不同接种条件对牙周膜干细胞体内外矿化的影响

Abstract

目的：分离培养人牙周膜干细胞(periodontal ligament stem cells，PDLSCs)，观察不同接种条件下成牙能力。方法：体外培养人牙周膜干细胞，细胞与胶原凝胶混合，实验组，接种至陶瓷化骨(ceramic bovine bone, CBB)，对照组接种至冻干胶原膜(lyophilization collagen membrane, LCM)，矿化液连续培养2 w后，移植入免疫缺陷小鼠皮下上，培养6周取材。扫描电镜观察与材料的贴附情况,细胞计数观察细胞在材料上的粘附和增殖情况，组织化学方法检测不同支架上矿化培养后PDLSCs的碱性磷酸酶(ALP)。扫描电镜观察体外培养条件下细胞附着，基质分泌，组织学观察牙周组织形成能力。结果：扫描电镜下可见PDLSCs与CBB相容性好，细胞生长密集，长入向BBC空隙内，体内条件下，CBB组形成牙周膜–牙骨质复合体结构，LCM组未形成牙周组织。结论：体内外培养条件下，CBB均具有较好诱导作用，是牙齿组织再生研究的优势支架材料。 Objective: To investigate the performance of combine collagen gen and ceramic bovine bone for human periodontal ligament stem cells. Methods: Human periodontal ligament stem cells were cultured in vitro. The subsequent cell passaging was conducted in conditioned medium containing dexamethasone, beta-sodium glycerophosphate and ascorbid acid. Then the human periodontal ligament stem cells were divided into two groups: one was cultured in BBC and the other was cul-tured in LCM. Attachment was observed by SEM and proliferation was evaluated by cell counting, ALP was measured in vitro histochemical method, and differentiation was observed by HE stain after transplant into nude mice. Results: BBC improved the proliferation and differentiation of cells cultured in vitro, showing no difference in attachment between two groups. Conclusion: BBC can improve the proliferation and differentiation of human periodontal stem cells.