, Facultat de Medicina I Odontologia, Universitat de València, 46010 Valencia, Spain; (J.L.S.); (C.L.); (M.M.)
| | - Sandro Rengo
- Department of Neurosciences, Reproductive and Odontostomatological Sciences, University of Naples “Federico II”, 80138 Napoli, Italy; (S.R.); (G.S.)
| | - Gianrico Spagnuolo
- Department of Neurosciences, Reproductive and Odontostomatological Sciences, University of Naples “Federico II”, 80138 Napoli, Italy; (S.R.); (G.S.)
- Institute of Dentistry, I. M. Sechenov First Moscow State Medical University, Moscow 119146, Russia
| | - Francisco Javier Rodríguez-Lozano
- Cellular Therapy and Hematopoietic Transplant Research Group, Biomedical Research Institute, Virgen de la Arrixaca Clinical University Hospital, IMIB-Arrixaca, University of Murcia, 30120 Murcia, Spain; (J.G.-G.); (F.J.R.-L.)
- Department of Dermatology, Stomatology, Radiology and Physical Medicine, Morales Meseguer Hospital, Faculty of Medicine, University of Murcia, 30100 Murcia, Spain
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Perczel-Kovách K, Hegedűs O, Földes A, Sangngoen T, Kálló K, Steward MC, Varga G, Nagy KS. STRO-1 positive cell expansion during osteogenic differentiation: A comparative study of three mesenchymal stem cell types of dental origin. Arch Oral Biol 2020; 122:104995. [PMID: 33278647 DOI: 10.1016/j.archoralbio.2020.104995] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/17/2020] [Revised: 10/31/2020] [Accepted: 11/16/2020] [Indexed: 02/08/2023]
Abstract
OBJECTIVE Although the osteogenic differentiation potential of mesenchymal stem cells of dental origin is well established, the roles of different marker proteins in this process remain to be clarified. Our aim was to compare the cellular and molecular changes, focusing in particular on mesenchymal stem cell markers, during in vitro osteogenesis in three dental stem cell types: dental follicle stem cells (DFSCs), periodontal ligament stem cells (PDLSCs) and dental pulp stem cells (DPSCs). DESIGN Human DFSCs, PDLSCs and DPSCs were isolated, cultured and their osteogenic differentiation was induced for 3 weeks. Mineralization was assessed by von Kossa staining and calcium concentration measurements. The expression of mesenchymal and osteogenic markers was studied by immunocytochemistry and qPCR techniques. Alkaline phosphatase (ALP) activity and the frequency of STRO-1 positive cells were also quantified. RESULTS The three cultures all showed abundant mineralization, with high calcium content by day 21. The expression of vimentin and nestin was sustained after osteogenic induction. The osteogenic medium induced a considerable elevation of STRO-1 positive cells. By day 7, the ALP mRNA level had increased more than 100-fold in DFSCs, PDLSCs, and DPSCs. Quantitative PCR results indicated dissimilarities of osteoblastic marker levels in the three dental stem cell cultures. CONCLUSIONS DFSCs, PDLSCs and DPSCs have similar functional osteogenic differentiation capacities although their expressional profiles of key osteogenic markers show considerable variations. The STRO-1 positive cell fraction expands during osteogenic differentiation while vimentin and nestin expression remain high. For identification of stemness, functional studies rather than marker expressions are needed.
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Affiliation(s)
- Katalin Perczel-Kovách
- Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary.
| | - Orsolya Hegedűs
- Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary.
| | - Anna Földes
- Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary.
| | - Thanyaporn Sangngoen
- Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary.
| | - Karola Kálló
- Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary
| | - Martin C Steward
- Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary; School of Medical Sciences, University of Manchester, Manchester, United Kingdom.
| | - Gábor Varga
- Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary.
| | - Krisztina S Nagy
- Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary.
