骨髓间充质干细胞与支架材料nHA/PA66复合培养的生物活性研究

目的：研究骨髓间充质干细胞与支架材料-纳米羟基磷灰石/聚酰胺66（nHA/PA66）复合培养时生物学特性。方法：体外培养兔骨髓间充质干细胞（MSCs）,并分为A、B、C、D四组。A组为对照;B组和D组进行骨向诱导;同时,C组和D组细胞与nHA/PA66复合培养。应用MTT实验、碱性磷酸酶（ALP）染色和生物化学分析、扫描电镜,分别对各组细胞的增殖速度、ALP活性及细胞在支架上的黏附情况进行检测。结果：MTT结果显示,A组细胞增殖曲线与C组相似,而B组和D组相似;A组和C组细胞增殖略高于B组和D组。结果表明,骨向诱导可使细胞增殖轻微下降,而与支架材料复合培养对MSCs增殖无影响。ALP染色及ALP活性生化分析进一步表明,MSCs与支架材料复合培养对其骨向分化能力无影响。SEM观察则表明,C组细胞和D组细胞与支架材料均粘附良好。结论：nHA/PA66适于MSCs的黏附、增殖及骨向分化,是一种极具价值的骨组织工程支架材料。

Biological Characteristics of Bone Marrow Mesenchymal Stem Cells Co-cultured with Nano-hydroxyapatite/Polyamide 66

Objective：To study the biological characteristics of bone marrow mesenchymal stem cells（MSCs） co-cultured with nano-hydroxyapatite/polyamide 66（nHA/PA66） . Methods：Rabbit bone marrow MSCs were isolated and cultured in vitro. Cells were divided into four groups：A,B,C and D. Group A served as control,while cells in group B and D underwent osteogenic induction;at the same time,cells in group C and D were cocultured with nHA/PA66. Cell proliferation,alkaline phosphatase（ALP） activity and cell adhension on scaffold,were detected by MTT test,ALP staning and biochemical analysis,and scanning electron microscope（SEM） ,respectively. Results：MTT test showed that cell proliferation in group A and group C were similar,so were in group B and D. However,cell proliferation of group A and C were higher than those of group B and D. This indicated that although osteogenic induction could slightly reduce MSCs proliferation,coculture of MSCs with nHA/PA66 had no effect on cell proliferation. ALP staining and biochemical analysis further demonstrated that the osteogenic differentiation of MSCs was not affected by co-culture of MSCs with bone scaffold. SEM observation verified that good cell adhesion was obtained in both C and D. Conclusion：nHA/PA66 was suitble for MSCs adhension,proliferation and osteogenic differentiation,and was a valuable scaffold for bone tissue engineering.