冻存骨髓基质细胞和胶原膜在裸鼠体内的成骨研究

目的：研究经超低温冻存后的骨髓基质细胞（bone marrow stromal cells,BMSCs）和支架材料胶原膜BME-10X复合体植入裸鼠体内的成骨情况.方法：体外分离培养Beagle犬 BMSCs,冻存二代生长状况良好的BMSCs.12个月后,复苏冻存的BMSCs,并在体外构建BMSCs和0.5 cm×0.5 cm大小的BME-10X复合体.将胶原膜＋BMSCs＋矿化诱导培养液、胶原膜＋BMSCs＋基础培养液、单纯胶原膜＋矿化诱导培养液培养5 d后,植入裸鼠体内,并于术后第4、8、12周取出标本,进行大体观察和组织病理学分析.结果：单纯胶原膜＋诱导培养液组在植入胶原膜后,胶原膜边界清晰,膜边缘及内部基本没有细胞生长;在胶原膜＋BMSCs＋基础培养液组,术后第4周可见,胶原膜内有细胞长入,并有细小的条索状新生胶原形成,随着时间的延长,支架胶原逐渐分解,新形成的条索状胶原纤维变粗大;在胶原膜＋BMSCs＋矿化诱导培养液组,植入后也可见支架胶原的分解和更多的细胞生长,大量新生的胶原形成类骨质样组织.结论：冻存BMSCs复苏后进行体外培养扩增与诱导分化,并在在体内环境下复合胶原支架材料,仍然具有较强成骨能力.

Osteogensis of the Construct Combined Collagenic Membrane and Cryopreserved Bone Marrow Stromal Cells in Nude Mice.

Objective:To investigate the osteogenic potentiality of the construct combine collagenic membrane and cryopreserved canine bone marrow stromal cells(BMSCs) under induction and non-induction condition in vivo.Methods:The BMSCs of beagle dog were cryopreserved for 12 months which were recoverd and cultured and subcultured in vitro.3 groups of compound materials were implanted into the nude mice.Group 1 was collagenic membrane with induced medium.Group 2 was BMSCs and collagenic membrane cultured with normal medium.Group 3 was BMSCs and collagenic membrane cultured with induced medium.4,8 and 12 weeks after implantation,the specimens were harvested for histological observation.Results:HE staining showed the group 3 had more cells proliferation,collagenic membrane degradation and collagen synthesis in implantation areas than that of group 2.Conclusion:Constructs combined cryopreserved BMSCs with collagenic membrane have strong activity of proliferation and osteogenic potentiality in vivo.This study revealed that the cryopreserved BMSCs could be used as the seed cells in the bone tissue engineering.