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Introduction: Mesenchymal stem cells (MSCs) are the adult stem cells with potential to differentiate into various tissues. Like in other tissues, MSCs also reside in dental pulp after toot development and help in repair and regeneration by differentiating into odontoblasts. Dental Pulp Stem Cells (DPSCs) and Stem cells from Apical Papilla (SCAP) are the type of MSCs from dental papilla and apical papilla respectively. Aim: The aim of this paper is to highlight the characteristics of DPSCs and SCAP. Method: Information was obtained and compiled from published literature and electronic database search engine from PubMed and Google Scholar. Results: In spite of both DPSCs and SCAP having similar cell population origin they possess some different characteristics. Conclusion: The Dental stem cells with different characteristics of similar origin can be utilized in the stem cell based tissue engineering.
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Objective@#To explore the effects of red LEDs on the proliferation and osteogenic differentiation of human stem cells from apical papilla (hSCAPs).@*Methods@#hSCAPs were obtained by isolation, culture and flow cytometry in vitro and irradiated with 1, 3, 5, and 7 J/cm2 red LEDs. The proliferation of hSCAPs was detected using a CCK-8 assay. The osteogenic differentiation of hSCAPs was evaluated using alkaline phosphatase (ALP) staining, ALP activity assay and Alizarin red quantitative detection. The effect of 5 J/cm2 red LEDs on the expression levels of the ALP, Runx2, OCN, OPN and BSP genes and proteins was detected by RT-PCR and western blot, respectively.@*Results@# Red LEDs at 1, 3, 5, and 7 J/cm2 promoted the proliferation of hSCAPs (P < 0.05). The effects of red LEDs with different light energies on the proliferation of hSCAPs were different at different time points (P < 0.05). On the 7th and 14th days after irradiation, red LEDs promoted the osteogenic differentiation of hSCAPs, and the effect of 5 J/cm2 red LEDs was the most obvious under osteogenic induction culture conditions (P<0.05). Red LEDs (5 J/cm2) promoted the expression of the ALP, Runx2, OCN, OPN and BSP genes and proteins (P < 0.05).@*Conclusion @#Red LEDs promoted the proliferation and osteogenic differentiation of hSCAPs.
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OBJECTIVE@#To investigate the effects of autophagy on osteogenic differentiation of stem cells from the apical papilla (SCAPs) in the presence of tumor necrosis factor- (TNF-) stimulation .@*METHODS@#SCAPs treated with TNF- (0, 5, and 10 ng/mL) with or without 5 mmol/L 3-MA were examined for the expression of autophagy marker LC3-Ⅱ using Western blotting. The cells were transfected with GFP-LC3 plasmid and fluorescence microscopy was used for quantitative analysis of intracellular GFP-LC3; AO staining was used to detect the acidic vesicles in the cells. The cell viability was assessed with CCK-8 assays and the cell apoptosis rate was analyzed using flow cytometry. The cells treated with TNF- or with TNF- and 3-MA were cultured in osteogenic differentiation medium for 3 to 14 days, and real- time PCR was used to detect the mRNA expressions of osteogenesis-related genes (ALP, BSP, and OCN) for evaluating the cell differentiation.@*RESULTS@#TNF- induced activation of autophagy in cultured SCAPs. Pharmacological inhibition of TNF--induced autophagy by 3-MA significantly decreased the cell viability and increased the apoptosis rate of SCAPs ( < 0.05). Compared with the cells treated with TNF- alone, the cells treated with both TNF- and 3-MA exhibited decreased expressions of the ALP and BSP mRNA on days 3, 7 and 14 during osteogenic induction ( < 0.05) and decreased expression of OCN mRNA on days 3 and 7 during the induction ( < 0.05).@*CONCLUSIONS@#Autophagy may play an important role during the osteogenic differentiation of SCAPs in the presence of TNF- stimulation.
