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Ishkitiev N, Micheva M, Miteva M, Gaydarova S, Tzachev C, Lozanova V, Lozanov V, Cheshmedzhieva D, Kandinska M, Ilieva S, Gargallo R, Baluschev S, Stoynov S, Dyankova-Danovska T, Nedelcheva-Veleva M, Landfester K, Mihaylova Z, Vasilev A. Nanoconfined Chlorine-Substituted Monomethine Cyanine Dye with a Propionamide Function Based on the Thiazole Orange Scaffold-Use of a Fluorogenic Probe for Cell Staining and Nucleic Acid Visualization. Molecules 2024; 29:6038. [PMID: 39770126 PMCID: PMC11677322 DOI: 10.3390/molecules29246038] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/15/2024] [Revised: 12/16/2024] [Accepted: 12/19/2024] [Indexed: 01/11/2025] Open
Abstract
The development of fluorescence-based methods for bioassays and medical diagnostics requires the design and synthesis of specific markers to target biological microobjects. However, biomolecular recognition in real cellular systems is not always as selective as desired. A new concept for creating fluorescent biomolecular probes, utilizing a fluorogenic dye and biodegradable, biocompatible nanomaterials, is demonstrated. The synthesis of a new dicationic asymmetric monomethine cyanine dye with benzo[d]thiazolium-N-propionamide and chloroquinoline end groups is presented. The photophysical properties of the newly synthesized dye were examined through the combined application of spectroscopic and theoretical methods. The applicability of the dye as a fluorogenic nucleic acid probe was proven by UV-VIS spectroscopy and fluorescence titration. The dye-nucleic acid interaction mode was investigated by UV-Vis and CD spectroscopy. The newly synthesized dicationic dye, like other similar fluorogenic structures, limited permeability, which restricts its use as a probe for RNA and DNA. To enhance cellular delivery, we utilized a patented technology that employs solid, insoluble lipid nanoparticles. This method ensures the complete introduction of the dye into cells while minimizing activity outside the cells. In our study involving two human cell lines, we observed improved penetration through the cell membrane and distinctive selectivity in visualizing nucleic acids within the cytoplasm and nucleus.
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Affiliation(s)
- Nikolay Ishkitiev
- Department of Medical Chemistry and Biochemistry, Medical University Sofia, 2 Zdrave Str., 1431 Sofia, Bulgaria; (N.I.); (M.M.); (V.L.); (V.L.)
| | - Maria Micheva
- Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany; (M.M.); (S.B.); (K.L.)
| | - Marina Miteva
- Department of Medical Chemistry and Biochemistry, Medical University Sofia, 2 Zdrave Str., 1431 Sofia, Bulgaria; (N.I.); (M.M.); (V.L.); (V.L.)
| | - Stefaniya Gaydarova
- Faculty of Chemistry and Pharmacy, Sofia University “St. Kliment Ohridski”, 1 James Bourchier Blvd., 1164 Sofia, Bulgaria; (S.G.); (C.T.); (D.C.); (M.K.); (S.I.)
- Lead Biotherapeutics Ltd., 24 Shipka Str., 1504 Sofia, Bulgaria
| | - Christo Tzachev
- Faculty of Chemistry and Pharmacy, Sofia University “St. Kliment Ohridski”, 1 James Bourchier Blvd., 1164 Sofia, Bulgaria; (S.G.); (C.T.); (D.C.); (M.K.); (S.I.)
- Lead Biotherapeutics Ltd., 24 Shipka Str., 1504 Sofia, Bulgaria
| | - Vesela Lozanova
- Department of Medical Chemistry and Biochemistry, Medical University Sofia, 2 Zdrave Str., 1431 Sofia, Bulgaria; (N.I.); (M.M.); (V.L.); (V.L.)
| | - Valentin Lozanov
- Department of Medical Chemistry and Biochemistry, Medical University Sofia, 2 Zdrave Str., 1431 Sofia, Bulgaria; (N.I.); (M.M.); (V.L.); (V.L.)
| | - Diana Cheshmedzhieva
- Faculty of Chemistry and Pharmacy, Sofia University “St. Kliment Ohridski”, 1 James Bourchier Blvd., 1164 Sofia, Bulgaria; (S.G.); (C.T.); (D.C.); (M.K.); (S.I.)
| | - Meglena Kandinska
- Faculty of Chemistry and Pharmacy, Sofia University “St. Kliment Ohridski”, 1 James Bourchier Blvd., 1164 Sofia, Bulgaria; (S.G.); (C.T.); (D.C.); (M.K.); (S.I.)
| | - Sonia Ilieva
- Faculty of Chemistry and Pharmacy, Sofia University “St. Kliment Ohridski”, 1 James Bourchier Blvd., 1164 Sofia, Bulgaria; (S.G.); (C.T.); (D.C.); (M.K.); (S.I.)
| | - Raimundo Gargallo
- Department of Chemical Engineering and Analytical Chemistry, University of Barcelona, Martí i Franqués 1-11, E-08028 Barcelona, Spain;
| | - Stanislav Baluschev
- Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany; (M.M.); (S.B.); (K.L.)
- Faculty of Physics, Sofia University “St. Kliment Ohridski”, 5 James Bourchier Blvd., 1164 Sofia, Bulgaria
| | - Stoyno Stoynov
- Institute of Molecular Biology ‘‘Roumen Tsanev,’’ Bulgarian Academy of Sciences, ‘‘Acad. George Bonchev’’ Str. 21, 1113 Sofia, Bulgaria (T.D.-D.)
| | - Teodora Dyankova-Danovska
- Institute of Molecular Biology ‘‘Roumen Tsanev,’’ Bulgarian Academy of Sciences, ‘‘Acad. George Bonchev’’ Str. 21, 1113 Sofia, Bulgaria (T.D.-D.)
| | - Marina Nedelcheva-Veleva
- Institute of Molecular Biology ‘‘Roumen Tsanev,’’ Bulgarian Academy of Sciences, ‘‘Acad. George Bonchev’’ Str. 21, 1113 Sofia, Bulgaria (T.D.-D.)
| | - Katharina Landfester
- Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany; (M.M.); (S.B.); (K.L.)
| | - Zornitsa Mihaylova
- Department of Oral and Maxillofacial Surgery, Medical University Sofia, 1 “G. Sofijski” Str., 1431 Sofia, Bulgaria;
| | - Aleksey Vasilev
- Faculty of Chemistry and Pharmacy, Sofia University “St. Kliment Ohridski”, 1 James Bourchier Blvd., 1164 Sofia, Bulgaria; (S.G.); (C.T.); (D.C.); (M.K.); (S.I.)
