不同保存方法对生物衍生骨支架材料细胞相容性的影响

目的了解不同保存方法对生物衍生骨支架材料细胞相容性的影响.方法冻干生物衍生骨用两种不同保存液同等条件4℃冷藏保存3个月,以保存相同时间冻干生物衍生骨为对照.实验分为A组:成骨细胞复合 1号保存液处理材料培养;B组:成骨细胞复合 2号保存液处理材料培养;C组:成骨细胞复合冻干骨培养;D组:单纯成骨细胞培养.以2×106个/ml密度的成骨细胞悬液复合材料培养1、3、5和7天.用倒置相差显微镜和扫描电镜观察细胞的黏附和生长情况、检测细胞活力、碱性磷酸酶(alkaline phosphatase,ALP)活性及流式细胞仪分析细胞周期.结果成骨细胞与三种不同方法保存的材料均可黏附,并在材料孔隙内生长、分化和增殖.复合培养1和3天,各组间无显著差异;复合培养5和7天,黏附于材料的细胞活力大小依次为 A组>C组>B组 (P＜0.01,P＜0.05).复合培养7天,黏附于材料的细胞ALP活性大小依次为 A组>C组>B组(P＜0.01) .各组细胞周期未见明显变化,未见异倍体细胞.结论选择适宜的保存液对生物衍生骨支架材料的细胞相容性有一定优化作用.

EFFECT OF VARIOUS STORAGE METHODS ON CELLULAR COMPATIBILITY OF BIO-DERIVED BONE

OBJECTIVE: To study the effect of various storage methods on cellular compatibility of bio-derived bone. METHODS: Freeze-dried biomaterials had been stored in two different preservation solutions for three months, while the biomaterials stored for the same time were observed as control group. The experiment was divided into groups A, B, C and D according to different storage methods (group A: with materials stored in preservation solution 1; group B: with materials stored in preservation solution 2; group C: with freeze-dried materials; and group D: simple osteoblasts). Osteoblasts at 2 x 10(6)/ml had been cocultured with materials for 1, 3, 5, and 7 days. The cell-material complex was observed under phase microscope and electronic scanning microscope to evaluate the adhesion and growth of osteoblasts; the cell viability and alkaline phosphatase(ALP) activity were measured, and the cell cycle was analysed by flow cytometer. RESULTS: Osteoblasts adhered to materials preserved by different methods, differentiated and proliferated in the hole of materials. The difference of cell viability was not significant between three groups on day 1 and day 3. The cell viability of osteoblasts adhered to three materials was A > C > B group on day 5 and day 7 (P < 0.01, P < 0.05). The ALP activity of osteoblasts adhered to three materials was A > C > B group on day 7 (P < 0.01). The cell cycle of different groups did not change significantly, the abnormal cells were not seen. CONCLUSION: The choice of proper preservation solution can optimize the cellular compatibility of bio-derived bone.