Apoptosis, a form of programmed cell death, is essential for growth and tissue homeostasis. Apoptotic bodies (ApoBDs) are a form of extracellular vesicles (EVs) released by dying cells in the last stage of apoptosis and were previously regarded as debris of dead cells. Recent studies unraveled that ApoBDs are not cell debris but the bioactive treasure left behind by the dying cells with an important role in intercellular communications related to human health and various diseases. Defective clearance of ApoBDs and infected-cells-derived ApoBDs are possible etiology of some diseases. Therefore, it is necessary to explore the function and mechanism of the action of ApoBDs in different physiological and pathological conditions. Recent advances in ApoBDs have elucidated the immunomodulatory, virus removal, vascular protection, tissue regenerative, and disease diagnostic potential of ApoBDs. Moreover, ApoBDs can be used as drug carriers enhancing drug stability, cellular uptake, and targeted therapy efficacy. These reports from the literature indicate that ApoBDs hold promising potential for diagnosis, prognosis, and treatment of various diseases, including cancer, systemic inflammatory diseases, cardiovascular diseases, and tissue regeneration. This review summarizes the recent advances in ApoBDs-related research and discusses the role of ApoBDs in health and diseases as well as the challenges and prospects of ApoBDs-based diagnostic and therapeutic applications. Graphical Abstract

Strategies to promote dental pulp stem cells (DPSCs) functions including proliferation, migration, pro-angiogenic effects, and odontogenic/osteogenic differentiation are in urgent need to restore pulpitis-damaged dentin/pulp regeneration and DPSCs-based bone tissue engineering applications. Cannabidiol (CBD), an active component of Cannabis sativa has shown anti-inflammation, chemotactic, anti-microbial, and tissue regenerative potentials. Based on these facts, this study aimed to analyze the effect of CBD on DPSCs proliferation, migration, and osteogenic/odontogenic differentiation in basal and inflammatory conditions. Highly pure DPSCs with characteristics of mesenchymal stem cells (MSCs) were successfully isolated, as indicated by the results of flowcytometry and multi-lineage (osteogenic, adipogenic, and chondrogenic) differentiation potentials. Among the concentration tested (0.1–12.5 µM), CBD (2.5 μM) showed the highest anabolic effect on the proliferation and osteogenic/odontogenic differentiation of DPSCs. Pro-angiogenic growth factor VEGF mRNA expression was robustly higher in CBD-treated DPSCs. CBD also prompted the migration of DPSCs and CBD receptor CB1 and CB2 expression in DPSCs. TNF-α inhibited the viability, migration, and osteogenic/odontogenic differentiation of DPSCs and CBD reversed these effects. CBD alleviated the TNF-α-upregulated expression of pro-inflammatory cytokines TNF-α, interleukin (IL)-1β, and IL-6 in DPSCs. In conclusion, our results indicate the possible application of CBD on DPSCs-based dentin/pulp and bone regeneration.

Periodontitis is the most prevalent oral disease worldwide, leading to inflammation, alveolar bone loss, and tooth loss. Mesenchymal stem cells (MSCs) transplantation has the potential to alleviate periodontitis and restore alveolar bone, but numerous studies have found that transplanted MSCs undergo apoptosis, releasing apoptotic extracellular vesicles (ApoEVs). Human periodontal ligament stem cells (hPDLSCs) ApoEVs have immunomodulatory and tissue regenerative potential. Still, their role in periodontitis treatment and alveolar bone restoration has not been investigated yet. The study isolated hPDLSCs-ApoEVs and investigated the modulatory effect on macrophaghe. We explored the specific molecular mechanisms of hPDLSC-ApoEVs in regulating macrophage polarization in vitro through mRNA-seq and inhibitors. Co-culture experiment demonstrated that hPDLSC-ApoEVs promoted hPDLSC osteogenic differentiation by regulating macrophage polarization. Finally, we verified that hPDLSC-ApoEVs inhibited periodontitis-induced periodontal tissue defects and promoted periodontal bone tissue regeneration in vivo. This study found that hPDLSCs-ApoEVs regulate M0 to anti-inflammatory M2 macrophage polarization, as indicated by the upregulation of CD163, IL-10, and ARG1. hPDLSCs-ApoEVs activated NF-κB signaling that upregulated S100A9 in macrophages. hPDLSCs-ApoEVs failed M2 macrophage polarization during inhibition of NF-κB or S100A9. Furthermore, hPDLSCs-ApoEVs-treated macrophage-conditioned medium robustly promoted osteogenic differentiation of hPDLSCs in vitro. Finally, we verified these results in the periodontitis model of male C57BL/6 mice. Local injection of hPDLSCs-ApoEVs facilitated macrophage M2 polarization, alleviated periodontitis, and restored alveolar bone. In conclusion, these findings provide new evidence for the clinical application of hPDLSCs-ApoEVs in periodontitis treatment via macrophage immune modulation-mediated alveolar bone restoration.