- Laboratory of Functional and Nanostructured Polymers, Institute of Polymers, Bulgarian Academy of Sciences, Akad. G. Bonchev Str., Bl. 103A, 1113 Sofia, Bulgaria
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Ishkitiev N, Miteva M, Micheva M, Stoyanova T, Lozanova VV, Lozanov VS, Mihaylova Z, Cheshmedzhieva DV, Kandinska M, Rangelov M, Todorova N, Ilieva S, Baluschev S, Gargallo R, Calenic B, Constantinescu I, Landfester K, Vasilev AA. Aggregation induced nucleic acids recognition by homodimeric asymmetric monomethyne cyanine fluorochromes in mesenchymal stem cells. Int J Biol Macromol 2023; 250:126094. [PMID: 37544569 DOI: 10.1016/j.ijbiomac.2023.126094] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/30/2023] [Revised: 07/13/2023] [Accepted: 07/21/2023] [Indexed: 08/08/2023]
Abstract
In the light of recent retrovirus pandemics, the issue of discovering new and diverse RNA-specific fluorochromes for research and diagnostics became of acute importance. The great majority of nucleic acid-specific probes either do not stain RNA or cannot distinguish between DNA and RNA. The versatility of polymethine dyes makes them suitable as stains for visualization, analysis, and detection of nucleic acids, proteins, and other biomolecules. We synthesized the asymmetric dicationic homodimeric monomethine cyanine dyes 1,1'-(1,3-phenylenebis(methylene))bis(4-((3-methylbenzo[d]thiazol-2(3H)-ylidene)methyl)pyridin-1-ium) bromide (Т1) and 1,1'-(1,3-phenylenebis(methylene))bis(4-((3-methylbenzo[d]thiazol-2(3H)-ylidene)methyl)quinolin-1-ium) bromide (M1) and tested their binding specificity, spectral characteristics, membrane penetration in living and fixed cells, cellular toxicity, and stability of fluorescent emission. Mesenchymal cells have diverse phenotypes and extensive proliferation and differentiation properties. We found dyes T1 and M1 to show high photochemical stability in living mesenchymal stem cells from apical papilla (SCAP) with a strong fluorescent signal when bound to nucleic acids. We found M1 to perform better than control fluorochrome (Hoechst 33342) for in vivo DNA visualization. T1, on the other hand, stains granular cellular structures resembling ribosomes in living cells and after permeabilization of the nuclear membrane stains the nucleoli and not the chromatin in the nucleus. This makes T1 suitable for the visualization of structures rich in RNA in living and fixed cells.
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Affiliation(s)
- Nikolay Ishkitiev
- Medical University Sofia, Dept. of Medical Chemistry and Biochemistry, 2 Zdrave str., Sofia 1431, Bulgaria
| | - Marina Miteva
- Medical University Sofia, Dept. of Medical Chemistry and Biochemistry, 2 Zdrave str., Sofia 1431, Bulgaria
| | - Maria Micheva
- Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany
| | - Tanya Stoyanova
- Medical University Sofia, Dept. of Medical Chemistry and Biochemistry, 2 Zdrave str., Sofia 1431, Bulgaria; Faculty of Chemistry and Pharmacy, Sofia University "St. Kliment Ohridski", 1 James Bourchier Blvd., 1164 Sofia, Bulgaria
| | - Vesela V Lozanova
- Medical University Sofia, Dept. of Medical Chemistry and Biochemistry, 2 Zdrave str., Sofia 1431, Bulgaria
| | - Valentin S Lozanov
- Medical University Sofia, Dept. of Medical Chemistry and Biochemistry, 2 Zdrave str., Sofia 1431, Bulgaria
| | - Zornitsa Mihaylova
- Medical University Sofia, Dept. of Oral and Maxillofacial Surgery, G. Sofijski 1 str., Sofia 1431, Bulgaria
| | - Diana V Cheshmedzhieva
- Faculty of Chemistry and Pharmacy, Sofia University "St. Kliment Ohridski", 1 James Bourchier Blvd., 1164 Sofia, Bulgaria
| | - Meglena Kandinska
- Faculty of Chemistry and Pharmacy, Sofia University "St. Kliment Ohridski", 1 James Bourchier Blvd., 1164 Sofia, Bulgaria
| | - Miroslav Rangelov
- Institute of Organic Chemistry with Center of Phytochemistry, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria
| | - Nadezhda Todorova
- Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria
| | - Sonia Ilieva
- Faculty of Chemistry and Pharmacy, Sofia University "St. Kliment Ohridski", 1 James Bourchier Blvd., 1164 Sofia, Bulgaria
| | - Stanislav Baluschev
- Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany; Faculty of Physics, Sofia University "St. Kliment Ohridski", 5 James Bourchier Blvd., 1164 Sofia, Bulgaria
| | - Raimundo Gargallo
- Departament d'Enginyeria Química i Química Analítica, University of Barcelona, Martí i Franqués 1-11, E-08028 Barcelona, Spain
| | - Bogdan Calenic
- Carol Davila University of Medicine and Pharmacy, 258 Fundeni Road, 022328 Bucharest, Romania.
| | - Ileana Constantinescu
- Carol Davila University of Medicine and Pharmacy, 258 Fundeni Road, 022328 Bucharest, Romania
| | - Katharina Landfester
- Max Planck Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany
| | - Aleksey A Vasilev
- Faculty of Chemistry and Pharmacy, Sofia University "St. Kliment Ohridski", 1 James Bourchier Blvd., 1164 Sofia, Bulgaria; Institute of Polymers, Bulgarian Academy of Sciences, Akad. G. Bonchev St., bl 103A, 1113 Sofia, Bulgaria.
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Inada R, Mendoza HY, Tanaka T, Horie T, Satomi T. Preclinical study for the treatment of diabetes mellitus using β-like cells derived from human dental pulp stem cells. Regen Med 2022; 17:905-913. [DOI: 10.2217/rme-2022-0092] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/05/2022] Open
Abstract
Aim: The current study assessed whether insulin-producing cells obtained from dental pulp stem cells (DPSCs) can be a new therapeutic approach in a rat model of diabetes mellitus (DM). Materials & methods: Stem cells were differentiated into pancreatic β cells under hydrogen sulfide exposure via 2D and 3D methods. Each β-like cell was immunostained and transplanted into DM rats, after which the in vivo therapeutic effect was determined. Results: Immunostaining revealed the expression of various β-cell markers in each β-like cell differentiated using the 3D method. DPSC-derived β-like cell differentiated via the 3D method promoted a sufficient therapeutic effect. Conclusion: The 3D method promoted islet differentiation, indicating that DPSC-derived β-like cell transplantation could be a new approach for DM treatment.
