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BACKGROUND: Less is reported about the effect of transforming growth factor-beta 3 and bone morphogenetic protein-2 on the proliferation and osteogenic differentiation of dental pulp stem cells. OBJECTIVE: To explore the effect of transforming growth factor-beta 3 and bone morphogenetic protein-2 on the osteogenic differentiation of dental pulp stem cells and its preliminary mechanism. METHODS: Dental pulp stem cells were isolated from the pulp tissue of New Zealand rabbit by the enzymatic digestion method. Dental pulp stem cells at passage 3 were divided into the blank group, bone morphogenetic protein-2 group (80 μg/L) and transforming growth factor-beta 3 group (80 μg/L). The cell proliferation activity in each group was tested by MTT assay. After 7 and 14 days of culture, alkaline phosphatase activities were measured. Runt-related transcription factor 2 and bone sialoprotein protein expression levels were conducted by immunocytochemical staining. Osteogenesis-related gene expression was tested through RT-qPCR method. Alizarin red S staining was conducted after 14 and 21 days of culture to observe mineralized nodules. RESULTS AND CONCLUSION: (1) Dental pulp stem cell proliferation activity and alkaline phosphatase activity were higher in the bone morphogenetic protein-2 and transforming growth factor-beta 3 groups than those in the blank group (P < 0.05). The alkaline phosphatase activity of the transforming growth factor-beta 3 group on day 7 was higher than that of the bone morphogenetic protein-2 group (P < 0.05). (2) The expression levels of Runt-related transcription factor 2 and bone sialoprotein in the transforming growth factor-beta 3 group and the bone morphogenetic protein-2 group were higher than those in the blank group (P < 0.05). The protein expression of Runt-related transcription factor 2 in the transforming growth factor-beta 3 group on day 7 was higher than that in the bone morphogenetic protein-2 group (P < 0.05). (3) The expression levels of alkaline phosphatase, type I collagen A1, Runt-related transcription factor 2, osteocalcin, osteopontin and Osx genes in the transforming growth factor-beta 3 group and bone morphogenetic protein-2 group were higher than those in the blank group (P < 0.05). On day 7, the expression levels of alkaline phosphatase, type I collagen A1, and Runt-related transcription factor 2 genes of transforming growth factor-beta 3 group were higher than those of bone morphogenetic protein-2 group (P < 0.05). (4) The mineralized nodules in the transforming growth factor-beta 3 group and bone morphogenetic protein-2 group were more obvious than those in the blank group. (5) The results show that transforming growth factor-beta 3 and bone morphogenetic protein-2 can improve the proliferation and osteogenic differentiation of dental pulp stem cells. The osteogenic induction ability of transforming growth factor-beta 3 on dental pulp stem cells was better than that of bone morphogenetic protein-2 at the early stage of osteogenesis. [ABSTRACT FROM AUTHOR]

Background: Because of the low regeneration efficiency and unclear underlying molecular mechanism, tooth regeneration applications are limited. In this study, we explored the influence of residual periodontal ligament on the dentin regeneration potential of dental pulp stem cells (DPSCs) in the jaw. Methods: To establish a tooth regeneration model, the incisors of New Zealand white rabbits were extracted while preserving residual periodontal ligament, followed by the implantation of DPSCs. After 3 months, micro-computed tomography (micro-CT), stereomicroscopy and scanning electron microscopy (SEM) were used to observe the volume, morphology and microstructure of regenerated tissue. Histological staining and immunostaining analyses were used to observe the morphological characteristics and expression of the dentin-specific proteins DMP1 and DSPP. To explore the mechanism, DPSCs and periodontal ligament stem cells (PDLSCs) were cocultured in vitro, and RNA was collected from the DPSCs for RNA-seq and bioinformatic analysis. Results: The results of micro-CT and stereomicroscopy showed that the number of sites with regeneration and the volume of regenerated tissue in the DPSCs/PDL group (6/8, 1.07 ± 0.93 cm3) were larger than those in the DPSCs group (3/8, 0.23 ± 0.41 cm3). The results of SEM showed that the regenerated dentin-like tissue in the DPSCs and DPSCs/PDL groups contained dentin tubules. Haematoxylin and eosin staining and immunohistochemical staining indicated that compared with the DPSCs group, the DPSCs/PDL group showed more regular regenerated tissue and higher expression levels of the dentin-specific proteins DMP1 and DSPP (DMP1: P = 0.02, DSPP: P = 0.01). RNA-seq showed that the coculture of DPSCs with PDLSCs resulted in the DPSCs differentially expressing 427 mRNAs (285 upregulated and 142 downregulated), 41 lncRNAs (26 upregulated and 15 downregulated), 411 circRNAs (224 upregulated and 187 downregulated), and 19 miRNAs (13 upregulated and 5 downregulated). Bioinformatic analysis revealed related Gene Ontology function and signalling pathways, including extracellular matrix (ECM), tumour necrosis factor (TNF) signalling and chemokine signalling pathways. Conclusions: Residual periodontal ligament in the extraction socket promotes the dentin regeneration potential of DPSCs in the jaw. RNA-seq and bioinformatic analysis revealed that ECM, TNF signalling and chemokine signalling pathways may represent the key factors and signalling pathways. [ABSTRACT FROM AUTHOR]