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Spatial Distributions, Characteristics, and Applications of Craniofacial Stem Cells. Stem Cells Int 2020; 2020:8868593. [PMID: 32908545 PMCID: PMC7475745 DOI: 10.1155/2020/8868593] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/20/2020] [Revised: 07/29/2020] [Accepted: 08/01/2020] [Indexed: 02/05/2023] Open
Abstract
Stem cells play an irreplaceable role in the development, homeostasis, and regeneration of the craniofacial bone. Multiple populations of tissue-resident craniofacial skeletal stem cells have been identified in different stem cell niches, including the cranial periosteum, jawbone marrow, temporomandibular joint, cranial sutures, and periodontium. These cells exhibit self-renewal and multidirectional differentiation abilities. Here, we summarized the properties of craniofacial skeletal stem cells, based on their spatial distribution. Specifically, we focused on the in vivo genetic fate mapping of stem cells, by exploring specific stem cell markers and observing their lineage commitment in both the homeostatic and regenerative states. Finally, we discussed their application in regenerative medicine.
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Thanasrisuebwong P, Kiattavorncharoen S, Surarit R, Phruksaniyom C, Ruangsawasdi N. Red and Yellow Injectable Platelet-Rich Fibrin Demonstrated Differential Effects on Periodontal Ligament Stem Cell Proliferation, Migration, and Osteogenic Differentiation. Int J Mol Sci 2020; 21:ijms21145153. [PMID: 32708242 PMCID: PMC7404021 DOI: 10.3390/ijms21145153] [Citation(s) in RCA: 26] [Impact Index Per Article: 5.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/04/2020] [Revised: 07/14/2020] [Accepted: 07/20/2020] [Indexed: 02/07/2023] Open
Abstract
The biological benefits of using two fractions derived from injectable platelet-rich fibrin (i-PRF) in bone regeneration remain unclear. Thus, the current study examined two fractionation protocols producing yellow i-PRF and red i-PRF on periodontal ligament stem cells (PDLSCs). The i-PRF samples from five donors were harvested from two different levels, with and without a buffy coat layer, to obtain red and yellow i-PRF, respectively. The PDLSCs were isolated and characterized before their experimental use. The culture medium in each assay was loaded with 20% of the conditioned medium containing the factors released from the red and yellow i-PRF. Cell proliferation and cell migration were determined with an MTT and trans-well assay, respectively. Osteogenic differentiation was investigated using alkaline phosphatase and Alizarin red staining. The efficiency of both i-PRFs was statistically compared. We found that the factors released from the red i-PRF had a greater effect on cell proliferation and cell migration. Moreover, the factors released from the yellow i-PRF stimulated PDLSC osteogenic differentiation earlier compared with the red i-PRF. These data suggest that the red i-PRF might be suitable for using in bone regeneration because it induced the mobilization and growth of bone regenerative cells without inducing premature mineralization.
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Affiliation(s)
- Prakan Thanasrisuebwong
- Dental Implant Center, Dental Hospital, Faculty of Dentistry, Mahidol University, Bangkok 10400, Thailand;
| | - Sirichai Kiattavorncharoen
- Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Mahidol University, Bangkok 10400, Thailand;
| | - Rudee Surarit
- Department of Oral Biology, Faculty of Dentistry, Mahidol University, Bangkok 10400, Thailand;
| | - Chareerut Phruksaniyom
- Department of Pharmacology, Faculty of Dentistry, Mahidol University, Bangkok 10400, Thailand;
| | - Nisarat Ruangsawasdi
- Department of Pharmacology, Faculty of Dentistry, Mahidol University, Bangkok 10400, Thailand;
- Correspondence:
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Cheng Q, Lin J, Chen Q, Zheng L, Tang Y, Wang F, Huang X, Zhang Y, Li S, Yang Z, Zhou P, He TC, Luo W, Zhang H. Role of Special AT-Rich Sequence-Binding Protein 2 in the Osteogenesis of Human Dental Mesenchymal Stem Cells. Stem Cells Dev 2020; 29:1059-1072. [PMID: 32484035 DOI: 10.1089/scd.2020.0013] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/06/2023] Open
Abstract
