The purpose of the present study was to evaluate the influence of systemic vitamin D (Vit D) administration with or without local photobiomodulation (PBM) on bone repair of surgically created critical-size defects (CSD) in rat calvaria.

Thirty-two rats were used. A 5 mm DTC was created on the calvaria of each animal. The animals were randomly distributed into four experimental groups (n = 8): Control: no treatment; Vit D: systemic Vit D administration through gavage; PBM: local irradiation with low-intensity laser therapy; Vit D/PBM: systemic Vit D administration through gavage and local irradiation with low-intensity laser therapy. The animals were euthanized 15 days after treatments. It was evaluated newly formed bone area (NFBA), percentage of mature and immature collagen fibers (picrosirius red staining and polarized light), immunohistochemical identification of tartrate-resistant acid phosphatase (TRAP), transforming growth factor (TGF), and osteocalcin (OCN). Data were statistically analyzed (ANOVA, Kruskal-Wallis, p < 0.05).

All experimental groups presented similar statistical results in the evaluated parameters. Some qualitative differences were observed, as follows. Groups PBM, Vit D, and Vit D/ PBM presented a slight amount of newly formed bone. Group Vit D presented a higher amount of mature collagen fibers. Groups Vit D and PBM presented higher immunoexpression of OCN than other experimental groups.

It can be concluded that Vit D and PBM did not significantly increase the amount of bone tissue formed but showed a trend towards enhanced bone maturation in the early healing stages of critical-sized defects surgically created in rat calvaria.

Periodontal disease, a significant global health burden, has encountered limited success with current therapeutic strategies to achieve full tissue regeneration. The emergence of Multilineage Differentiation and Stress-Enduring (Muse) cells presents a promising avenue for periodontal tissue regeneration. This study introduced a novel three-dimensional (3D) culture system utilizing Magnolia leaf vein scaffolds, characterized for their biocompatibility and evaluated for their impact on Muse cells' proliferation, adhesion, osteogenic differentiation, and exosome secretion. The isolation of Muse cells from Periodontal Ligament Stem Cells (PDLSCs) was successfully accomplished, with excellent compatibility observed with the plant-derived scaffolds. Notably, the 3D culture substantially upregulated osteogenic markers and promoted the formation of mineralized nodules, signifying enhanced osteogenic potential. Additionally, Muse cells in 3D culture exhibited a significant increase in exosome secretion, which were more effective in stimulating PDLSCs proliferation. The study concluded that plant leaf vein scaffolds provide a sustainable and effective platform for 3D stem cell culture, with the potential to significantly enhance the therapeutic efficacy of Muse cells in periodontal tissue engineering.

Periodontitis is characterized by the activity of neutrophil elastase, a host defense factor that leads to the destruction of the epithelial barrier and bacterial invasion of the periodontal tissue. Secretory leukocyte protease inhibitors (SLPI), predominantly secreted by epithelial cells, diffuse into the mucosal surface and inhibit excessive tissue loss caused by elastase during inflammation. The SLPI level is high in healthy gingiva and low in severe periodontitis. In this study, we hypothesized that intragingival administration of SLPI inhibits periodontal tissue destruction caused by periodontitis. Administration of SLPI significantly reduced neutrophil elastase activity in periodontal tissue and alleviated alveolar bone loss in mice. Real-time PCR analysis revealed that SLPI administration downregulated the transcription of pro-inflammatory cytokines and osteoclast-related factors in the gingival tissue. Furthermore, in vitro treatment of bone marrow macrophages with SLPI resulted in the downregulation of osteoclast differentiation. SLPI treatment of MC3T3-E1 cells promoted osteoblast differentiation and bone formation. These findings suggest that SLPI protects against periodontal tissue damage by suppressing inflammation and bone resorption and promoting bone regeneration.

Harnessing natural developmental programs to repair and replace damaged organs represents promising approaches in regenerative medicine. However, effective strategies are still lacking for tissue regeneration in complicated conditions, such as the periodontal bone defect. Here, human dental follicle stem cells (hDFSCs) and their aggregates (hDFSCA) are cultured and characterized, which are formed based on the inherent property of these stem cells self-assembly into compact spheroid-like structures, mimicking mesenchymal condensation in development. A periodontal tissue-specific microenvironment simulation material is then established, human decellularized alveolar bone matrix particles (hDABMPs), which possess favorable physicochemical and biological properties for regenerative use. hDFSCs co-cultured with hDABMPs exhibit improved cell function, and hDFSCA-hDABMP co-aggregates are subsequently constructed, which activate the developmental gene expression in hDFSCA and initiate hypoxic adaptation mechanisms for tissue regeneration. Indeed, hDFSCA-hDABMP co-aggregates significantly promote regeneration after implantation in alveolar bone defects with good biosafety. Interestingly, during the early stages of implantation, hDABMPs enhance hDFSC survival and expansion, thereby providing a sufficient source of cells for tissue regeneration. Collectively, this study reveals a development-inspired, engineered cell-niche co-aggregation strategy for enhancing CA therapeutic potential by simulating tissue-specific microenvironments, offering novel insights for functional tissue regeneration.

Dental stem cells are widely viewed as good options for bone regeneration because of their ease of acquisition, innate ability to renew themselves, and ability to differentiate into different types of cells. However, the process of osteogenic differentiation of dental stem cells is orchestrated by an intricate system of regulatory mechanisms. Recent studies have demonstrated the critical impacts of circular RNAs (circRNAs) on osteogenic differentiation of dental stem cells. Exploring the roles and regulatory pathways of circRNAs in dental stem cells could identify novel targets and approaches for utilizing dental stem cell therapy in clinical settings. This review provides a comprehensive overview of the functions and mechanisms of circRNAs, with a particular focus on their expression patterns and regulatory roles in osteogenic differentiation of various dental stem cell types. Furthermore, this review discusses current research challenges in this field and proposes future directions for advancing our understanding of circRNA-mediated regulation in dental stem cell biology.

In recent years, lactylation, a novel post-translational modification, has demonstrated a unique role in bridging cellular metabolism and epigenetic regulation. This modification exerts a dual-edged effect in both cancer and non-cancer diseases by dynamically integrating the supply of metabolic substrates and the activity of modifying enzymes: on one hand, it promotes tissue homeostasis and repair through the activation of repair genes; on the other, it exacerbates pathological progression by driving malignant phenotypes. In the field of oncology, lactylation regulates key processes such as metabolic reprogramming, immune evasion, and therapeutic resistance, thereby shaping the heterogeneity of the tumor microenvironment. In non-cancerous diseases, including neurodegeneration and cardiovascular disorders, its aberrant activation can lead to mitochondrial dysfunction, fibrosis, and chronic inflammation. Existing studies have revealed a dynamic regulatory network formed by the cooperation of modifying and demodifying enzymes, and have identified mechanisms such as subcellular localization and RNA metabolism intervention that influence disease progression. Nevertheless, several challenges remain in the field. This article comprehensively summarizes the disease-specific regulatory mechanisms of lactylation, with the aim of providing a theoretical foundation for its targeted therapeutic application.