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Affiliation(s)
- Ryo Inada
- Department of Oral & Maxillofacial Surgery, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Hiromi Yagi Mendoza
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Tomoko Tanaka
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Tetsuro Horie
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Takafumi Satomi
- Department of Oral & Maxillofacial Surgery, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
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Y Baena AR, Casasco A, Monti M. Hypes and Hopes of Stem Cell Therapies in Dentistry: a Review. Stem Cell Rev Rep 2022; 18:1294-1308. [PMID: 35015212 PMCID: PMC8748526 DOI: 10.1007/s12015-021-10326-4] [Citation(s) in RCA: 11] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 12/29/2021] [Indexed: 12/20/2022]
Abstract
One of the most exciting advances in life science research is the development of 3D cell culture systems to obtain complex structures called organoids and spheroids. These 3D cultures closely mimic in vivo conditions, where cells can grow and interact with their surroundings. This allows us to better study the spatio-temporal dynamics of organogenesis and organ function. Furthermore, physiologically relevant organoids cultures can be used for basic research, medical research, and drug discovery. Although most of the research thus far focuses on the development of heart, liver, kidney, and brain organoids, to name a few, most recently, these structures were obtained using dental stem cells to study in vitro tooth regeneration. This review aims to present the most up-to-date research showing how dental stem cells can be grown on specific biomaterials to induce their differentiation in 3D. The possibility of combining engineering and biology principles to replicate and/or increase tissue function has been an emerging and exciting field in medicine. The use of this methodology in dentistry has already yielded many interesting results paving the way for the improvement of dental care and successful therapies.
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Affiliation(s)
- Alessandra Rodriguez Y Baena
- Program in Biomedical Sciences and Engineering, Department of Molecular, Cell, and Developmental Biology, University of California-Santa Cruz, Santa Cruz, CA, 95064, USA
| | - Andrea Casasco
- Department of Public Health, Experimental and Forensic Medicine, Histology and Embryology Unit, University of Pavia, Pavia, Italy.,Dental & Face Center, CDI, Milan, Italy
| | - Manuela Monti
- Department of Public Health, Experimental and Forensic Medicine, Histology and Embryology Unit, University of Pavia, Pavia, Italy. .,Research Center for Regenerative Medicine, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
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Shimamura N, Fujii K, Ohkoshi S. Detection and quantification of human-specific mRNA from hepatocyte-like cells derived from dental pulp using real-time polymerase chain reaction. J Oral Biosci 2021; 63:298-305. [PMID: 34311038 DOI: 10.1016/j.job.2021.07.005] [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: 06/15/2021] [Revised: 07/08/2021] [Accepted: 07/18/2021] [Indexed: 11/25/2022]
Abstract
OBJECTIVES We quantified viable hepatocyte-like cells (HLCs) administered via portal or tail veins in the livers and lungs of immunodeficient rats using real-time reverse transcription polymerase chain reaction (RT-PCR) and human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) primers. METHODS Immunodeficient rats were infused with HLCs via portal or tail veins. mRNA was quantified based on the route of cell administration and the presence of liver injury. RESULTS Human-specific GAPDH mRNA primers detected 0.1 pg human RNA in 100 ng (1:106) of rat liver RNA. When infused into the portal vein, the quantity of HLC mRNA reduced to 5% 3 h after infusion. Most HLCs were entrapped in the lungs when infused via the tail vein and decreased to approximately 10% 6 h after infusion. A small number of HLCs made it to the liver but disappeared rapidly, regardless of liver injury. 24 h after infusion, viable HLCs were detected only in the lungs of rats with liver injury (P < 0.05). CONCLUSIONS The quantity of viable human cells in immunodeficient rats was estimated using real-time RT-PCR and primers specific to human mRNA.
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Affiliation(s)
- Naohiro Shimamura
- Clinical Examination, The Nippon Dental University Graduate School of Life Dentistry at Niigata, Niigata, Japan.
| | - Kazuyuki Fujii
- Department of Dental Anesthesiology, The Nippon Dental University School of Life Dentistry at Niigata, Niigata, Japan.
| | - Shogo Ohkoshi
- Clinical Examination, The Nippon Dental University Graduate School of Life Dentistry at Niigata, Niigata, Japan; Department of Internal Medicine, The Nippon Dental University School of Life Dentistry at Niigata, Niigata, Japan.
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Yin JY, Luo XH, Feng WQ, Miao SH, Ning TT, Lei Q, Jiang T, Ma DD. Multidifferentiation potential of dental-derived stem cells. World J Stem Cells 2021; 13:342-365. [PMID: 34136070 PMCID: PMC8176842 DOI: 10.4252/wjsc.v13.i5.342] [Citation(s) in RCA: 13] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/21/2020] [Revised: 03/10/2021] [Accepted: 04/05/2021] [Indexed: 02/06/2023] Open
Abstract
Tooth-related diseases and tooth loss are widespread and are a major public health issue. The loss of teeth can affect chewing, speech, appearance and even psychology. Therefore, the science of tooth regeneration has emerged, and attention has focused on tooth regeneration based on the principles of tooth development and stem cells combined with tissue engineering technology. As undifferentiated stem cells in normal tooth tissues, dental mesenchymal stem cells (DMSCs), which are a desirable source of autologous stem cells, play a significant role in tooth regeneration. Researchers hope to reconstruct the complete tooth tissues with normal functions and vascularization by utilizing the odontogenic differentiation potential of DMSCs. Moreover, DMSCs also have the ability to differentiate towards cells of other tissue types due to their multipotency. This review focuses on the multipotential capacity of DMSCs to differentiate into various tissues, such as bone, cartilage, tendon, vessels, neural tissues, muscle-like tissues, hepatic-like tissues, eye tissues and glands and the influence of various regulatory factors, such as non-coding RNAs, signaling pathways, inflammation, aging and exosomes, on the odontogenic/osteogenic differentiation of DMSCs in tooth regeneration. The application of DMSCs in regenerative medicine and tissue engineering will be improved if the differentiation characteristics of DMSCs can be fully utilized, and the factors that regulate their differentiation can be well controlled.
