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组间差异无统计学意义(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.

abstractsObjective 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 MTI 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 ＞ O.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 ＜ O.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.