目的:研究血小板裂解液(Platelet Lysate,PL)在体内外对人根尖牙乳头干细胞(SCAP)和牙周膜干细胞(PDLSCs)增殖、矿化的干预效应,以期找到PL应用于这两种细胞的最佳方式。方法:在矿化诱导培养体系中按一定体积比加入PL,分别作用于体内、外复合HA-TCP支架材料三维生长的SCAP和PDLSCs,扫描电镜(SEM)以及组织学观察细胞生长、分化情况。结果:体外三维培养模式下,PL能够促进SCAP和PDLSCs的增殖以及矿化细胞外基质的形成,并利于在支架材料上形成完整的细胞膜片(cell sheet)样结构,以上效应对于PDLSCs作用更为显著。而在体内移植模式下,经50 mL/L、10 mL/L PL培养体系预培养的SCAP能形成包含成牙本质细胞样细胞、牙本质样基质和牙髓组织的牙本质牙髓复合体样结构;而PDLSCs经50 mL/LPL干预后,可分化为成牙骨质细胞样细胞,并生成牙骨质样基质、牙周膜样组织。结论:一定体积浓度比的PL能促进SCAP和PDLSCs在体内外的增殖以及成骨、成牙本质分化。
AIM: To explore the effects of platelet lysate (PL) on the osteogenic/odontogenic differentia- tion of human stem cells from the apical papilla (SCAP) and periodontal ligament stem cells (PDLSCs) under different culture conditions in vitro and in vivo. METHODS : Certain concentrations of PL preparation was added in mineraliza- tion medium to support SCAP and PDLSCs cultured on HA/TCP scaffolds in vitro and in vivo, the cell attachment on HA-TCP scaffolds treated with the mineralization medium added with PL preparation was observed by scanning electron microscope and histological analysis. RESULTS: The SCAP and PDLSCs seeded on HA-TCP scaffolds in osteogenic/ odontogenie differentiation medium added with PL showed increased proliferation, more calcium deposit and formed a more extensive sheet-like structure in the time course in vitro, and these effects were more significant in the PDLSCs groups. In vivo, the SCAP samples treated with 50 mL/L PL and 10 mL/L PL could form dentin/pulp-likestructure containing odontoblast-like cells, dentin-like matrix and pulp-like tissue. The PDLSC samples treated with 50 mL/L PL could differentiate into cementoblast-like cells that formed a cementum-like structure, cementocyte-like cells, andPDL-like tissue was also generated. CONCLUSION : Appropriate concentration of PL treatment may enhance prolifer- ation and osteogenic/odontogenic differentiation of SCAP and PDL stem cells in vitro and in vivo.