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Affiliation(s)
- Jing-Yao Yin
- Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China
| | - Xing-Hong Luo
- Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China
| | - Wei-Qing Feng
- Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China
| | - Sheng-Hong Miao
- Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China
| | - Ting-Ting Ning
- Department of Endodontics, Stomatological Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
| | - Qian Lei
- Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China
| | - Tao Jiang
- Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, Guangdong Province, China
| | - Dan-Dan Ma
- Department of Endodontics, Stomatological Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
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Zhou YK, Zhu LS, Huang HM, Cui SJ, Zhang T, Zhou YH, Yang RL. Stem cells from human exfoliated deciduous teeth ameliorate concanavalin A-induced autoimmune hepatitis by protecting hepatocytes from apoptosis. World J Stem Cells 2020; 12:1623-1639. [PMID: 33505604 PMCID: PMC7789126 DOI: 10.4252/wjsc.v12.i12.1623] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/31/2020] [Revised: 09/20/2020] [Accepted: 10/12/2020] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND Autoimmune hepatitis is a serious autoimmune liver disease that threatens human health worldwide, which emphasizes the urgent need to identify novel treatments. Stem cells from human exfoliated deciduous teeth (SHED), which are easy to obtain in a non-invasive manner, show pronounced proliferative and immunomodulatory capacities. AIM To investigate the protective effects of SHED on concanavalin A (ConA)-induced hepatitis in mice, and to elucidate the associated regulatory mechanisms. METHODS We used a ConA-induced acute hepatitis mouse model and an in vitro co-culture system to study the protective effects of SHED on ConA-induced autoimmune hepatitis, as well as the associated underlying mechanisms. RESULTS SHED infusion could prevent aberrant histopathological liver architecture caused by ConA-induced infiltration of CD3+, CD4+, tumor necrosis-alpha+, and interferon-gamma+ inflammatory cells. Alanine aminotransferase and aspartate aminotransferase were significantly elevated in hepatitis mice. SHED infusion could therefore block ConA-induced alanine aminotransferase and aspartate aminotransferase elevations. Mechanistically, ConA upregulated tumor necrosis-alpha and interferon-gamma expression, which was activated by the nuclear factor-kappa B pathway to induce hepatocyte apoptosis, resulting in acute liver injury. SHED administration protected hepatocytes from ConA-induced apoptosis. CONCLUSION SHED alleviates ConA-induced acute liver injury via inhibition of hepatocyte apoptosis mediated by the nuclear factor-kappa B pathway. Our findings could provide a potential treatment strategy for hepatitis.
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Affiliation(s)
- Yi-Kun Zhou
- Department of Orthodontics, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
| | - Ling-Su Zhu
- Department of Orthodontics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
| | - Hua-Ming Huang
- Department of Orthodontics, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
| | - Sheng-Jie Cui
- Department of Orthodontics, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
| | - Ting Zhang
- Department of Orthodontics, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
| | - Yan-Heng Zhou
- Department of Orthodontics, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
| | - Rui-Li Yang
- Department of Orthodontics, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
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Mochizuki M, Sagara H, Nakahara T. Type I collagen facilitates safe and reliable expansion of human dental pulp stem cells in xenogeneic serum-free culture. Stem Cell Res Ther 2020; 11:267. [PMID: 32660544 PMCID: PMC7359624 DOI: 10.1186/s13287-020-01776-7] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/08/2020] [Revised: 05/26/2020] [Accepted: 06/12/2020] [Indexed: 12/12/2022] Open
Abstract
BACKGROUND Human dental pulp stem cells (DPSCs) are a readily accessible and promising cell source for regenerative medicine. We recently reported that a xenogeneic serum-free culture medium (XFM) is preferable to fetal bovine serum-containing culture medium for ex vivo expansion of DPSCs; however, we observed that, upon reaching overconfluence, XFM cells developed a multilayered structure and frequently underwent apoptotic death, resulting in reduced cell yield. Therefore, we focused on optimization of the XFM culture system to avoid the undesirable death of DPSCs. METHODS We selected type I collagen (COL) as the optimal coating substrate for the cultureware and compared DPSCs cultured on COL in XFM (COL-XFM cells) to the conventional XFM cultures (XFM cells). RESULTS Our results demonstrated that COL coating facilitated significantly higher rates of cell isolation and growth; upon reaching overconfluence, cell survival and sustained proliferative potential resulted in two-fold yield compared to the XFM cells. Surprisingly, after subculturing the overconfluent COL-XFM cultures, the cells retained stem cell behavior including stable cell growth, multidifferentiation potential, stem cell phenotype, and chromosomal stability, which was achieved through HIF-1α-dependent production and uniform distribution of collagen type I and its interactions with integrins α2β1 and α11β1 at overconfluency. In contrast, cells undergoing apoptotic death within overconfluent XFM cultures had disorganized mitochondria with membrane depolarization. CONCLUSION The use of COL as a coating substrate promises safe and reliable handling of DPSCs in XFM culture, allowing translational stem cell medicine to achieve stable isolation, expansion, and banking of donor-derived stem cells.
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Affiliation(s)
- Mai Mochizuki
- Department of Life Science Dentistry, The Nippon Dental University, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
- Department of Developmental and Regenerative Dentistry, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Hiroshi Sagara
- Medical Proteomics Laboratory, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan
| | - Taka Nakahara
- Department of Developmental and Regenerative Dentistry, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan.
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Xie F, He J, Chen Y, Hu Z, Qin M, Hui T. Multi-lineage differentiation and clinical application of stem cells from exfoliated deciduous teeth. Hum Cell 2020; 33:295-302. [PMID: 32006349 DOI: 10.1007/s13577-020-00323-z] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/24/2019] [Accepted: 01/15/2020] [Indexed: 12/12/2022]
Abstract
Stem cells from human exfoliated deciduous teeth (SHED) have now been considered one of the most promising sources of stem cells for tissue engineering and stem cell therapies due to their stemness and potential to differentiate into other cell lines. The high proliferation rate, the differentiation capacity, the easy access and less ethical concerns make SHED a brilliant solution for many diseases. The purpose of this review is to describe current knowledge of SHED's capability of differentiation, applications and immune status and to draw attention to further research on the mechanism and the dependability of stem cell therapy with SHED.
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Affiliation(s)
- Fei Xie
- Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, No.22, Zhongguancun South Avenue, Haidian District, Beijing, People's Republic of China
| | - Jie He
- Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, No.22, Zhongguancun South Avenue, Haidian District, Beijing, People's Republic of China
| | - Yingyi Chen
- Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, No.22, Zhongguancun South Avenue, Haidian District, Beijing, People's Republic of China
| | - Ziqi Hu
- Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, No.22, Zhongguancun South Avenue, Haidian District, Beijing, People's Republic of China
| | - Man Qin
- Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, No.22, Zhongguancun South Avenue, Haidian District, Beijing, People's Republic of China.
| | - Tianqian Hui
- Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, No.22, Zhongguancun South Avenue, Haidian District, Beijing, People's Republic of China.