Understanding the complexities of periodontal regeneration, particularly the unpredictable osteogenic/cementogenic differentiation of low-potential PDLSCs (LOP-PDLSCs), remains challenging. Identifying new therapeutic targets is crucial for enhancing regeneration. This study investigates the modulation of the Cholecystokinin (CCK) pathway, a key signaling cascade with roles in the gastrointestinal system, as a potential osteogenic/cementogenic pathway in PDLSCs.

Gastrointestinal CCK-related drugs, Lorglumide and Sincalide, were tested for their effects on mineralization in PDLSCs. Lorglumide blocked the CCK pathway in high-potential PDLSCs (HOP-PDLSCs), while Sincalide enhanced mineralization in low-potential PDLSCs (LOP-PDLSCs). Cellular viability was tested under different drug concentrations, followed by a mineralization assay (AR-S) using non-toxic doses. RT-qPCR for osteogenic-related genes (IGF1, OCN, RUNX2) and CCK pathway-related genes (CCK, CCKAR, CCKBR, COX2, FOS, JNK3, RGS2) assessed gene modulation. Alkaline phosphatase (ALP) activity, Ca²⁺ quantification, and IP3 receptor phosphorylation were also evaluated.

Lorglumide reduced mineralization, ALP activity, and RUNX2, OCN, and IGF1 transcripts in HOP-PDLSCs (p < 0.05). It decreased CCK and CCKAR expression, modulated COX2, FOS, JNK3, and RGS2 genes, reduced IP3 receptor phosphorylation, and lowered calcium levels (p < 0.05). Conversely, Sincalide enhanced mineralization in LOP-PDLSCs, increasing ALP activity and OCN and IGF1 expression (p < 0.05). It upregulated COX2, FOS, JNK3, and RGS2 genes, phosphorylated IP3 receptors in LOP1, and increased calcium levels in all LOP-PDLSCs (p < 0.05).

Sincalide and Lorglumide modulate PDLSCs' osteogenesis/cementogenesis, revealing the complex interplay of gastrointestinal drugs in periodontal tissue regeneration and offering insights for innovative therapies.

This study demonstrates the potential of gastrointestinal drugs targeting the CCK signaling pathway as innovative modulators for periodontal regeneration. By regulating osteogenic/cementogenic differentiation in hPDLSCs, these findings may pave the way for the development of novel biomaterials and therapies, promising improved outcomes in periodontal tissue regeneration for clinical applications.

This study aimed to investigate the specific role and mechanistic actions of tumor necrosis factor receptor-associated factor 4 (TRAF4) in periodontitis.

Human periodontal ligament stem cells (PDLSCs) were exposed to lipopolysaccharide (LPS). Then, real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting (WB) were carried out to determine the mRNA and protein expression levels of Smad ubiquitination regulator 1 (SMURF1). The relationship between TRAF4 and SMURF1, as predicted by the STRING and GeneMANIA databases, was verified by co-immunoprecipitation (Co-IP). Finally, both TRAF4 and SMURF1 were inhibited in PDLSCs by cell transfection, and the regulatory mechanisms involved were investigated by cell counting kit-8 assays, enzyme linked immunosorbent assay, WB, alkaline phosphatase, and alizarin red staining.

The gene and protein expression levels of SMURF1 in PDLSCs increased following LPS induction (p < 0.001); cell viability was decreased (p < 0.001), TRAF4 expression was decreased (p < 0.001), and cell-mineralized nodules were inhibited. Inhibition of SMURF1 expression increased PDLSCs activity and TRAF4 expression levels (p < 0.001), increased the number of cell-mineralized nodules, and enhanced cellular osteogenic capacity (p < 0.001).

SMURF1 regulates LPS-stimulated injury and improves the capacity for osteogenic differentiation in PDLSCs by downregulating the expression of TRAF4.

Dydrogesterone (DG), a synthetic isomer of progesterone, plays a potential regulatory role in the periodontal environment. The aim of this study was to investigate the potential effects of DG on periodontitis under periodontal therapy (PT) and the underlying mechanisms related to oral microbiota.

As a cohort study, perimenopausal women with periodontitis and abnormal uterine bleeding associated with ovulatory dysfunction were screened. A total of 30 women received PT (PT group) and 30 women received PT and oral DG 10 mg twice/day for 10 days/month (PT + DG group). At baseline and 3 months after treatment, pocket probing depth (PPD), bleeding index (BI), bleeding on probing (BOP), plaque index, CRP, IL-6, and TNF-α were measured. Additionally, 16S rDNA sequencing was performed to determine the characteristics of oral microbiota, mainly in terms of abundance, diversity, composition, and community structure.

Three months after treatment, the levels of PPD, BI, and BOP, as well as the levels of CRP, IL-6, and TNF-α in gingival crevicular fluid were significantly lower in the PT + DG group than those in the PT group. After treatment, a relatively lower microbial abundance, and some differences in microbial composition were revealed between the PT and PT + DG groups. At the genus level, significantly fewer Escherichia-Shigella, Porphyromonas, and Absconditabacteriales (SR1), and more Lactobacillus, Gordonia, Bifidobacterium, and Oribacterium were found in the PT group than in the PT + DG group.

DG enhances the effect of PT on inhibiting inflammatory response in women with periodontitis by mediating oral microbiota.

The bidirectional correlation between diabetes and periodontitis positions the latter as the most prevalent complication of the former. Rehabilitation of the periodontal tissues damaged by diabetic periodontitis presents a significant clinical challenge. The multifaceted nature of the pathogenesis of diabetic periodontitis necessitates a comprehensive approach in its treatment to mitigate its adverse effects. To address this, a temperature-sensitive hydrogel containing phlorotannins (PL) and antimicrobial peptide LL-37 was developed to shift the microenvironment of diabetic periodontitis from an exacerbated high-glycemic inflammatory state to a regenerative one. The addition of PL significantly enhanced the antimicrobial properties, stability, and safety of LL-37. Vitro experiments confirmed that PL/LL-37 had good biocompatibility and promoted osteogenic differentiation of bone. PL/LL-37 demonstrated antioxidant properties by scavenging DPPH free radicals and inhibiting NO production. Furthermore, PL/LL-37 effectively modulated macrophage polarization from a M1 phenotype to an M2 phenotype through NF-κB P-p65 inflammatory pathway, thereby reducing the release of pro-inflammatory cytokines and promoting the secretion of anti-inflammatory cytokines. Interestingly, it could downregulate the AGE-RAGE signaling pathway, exerting a protective effect against diabetes. In addition, PL/LL-37 could attenuate inflammation levels, inhibit osteoclast production, promote bone regeneration, inhibit apoptosis and decrease RAGE levels in a rat model of diabetic periodontitis. These combined features synergistically accelerate diabetic periodontal bone regeneration. Consequently, PL/LL-37 emerges as a promising candidate for clinical treatment of diabetic periodontitis.