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Humans , Autophagy , Physiology , Cell Differentiation , Physiology , Cell Survival , Cells, Cultured , Dental Papilla , Cell Biology , Green Fluorescent Proteins , Osteogenesis , Physiology , Stem Cells , Physiology , Transfection , Tumor Necrosis Factor-alpha , PharmacologyABSTRACT
Objective @#To investigate the effect of different concentrations of MTA on the proliferation and differentiation of stem cells from the apical papilla (SCAP) and the potential of the SCAP to differentiate into odontoblasts.@*Methods@#SCAP were cultured in different concentrations of mineral trioxide aggregate（MTA）. MTA experimental group with concentration of 0.01 mg/mL, 0.02 mg/mL, 0.1 mg/mL, 0.2 mg/mL, 1 mg/mL, 2 mg/mL, 10 mg/mL and 20 mg/mL were prepared. The number of cells at 1 day, 3 days, 5 days and 7 days were measured via a CCK-8 assay to observe the effect of MTA on SCAP proliferation. Real-time PCR was used to detect the gene expression changes. Cells cultured in alpha MEM culture containing 15% FBS without MTA were set as the control group.@*Results @#When cultured for 1 d, statistically significant differences in the promotion of in vitro proliferation of SCAP were not observed between each MTA experimental group and the control group (P>0.05). When cultured for 3 d, 5 d and 7 d, the 0.01 mg/mL MTA group presented obvious promotion of SCAP proliferation compared with the control group (P<0.05), whereas the 0.02 mg/mL, 0.1 mg/mL, 0.2 mg/mL, 1 mg/mL groups did not presented differences with the control group (P>0.05). The in vitro proliferation of the 2 mg/mL, 10 mg/mL and 20 mg/mL groups was lower than that of the control group (P<0.05). Real-time PCR detection showed that the expression levels of DSPP (t=-11.12, P < 0.05) and Runx2 (t=-10.62, P < 0.05) in the experimental group treated with 0.01 mg/mL MTA for 7 days were higher than those in the control group. @*Conclusion @#The 0.01 mg/mL concentration of MTA significantly promotes the proliferation of SCAP and shows the best ability to induce osteogenic and odontoblast differentiation in the SCAP, whereas high concentrations of MTA inhibited the proliferation of SCAP.
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Objective To compare the reprogramming effects and biological characteristics of two types of human odontogen-ic induced pluripotent stem cells(iPSCs).Methods Human dental pulp stem cells(DPSCs)and stem cells from apical papilla (SCAP)were isolated and primarily cultured.The Sendai reprogramming system was utilized to induce DPSCs and SCAP into iP-SCs.The morphology,reprogramming efficiency,reprogramming and time were compared between human DPSCs-iPSCs and SCAP-iPSCs.The SeV and exogenous transcriptional gene expression were detected by RT-PCR.Results Human DPSCs and SCAP were reprogrammed as iPSCs with classical ES-like clonal morphology.The reprogramming efficiencies were(0.68 ± 0.02)% and(0.7 ± 0.01)% respectively,the difference was not statistically significant(P>0.05).The reprogramming time was(26.0 ± 2.1)d and (27.0 ± 1.4)d respectively,the difference was not statistically significant(P>0.05).The RT-PCR results showed that no expres-sion of exogenous virus or transcriptional gene sequence in both iPSCs.Conclusion Human DPSCs and SCAP can be reprogrammed as virus-free and transgene-free iPSCs,which are the ideal sources of iPSCs.
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Objective To comparatively study the features of two reprogramming systems of induced pluripotent stem cells (iPSCs)from human dental origin.Methods Two kinds of reprogramming system,i.e.STEMCCA lentivirus /feed layer and Sen-dai virus /matrigel were used to induce human stem cells from apical papilla(SCAP)into iPSCs,respectively.The induction efficien-cies,workload of generating iPSCs,aneuploidy karyotype ratio,complexities of eliminating exogenous transcription factors and spe-cific markers expression were compared between these two systems.Results The STEMCCA reprogramming system required to prepare the feeder cell MEF.The reprogramming efficiency was 0.1%.Transcription gene-free iPSCs cells were obtained by the Cre-loxp enzyme digestion technique at the later stage.Sendai virus reprogramming system was feeder-free and the preparation of matrigel was quite simple with unified standard.The reprogramming efficiency was 0.7%,which was much higher than that of STEMCCA system(P <0.05).The exogenous virus and transgenes could be gradually eliminated after several passages of natural subclone.Conclusion The Sendai virus/matrigle reprogramming system is much more applicable for the induction of iPSCs from dental origin than the STEMCCA system.