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10
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Hara H, Sano K, Ishikawa H, Ohkoshi S. Differentiation of Dental Pulp-Derived MSCs into Hepatocyte-Like Cells and Their Therapeutic Use for Chemical Liver Injuries of Rats. J HARD TISSUE BIOL 2020; 29:215-222. [DOI: 10.2485/jhtb.29.215] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/23/2022]
Affiliation(s)
- Hajime Hara
- Department of Internal Medicine, School of Life Dentistry at Niigata, The Nippon Dental University
| | - Kimito Sano
- Department of Dental Anesthesiology, School of Life Dentistry at Niigata, The Nippon Dental University
| | - Hiroshi Ishikawa
- Laboratory of Clinical Regenerative Medicine, Faculty of Medicine, University of Tsukuba
| | - Shogo Ohkoshi
- Department of Internal Medicine, School of Life Dentistry at Niigata, The Nippon Dental University
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11
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Chen CA, Chen YL, Huang JS, Huang GTJ, Chuang SF. Effects of Restorative Materials on Dental Pulp Stem Cell Properties. J Endod 2019; 45:420-426. [PMID: 30819529 DOI: 10.1016/j.joen.2018.10.021] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/07/2018] [Revised: 07/21/2018] [Accepted: 10/08/2018] [Indexed: 01/09/2023]
Abstract
INTRODUCTION Dental pulp stem cells (DPSCs) are multipotent progenitors for biotechnological practices, but the influences of existing restorations on their viability and differentiation are not well-known. This study was aimed to investigate in vivo and in vitro responses of DPSCs to restorative materials. METHODS Class I cavities were prepared on molars scheduled to be extracted and then restored with a resin-based composite (RBC), a glass ionomer cement, or zinc oxide eugenol. Intact teeth were used as controls. Twelve molars in each group were extracted on day 7 or day 30 after restorations to assess the early or intermediate pulp responses and were then cut in half. One half was processed for histopathological analysis, and the other was used to isolate DPSCs for a colony-forming unit assay and real-time polymerase chain reaction for NANOG, OCT4, and CD44 expression. RESULTS All restored teeth showed pulp damage at various levels, whereas mild to moderate inflammation persisted in the RBC group until day 30. The existence of DPSCs in the pulp cores of all groups was revealed based on CD44 immunoreactivity. Glass ionomer cement and zinc oxide eugenol did not affect the relative percentages of DPSCs in either early or intermediate stages, whereas RBCs reduced the percentage. The colony-forming units in all restoration groups were comparable with those in the control. Nevertheless, the restorations significantly enhanced OCT4 expression, especially in RBC/day 30. CONCLUSIONS Dental restorations cause mild pulp damage but do not affect DPSC viability. RBC decreases DPSC densities but might increase the stemness of surviving DPSCs through an inflammation-stimulation process.
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Affiliation(s)
- Chao-An Chen
- Department of Endodontics, Chi Mei Medical Center, Liouying, Tainan, Taiwan
| | - Yuh-Ling Chen
- Institute of Oral Medicine, National Cheng Kung University, Tainan, Taiwan
| | - Jehn-Shyun Huang
- Institute of Oral Medicine, National Cheng Kung University, Tainan, Taiwan; Department of Stomatology, National Cheng Kung University Hospital, Tainan, Taiwan
| | - George T-J Huang
- Department of Bioscience Research, University of Tennessee Health Science Center, Memphis, Tennessee
| | - Shu-Fen Chuang
- Institute of Oral Medicine, National Cheng Kung University, Tainan, Taiwan; Department of Stomatology, National Cheng Kung University Hospital, Tainan, Taiwan.
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12
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Yokoyama T, Yagi Mendoza H, Tanaka T, Ii H, Takano R, Yaegaki K, Ishikawa H. Regulation of CCl 4-induced liver cirrhosis by hepatically differentiated human dental pulp stem cells. Hum Cell 2019; 32:125-140. [PMID: 30637566 DOI: 10.1007/s13577-018-00234-0] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/01/2018] [Accepted: 12/09/2018] [Indexed: 02/07/2023]
Abstract
Liver transplantation is the most effective treatment for treating liver cirrhosis. However, a limited number of donors, graft rejection, and other complications can undermine transplant success. It is considered that cell transplantation is an alternative approach of liver transplantation. We previously developed a protocol for hepatic differentiation of cluster of differentiation 117+ stem cells isolated from human exfoliated deciduous tooth pulp (SHEDs) under hydrogen sulfide exposure. These cells showed excellent hepatic function. Here, we investigated whether hepatocyte-like cell transplantation is effective for treating carbon tetrachloride (CCl4)-induced liver cirrhosis. SHEDs were hepatically differentiated, which was confirmed via immunological analyses and albumin concentration determination in the medium. Rats were intraperitoneally injected with CCl4 for and the differentiated cells were injected into rat spleen. Histopathological and immunohistochemical analyses were performed. Liver functions were serologically and pathologically determined. Quantitative real-time-polymerase chain reaction was implemented to clarify the treatment procedure of liver cirrhosis. In vitro-differentiated hepatocyte-like cells were positive for all examined hepatic markers. SHED-derived hepatocyte transplantation eliminated liver fibrosis and restored liver structure in rats. Liver immunohistochemical analyses showed the presence of human-specific hepatic markers, i.e., a large amount of human hepatic cells were very active in the liver and spleen. Serological tests revealed significant liver function recovery in the transplantation group. Expression of genes promoting fibrosis increased after cirrhosis induction but was suppressed after transplantation. Our results suggest that xenotransplantation of hepatocyte-like cells of human origin can treat cirrhosis. Moreover, cell-based therapy of chronic liver conditions may be an effective option.
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Affiliation(s)
- Tomomi Yokoyama
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Hiromi Yagi Mendoza
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Tomoko Tanaka
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Hisataka Ii
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Riya Takano
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Ken Yaegaki
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan.
| | - Hiroshi Ishikawa
- Department of Oral Health, The Nippon Dental University School of Life Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan.,Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, Laboratory of Advanced Research D # 326, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan
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13
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Mendi A, Ulutürk H, Ataç MS, Yılmaz D. Stem Cells for the Oromaxillofacial Area: Could they be a promising source for regeneration in dentistry? ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2019; 1144:101-121. [PMID: 30725365 DOI: 10.1007/5584_2018_327] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
Oromaxillofacial tissues (OMT) are composed of tooth and bone, together with nerves and blood vessels. Such a composite material is a huge source for mesenchymal stem cells (MSCs) that can be obtained with ease from extracted teeth, teeth structures and socket blood, flapped gingiva tissue, and mandibular/maxillar bone marrow. They offer a biological answer for restoring damaged dental tissues such as the regeneration of alveolar bone, prevention of pulp tissue defects, and dental structures. Dental tissue-derived mesenchymal stem cells share properties with bone marrow-derived mesenchymal stem cells and there is a considerable potential for these cells to be used in different stem cell-based therapies, such as bone and nerve regeneration. Dental pulp tissue might be a very good source for neurological disorders whereas gingiva-derived mesenchymal stem cells could be a good immune modulatory/suppressive mediators. OMT-MSCs is also promising candidates for regeneration of orofacial tissues from the perspective of developmental fate. Here, we review the fundamental biology and potential for future regeneration strategies of MSCs in oromaxillofacial research.