Developing biomaterials capable of promoting bone regeneration in bacteria-infected sites is of utmost urgency for periodontal disease therapies. Here we produce a hybrid hydrogel by integrating CuS nanoparticles (CuSNPs), which could kill bacteria through photothermal therapy (PTT) triggered by a near infrared (NIR) light, and a gelatin methacryloyl (GelMA) hydrogel, which is injectable and biocompatible. Specifically, CuSNPs were precipitated by chitosan (CS) firstly, then grafted with methacrylic anhydride (MA) to form CuSNP@CS-MA, which was photo-crosslinked with GelMA to synthesize hybrid hydrogels (GelMA/CuSNP). The hybrid hydrogels exhibited a broad-spectrum antibacterial property that could be spatiotemprorally manipulated through applying a NIR light. Their mechanical properties were adjustable by controlling the concentration of CuSNPs, enabling the hydrogels to become more adapted to the oral diseases. Meanwhile, the hybrid hydrogels showed good cytocompatibility in vitro and improved hemostasis in vivo. Moreover, they accelerated alveolar osteogenesis and vascular genesis, successfully treating periodontis in four weeks in a rat model. GelMA/CuSNP hydrogels showed a broad-spectrum sterilization ability via PTT in vitro and outstanding antibacterial property in vivo, suggesting that the hybrid hydrogels could function in the challenging, bacteria-rich, oral environment. Such injectable hybrid hydrogels, capable of achieving both facilitated osteogenesis and NIR-inducible sterilization, represent a new biomaterial for treating periodontitis.

The keystone pathogen Porphyromonas gingivalis (P.g.) is responsible for cementum resorption in periodontitis; however, the mechanism involved in it remains unclear. Sirtuin 3 (Sirt3) is a NAD+-dependent protein deacetylase contributing to mitochondrial homeostasis and various cell functions. In this study, the expression of Sirt3 in cementoblasts was found to be increased during cementoblast mineralisation and cementum development, while it decreased gradually under P.g. infection in a multiplicity of infection-dependent manner. Compared with wild type mice, the Sirt3 knockout mice showed less cellular cementum and lower mineralisation capacity with decreased expression of Runx2 and OCN in cementoblasts. Sirt3 inhibition by 3-TYP or Sirt3 silencing by lentivirus infection both confirmed the impaired cementogenesis. Conversely, honokiol (HKL) was simulated to bind Sirt3 and was applied to activate Sirt3 in cementoblasts. HKL-mediated Sirt3 activation facilitated cementoblast mineralisation and rescued P.g.-suppressed cementoblast mineralisation markedly. Superoxide dismutase 2 (SOD2), the downstream molecule of Sirt3, showed a similar expression pattern to Sirt3 under different conditions. Silencing of SOD2 was demonstrated to restrain cementoblast mineralisation. The pan acetylation was detected to decrease under Sirt3-upregulating conditions and increase under Sirt3-downregulating conditions. The binding of Sirt3 and SOD2 in cementoblasts was also verified. Furthermore, SOD2 acetylation and specific SOD2-K68 acetylation were found to be upregulated under P.g. or Sirt3 silencing conditions and downregulated by HKL stimulation. Moreover, K68Q mutation simulating acetylation decreased cementoblast mineralisation, while K68R mutation simulating deacetylation increased it. Altogether, Sirt3 deacetylates SOD2 via K68 to orchestrate P.g.-perturbed cementogenesis, and HKL is a Sirt3-targeted treatment candidate.

Shilajit is a pale-brown to blackish-brown fluid that varies in consistency and is released from rock layers in various mountain ranges across the world. For thousands of years, traditional medical systems in several nations have included shilajit in one form or another as a rejuvenator and adaptogen. Numerous medicinal qualities have been attributed to it, several of which have been confirmed by contemporary scientific analysis. This in vitro study was established to investigate the effect of shilajit on human Periodontal ligament (hPDL) cell wound closure.

The cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium reduction (MTT) test following a 24-hour exposure of shilajit. With the use of an inverted phase contrast microscope, morphological alterations were noted. Acridine orange/ethidium bromide dual labeling, to evaluate the apoptotic cell death in shilajit treated cells. An in vitro wound healing test was utilized to evaluate wound healing in wounded hPDL cell monolayers for 24 h in the presence or absence of shilajit. The Matrix metalloproteinases-2 and 9 (MMP-2 and MMP-9) in hPDL cells treated with shilajit were measured using the enzyme-linked immunosorbent assay (ELISA) technique, and real-time PCR was used to examine gene expression linked to wound healing and apoptosis.

Shilajit's cytotoxicity evaluation on hPDL cells showed that dosages as high as 3 mg/mL had no adverse effects and maintains the cell viability, suggesting a possible stimulatory effect on cell growth. Cell viability was reduced significantly (p < 0.05) by dosages more than 4 mg/mL, indicating cytotoxicity at higher concentrations. According to the scratch wound healing assay, shilajit administration at doses of 2 and 3 mg/mL accelerated wound healing and improved cell migration in hPDL cells. Shilajit promoted a controlled inflammatory response and supported periodontal ligament healing by upregulating the expression of genes involved in collagen synthesis, collagen type I alpha 1 chain (COL1A1) and pro-inflammatory cytokines, tumor necrosis factor alpha (TNF-α) and, interleukin-1-beta (IL-1β), according to real-time PCR data. In addition, Shilajit raised the protein levels of MMP2 and MMP9, two important enzymes involved in tissue remodeling. Shilajit-treated hPDL cells showed a substantial increase of cell proliferation and no discernible apoptotic activity.

Our research offers novel proof that shilajit promotes hPDL cell migration and proliferation, which in turn promotes wound closure. According to these results, Shilajit may improve tissue regeneration, accelerate wound healing, and encourage the growth of periodontal ligament cells.

Human periodontal ligament stem cells (PDLSCs) are widely available and have strong osteogenic differentiation ability, which makes them promising tools for bone regeneration. Circular RNAs (circRNAs) play a variety of functions in the process of cell differentiation and are potential therapeutic targets. Here, we identified a new circRNA, circ-SPATA13, and found that it was highly positively correlated with the osteogenic differentiation of PDLSCs. Therefore, in this study, we revealed the significance and mechanism of circ-SPATA13 in the osteogenic differentiation of PDLSCs.

PDLSCs were isolated from third molars with incomplete apical development and induced to undergo chondrogenic, adipogenic, or osteogenic differentiation. Surface markers were detected via flow cytometry. Proliferation was assessed with EdU and CCK-8 assays. The circ-SPATA13 and miR-485-5p_R + 1-mediated control of mineral deposition was evaluated through alizarin red and alkaline phosphatase staining. Osteogenesis-related factor expression was detected through western blotting, immunofluorescence, and qRT-PCR. Fluorescence in situ hybridization was used to examine circ-SPATA13 localization within PDLSCs. The relationships among circ-SPATA13, miR-485-5p_R + 1, and BMP7 during PDLSCs osteogenesis were assessed through western blotting, qRT-PCR, dual-luciferase assay, rescue experiment, and bioinformatics approaches.