Dental mesenchymal stem cells (MSCs) are recognized as a critical factor in repair of defective craniofacial bone owing to the multiple differentiation potential, the ability to regenerate distinct tissues, and the advantage that they can be easily obtained by relatively noninvasive procedures. Special AT-rich sequence-binding protein 2 (SATB2) is a nuclear matrix protein, involved in chromatin remodeling and transcriptional regulation, and has been reported to be as a positive regulator of osteoblast differentiation, bone formation, and bone regeneration in MSCs. In this study, we systematically investigated the capability of SATB2 to promote the osteogenic differentiation of periodontal ligament stem cells (PDLSCs), dental pulp stem cells (DPSCs), and stem cells from human exfoliated deciduous teeth (SHED). RNA-seq analysis and quantitative real-time PCR (RT-PCR) revealed that genes regulating osteogenic differentiation were differentially expressed among three cell types and SATB2 was found to be expressed at a relatively high level. When the three cell types overexpressed SATB2 with AdSATB2 infection, alkaline phosphatase (ALP) staining, ALP activity, Alizarin Red S staining, and quantification tended to increase with an increasing infection rate. It showed opposite results after infection with AdsiSATB2. RNA-seq analysis indicated that the expression of downstream osteogenic genes was affected by AdSATB2 infection and quantitative RT-PCR confirmed that nine osteogenic genes (Spp1, Sema7a, Atf4, Ibsp, Col1a1, Sp7, Igfbp3, Dlx3, and Alpl) were upregulated, to various extents, following SATB2 overexpression. In addition, quantitative PCR results indicated that SATB2 affected the expression of MSC markers. These results suggested an important role of SATB2 in the osteogenesis of PDLSCs, DPSCs, and SHED. Further research is warranted to investigate SATB2-mediated regulation of osteogenic differentiation and to evaluate the therapeutic use of SATB2 for the regeneration of defective craniofacial bone tissue.
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Affiliation(s)
- Qianyu Cheng
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
| | - Juhong Lin
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
| | - Qiuman Chen
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
| | - Liwen Zheng
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
| | - Yingying Tang
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
| | - Feilong Wang
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
| | - Xia Huang
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing, China
| | - Yuxin Zhang
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
| | - Shuang Li
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
| | - Zhuohui Yang
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
| | - Pengfei Zhou
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing, China
| | - Tong-Chuan He
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, Illinois, USA
| | - Wenping Luo
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing, China
| | - Hongmei Zhang
- Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, The Affiliated Hospital of Stomatology of Chongqing Medical University, Chongqing, China.,Department of Pediatric Dentistry, The Affiliated Stomatology Hospital, Chongqing Medical University, Chongqing, China
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Sabbagh J, Ghassibe-Sabbagh M, Fayyad-Kazan M, Al-Nemer F, Fahed JC, Berberi A, Badran B. Differences in osteogenic and odontogenic differentiation potential of DPSCs and SHED. J Dent 2020; 101:103413. [PMID: 32585262 DOI: 10.1016/j.jdent.2020.103413] [Citation(s) in RCA: 30] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/28/2020] [Revised: 06/15/2020] [Accepted: 06/18/2020] [Indexed: 01/06/2023] Open
Abstract
OBJECTIVE Dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED) are types of human dental tissue-derived mesenchymal stem cells (MSCs) that have emerged as an interesting and promising source of stem cells in the field of tissue engineering. The aim of this work is to isolate stem cells from DPSCs and SHED, cultivate them in vitro and compare their odontogenic differentiation potential. METHODS DPSCs and SHED were extracted from molars, premolars and canines of six healthy subjects aged 5-29 years. The cells were characterized, using flow cytometry, for mesenchymal stem cell surface markers. MTT colorimetric assay was applied to assess cell viability. Alizarin red staining, alkaline phosphatase (ALP) activity, quantitative real-time PCR (qRT-PCR) and western blot were carried out to determine DPSCs and SHED osteogenic/odontogenic differentiation. RESULTS DPSCs express higher STRO-1 and CD44 levels compared to SHED. Moreover, the cells differentiate and acquire columnar shape with a level of calcium deposition and mineralization that is the same between DPSCs and SHED. ALP activity, ALP, COLI, DMP-1, DSPP, OC, and RUNX2 (osteogenic/odontogenic differentiation markers) expression levels were higher in DPSCs. CONCLUSIONS DPSCs and SHED express MSCs markers. Although both cell types had calcium deposits, DPSCs presented a higher ALP activity level. In addition, DPSCs showed higher levels of osteogenic and odontogenic differentiation markers such as COLI, DSPP, OC, RUNX2, and DMP-1. These results suggest that DPSCs are closer to the phenotype of odontoblasts than SHED and may improve the efficacy of human dental tissue-derived mesenchymal stem cells therapeutic protocols. 'CLINICAL SIGNIFICANCE' DPSCs are closer than t SHED to the phenotype of odontoblasts. This would be helpful to enable better therapeutic decisions when applying MSCs-based therapy in the field of dentistry.