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Affiliation(s)
- Ayşegül Mendi
- Faculty of Dentistry, Department of Basic Sciences, Gazi University, Ankara, Turkey.
| | - Hacer Ulutürk
- Faculty of Dentistry, Department of Oral and Maxillofacial Surgery, Gazi University, Ankara, Turkey
| | - Mustafa Sancar Ataç
- Faculty of Dentistry, Department of Oral and Maxillofacial Surgery, Gazi University, Ankara, Turkey
| | - Derviş Yılmaz
- Faculty of Dentistry, Department of Oral and Maxillofacial Surgery, Gazi University, Ankara, Turkey
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14
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Ohkoshi S, Hirono H, Nakahara T, Ishikawa H. Dental pulp cell bank as a possible future source of individual hepatocytes. World J Hepatol 2018; 10:702-707. [PMID: 30386463 PMCID: PMC6206155 DOI: 10.4254/wjh.v10.i10.702] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/29/2018] [Revised: 05/30/2018] [Accepted: 07/10/2018] [Indexed: 02/06/2023] Open
Abstract
Mesenchymal stem cells (MSCs) as a source for regenerative medicine are now the subject of much clinical attention. There are high expectations due to their safety, low tumorigenic risk, and low ethical concerns. MSC therapy has been approved for acute graft-versus host diseases since 2015. Tooth-derived MSCs are known to have a great potential in their proliferation and differentiation capacities, even when compared with bone-marrow-derived MSCs. In particular, stem cells from human exfoliated deciduous teeth (SHEDs) are the best candidates for personal cell banking (dental pulp cell bank), because they can be obtained less invasively in the natural process of individual growth. SHEDs are known to differentiate into hepatocytes. There have been several studies showing the effectiveness of SHEDs on the treatment of liver failure in animal models. They may exert their effects either by repopulation of cells in injured liver or by paracrine mechanisms due to their immune-regulatory functions. Moreover, it may be possible to use each individuals' dental pulp cells as a future source of tailor-made differentiated hepatocytes in the context of a bioartificial liver or liver-on-a-chip to screen for drug toxicity.
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Affiliation(s)
- Shogo Ohkoshi
- Department of Internal Medicine, School of Life Dentistry at Niigata, the Nippon Dental University, Niigata 951-8580, Japan.
| | - Haruka Hirono
- Department of Internal Medicine, School of Life Dentistry at Niigata, the Nippon Dental University, Niigata 951-8580, Japan
| | - Taka Nakahara
- Department of Developmental and Regenerative Dentistry, School of Life Dentistry at Tokyo, the Nippon Dental University, Chiyoda-ku 102-8159, Japan
| | - Hiroshi Ishikawa
- Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, Laboratory of Advanced Research D #326, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan
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15
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Yagi Mendoza H, Yokoyama T, Tanaka T, Ii H, Yaegaki K. Regeneration of insulin-producing islets from dental pulp stem cells using a 3D culture system. Regen Med 2018; 13:673-687. [PMID: 30028236 DOI: 10.2217/rme-2018-0074] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022] Open
Abstract
AIM In this study, we aimed to establish the differentiation protocol of dental pulp stem cells (DPSCs) into pancreatic islets using a 3D structure. MATERIALS & METHODS DPSCs were differentiated in a 3D culture system using a stepwise protocol. Expression of β-cell markers, glucose-stimulated insulin secretion, and PI3K/AKT and WNT pathways were compared between monolayer-cultured pancreatic cells and islets. RESULTS Islet formation increased insulin and C-peptide production, and enhanced the expression of pancreatic markers. Glucose-dependent secretion of insulin was increased by islets. Pancreatic endocrine markers, transcriptional factors, and the PI3K/AKT and WNT pathways were also upregulated. CONCLUSION Pancreatic islets were generated from DPSCs in a 3D culture system. This system could provide novel strategies for controlling diabetes through regenerative medicine.
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Affiliation(s)
- Hiromi Yagi Mendoza
- Department of Oral Health, School of Life Dentistry at Tokyo, Nippon Dental University, 1-9-20, Fujimi, Chiyoda ku, 102-8159 Tokyo, Japan
| | - Tomomi Yokoyama
- Department of Oral Health, School of Life Dentistry at Tokyo, Nippon Dental University, 1-9-20, Fujimi, Chiyoda ku, 102-8159 Tokyo, Japan
| | - Tomoko Tanaka
- Department of Oral Health, School of Life Dentistry at Tokyo, Nippon Dental University, 1-9-20, Fujimi, Chiyoda ku, 102-8159 Tokyo, Japan
| | - Hisataka Ii
- Department of Oral Health, School of Life Dentistry at Tokyo, Nippon Dental University, 1-9-20, Fujimi, Chiyoda ku, 102-8159 Tokyo, Japan
| | - Ken Yaegaki
- Department of Oral Health, School of Life Dentistry at Tokyo, Nippon Dental University, 1-9-20, Fujimi, Chiyoda ku, 102-8159 Tokyo, Japan
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16
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Mochizuki M, Nakahara T. Establishment of xenogeneic serum-free culture methods for handling human dental pulp stem cells using clinically oriented in-vitro and in-vivo conditions. Stem Cell Res Ther 2018; 9:25. [PMID: 29394956 PMCID: PMC5797401 DOI: 10.1186/s13287-017-0761-5] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/29/2017] [Revised: 12/04/2017] [Accepted: 12/20/2017] [Indexed: 12/13/2022] Open
Abstract
Background Currently, ex-vivo handling of stem cells, including transport after harvest and therapeutic preparation, is generally done in culture media containing fetal bovine serum (FBS), which promotes cell attachment, proliferation, and differentiation. However, because of safety concerns associated with the use of FBS, including potential transmission of zoonotic agents and transplant rejection because of the incorporation of foreign proteins into the stem cells, there is a need for xenogeneic serum-free culture media for clinical handling of stem cells. Methods Dental pulp stem cells were derived from wisdom teeth donated by eight healthy volunteers and cultured in xenogeneic serum-free culture medium (XFM) or xenogeneic serum-containing culture medium (SCM). Cells were subjected to morphological, proliferation, karyotype, differentiation, marker expression, cryopreservation, and cytotoxic susceptibility analyses in vitro, as well as transplantation in vivo. Results In primary culture, XFM cells showed lower adhesion and slightly different morphology, although the single-cell size was similar to that of SCM cells. XFM cells exhibited typical mesenchymal stem cell (MSC) characteristics in vitro and in vivo, including marker gene/protein expression, trilineage differentiation potential, and hard, osteo-dentin tissue formation. Additionally, XFM cells maintained a normal karyotype in vitro and nontumorigenic potential in vivo; however, XFM cells were more susceptible to H2O2 and ultraviolet cytotoxic stimuli. XFM cells formed a multilayered structure showing excessive cell death/division in contrast to the monolayered structure of SCM cells when reaching overconfluence. Proliferation was disrupted in overconfluent XFM cells, and these cells could not be subcultured. Dimethyl sulfoxide-free cryopreserved XFM cells yielded similar results in all of the experiments. Conclusions This study is the first reporting successful isolation and expansion of an MSC population from donor-derived tissue (dental pulp) under xenogeneic serum-free culture conditions, as well as the application of cryopreservation, using a research strategy based on clinically oriented in-vitro and in-vivo experiments. Electronic supplementary material The online version of this article (doi:10.1186/s13287-017-0761-5) contains supplementary material, which is available to authorized users.
