预制作生物工程性肌骨瓣的实验研究

Experimental investigation on construction of tissue-engineered osteo-musculo-cutaneous flaps

目的探索用羟基磷灰石/磷酸三钙(HA/TCP)的支架材料与成骨细胞复合构成组织工程性肌骨瓣的方法,为修复骨缺损奠定基础。方法自2004年12月至2006年1月,将36只新西兰大耳白兔髂后上棘抽取骨髓基质干细胞并诱导分化为成骨细胞,与HA/TCP结合培养2周,通过相差显微镜、扫描电镜观察体外细胞生长情况。将复合物埋入兔的背阔肌下构成组织工程性肌骨瓣。术后6周进行大体解剖观察,HE染色,研究组织工程性肌骨瓣的生长情况。结果扫描电镜显示支架材料表面和孔隙内均有成骨细胞生长,有良好的增殖活动性和稳定细胞表型,材料中心区未见细胞生长,植入体内6周后取材见内植物有新骨形成,多位于内植物表面,可见成骨细胞、骨细胞、髓腔样结构、板层样骨基质等正常骨组织结构。结论骨髓来源的成骨细胞可用作骨组织工程的种子细胞,与HA/TCP结合构建出组织工程性肌骨瓣。

Objective To construct a tissue-engineering muscle pedicel bone graft by culture marrow cells combined with biodegradable polymeric scoffolds （HA/TCP） which be used to repairing a segmental of long bone. Methods From December 2004 to January 2006, bone marrow stromal cells （BMSC） being aspirated from spina iliaca posterior superior of young rabbits were induced into osteoblasts in vitro. Osteoblasts which were isolated from bone marrow stromal cells of New Zealand rabbits were cultured as the seeding cells, and the cells were cultured with HA/TCP in virto. The complex was observed under phase microscope and histology to evaluate the interaction between osteoblasts and HA/TCP. The compound co-cultured for 14 days was implanted intermuscularly into muscle latissimus dorsi of rabbits which were investigated at 6 weeks after postoperatively. The muscle pedicel bone graft were investigated by means of gross observation, histology of HE. Results Scanning electronic micrographs showed that the pores of HA/TCP and its surface were filled with cultured osteoblasts, especially in the longer cultured block, no cell growth was found in the central area, new bone formation was observed in all blocks at 6 weeks after implantation. Conclusion The osteoblasts from bone marrow stromal cells can be used as the seeded cells for bone tissue engineering, bio-derived materials being constituted with BMSC may be a good option in construction of bone tissue engineering.