Primary PDLSCs expressing mesenchymal stem cell surface markers were isolated. Circ-SPATA13 was identified and found to have no impact on PDLSC proliferation, whereas it was a positive regulator of their osteogenic differentiation, a process which was antagonized by miR-485-5p_R + 1. Dual-luciferase reporter assays revealed that circ-SPATA13 was able to function as a molecular sponge to sequester miR-485-5p_R + 1 within PDLSCs, while this miRNA was able to bind to the 3'-UTR of the target mRNA BMP7. In rescue experiments, circ-SPATA13 was confirmed to regulate the osteogenic differentiation of PDLSCs via this miR-485-5p_R + 1/BMP7 axis. Moreover, in vivo experiments in rats demonstrated that the overexpression of circ-SPATA13 in PDLSCs was associated with the promotion of bone formation in a skull defect model system.

These data supported the osteogenic functions of circ-SPATA13 in PDLSCs. Mechanistically, this circRNA was found to function as a molecular sponge for miR-485-5p_R + 1, in turn targeting BMP7 to promote the osteogenic differentiation of PDLSCs. This circ-SPATA13/miR-485-5p_R + 1/BMP7 axis may be a novel target for treatments promoting PDLSCs osteogenic differentiation.

Periodontal bony defects pose a significant challenge in periodontology, necessitating advanced regenerative approaches to restore the lost structures. Stem cell-based therapies have emerged as a promising solution due to their ability to differentiate into various cells, modulating the regenerative microenvironment. This narrative review explores the potential of stem cells derived from multiple sources in treating periodontal bony defects. Additionally, we examine evidence from both animal and human studies, highlighting advancements, clinical outcomes, and limitations. By investigating these findings, this article provides a comprehensive overview of the advantages of stem cell-based therapies compared to other regenerative techniques in addressing periodontal bony defects and discusses the limitations of their translation into routine clinical practice.

Periodontitis is a chronic inflammatory disease affecting the supporting tissues of the teeth and has emerged as a global public health issue. Current therapies primarily address pathogenic factors and alleviate symptoms, with limited options available for complete restoration and reconstruction of already absorbed periodontal bone tissue. In this study, we developed a nanotherapeutic strategy utilizing fusion nanovesicles (FVs) to modulate the inflammatory microenvironment and create a regenerative niche for periodontal ligament stem cells (PDLSCs), which play a crucial role in periodontal tissue repair. The FVs are composed of Scutellaria baicalensis nanovesicles (SBNVs) with anti-Porphyromonas gingivalis (P. gingivalis) and anti-inflammatory properties, combined with PDLSC membrane-derived nanovesicles genetically engineered to express TNFR1. These FVs preserved the biological activity of SBNVs and the immunomodulatory function of PDLSCs. Additionally, FVs effectively captured and cleared TNF-α from the microenvironment through TNFR1. Moreover, FVs alleviated the inflammatory response of PDLSCs induced by P. gingivalis-LPS (Pg-LPS) and TNF-α, restoring their proliferation, migration, and osteogenic differentiation capabilities. Hence, this nanotherapeutic strategy holds great potential for treating periodontitis.

Periodontal disease is a common chronic inflammatory condition affecting the tissues that support teeth, leading to their destruction. Mitophagy, a specialized form of autophagy responsible for degrading damaged mitochondria, plays a crucial role in maintaining cellular homeostasis. However, its role in periodontal disease progression remains poorly understood. This review aims to summarize recent research on mitophagy's role in periodontal disease pathogenesis.

A comprehensive literature review on mitophagy was conducted using PubMed, Scopus, and Web of Science databases, employing keywords related to periodontal disease such as "periodontal," "periodontitis," "gingiva," and "gingivitis."

A review of 18 original studies revealed that mitophagy plays a crucial role in periodontal disease by regulating key pathophysiological mechanisms. Specifically, mitophagy modulates periodontal inflammation by influencing pro-inflammatory cytokines and mitochondrial reactive oxygen species. Additionally, it is essential for alveolar bone remodeling, impacting both bone resorption and regeneration. Mitophagy also regulates cell apoptosis within periodontal tissues, helping to preserve cellular function and tissue integrity during periodontal disease progression.

Mitophagy regulates periodontal disease pathogenesis by modulating inflammation, bone remodeling, and cell death in periodontal tissues. Further research is needed to explore its therapeutic potential in periodontal disease treatment and improve targeted interventions.

Macrophages are vital for regulating periodontal health, and numerous studies have extensively investigated their role in periodontitis progression. This study employs bibliometric analysis to identify research trends and hotspots related to macrophages in periodontitis from 2004 to 2024, thereby guiding future investigations and exploring potential clinical applications. Literature retrieval and dataset export were conducted via the Web of Science Core Collection database. Subsequently, bibliometric analysis was performed and the results were visualized using Microsoft Office Excel, VOSviewer, CiteSpace, and GraphPad Prism. A total of 1542 papers from 2004 to 2024 on the macrophages associated with periodontitis were identified. The annual number of publications and citations has steadily increased. China and the United States were the primary collaborators and drivers of research. Sichuan University had the largest number of published studies. The Journal of Periodontal Research published the most papers, while the Journal of Periodontology had the most citations. Daniel Grenier was the most prolific author and George Hajishengallis was the most frequently co-cited author. Co-citation analysis indicated that researchers primarily investigate the mechanisms of macrophages in periodontitis through both in vivo and in vitro experiments. Emerging research hotspots encompass keywords such as "macrophage polarization," "extracellular vesicles," and "regeneration." Research on macrophages in periodontitis is progressing quickly, and their strategic targeting offers a new and promising direction for future studies on regenerating periodontal tissues. The utilization of extracellular vesicles derived from diverse sources to regulate M1/M2 macrophage polarization presents a promising strategy for the treatment of periodontitis.

DNA methyltransferase 3A (Dnmt3a) is an enzyme that catalyzes the de novo methylation of DNA, and plays essential roles in a wide range of physiological and pathological processes. However, it remains unclear whether Porphyromonas gingivalis affects cementoblasts, the cells responsible for cementum formation, through Dnmt3a.

The samples were collected from models of mouse periapical lesions and mice of different ages, and the expression of Dnmt3a was detected through immunofluorescence. Porphyromonas gingivalis was co-cultured with cementoblasts that simultaneously overexpressed Dnmt3a. Additionally, cementoblasts were subjected to either Dnmt3a knockout or DNA methylation inhibition. Changes in global DNA methylation were analyzed, and quantitative PCR, western blotting, alkaline phosphatase (ALP) activity assays, and Alizarin Red staining were employed to evaluate alterations in the mineralization capacity of cementoblasts.RNA sequencing further showed the mechanisms by which Dnmt3a regulated mineralization. Flow cytometry, MitoSox, and TRMR staining were used to verify the participation of mitochondria-dependent apoptosis.

The effect of P. gingivalis on Dnmt3a and global DNA methylation in cementoblasts was first verified. Dnmt3a expression and global DNA methylation were upregulated during cementoblast mineralization. Samples with periapical inflammation exhibited reduced Dnmt3a expression. P. gingivalis stimulation reduced the global DNA methylation and the mineralization ability of cementoblasts. Both the knockdown of Dnmt3a and using DNA methylation inhibitors suppressed cementoblast mineralization. In addition, Dnm3a depletion was significantly correlated with the mitochondria-dependent apoptosis pathway in cementoblasts.