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Affiliation(s)
- Mai Mochizuki
- Department of Developmental and Regenerative Dentistry, School of Life Dentistry at Tokyo, The Nippon Dental University, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan
| | - Taka Nakahara
- Department of Developmental and Regenerative Dentistry, School of Life Dentistry at Tokyo, The Nippon Dental University, 1-9-20 Fujimi, Chiyoda-ku, Tokyo, 102-8159, Japan.
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17
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Ebrahimi M, Botelho M. Adult Stem Cells of Orofacial Origin: Current Knowledge and Limitation and Future Trend in Regenerative Medicine. Tissue Eng Regen Med 2017; 14:719-733. [PMID: 30603522 PMCID: PMC6171671 DOI: 10.1007/s13770-017-0078-6] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/23/2017] [Revised: 07/19/2017] [Accepted: 08/04/2017] [Indexed: 12/21/2022] Open
Abstract
Stem cell research is one of the most rapidly expanding field of medicine which provides significant opportunities for therapeutic and regenerative applications. Different types of stem cells have been isolated investigating their accessibility, control of the differentiation pathway and additional immunomodulatory properties. Bulk of the literature focus has been on the study and potential applications of adult stem cells (ASC) because of their low immunogenicity and reduced ethical considerations. This review paper summarizes the basic available literature on different types of ASC with special focus on stem cells from dental and orofacial origin. ASC have been isolated from different sources, however, isolation of ASC from orofacial tissues has provided a novel promising alternative. These cells offer a great potential in the future of therapeutic and regenerative medicine because of their remarkable availability at low cost while allowing minimally invasive isolation procedures. Furthermore, their immunomodulatory and anti-inflammatory potential is of particular interest. However, there are conflicting reports in the literature regarding their particular biology and full clinical potentials. Sound knowledge and higher control over proliferation and differentiation mechanisms are prerequisites for clinical applications of these cells. Therefore, further standardized basic and translational studies are required to increase the reproducibility and reduce the controversies of studies, which in turn facilitate comparison of related literature and enhance further development in the field.
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Affiliation(s)
- Mehdi Ebrahimi
- Department of Oral Rehabilitation, Faculty of Dentistry, Prince Philip Dental Hospital, The University of Hong Kong, 34 Hospital Road, Sai Ying Pun, Hong Kong
| | - Michael Botelho
- Department of Oral Rehabilitation, Faculty of Dentistry, Prince Philip Dental Hospital, The University of Hong Kong, 34 Hospital Road, Sai Ying Pun, Hong Kong
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18
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Ohkoshi S, Hara H, Hirono H, Watanabe K, Hasegawa K. Regenerative medicine using dental pulp stem cells for liver diseases. World J Gastrointest Pharmacol Ther 2017; 8:1-6. [PMID: 28217369 PMCID: PMC5292602 DOI: 10.4292/wjgpt.v8.i1.1] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/31/2016] [Revised: 11/16/2016] [Accepted: 12/16/2016] [Indexed: 02/06/2023] Open
Abstract
Acute liver failure is a refractory disease and its prognosis, if not treated using liver transplantation, is extremely poor. It is a good candidate for regenerative medicine, where stem cell-based therapies play a central role. Mesenchymal stem cells (MSCs) are known to differentiate into multiple cell lineages including hepatocytes. Autologous cell transplant without any foreign gene induction is feasible using MSCs, thereby avoiding possible risks of tumorigenesis and immune rejection. Dental pulp also contains an MSC population that differentiates into hepatocytes. A point worthy of special mention is that dental pulp can be obtained from deciduous teeth during childhood and can be subsequently harvested when necessary after deposition in a tooth bank. MSCs have not only a regenerative capacity but also act in an anti-inflammatory manner via paracrine mechanisms. Promising efficacies and difficulties with the use of MSC derived from teeth are summarized in this review.
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19
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Han YJ, Kang YH, Shivakumar SB, Bharti D, Son YB, Choi YH, Park WU, Byun JH, Rho GJ, Park BW. Stem Cells from Cryopreserved Human Dental Pulp Tissues Sequentially Differentiate into Definitive Endoderm and Hepatocyte-Like Cells in vitro. Int J Med Sci 2017; 14:1418-1429. [PMID: 29200956 PMCID: PMC5707759 DOI: 10.7150/ijms.22152] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/30/2017] [Accepted: 10/13/2017] [Indexed: 01/03/2023] Open
Abstract
We previously described a novel tissue cryopreservation protocol to enable the safe preservation of various autologous stem cell sources. The present study characterized the stem cells derived from long-term cryopreserved dental pulp tissues (hDPSCs-cryo) and analyzed their differentiation into definitive endoderm (DE) and hepatocyte-like cells (HLCs) in vitro. Human dental pulp tissues from extracted wisdom teeth were cryopreserved as per a slow freezing tissue cryopreservation protocol for at least a year. Characteristics of hDPSCs-cryo were compared to those of stem cells from fresh dental pulps (hDPSCs-fresh). hDPSCs-cryo were differentiated into DE cells in vitro with Activin A as per the Wnt3a protocol for 6 days. These cells were further differentiated into HLCs in the presence of growth factors until day 30. hDPSCs-fresh and hDPSCs-cryo displayed similar cell growth morphology, cell proliferation rates, and mesenchymal stem cell character. During differentiation into DE and HLCs in vitro, the cells flattened and became polygonal in shape, and finally adopted a hepatocyte-like shape. The differentiated DE cells at day 6 and HLCs at day 30 displayed significantly increased DE- and hepatocyte-specific markers at the mRNA and protein level, respectively. In addition, the differentiated HLCs showed detoxification and glycogen storage capacities, indicating they could share multiple functions with real hepatocytes. These data conclusively show that hPDSCs-cryo derived from long-term cryopreserved dental pulp tissues can be successfully differentiated into DE and functional hepatocytes in vitro. Thus, preservation of dental tissues could provide a valuable source of autologous stem cells for tissue engineering.