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<p><b>OBJECTIVE</b>To compare characterization of microRNAs (miRNAs) expression profiles of induced pluripotent stem cells (iPSCs) reprogrammed from human dental pulp stem cells (DPSCs) and stem cells from apical papilla (SCAP) and screen-specific microRNA.</p><p><b>METHODS</b>Human DPSCs and SCAP were reprogrammed into iPSCs using a Sendai virus vector. Total RNA of human DPSCs-iPSCs and SCAP-iPSCs were extracted. miRNAs were labeled and hybridized. Slides were scanned, and images were imported into GenePix Pro 6.0 for grid alignment and data extraction. Significant differentially expressed miRNAs between the two groups were identified using fold change and P-value and were analyzed.</p><p><b>RESULTS</b>Both human DPSCs and SCAP were successfully reprogrammed into iPSCs. Among miRNA genes analyzed by miRNA microarray, 68 were differentially expressed by more than 10-fold in DPSCs-iPSCs; 37 of these genes were up-regulated, and 31 were down-regulated. In SCAP-iPSCs, 107 genes were differentially expressed by more than 10-fold; 68 were up-regulated, and 39 were down-regulated. In both cells, only miR-302e was up-regulated, whereas 9 miRNAs were down-regulated: miR-29b-3p, miR-181b-5p, miR-4328, miR-22-5p, miR-145-5p, miR-4324, let-7b-5p, miR-181a-5p, and miR-27b-3p.</p><p><b>CONCLUSIONS</b>Multiple miRNAs participated in reprogramming of human DPSCs and SCAP into iPSCs. Most miRNAs are related to cell cycle, transforming growth factor-β signaling pathways and epithelial-mesenchymal transition.</p>
Subject(s)
Humans , Cell Cycle , Cell Division , Dental Pulp , Down-Regulation , Electrodes , Epithelial Cells , Induced Pluripotent Stem Cells , MicroRNAs , Taste Buds , Up-RegulationABSTRACT
Autologous bone transplantation is the current gold standard for reconstruction of jawbone defects. Bone regeneration usingmesenchymal stem cells (MSC) is an interesting alternative to improve the current techniques,which necessitate a second site of surgery resulting in donor site morbidity. In this study,we compared the osteogenic ability of jawboneMSC(JB-MSC) withMSC from tissues with neural crest origin, namely, the dental pulp, apical papilla and periodontal ligament. All four types ofMSC were isolated from the same patient (n = 3 donors) to exclude inter-individual variations.TheMSCgrowth and differentiation properties were characterized. The osteogenic differentiation potential in each group of cells was assessed quantitatively to determine if there were any differences between the cell types. All cells expressed the MSC-associated surface markers CD73, CD90, CD105, and CD146 and were negative for CD11b, CD19, CD34, CD45 and HLA-DR. All cell types proliferated at similar rates, exhibited similar clonogenic activity and could differentiate into adipocytes and osteoblasts. An alkaline phosphatase assay, OsteoImage™ assay for mineralization and qRT-PCR measuring the genes runx2, ALP and OCN, indicated that there were no significant differences in the osteogenic differentiation ability between the variousMSCs. In conclusion,we show that from a small segment of jawbone it is possible to isolate sufficient quantities of MSC and that these cells can easily be expanded and differentiated into osteoblasts. JB-MSC appear to be good candidates for future bone regeneration applications in the craniofacial region.