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Affiliation(s)
- Joseph Sabbagh
- Department of Restorative Dentistry and Endodontics, Faculty of Dental Medicine, Lebanese University, Beirut, Lebanon.
| | - Michella Ghassibe-Sabbagh
- Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, Beirut, Lebanon.
| | - Mohammad Fayyad-Kazan
- Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, Beirut, Lebanon; Laboratory of Cancer Biology and Molecular Immunology, Faculty of Sciences-I, Lebanese University, Hadath- Beirut, Lebanon.
| | - Fatima Al-Nemer
- Laboratory of Cancer Biology and Molecular Immunology, Faculty of Sciences-I, Lebanese University, Hadath- Beirut, Lebanon.
| | - Jean Claude Fahed
- Department of Restorative Dentistry and Endodontics, Faculty of Dental Medicine, Lebanese University, Beirut, Lebanon.
| | - Antoine Berberi
- Department of Oral and Maxillofacial Surgery, Faculty of Dental Medicine, Lebanese University, Beirut, Lebanon.
| | - Bassam Badran
- Laboratory of Cancer Biology and Molecular Immunology, Faculty of Sciences-I, Lebanese University, Hadath- Beirut, Lebanon.
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Lv H, Yang H, Wang Y. Effects of miR-103 by negatively regulating SATB2 on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells. PLoS One 2020; 15:e0232695. [PMID: 32379794 PMCID: PMC7205233 DOI: 10.1371/journal.pone.0232695] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/30/2019] [Accepted: 04/19/2020] [Indexed: 12/12/2022] Open
Abstract
Background The proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells (HBMScs) are modulated by a variety of microRNAs (miRNAs). SATB homeobox 2 (SATB2) is a critical transcription factor that contributes to maintain the balance of bone metabolism. However, it remains unclear how the regulatory relationship between miR-103 and SATB2 on HBMScs proliferation and osteogenic differentiation. Methods HBMScs were obtained from Cyagen Biosciences and successful induced osteogenic differentiation. The proliferation abilities of HBMScs after treatment with agomiR-103 and antagomiR-103 were assessed using a cell counting Kit-8 (CCK-8) assay, and osteogenic differentiation was determined using alizarin red S staining and alkaline phosphatase (ALP) activity assay. The expression levels of miR-103, SATB2, and associated osteogenic differentiation biomarkers, including RUNX family transcription factor 2 (RUNX2), bone gamma-carboxyglutamate protein (BGLAP), and secreted phosphoprotein 1 (SPP1), were evaluated using real-time qPCR and Western blot. The regulatory sites of miR-103 on SATB2 were predicted using bioinformatics software and validated using a dual luciferase reporter assay. The underlying mechanism of miR-103 on SATB2-medicated HBMScs proliferation and osteogenic differentiation were confirmed by co-transfection of antagomiR-103 and SATB2 siRNA. Results The expression of miR-103 in HBMScs after induction of osteogenic differentiation was reduced in a time-dependent way. Overexpression of miR-103 by transfection of agomiR-103 suppressed HBMScs proliferation and osteogenic differentiation, while silencing of miR-103 by antagomiR-103 abolished these inhibitory effects. Consistently, RUNX2, BGLAP and SPP1 mRNA and protein expression were decreased in agomiR-103 treated HBMScs compared with those in agomiR-NC group. Meanwhile, antagomiR-103 upregulated the mRNA and protein expression levels of RUNX2, BGLAP and SPP1 in HBMScs. Further studies revealed that SATB2 was a direct target gene of miR-103. BMSCs transfected with agomiR-103 exhibited significantly downregulated protein expression level of SATB2, whereas knockdown of miR-103 promoted it. Additionally, rescue assays confirmed that silencing of SATB2 partially reversed the effects of antagomiR-103 induced HBMScs proliferation and osteogenic differentiation. Conclusions The present results suggested that miR-103 negatively regulates SATB2 to serve an inhibitory role in the proliferation and osteogenic differentiation of HBMScs, which sheds light upon a potential therapeutic target for treating bone-related diseases.
