表面修饰对纳米晶胶原基骨细胞相容性的影响

Effect of surface modification on cellular compatibility of nanoHydroxyapatite/collagen

目的:探索纤维蛋白(fibrin,FB)修饰的纳米晶胶原基骨(nanoHydroxyapatite/collagen,nHAC)支架材料上成骨诱导的骨髓间充质干细胞(marrow mesenchymal stem cells,MSCs)黏附、增殖及分化的情况。方法:实验分为两组:实验组,纤维蛋白修饰的纳米晶胶原基骨(FB-nHAC);对照组,单纯的纳米晶胶原基骨(nHAC)。将SD大鼠骨髓间充质干细胞定向诱导为成骨细胞,种植于支架材料上,体外复合培养。通过检测支架材料的细胞黏附率、不同时间点(3,7,10,14 d)支架材料中细胞数、碱性磷酸酶表达量以及扫描电镜观察细胞在支架材料上的生长状况,比较分析不同支架材料与细胞生物相容性差异。结果:大鼠MSCs经诱导培养14 d后,I型胶原免疫荧光染色为阳性;实验组支架材料的细胞黏附率显著高于对照组(P<0.05);且相同时间点实验组支架材料中的细胞数及碱性磷酸酶表达量也明显高于对照组(P<0.05);电镜观察发现两组材料上均有细胞生长,但实验组的细胞生长状况明显好于对照组。结论:表面修饰纤维蛋白后的nHAC支架材料具有更好的细胞黏附、增殖及促成骨分化能力。

Objective ： NanoHydroxyapatite/collagen （nHAC） modified with fibrin was prepared, and the adhesion, proliferation and differentiation of marrow mesenchymal stem cells on the scaffolds were investigated. Methods： MSCs obtained from SD rat bone marrow were induced and proliferated. After their osteoblastic phenotypes were demonstrated, MSCs were seeded onto nHAC modified with fibrin （experiment group） and common nHAC （control group）. The adhesive rates were calculated respectively. At different time points（3, 7, 10,14 d）, the number of the cells in the scaffolds was counted and alkaline phosphatase （ALP） activity was detected. The cells growth on scaffolds also was observed by scanning electron micrograph （SEM）. The differences between groups were compared and analyzed. Results： The differentiation of MSCs to osteoblastic phenotype were demonstrated by the positive staining of collagen type I. The efficiency of cell adhesion in the modified nHAC was higher than the unmodified nHAC （P 〈 0.05 ）. The cell number and ALP activity of MSCs adhered to nHAC modified by fibrin were obviously higher than that of MSCs adhered to simple material （P 〈 0.05 ）. Cells could be observed on every scaffold by SEM, but cells on the modified nHAC grew obviously better than that on the unmodified nHAC. Conclusion： nHAC modified with fibrin had a better cell adhesion, proliferation and facilitate bone formation ability.