人骨髓基质干细胞与支架材料纳米羟基磷灰石／聚酰胺66的生物相容性研究  

Biocompatibility of nano-hydroxyapatite/polyamide 66 composite with human bone mesenchymal stem cells

作　　者：李昂[1] 王晓宇[1] 李泽成[2] 许茜楠 毕郑刚[1] 

机构地区：[1]哈尔滨医科大学附属第一医院骨科,150001 [2]哈尔滨医科大学附属第二医院骨科

出　　处：《中华创伤骨科杂志》2016年第3期241-246,共6页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金（81171692）

摘　　要：目的研究人骨髓基质干细胞（hBMSCs）与纳米羟基磷灰石／聚酰胺66（nHA／PA66）复合后的生物学特性。方法体外分离培养hBMSCs后将其分为A、B、C3组：A组为未经骨向诱导的hBMSCs，B组为经骨向诱导的hBMSCs，C组为与nHA／PA66复合并骨向诱导的hBMSCs。通过噻唑蓝（MTT）检测B、c组细胞在第1、2、3周的增殖情况；电镜扫描观察C组细胞在支架上的黏附情况；碱性磷酸酶（ALP）活性检测以及矿化结节染色验证hBMSCs的体外成骨分化能力，并通过检测第6、12天ALP的活性研究其与nHA／PA66复合后的骨生成作用。结果c组细胞在nHA／PA66上黏附良好；B组与C组细胞生长增殖活性差异无统计学意义（P〉0．05），说明骨向诱导后的hBMSCs增殖活性不受支架材料nHA／PA66影响；B组矿化结节数量明显多于A组，第6、12天A组ALP活性值均低于B、c组，差异有统计学意义（P〈0．05），而B、c组问差异无统计学意义（P〉0．05），说明hBMSCs体外骨向诱导后ALP表达增强并骨向分化，nHA／PA66并没有影响hBMSCs的成骨潜能。B、c组细胞在第12天的ALP活性值显著高于第6天，差异均有统计学意义（P〈0．05），说明AKP活性随骨向诱导时间增长而逐渐增强。结论hBMSCs适于在nHA／PA66上黏附、增殖与骨向分化，nHA／PA66可以作为hBMSCs的载体应用于骨组织工程。Objective To explore the biocompatibility of nano-hydroxyapatite/polyamide 66 （nHA/PA66） with human bone mesenchymal stem cells （hBMSCs） after osteogenic induction. Methods After hBMSCs were isolated and cultured in vitro, the experiment was conducted in 3 groups. Group A were hBMSCs subjected to no osteogenic induction, group B hBMSCs subjected to osteogenic induction, and group C was the composite of nHA/PA66 with hBMSCs subjected to osteogenic induction. Adhesion of the cells onto the nHA/PA66 in group C was observed by electron microscope scanning. Growth and proliferation of the cells in groups B and C were detected by MTY test at 1, 2 and 3 weeks. The ability of osteogenic differentiation of hBMSCs in vitro was analyzed by alkaline phosphatase （ALP） activity and alizarin red staining. The ability of osteogenic differentiation of hBMSCs on nHA/PA66 was tested by ALP activity. Results Electron microscope scanning showed that the cells spread and attached well on the surface of the composite scaffold in group C; the proliferation of the cells in groups B and C showed no significant difference （ P 〉 0. 05）. These suggested that the proliferation of hBMSCs was not affected by nHA/PA66. The number of mineralized nodules in group B was significantly larger than in group A （ P 〈 0. 05 ） ; the ALP activity of the cells in group A was significantly lower than in group B at 6 and 12 days （ P 〈 0. 05）; no significant differences were observed between groups B and C （ P 〉 0. 05）. These indicated that the hBMSCs were capable of osteogenic differentiation which was not affected by nHA/PA66. In groups B and C, the ALP activity of the cells at 12 days was significantly higher than at 6 days, indicating the ALP activity increased with increased induction time （ P 〈 0. 05）. Conclusion nHA/PA66 can be used as a carrier of hBMSCs in bone tissue engineering because hBMSCs can well adhere to, proliferate, and differentiate into bone on nHA/PA66 scaffolds.

关 键 词：骨髓细胞 羟基磷灰石 聚酰胺类 生物相容性材料 

分 类 号：R318.08[医药卫生—生物医学工程] R68[医药卫生—基础医学]