P. gingivalis blocks DNA methylation by silencing Dnmt3a in cementoblasts, thereby inducing mitochondrial-dependent apoptosis and, ultimately, impaired cementogenesis.

To investigate the role of lncRNA Lockd in mandibular mesenchymal stem cell (M-MSC) proliferation and osteogenic capability in the inflammatory microenvironment, focusing on its interaction with SUZ12.

Using lncR Lockd knockdown/overexpression cell models and a murine periodontitis model, we explored Lockd's effects on M-MSC proliferation and osteogenic capability in the inflammatory microenvironment. Predictions from multiple databases and a series of rescue experiments revealed the regulatory role of the Lockd/SUZ12 signalling axis of M-MSC in the inflammatory microenvironment.

Lockd was found to stimulate M-MSC proliferation but impair osteogenic differentiation. The in vitro studies suggested that the activation of Lockd negatively inhibited the osteogenic differentiation process and may ultimately impact bone formation in periodontitis. Mechanistically, it was elucidated that Lockd interacts with SUZ12, a core component of the polycomb repressive complex 2 (PRC2), and may affect the PRC2 complex's role in osteogenic gene expression.

Lockd boosts the proliferation of M-MSCs but inhibits their osteogenic differentiation by interacting with SUZ12, potentially inhibiting osteogenic capability in the inflammatory microenvironment.

Cementum, a bone-like tissue, is an essential component of periodontium, and periodontitis can lead to degenerative changes in the cementum, eventually resulting in tooth loss. The therapeutic strategy for advanced periodontitis is to achieve periodontal regeneration, of which cementum regeneration is a key criterion. Cementoblasts are responsible for cementogenesis, and their mineralization counts in cementum regeneration. However, research is still limited. Thus, novel treatment targets are required. The expression levels of lysine (K)-specific demethylase 6B (KDM6B), fatty acid oxidation (FAO), and cementogenic markers were detected by quantitative polymerase chain reaction, Western blot, immunofluorescence, and immunohistochemical assays. FAO levels were analyzed by assay kit. In vivo, injection of GSK-J4 into mice detected the influence of KDM6B on cementum formation. Chromatin immunoprecipitation sequencing, transcriptomic RNA sequencing, subsequent chromatin immunoprecipitation-quantitative polymerase chain reaction and overexpression of HADHA (hydroxyacyl-coA dehydrogenase trifunctional multienzyme complex subunit alpha) elucidated the KDM6B-Hadha axis. Global lactylation was detected by Western blot. Lactylation proteomics clarified the modified sites of HADHA. Mutating these sites and applying coimmunoprecipitation confirmed their significance. Knockdown of Kdm6b was utilized to assess its regulation on the lactylation of HADHA, FAO, and mineralization levels. FAO and KDM6B expression was elevated during cementoblast mineralization. KDM6B targeted Hadha and activated its transcription, thereby increasing FAO levels and promoting mineralization. Lactylation occurred in the process of mineralization, and KDM6B could regulate the lactylation of HADHA to promote FAO and mineralization. Overexpression of Hadha and the addition of lactate sodium could rescue the inhibition of mineralization by knockdown of Kdm6b. In summary, during cementoblast mineralization, KDM6B regulates HADHA by mediating histone demethylation and lactylation, thereby upregulating FAO and thus promoting mineralization.

Intact cementum is vital for tooth stability and health. Cementoblasts, which line the root surface, are responsible for cementum formation. Recent evidence suggests that circular RNAs (circRNAs) are involved in various cellular functions and may have clinical applications. Although circHIPK3 has been shown to participate in osteogenesis, its role in cementoblast differentiation and mineralization is not well understood.

The ring structure of circHIPK3 was confirmed using Sanger sequencing and an actinomycin D assay. Subcellular localization of circHIPK3 was assessed using a nucleus-cytoplasm separation assay. RT-qPCR was employed to analyze circHIPK3 expression during cementoblast differentiation and following TNF-α treatment. In vivo, periapical lesions were induced in mouse mandibular first molars to mimic an inflammatory environment, and circHIPK3 expression was evaluated. The interaction of the circHIPK3/miR-10b-5p/DOHH axis was explored through RNA pull-down assays, bioinformatics analysis, and dual-luciferase reporter assays. The effects on cementoblast differentiation and mineralization were assessed by measuring osteogenic markers, alkaline phosphatase (ALP) activity, ALP staining, and alizarin red S staining.

CircHIPK3 was predominantly located in the cytoplasm of cementoblasts, and its expression was significantly upregulated during cementoblast differentiation. Knockdown of circHIPK3 inhibited cementoblast differentiation and mineralization, whereas its overexpression promoted these processes. Mechanistically, circHIPK3 upregulated DOHH expression by sponging miR-10b-5p, thereby enhancing cementoblast differentiation and mineralization. The NF-κB pathway was found to act downstream of the circHIPK3/miR-10b-5p/DOHH axis in these processes. Additionally, circHIPK3 expression was significantly downregulated in inflammatory environments both in vitro and in vivo. Forced overexpression of circHIPK3 mitigated the inhibitory effects of TNF-α on cementoblast differentiation and mineralization.

CircHIPK3 acts as a positive regulator of cementoblast differentiation and mineralization through the miR-10b-5p/DOHH/NF-κB axis, playing a crucial role in both normal and pathological cementogenesis.

Dental pulp stem cells (DPSCs) have been extensively used for tissue regeneration owing to their notable capabilities. Insulin-like growth factor-binding protein 5 (IGFBP5) regulates osteogenic differentiation of mesenchymal stem cells (MSCs); however, the underlying regulatory mechanisms require further investigation.

Carboxyfluorescein succinimidyl ester, an alkaline phosphatase (ALP) activity assay and Alizarin Red staining were used to reveal the role of IGFBP5 in DPSCs. Protein expression levels were determined using western blotting. Immunofluorescence was used to observe cell sub-localisation. Subcutaneous transplantation in nude mice was used to observe the osteogenesis of DPSCs in vivo.

IGFBP5 enhanced the proliferation and osteogenic differentiation of DPSCs. Deletion of the nuclear localisation sequence (NLS) of IGFBP5 prevented its nuclear import and abolished all its promoting effects on DPSCs; ivermectin stimulation attenuated the enhancement of ALP activity by IGBFP5. Bone-like tissue formation promoted by IGFBP5 in vivo vanishes when the NLS is deleted. Inhibition of IGFBP5 nuclear import attenuated the IGFBP5-induced phosphorylation of JNK (p-JNK) and phosphorylated ERK (p-ERK) in DPSCs.

Our findings suggest that cell proliferation and osteogenic differentiation effects exerted by IGFBP5 on DPSCs are closely associated with their entry into the nucleus, thereby providing a novel potential target for tissue regeneration.