BACKGROUND: Less is reported about the effect of transforming growth factor-beta 3 and bone morphogenetic protein-2 on the proliferation and osteogenic differentiation of dental pulp stem cells.OBJECTIVE: To explore the effect of transforming growth factor-beta 3 and bone morphogenetic protein-2 on the osteogenic differentiation of dental pulp stem cells and its preliminary mechanism.METHODS: Dental pulp stem cells were isolated from the pulp tissue of New Zealand rabbit by the enzymatic digestion method. Dental pulp stem cells at passage 3 were divided into the blank group, bone morphogenetic protein-2 group（80 μg/L） and transforming growth factor-beta 3 group（80 μg/L）. The cell proliferation activity in each group was tested by MTT assay. After 7 and 14 days of culture, alkaline phosphatase activities were measured. Runt-related transcription factor 2 and bone sialoprotein protein expression levels were conducted by immunocytochemical staining. Osteogenesis-related gene expression was tested through RT-qPCR method. Alizarin red S staining was conducted after 14 and 21 days of culture to observe mineralized nodules. RESULTS AND CONCLUSION:（1） Dental pulp stem cell proliferation activity and alkaline phosphatase activity were higher in the bone morphogenetic protein-2 and transforming growth factor-beta 3 groups than those in the blank group（P < 0.05）. The alkaline phosphatase activity of the transforming growth factor-beta 3 group on day 7 was higher than that of the bone morphogenetic protein-2 group（P < 0.05）.（2） The expression levels of Runt-related transcription factor 2 and bone sialoprotein in the transforming growth factor-beta 3 group and the bone morphogenetic protein-2 group were higher than those in the blank group（P < 0.05）. The protein expression of Runt-related transcription factor 2 in the transforming growth factor-beta 3 group on day 7 was higher than that in the bone morphogenetic protein-2 group（P < 0.05）.（3） The expression levels of alkaline phosphatase, type I collagen A1, Runt-related transcription factor 2, osteocalcin, osteopontin and Osx genes in the transforming growth factor-beta 3 group and bone morphogenetic protein-2 group were higher than those in the blank group（P < 0.05）. On day 7, the expression levels of alkaline phosphatase, type I collagen A1, and Runt-related transcription factor 2 genes of transforming growth factor-beta 3 group were higher than those of bone morphogenetic protein-2 group（P < 0.05）.（4） The mineralized nodules in the transforming growth factor-beta 3 group and bone morphogenetic protein-2 group were more obvious than those in the blank group.（5） The results show that transforming growth factor-beta 3 and bone morphogenetic protein-2 can improve the proliferation and osteogenic differentiation of dental pulp stem cells. The osteogenic induction ability of transforming growth factor-beta 3 on dental pulp stem cells was better than that of bone morphogenetic protein-2 at the early stage of osteogenesis. [ABSTRACT FROM AUTHOR]

Introduction: Dental pulp remains one of the important sources of mesenchymal stem cells for most preclinical and clinical studies. Aim and Objectives: To assess the safety after injecting human dental pulp-derived mesenchymal stem cells by intramucosal and intrabony routes in rabbits for clinical application. Materials and Methods: Animal studies were carried out among 30 New Zealand male white rabbits (3-5 months old), weighing 1.5-2 kgs, which were divided into three groups with 10 animals in each group. Group 1: control group, Group 2: intramucosal route, Group 3: intrabony route. Data were analyzed using Student's t-test, and any P = 0.05 was statistically significant. Results: A total of 30 rabbits were selected for the study, among which significant statistical difference for Packed cell volume (PCV) (P < 0.05), MCHC (P < 0.05), platelet count (P < 0.05), and ESR (p < 0.001) has been reported in the hematological parameters. The results of the present study indicate that the transplantation of hDPSCs by intramucosal and intrabony routes into a rabbit is non-toxic without any detectable side effects or local or systemic rejection. The pre-clinical safety and toxicity of the hDPSCs in various human disease models need to be determined in future studies. Various pre-clinical studies to determine the safety and toxicity of hDPSCs in human disease models have to be done in the future. Conclusion: This study showed that the intramucosal route and intrabony route of administration of stem cells were found to be non-toxic at 10 million per mL concentration. A further evaluation must be done for more definitive results. [ABSTRACT FROM AUTHOR]

