脱细胞天然骨基质与诱导性成骨细胞的体外相容性

摘要 背景：前期工作表明TritonX-100处理的脱细胞骨基质已满足组织学和免疫学方面的修复要求。如果细胞能在材料表面很好地生长，将利于进一步进行体内动物实验。 目的：采用细胞培养法在体外评估脱细胞骨基质与诱导后成骨细胞的生物相容性。 方法：第3代骨髓基质干细胞经成骨诱导分化培养液诱导分化为成骨细胞，接种于TritonX-100处理的脱细胞骨基质及羟基磷灰石表面，检测成骨细胞的碱性磷酸酶表达并用扫描电镜观察材料表面的细胞生长情况。 结果与结论：碱性磷酸酶活性分析均表明，TritonX-100处理的脱细胞骨基质在培养48 h之后比羟基磷灰石更利于诱导成骨细胞生长；扫描电镜下可见，成骨细胞在脱细胞骨基质表面呈现立体生长方式，细胞呈球形，并且聚集成簇。体外实验结果显示成骨细胞与脱细胞天然骨基质有较好的生物相容性。 关键词：天然脱细胞骨基质；骨髓基质干细胞；成骨细胞；体外培养；生物相容性 doi:10.3969/j.issn.1673-8225.2011.12.013

Biocompatibility of acellular natural bone matrix with induced osteoblasts in vitro

Abstract BACKGROUND: The repair requirements of histology and immunology can be met by acellular bone matrix treated by TritonX-100. The test cells will be able to grow nicely on the surface of test materials, which will benefit further evaluating them in animals in vivo. OBJECTIVE: To evaluate the biocompatibility of acellular natural bone matrix prepared with TritonX-100 to osteoblasts by cell culture assay. METHODS: The third passage of bone marrow stromal stem cells were induced to differentiate into osteoblasts, then the identified osteoblasts were seeded on surface of acellular natural bone matrix prepared with TritonX-100 and calcium hydroxyapatite. Alkaline phosphatase vitality was measured by alkaline phosphatase detection kit and the growth of osteoblast-like cells on surface of the material was observed with scanning electron microscopy. RESULTS AND CONCLUSION: Alkaline phosphatase detection kit analysis indicated that acellular natural bone matrix prepared with TritonX-100 were able to promote the growth of osteoblast-like cells compared with calcium hydroxyapatite after 48 hours of culture. Scanning electron microscopy displayed that osteoblast-like cells on the surface of acellular natural bone matrix aggregated together and grew as three-dimensional style. In vitro experimental proved good biocompatibility of osteoblast-like cells and acellular natural bone matrix.