Improving the microenvironment to augment endogenous regenerative potential has emerged as a fundamental concept for stimulating and expediting periodontal tissue repair and regeneration. Previous studies have demonstrated that TPPU, a soluble epoxide hydrolase inhibitor (sEHi), mediates the suppression of inflammatory bone loss in periodontitis models. However, the underlying mechanisms remain largely elusive.

In this study, we constructed a human umbilical vein endothelial cell (HUVEC) and periodontal ligament stem cell (PDLSC) coculture system in vitro and tested the anti-inflammatory effect of TPPU under inflammatory conditions. The roles of HIF-1α and Endomucin (EMCN) in the anti-inflammatory effects of TPPU were analyzed. The effects of TPPU on osteogenesis and osteoclastogenesis in cocultured cells were examined. The in vivo periodontitis model further verified the effects of TPPU on inhibiting neutrophil adhesion and inflammation and inhibiting osteoclasts.

Our in vitro experiments demonstrated that TPPU enhances the interaction between mesenchymal stem cells and vascular endothelial cells to enhance anti-inflammatory and osteogenic differentiation effects and revealed a new anti-inflammatory mechanism of TPPU involving the upregulation of EMCN in endothelial cells to prevent lymphocyte recruitment. We also confirmed that TPPU inhibits osteoclast activity. Our in vivo findings showed that TPPU inhibits osteoclast activity and neutrophil adhesion and enhances periodontal tissue repair and regeneration.

TPPU promotes local regeneration in periodontitis by inhibiting inflammation and bone resorption. Thus, targeting soluble epoxide hydrolase represents an endogenous regenerative strategy for periodontitis treatment.

This narrative review delves into the molecular mechanisms of hyaluronic acid (HA) and re-epithelializing agents in the context of periodontal regeneration. Periodontitis, characterized by chronic inflammation and the destruction of tooth-supporting tissues, presents a significant challenge in restorative dentistry. Traditional non-surgical therapies (NSPTs) sometimes fail to fully manage subgingival biofilms and could benefit from adjunctive treatments. HA, with its antibacterial, antifungal, anti-inflammatory, angiogenic, and osteoinductive properties, offers promising therapeutic potential. This review synthesizes the current literature on the bioactive effects of HA and re-epithelializing agents, such as growth factors and biomaterials, in promoting cell migration, proliferation, and extracellular matrix (ECM) synthesis. By modulating signaling pathways like the Wnt/β-catenin, TGF-β, and CD44 interaction pathways, HA enhances wound healing processes and tissue regeneration. Additionally, the role of HA in facilitating cellular crosstalk between epithelial and connective tissues is highlighted, as it impacts the inflammatory response and ECM remodeling. This review also explores the combined use of HA with growth factors and cytokines in wound healing, revealing how these agents interact synergistically to optimize periodontal regeneration. Future perspectives emphasize the need for further clinical trials to evaluate the long-term outcomes of these therapies and their potential integration into periodontal treatment paradigms.

Bacterial infection severely limits the effectiveness of biomaterials for tissue repair, posing a major challenge to modern medicine. Despite advances in novel antibiotics and their application in treatment, challenges remain in clinical practice. To address this issue, biomaterials are engineered to achieve desirable anti-infective performance and compatibility via adjusting their surface physicochemical properties. Recently, numerous studies on piezoelectric materials have been performed for anti-infective and regenerative therapies, but a comprehensive review is still lacking. This article provides a brief overview of the different types of piezoelectric materials and their characteristics. Building on this understanding, this review highlights the antibacterial mechanisms including orchestrating electric field and optimizing piezoelectric catalysis, which promote infective tissue regeneration, as well as discusses the anti-infective bioapplication of piezoelectric materials. Furthermore, this review concludes with perspectives into the challenges and future research directions of piezoelectric biomaterials.

Cementum is a vital component of periodontium, yet its regeneration remains a challenge. Pentraxin 3 (PTX3) is a multifunctional glycoprotein involved in extracellular matrix remodeling and bone metabolism regulation. However, the role of PTX3 in cementum formation and cementoblast differentiation has not been elucidated. In this study, we initially observed an increase in PTX3 expression during cementum formation and cementoblast differentiation. Then, overexpression of PTX3 significantly enhanced the differentiation ability of cementoblasts. While conversely, PTX3 knockdown exerted an inhibitory effect. Moreover, in Ptx3-deficient mice, we found that cementum formation was hampered. Furthermore, we confirmed the presence of PTX3 within the hyaluronan (HA) matrix, thereby activating the ITGB1/FAK/YAP1 signaling pathway. Notably, inhibiting any component of this signaling pathway partially reduced the ability of PTX3 to promote cementoblast differentiation. In conclusion, our study indicated that PTX3 promotes cementum formation and cementoblast differentiation, which is partially dependent on the HA/ITGB1/FAK/YAP1 signaling pathway. This research will contribute to our understanding of cementum regeneration after destruction.

One of the most promising approaches to correct periodontal bone defects and achieve periodontal regeneration is platelet-rich fibrin (PRF). This systematic review and meta-analysis aimed to evaluate the regeneration of periodontal bone defects using PRF compared to other regenerative treatments. The data search and retrieval process followed the PRISMA guidelines. An electronic search of MEDLINE, Cochrane, and PubMed databases was performed, selecting exclusively randomized clinical trials where the following were measured: probing depth reduction (PD), clinical attachment level gain (CAL), and radiographic bone fill (RBF). Out of 284 selected articles, 32 were chosen based on inclusion criteria. The use of platelet-rich fibrin (PRF) + open flap debridement (OFD), PRF + metformin, PRF + platelet-rich plasma (PRP), and PRF + OFD/bone graft (BG) significantly reduced PD and improved CAL and RBF. However, the combination of PRF + BG, PRF + metformin, and PRF + STATINS reduced CAL. The intervention of PRF combined with different treatments such as metformin, OFD, PRP, BG, and STATINS has a significant impact on improving PD and CAL. The use of PRF significantly improved the regeneration of periodontal bone defects compared to other treatments.

Treatment strategies for hard tissue defects aim to establish a mineralized microenvironment that facilitates tissue remodeling. As a mineralized tissue, cementum shares a similar structure with bone and exhibits an excellent capacity to resist resorption under compression. Macrophages are crucial for mineralized remodeling; however, their functional alterations in the microenvironment of cementum remain poorly understood. Therefore, this study explores the mechanisms by which cementum resists resorption under compression and the regulatory roles of cementoblasts in macrophage functions. As a result, extracellular vesicles from compression-loaded cementoblasts (Comp-EVs) promote macrophage M2 polarization and enhance the clearance of apoptotic cells (efferocytosis) by 2- to 3-fold. Local injection of Comp-EVs relieves cementum destruction in mouse root resorption model by activating the tissue repair function of macrophages. Moreover, Comp-EV-loaded hydrogels achieve significant bone healing in calvarial bone defect. Unexpectedly, under compression, EV secretion in cementoblasts is reduced by half. RNA-Seq analysis and verification reveal that Rab35 expression decreases by 60% under compression, thereby hampering the release of EVs. Rab35 overexpression is proposed as a modification of cementoblasts to boost the yield of Comp-EVs. Collectively, Comp-EVs activate the repair function of macrophages, which will be a potential therapeutic strategy for hard tissue repair and regeneration.