BACKGROUND: The dental pulp stem cells have strong proliferation ability and multi differentiation potential ability, which plays an important role in bone and cartilage tissue engineering. Therefore, to study the therapeutic effect of dental pulp stem cells on femoral head necrosis will bring new treatment strategies and hope for patients with femoral head necrosis. OBJECTIVE: To observe the therapeutic effect of core decompression combined with dental pulp stem cells on early steroid-associated femoral head necrosis in rabbits. METHODS: Fifty-two adult healthy New Zealand white rabbits were randomly divided into four groups: normal control group, model group, solvent group, and dental pulp stem cell group. Except the normal control group, the model of steroid induced osteonecrosis of femoral head was established in the other three groups. After successful modeling (the 4th week), core decompression was performed on the right side in the solvent group and dental pulp stem cell group. The dental pulp stem cell group was injected with dental pulp stem cells in the decompression hole of the femoral head, and the solvent group was injected with the same volume of sodium chloride injection at the same time. There were three injections in the 4th, 5th and 6th weeks. At the 12th week, indicators such as bone density, bone morphology parameters, empty bone lacuna rate, and bone trabecular area ratio were measured. RESULTS AND CONCLUSION: (1) Imaging examination showed that the treatment effects of solvent group and dental pulp stem cell group were significantly improved compared with the model group. Bone volume fraction, trabecular number, and bone mineral density of femoral head in the dental pulp stem cell group were better than those in the solvent group, and the difference was statistically significant (P < 0.05). (2) Compared with the normal control group, the empty bone lacuna rate and trabecular area ratio of the dental pulp stem cell group were closer; the empty bone lacuna rate of the solvent group was higher than that of the normal control group (P < 0.05); and the trabecular area ratio was lower than that of the normal control group (P < 0.05). The empty bone lacuna rate of the solvent group and dental pulp stem cell group was significantly lower than that of the model group (P < 0.05), and the bone trabecular area ratio was significantly higher than that of the model group (P < 0.05). (3) The results indicate that the treatment of early femoral head necrosis by using core decompression combined with dental pulp mesenchymal stem cells is better than that of simple core decompression. [ABSTRACT FROM AUTHOR]

Purpose: This research evaluated clinical, histological, and radiological osseous regeneration in a critical-sized bilateral cortico-medullary osseous defect in model rabbits from New Zealand after receiving a hydroxyapatite matrix and polylactic polyglycolic acid (HA/PLGA) implanted with human dental pulp stem cells (DPSCs). Methods: Eight New Zealand rabbits with bilateral mandibular critical-sized defects were performed where one side was treated with an HA/PLGA/DPSC matrix and the other side only with an HA/PLGA matrix for 4 weeks. Results: An osseointegration was clinically observed as well as a reduction of 70% of the surgical lumen on one side and a 35% on the other. Histologically, there was neo-bone formation in HA/PLGA/DPSC scaffold and angiogenesis. A bone radiodensity (RD) of 80% was radiologically observed achieving density levels similar to mandibular bone, while the treatment with HA/PLGA matrix achieves RD levels of 40% on its highest peaks. Conclusions: HA/PLGA/DPSC scaffold was an effective in vivo method for mandibular bone regeneration in critical-sized defects induced on rabbit models. [ABSTRACT FROM AUTHOR]

This study examined and compared wound healing between Thai propolis product and calcium hydroxide paste as pulp-capping agents after partial pulpotomy in New Zealand white rabbits. Forty incisor teeth from 10 rabbits were treated. Thirty-six teeth received class V cavity preparations with partial pulpotomy and application of either propolis or calcium hydroxide paste. Similar cavity preparations were performed in 2 teeth without any capping material as a positive control, whereas 2 teeth without the cavity preparation served as a negative control. Histological evaluation showed that both groups had dentin bridge formation. Dentinal tubules in the dentin bridge were more orderly arranged in the Thai propolis group than in the calcium hydroxide group. Wound healing and the median number of hyperemic blood vessels were not statistically significant different between the 2 groups. Thai propolis product may be used as a pulp-capping agent. [ABSTRACT FROM AUTHOR]

Disease and trauma leading to tooth loss and destruction of supporting bone is a significant oral handicap, which may be addressed through surgical therapies that aim to regenerate the lost tissue. Whilst complete regeneration of teeth is still aspirational, regeneration of supporting structures (dental pulp, cementum, periodontal ligament, bone) is becoming commonplace, both for teeth and for titanium dental implants that are used to replace teeth. Most grafting materials are essentially passive, however the next generation of oral regenerative devices will combine non-antibiotic antimicrobials and/or osteogenic or inductive factors and/or appropriate multipotential stem cells. The review gives an overview of the approaches taken, including fabrication of novel scaffolds, incorporation of growth factors and cell-based therapies, and discusses the preclinical animal models we employ in the development pathway. [Display omitted] • Regenerative therapies commonly used for oral disease and trauma • Most regenerative approaches employ grafting with passive scaffolds • Development needs expertise in biomaterials, cell culture and pre-clinical models • Active scaffolds combine novel non-antibiotic anti-microbials and bioactives • 3-D printing of active scaffolds is challenging but holds promise • Various oral stem-cell reservoirs may be utilised [ABSTRACT FROM AUTHOR]