Regeneration characteristics of different dental derived stem cell sheets.
Hu Lei,Zhao Bin,Gao Zhenhua,Xu Junji,Fan Zhipeng,Zhang Chunmei,Wang Jinsong,Wang Songlin
Journal of oral rehabilitation
BACKGROUND:Although cell sheets have gained much interest as a non-scaffold strategy for tissue regeneration, the regenerative features of different cell sheets remain unclear. OBJECTIVE:In this study, we aimed to compare the regeneration characteristics of cell sheets derived from dental pulp stem cells (DPSCs), periodontal ligament stem cells (PDLSCs) and stem cells of the apical papilla (SCAPs). METHODS:Dental pulp stem cells, PDLSCs and SCAPs from the same individual were acquired and induced to form sheets using 20 μg/mL vitamin C. Immunofluorescence staining was used to detect the expression of collagen I, fibronectin, integrin β1 and vimentin. Real-time PCR was used to determine NANOG, OCT4, SOX2 and TERT gene expression. The cell sheets with hydroxyapatite/tricalcium phosphate were transplanted into nude mice subcutaneously to evaluate tissue regeneration characteristics. RESULTS:No obvious differences were found in the histological structure and extracellular matrix protein expression between DPSC, PDLSC and SCAP sheets. Dental pulp stem cell sheet showed higher expression of OCT4 and TERT than PDLSC and SCAP sheets. All three cell sheets displayed the ability of mineral tissue formation and highly expressed periostin. The tissue derived from DPSC sheet showed higher CD31 expression and porous fibres compared with that from the others. The tissue fibres formed from PDLSC sheet were directionally arranged, while the tissue derived from SCAP sheet showed highest mineral tissue formation. CONCLUSION:Although in vitro DPSC, PDLSC and SCAP cell sheets have similar characteristics, their regenerative characteristics in vivo are different, with each showing potential application for regeneration of different tissues.
[Periodontal tissue in a bio-implant by periodontal ligament cells sheet and bone marrow stromal cells sheet].
Sun Chuanming,Liu Hongwei
Zhonghua kou qiang yi xue za zhi = Zhonghua kouqiang yixue zazhi = Chinese journal of stomatology
OBJECTIVE:To fabricate the bio-implant supported by regenerated periodontal tissue utilizing periodontal ligament cells (PDLC)-bone marrow stromal cells (BMSC) sheet and natural root. METHODS:Premolars of 2 beagle dogs were extracted to prepare the implanted area. Autologous tooth roots were carved into cylinders. PDLC and BMSC separated from beagle dogs were cultured into cell sheets in medium. Tooth roots were wrapped by one type of cell sheets or both to fabricate bio-implant and divided into four groups, tooth roots were wrapped by PDLC sheets and BMSC sheets successively (2 samples each dog), tooth roots were wrapped by PDLC sheets alone (2 samples each dog), tooth roots were wrapped by BMSC sheet alone (2 samples each dog), tooth roots without cell sheet (1 sample each dog). The implants were implanted into the mandibles. The mandibles were dissected 12 weeks later, sliced and stained by HE and Masson dyes for histological examination. RESULTS:In the PDLC cell sheet/root implants, histological examination revealed that new periodontal-like tissue including cementum, periodontium and alveolar bone was regenerated.In the BMSC implants, tooth ankylosis was observed. CONCLUSIONS:PDLC sheet and natural root can be used to fabricate bio-implant. PDLC sheet could promote periodontal regeneration.
Analysis of gene expression profiles between apical papilla tissues, stem cells from apical papilla and cell sheet to identify the key modulators in MSCs niche.
Diao Shu,Lin Xiao,Wang Liping,Dong Rui,Du Juan,Yang Dongmei,Fan Zhipeng
OBJECTIVES:The microenvironmental niche plays the key role for maintaining the cell functions. The stem cells from apical papilla (SCAPs) are important for tooth development and regeneration. However, there is limited knowledge about the key factors in niche for maintaining the function of SCAPs. In this study, we analyse the gene expression profiles between apical papilla tissues, SCAPs and SCAPs cell sheet to identify the key genes in SCAPs niche. MATERIALS AND METHODS:Microarray assays and bioinformatic analysis were performed to screen the differential genes between apical papilla tissues and SCAPs, and SCAPs and SCAPs cell sheet. Recombinant human BMP6 protein was used in SCAPs. Then CCK-8 assay, CFSE assay, alkaline phosphatase activity, alizarin red staining, quantitative calcium analysis and real-time reverse transcriptase-polymerase chain reaction were performed to investigate the cell proliferation and differentiation potentials of SCAPs. RESULTS:Microarray analysis found that 846 genes were up-regulated and 1203 genes were down-regulated in SCAPs compared with apical papilla tissues. While 240 genes were up-regulated and 50 genes were down-regulated in SCAPs compared to in SCAPs cell sheet. Moreover, only 31 gene expressions in apical papilla tissues were recovered in cell sheet compared with SCAPs. Bioinformatic analysis identified that TGF-β, WNT and MAPK signalling pathways may play an important role in SCAPs niche. Based on the analysis, we identified one key growth factor in niche, BMP6, which could enhance the cell proliferation, the osteo/dentinogenic, neurogenic and angiogenic differentiation potentials of SCAPs. CONCLUSIONS:Our results provided insight into the mechanisms of the microenvironmental niche which regulate the function of SCAPs, and identified the key candidate genes in niche to promote mesenchymal stem cells-mediated dental tissue regeneration.