This study examined how range concentrations of Fibroblast Growth Factor-2 (FGF-2) influence the differentiation and activity of human-derived periodontal ligament (hPDLSCs) and alveolar bone-derived stem cells (haBMSCs).

hPDLSCs and haBMSCs were cultured with varying concentrations of FGF-2 (0, 1, 2.5, 5, 10, 20 ng/mL) and monitored for osteogenic differentiation through alkaline phosphatase (ALP) activity and quantification of gene expression (qRT-PCR) for osteogenesis markers. Additionally, alizarin red staining and a hydroxyproline colorimetric assay evaluated and quantified osteogenic matrix mineralization and collagen deposition. Statistical analyses were performed using one-way ANOVA or two-way ANOVA for multiple comparisons between groups.

At low FGF-2 concentrations, hPDLSCs differentiated toward an osteogenic lineage, whereas higher concentrations of FGF-2 inhibited osteogenesis and promoted fibroblastic differentiation. The effect of FGF-2 at the lowest concentration tested (1 ng/mL) led to significantly higher ALP activity than osteogenically induced positive controls at early time points and equivalent RUNX2 expression at early and later time points. FGF-2 supplementation of haBMSC cultures was sufficient, at all concentrations, to increase ALP activity at an earlier time point. Mineralization of haBMSC cultures increased significantly within 5-20 ng/mL FGF-2 concentrations under basal growth media conditions (α-minimal essential medium supplemented with 15 % fetal bovine serum and 1 % penicillin/streptomycin).

FGF-2 has a dual capacity in promoting osteogenic and fibroblastic differentiation within hPDLSCs contingent upon the dosage and timing of administration, alongside supporting osteogenic differentiation in haBMSCs. These findings underscore the need for precision growth factors dosing when considering the design of biomaterials for periodontal regeneration.

Periodontal tissue loss is the main reason for tooth mobility and loss caused by periodontal disease. Dental follicle stem cells (DFSCs) have significant therapeutic potential in periodontal regeneration, which maybe mainly depends on their potent immunomodulatory capacity. Consequently, this study aims to elucidate the impact of implanted xenogenous DFSCs on innate immune responses during early and late stages in the periodontal defect repair period.

To trace and investigate the immunomodulation mechanisms of DFSCs in vivo, DFSCs were engineered (E-DFSCs) using lentiviral vectors expressing CD63-enhanced green fluorescent protein (CD63-EGFP) and β-Actin-mCherry protein (ACTB-mCherry) to exhibit green and red fluorescence. The biological characteristics and functions of E-DFSCs were verified by proliferation, differentiation, and co-culture experiments in vitro. In vivo, the periodontal regeneration capacity of E-DFSCs was detected by implantation of murine periodontal defect model, and the response of innate immune cells was detected at the 1st, 3rd, and 5th days (early stage) and 4th week (late stage) after implantation.

In vitro assessments showed that E-DFSCs retain similar properties to their non-engineered counterparts but exhibit enhanced macrophage immunomodulation capability. In mice models, four-week micro-CT and histological evaluations indicated that E-DFSCs have equivalent efficiency to DFSCs in periodontal defect regeneration. At the early stage of repair in mice periodontal defect, fluorescence tracking showed that implanted E-DFSCs might primarily activate endogenous cells through direct contact and indirect actions, and most of these cells are myeloperoxidase-positive neutrophils. Additionally, compared with the control group, the neutrophilic infiltration and conversion of N2-type were significantly increased in the E-DFSC group. At the late stage of defect regeneration, more M2-type macrophages, fewer TRAP + osteoclasts, and an upregulated OPG/RANKL ratio were detected in the E-DFSC group compared to the control group, which indicated that immune balance tilts towards healing and bone formation.

The xenogenous implanted DFSCs can induce the N2 phenotype of neutrophils in the early stage, which can activate the innate immune mechanism of the host to promote periodontal tissue regeneration.

This report aims to present a treatment of retrograde peri-implantitis originating from apical periodontitis of an adjacent tooth in an 84-year-old male. Apical periodontitis of the maxillary left central incisor (#9) extended to the apex of the maxillary left lateral incisor implant (#10), which had been functioning for 16 years. Root canal treatment for #9 was performed, followed by root end surgery to treat the apical periodontitis, which showed a periapical radiolucency measured 1 cm in its greatest dimension. After the root end filling was placed, neither bone substitute materials nor barrier membranes were used to fill and cover the bony defect area. A 2-year postoperative radiograph confirmed the osseous healing around the apices of #9 and #10.

Prevention of periodontal bone resorption triggered by Porphyromonas gingivalis (P. gingivalis) is crucial for dental stability. Capsaicin, known as the pungent ingredient of chili peppers, can activate key signaling molecules involved in osteogenic process. However, the effect of capsaicin on osteogenesis of periodontal ligament stem cells (PDLSCs) under inflammation remains elusive.

P. gingivalis culture suspension was added to mimic the inflammatory status after capsaicin pretreatment. The effects of capsaicin on the osteogenesis of PDLSCs, as well as mitochondrial morphology, Ca2+ level, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and osteogenesis-regulated protein expression levels were analyzed. Furthermore, a mouse experimental periodontitis model was established to evaluate the effect of capsaicin on alveolar bone resorption and the expression of osteogenesis-related proteins.

Under P. gingivalis stimulation, capsaicin increased osteogenesis of PDLSCs. Not surprisingly, capsaicin rescued the damage to mitochondrial morphology, decreased the concentration of intracellular Ca2+ and ROS, enhanced MMP and activated phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway. The in vivo results showed that capsaicin significantly attenuated alveolar bone loss and augmented the expression of bone associated proteins.

Capsaicin increases osteogenesis of PDLSCs under inflammation and reduces alveolar bone resorption in mouse experimental periodontitis.

Periodontitis is a widely prevalent oral disease around the world characterized by the disruption of the periodontal ligament and the subsequent development of periodontal pockets, as well as the loss of alveolar bone, and may eventually lead to tooth loss. This research aims to assess the suppressive impact of Eupatilin, a flavone obtained from Artemisia argyi, on osteoclastogenesis in vitro and periodontitis in vivo. We found that Eupatilin can efficiently obstruct the differentiation of Raw264.7 and bone marrow-derived macrophages (BMDMs) induced by RANKL, leading to the formation of mature osteoclasts. Consistently, bone slice resorption assay showed that Eupatilin significantly inhibited osteoclast-mediated bone resorption in a dose-dependent manner. Eupatilin also downregulated the expression of osteoclast-specific genes and proteins in Raw264.7 and BMDMs. RNA sequencing showed that Eupatilin notably downregulated the expression of Siglec-15. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses identified significantly enriched pathways in DEGs, including MAPK signaling pathway. And further mechanistic investigations confirmed that Eupatilin repressed MAPKs/NF-κBsignaling pathways. It was found that Siglec-15 overexpression reversed the inhibitory impact of Eupatilin on the differentiation of osteoclasts. Furthermore, activating MAPK signaling pathway reversed the downregulation of Siglec-15 and the inhibition of osteoclastogenesis by Eupatilin. To sum up, Eupatilin reduced the expression of Siglec-15 by suppressing MAPK signaling pathway, ultimately leading to the inhibition of osteoclastogenesis. Meanwhile, Eupatilin suppressed the alveolar bone resorption caused by experimentalperiodontitis in vivo. Eupatilin exhibits potential therapeutic effects in the treatment of periodontitis, rendering it a promising pharmaceutical agent.