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Affiliation(s)
- Young-Jin Han
- Department of Dentistry, Gyeongsang National University School of Medicine and Institute of Health Science, Jinju, Republic of Korea
| | - Young-Hoon Kang
- Department of Dentistry, Gyeongsang National University School of Medicine and Institute of Health Science, Jinju, Republic of Korea.,Department of Oral and Maxillofacial Surgery, Changwon Gyeongsang National University Hospital, Changwon, Republic of Korea
| | - Sarath Belame Shivakumar
- Department of Theriogenology and Biotechnology, College of Veterinary Medicine and Research Institute of Life Science, Gyeongsang National University, Jinju, Republic of Korea
| | - Dinesh Bharti
- Department of Theriogenology and Biotechnology, College of Veterinary Medicine and Research Institute of Life Science, Gyeongsang National University, Jinju, Republic of Korea
| | - Young-Bum Son
- Department of Theriogenology and Biotechnology, College of Veterinary Medicine and Research Institute of Life Science, Gyeongsang National University, Jinju, Republic of Korea
| | - Yong-Ho Choi
- Department of Theriogenology and Biotechnology, College of Veterinary Medicine and Research Institute of Life Science, Gyeongsang National University, Jinju, Republic of Korea
| | - Won-Uk Park
- Department of Dental Technology, Jinju Health College, Jinju, Republic of Korea
| | - June-Ho Byun
- Department of Dentistry, Gyeongsang National University School of Medicine and Institute of Health Science, Jinju, Republic of Korea
| | - Gyu-Jin Rho
- Department of Theriogenology and Biotechnology, College of Veterinary Medicine and Research Institute of Life Science, Gyeongsang National University, Jinju, Republic of Korea
| | - Bong-Wook Park
- Department of Dentistry, Gyeongsang National University School of Medicine and Institute of Health Science, Jinju, Republic of Korea.,Department of Oral and Maxillofacial Surgery, Changwon Gyeongsang National University Hospital, Changwon, Republic of Korea
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Paduano F, Marrelli M, Palmieri F, Tatullo M. CD146 Expression Influences Periapical Cyst Mesenchymal Stem Cell Properties. Stem Cell Rev Rep 2016; 12:592-603. [DOI: 10.1007/s12015-016-9674-4] [Citation(s) in RCA: 46] [Impact Index Per Article: 5.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/26/2022]
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21
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Generation of functional hepatocyte-like cells from human deciduous periodontal ligament stem cells. Naturwissenschaften 2016; 103:62. [PMID: 27379400 DOI: 10.1007/s00114-016-1387-7] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/17/2015] [Revised: 06/14/2016] [Accepted: 06/16/2016] [Indexed: 12/22/2022]
Abstract
Human deciduous periodontal ligament stem cells have been introduced for as an easily accessible source of stem cells from dental origin. Although recent studies have revealed the ability of these stem cells in multipotential attribute, their efficiency of hepatic lineage differentiation has not been addressed so far. The aim of this study is to investigate hepatic lineage fate competence of periodontal ligament stem cells through direct media induction. Differentiation of periodontal ligament stem cells into hepatocyte-like cells was conducted by the exposure of two phase media induction. First phase was performed in the presence of hepatocyte growth factors to induce a definitive endoderm formation. In the subsequent phase, the cells were treated with oncostatin M and dexamethosone followed by insulin and transferrin to generate hepatocyte-like cells. Hepatic-related characters of the generated hepatocyte-like cells were determined at both mRNA and protein level followed by functional assays. Foremost changes observed in the generation of hepatocyte-like cells were the morphological features in which these cells were transformed from fibroblastic shape to polygonal shape. Temporal expression of hepatic markers ranging from early endodermal up to late markers were detected in the hepatocyte-like cells. Crucial hepatic markers such as glycogen storage, albumin, and urea secretion were also shown. These findings exhibited the ability of periodontal ligament stem cells of dental origin to be directed into hepatic lineage fate. These cells can be regarded as an alternative autologous source in the usage of stem cell-based treatment for liver diseases.
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Rosa V, Dubey N, Islam I, Min KS, Nör JE. Pluripotency of Stem Cells from Human Exfoliated Deciduous Teeth for Tissue Engineering. Stem Cells Int 2016; 2016:5957806. [PMID: 27313627 PMCID: PMC4904107 DOI: 10.1155/2016/5957806] [Citation(s) in RCA: 46] [Impact Index Per Article: 5.1] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/29/2016] [Accepted: 05/04/2016] [Indexed: 12/11/2022] Open
Abstract
Stem cells from human exfoliated deciduous teeth (SHED) are highly proliferative pluripotent cells that can be retrieved from primary teeth. Although SHED are isolated from the dental pulp, their differentiation potential is not limited to odontoblasts only. In fact, SHED can differentiate into several cell types including neurons, osteoblasts, adipocytes, and endothelial cells. The high plasticity makes SHED an interesting stem cell model for research in several biomedical areas. This review will discuss key findings about the characterization and differentiation of SHED into odontoblasts, neurons, and hormone secreting cells (e.g., hepatocytes and islet-like cell aggregates). The outcomes of the studies presented here support the multipotency of SHED and their potential to be used for tissue engineering-based therapies.
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Affiliation(s)
- Vinicius Rosa
- Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore 119083
| | - Nileshkumar Dubey
- Oral Sciences, Faculty of Dentistry, National University of Singapore, Singapore 119083
| | - Intekhab Islam
- Discipline of Oral and Maxillofacial Surgery, Faculty of Dentistry, National University of Singapore, Singapore 119083
| | - Kyung-San Min
- Department of Conservative Dentistry, School of Dentistry, Chonbuk National University, Jeonju 54596, Republic of Korea
| | - Jacques E. Nör
- Department of Cariology, Restorative Sciences and Endodontics, School of Dentistry, University of Michigan, Ann Arbor, MI 48109, USA
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Regenerative Applications Using Tooth Derived Stem Cells in Other Than Tooth Regeneration: A Literature Review. Stem Cells Int 2015; 2016:9305986. [PMID: 26798366 PMCID: PMC4699044 DOI: 10.1155/2016/9305986] [Citation(s) in RCA: 45] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/05/2015] [Revised: 09/03/2015] [Accepted: 09/08/2015] [Indexed: 12/13/2022] Open
Abstract
Tooth derived stem cells or dental stem cells are categorized according to the location from which they are isolated and represent a promising source of cells for regenerative medicine. Originally, as one kind of mesenchymal stem cells, they are considered an alternative of bone marrow stromal cells. They share many commonalties but maintain differences. Considering their original function in development and the homeostasis of tooth structures, many applications of these cells in dentistry have aimed at tooth structure regeneration; however, the application in other than tooth structures has been attempted extensively. The availability from discarded or removed teeth can be an innate benefit as a source of autologous cells. Their origin from the neural crest results in exploitation of neurological and numerous other applications. This review briefly highlights current and future perspectives of the regenerative applications of tooth derived stem cells in areas beyond tooth regeneration.
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