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Affiliation(s)
- Hao Lv
- Department of Trauma Center, Jinan Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong Province, P.R. China
| | - Huashan Yang
- Department of Trauma Center, Jinan Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong Province, P.R. China
| | - Yuanrui Wang
- Department of Trauma Center, Jinan Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong Province, P.R. China
- * E-mail:
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Aljohani H, Senbanjo LT, Chellaiah MA. Methylsulfonylmethane increases osteogenesis and regulates the mineralization of the matrix by transglutaminase 2 in SHED cells. PLoS One 2019; 14:e0225598. [PMID: 31805069 PMCID: PMC6894810 DOI: 10.1371/journal.pone.0225598] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/16/2019] [Accepted: 11/07/2019] [Indexed: 01/09/2023] Open
Abstract
Methylsulfonylmethane (MSM) is a naturally occurring, sulfate-containing, organic compound. It has been shown to stimulate the differentiation of mesenchymal stem cells into osteoblast-like cells and bone formation. In this study, we investigated whether MSM influences the differentiation of stem cells from human exfoliated deciduous teeth (SHED) into osteoblast-like cells and their osteogenic potential. Here, we report that MSM induced osteogenic differentiation through the expression of osteogenic markers such as osterix, osteopontin, and RUNX2, at both mRNA and protein levels in SHED cells. An increase in the activity of alkaline phosphatase and mineralization confirmed the osteogenic potential of MSM. These MSM-induced effects were observed in cells grown in basal medium but not osteogenic medium. MSM induced transglutaminase-2 (TG2), which may be responsible for the cross-linking of extracellular matrix proteins (collagen or osteopontin), and the mineralization process. Inhibition of TG2 ensued a significant decrease in the differentiation of SHED cells and cross-linking of matrix proteins. A comparison of mineralization with the use of mineralized and demineralized bone particles in the presence of MSM revealed that mineralization is higher with mineralized bone particles than with demineralized bone particles. In conclusion, these results indicated that MSM could promote differentiation and osteogenic potential of SHED cells. This osteogenic property is more in the presence of mineralized bone particles. TG2 is a likely cue in the regulation of differentiation and mineral deposition of SHED cells in response to MSM.
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Affiliation(s)
- Hanan Aljohani
- Department of Oncology and Diagnostic Sciences, University of Maryland School of Dentistry, Baltimore, MD, United States of America
- Department of Oral Medicine and Diagnostics Sciences, King Saud University School of Dentistry, Riyadh, KSA
| | - Linda T. Senbanjo
- Department of Oncology and Diagnostic Sciences, University of Maryland School of Dentistry, Baltimore, MD, United States of America
| | - Meenakshi A. Chellaiah
- Department of Oncology and Diagnostic Sciences, University of Maryland School of Dentistry, Baltimore, MD, United States of America
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