This narrative review illuminates on the application of single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics (ST) in periodontitis and highlights the probability of relating cell population and gene signatures to the pathogenesis of the disease for a better diagnosis.

An electronic search of the literature in the PubMed database for the keywords, "single cell sequencing" OR "spatial transcriptomics" and "periodontitis" OR "gingiva" OR "oral mucosa" yielded 486 research articles and reviews. After filtering duplicates and careful curation, 22 papers conducted in humans were retained.

The molecular mechanisms underlying periodontitis are complex and involve the interaction of multiple cells and various gene expressions. Most residing cells in periodontal tissues participate in maintaining homeostasis and health, while in addition to infiltrating immune cells contribute to the fight against the bacterial insult.

scRNA-seq and ST have provided new insights into the cellular and molecular changes associated with periodontitis for a better diagnosis and clinical outcome. New functions of cells and genes are revealed with these techniques; however, no cells or gene signatures are attributed to periodontitis so far.

Maxillofacial bone defect is one of the common symptoms in maxillofacial, which affects the function and aesthetics of maxillofacial region. Periodontal ligament stem cells (PDLSCs) are extensively used in bone tissue engineering. The mechanism that regulates the osteogenic differentiation of PDLSCs remains not fully elucidated. Previous studies demonstrated that l-Caldesmon (l-CALD, or CALD1) might be involved in the osteogenic differentiation of PDLSCs. Here, the mechanism by which CALD1 regulates the osteogenic differentiation of PDLSCs is investigated. The osteogenic differentiation of PDLSCs is enhanced with Cald1 knockdown. Whole transcriptome sequencing (RNA-seq) analysis shows that bone morphogenetic proteins (BMP) signaling pathway and Wingless type (Wnt) pathway have significant change with Cald1 knockdown, and the expressions of Wnt-induced secreted protein 1 (WISP1), BMP2, Smad1/5/9, and p-Smad1/5/9 are significantly upregulated, while Glycogen synthase kinase 3β (GSK3β) and p-GSK3β are downregulated. In addition, subcutaneous implantation in nude mice shows that knockdown of Cald1 enhances the osteogenic differentiation of PDLSCs in vivo. Taken together, this study demonstrates that knockdown of Cald1 enhances the osteogenic differentiation of PDLSCs by BMP and Wnt signaling pathways, and provides a novel approach for subsequent clinical treatment.

Inflammation-related bone defects often lead to poor osteogenesis. Therefore, it is crucial to reduce the inflammation response and promote the osteogenic differentiation of stem/progenitor cells to revitalize bone physiology. Here, a kind of hybrid nano-hydroxyapatite was prepared using the confined phosphate ion release method with the participation of fucoidan, a marine-sourced polysaccharide with anti-inflammation property. The physicochemical analyses confirmed that the fucoidan hybrid nano-hydroxyapatite (FC/n-HA) showed fine needle-like architectures. With a higher amount of fucoidan, the crystal size and crystallinity of the FC/n-HA reduced while the liquid dispersibility was improved. Cell experiences showed that FC/n-HA had an optimal cytocompatibility at concentration of 50 μg/mL. Moreover, the lipopolysaccharide-induced cellular inflammatory model with PDLSCs was established and used to evaluate the anti-inflammatory and osteogenic properties. For the 1%FC/n-HA group, the expression levels of TNF-α and IL-1β were significantly reduced at 24 h, while the expression of alkaline phosphatase of PDLSCs was significantly promoted at days 3 and 7, and calcium precipitates was enhanced at 21 days. In this study, the FC/n-HA particles showed effective anti-inflammatory properties and facilitated osteogenic differentiation of PDLSCs, indicating which has potential application in treating bone defects associated with inflammation, such as periodontitis.

Periodontal disease is a common oral infection which affects the tooth-supportive tissues directly. Considering the limitation of present regenerative treatments for severe periodontal cases, cytotherapies have been gradually introduced. Human periodontal ligament-derived mesenchymal stromal cells (hPDLMSCs), while identified as one of the promising cell sources for periodontal regenerative therapy, still hold some problems in the clinical application especially their limited life span. To solve the problems, human induced pluripotent stem cells (hiPSCs) are taken into consideration as a robust supply for hPDLMSCs.

The induction of hPDLMSCs was performed based on the generation of neural crest-like cells (NCLCs) from hiPSCs. Fibronectin and laminin were tested as coating materials for NCLCs differentiation when following previous protocol, and the characteristics of induced cells were identified by flow cytometry and RT-qPCR for evaluating the induction efficiency. Subsequently, selected dental ectoderm signaling-related cytokines were applied for hPDLMSCs induction for 14 days, and dental mesenchyme-related genes, dental follicle-related genes and hPDL-related genes were tested by RT-qPCR for the evaluation of differentiation.

Compared to the 58% in laminin-coated condition, fibronectin-coated condition had a higher induction efficiency of CD271high cells as 86% after 8-day induction, while the mesenchymal potential of induced NCLCs was similar between two coating materials.It was shown that the gene expressions of dental mesenchyme, dental follicles and hPDL cells were significantly enhanced with the stimulation of the combination with fibroblast growth factor 8b (FGF8b), FGF2, and bone morphogenetic protein 4 (BMP4).

FN coating was more effective in NCLCs induction, and the FGF8b+FGF2+BMP4 growth factor cocktail was effective in hPDLMSC-like cell generation. These findings underscored the likely regenerative potential of hiPSCs as an applicable and promising curative strategy for periodontal diseases.

Periodontitis is an inflammatory process that starts with soft tissue inflammation caused by the intervention of oral bacteria. By modulating local immunity, it is possible to supplement or replace current therapeutic methods. The aim of this study was to compare the effects of an immunostimulatory treatment with the antibiotherapy usually applied to periodontitis patients. On a model of periodontitis induced in 30 rats (divided into three equal groups) with bacterial strains selected from the human oral microbiome (Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum and Streptococcus oralis), we administered antibiotics, bacterial lysates and saline for 10 days. Clinically, no significant lesions were observed between the groups, but hematologically, we detected a decrease in lymphocyte and neutrophil counts in both the antibiotic and lysate-treated groups. Immunologically, IL-6 remained elevated compared to the saline group, denoting the body's effort to compensate for bone loss due to bacterial action. Histopathologically, the results show more pronounced oral tissue regeneration in the antibiotic group and a reduced inflammatory reaction in the lysate group. We can conclude that the proposed bacterial lysate has similar effects to antibiotic therapy and can be considered an option in treating periodontitis, thus eliminating the unnecessary use of antibiotics.