Background: The photodynamic therapy (PDT) showed promising potential in treating tongue squamous cell carcinoma (TSCC). The Food and Drug Administration approved Verteporfin (Ver) is a powerful alternative in this field for its penetrating power and high production of reactive oxygen species (ROS). However, its applications in the treatment of TSCC are still rare. Methods: Ver was loaded onto Poly (lactic-co-glycolic acid) (PLGA) nanoparticles, followed by the modification with RGD peptide as the ligand. The nanostructured was named as RPV. In vitro assessments were conducted to evaluate the cytotoxicity of RPV through the Live/Dead assay analysis and Cell Counting Kit-8 (CCK-8) assay. Using the reactive oxygen species assay kit, the potential for inducing targeted tumor cell death upon laser irradiation by promoting ROS production was investigated. In vivo experiments involved with the biological distribution of RPV, the administration with RPV followed by laser irradiation, and the measurement of the tumor volumes. Immunohistochemical analysis was used to detect the Ki-67 expression, and apoptosis induced by RPV-treated group. Systemic toxicity was evaluated through hematoxylin-eosin staining and blood routine analysis. Real-time monitoring was employed to track RPV accumulation at tumor sites. Results: The in vitro assessments demonstrated the low cytotoxicity of RPV and indicated its potential for targeted killing TSCC cells under laser irradiation. In vivo experiments revealed significant tumor growth inhibition with RPV treatment and laser irradiation. Immunohistochemical analysis showed a notable decrease in Ki-67 expression, suggesting the effective suppression of cell proliferation, and TUNEL assay indicated the increased apoptosis in the RPV-treated group. Pathological examination and blood routine analysis revealed no significant systemic toxicity. Real-time monitoring exhibited selective accumulation of RPV at tumor sites. Conclusion: The findings collectively suggest that RPV holds promise as a safe and effective therapeutic strategy for TSCC, offering a combination of targeted drug delivery with photodynamic therapy.

Apoptotic extracellular vesicles (ApoEVs) derived from human dental pulp stem cells (hDPSCs) are subcellular structures released during programmed cell death. These vesicles carry rich biological information and regulatory potential, and have demonstrated significance in regulating cell behavior and tissue repair. Human periodontal ligament stem cells (hPDLSCs) are key to regenerating periodontal tissue, and their osteogenic differentiation ability directly influences the repair and reconstruction of periodontal bone tissue. However, the potential of ApoEVs derived from human dental pulp stem cells (hDPSCs-ApoEVs) to regulate the osteogenic differentiation of hPDLSCs remains unexplored. This study aims to investigate the effects of hDPSCs-ApoEVs on the osteogenic differentiation of hPDLSCs, offering new insights and methods for treating periodontal bone tissue injuries, with important scientific and clinical implications. hDPSCs-ApoEVs were isolated via ultracentrifugation and characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), and Western blot. In vitro, hPDLSCs were treated with hDPSCs-ApoEVs to determine whether the vesicles could enter hPDLSCs and to assess the proliferation, migration, and osteogenic potential of hPDLSCs. In vivo, a rat periodontal bone defect model was established, with local injections of hDPSCs-ApoEVs or PBS administered every other day. After 6 weeks, the maxillae were collected and analyzed using micro-CT and histological staining. In vitro, hDPSCs-ApoEVs promoted the migration and osteogenic differentiation of hPDLSCs. In vivo, treatment with hDPSCs-ApoEVs can enhance the recovery of periodontal bone defects in rats. hDPSCs-ApoEVs play a significant role in promoting bone regeneration, suggesting their potential as a promising cell-free therapeutic strategy for periodontal bone tissue regeneration.

Objective To investigate the prevalence of deciduous dental caries in preschool children aged 36–71 months as well as the eruption and caries status of permanent teeth in children aged 60–71 months in Guangzhou city, providing references for oral health work. Design This study was a cross-sectional study design. Setting The study was conducted in 34 kindergartens in the central and non-central urban areas of Guangzhou city. Participants 14 883 preschool children aged 36–71 months in Guangzhou city were included in the ultimate analysis. Main outcome measures The prevalence of dental caries and other indicators of deciduous teeth, eruption and caries status of permanent teeth in children aged 60–71 months. Results The prevalence of dental caries in children aged 36–47 months, 48–59 months and 60–71 months were 41.6% (95% CI: 40.1%, 43.1%), 57.1% (95% CI: 55.8%, 58.5%) and 65.2% (95% CI: 64.0%, 66.4%), respectively. Mean decayed–missing–filled teeth increased with age, with differences between genders and areas. The percentage of permanent first molars and central incisors eruption in children aged 60–71 months differed, and caries conditions varied. Conclusion The problem of early childhood caries in preschool children in Guangzhou city is prominent. Prevention should be strengthened from early life, and oral health education and publicity enhanced to narrow regional oral health gaps.