摘要
目的探讨骨形成蛋白4/基质细胞衍生因子1(BMP-4/SDF-1)释体联合骨髓间充质干细胞(BMSCs)修复骨缺损的可行性。方法冷冻干燥和乳化交联法制备BMP-4/SDF-1壳聚糖缓释体,通过电镜扫描观察缓释体形态,酶联免疫吸附测定法检测BMP-4和SDF-1的缓释作用;体外分离培养兔BMSCs,Transwell检测缓释体对BMSCs的趋化作用,单核细胞直接细胞毒性实验检测细胞因子缓释体对兔BMSCs的增殖作用,定量聚合酶链反应检测成骨相关指标[碱性磷酸酶(ALP)/骨钙素(OCN)]茜素红染色观察带有细胞因子的缓释体对BMSCs矿化结节形成能力。结果空白组、BMP-4组、SDF-1组、BMP-4/SDF-1组的兔BMSCs增殖随着时间的增长而逐渐升高,且BMP-4/SDF-1组的兔BMSCs增殖率显著高于其他各组,差异有统计学意义(P<0.05)。将共培养的兔BMSCs小室进行结晶紫染色发现,BMP-4/SDF-1组细胞穿膜数目相对空白组、BMP-4组、SDF-1组显著增多(P<0.05)。复合缓释体与兔BMSCs共培养7 d后BMP-4/SDF-1组ALP和OCN表达均明显高于其他组,差异有统计学意义(P<0.05)。BMP-4/SDF-1矿化结节较空白组、BMP-4组和SDF-1组明显增多。结论制备兔骨缺损模型,将SDF-1/BMP-2壳聚糖换实体与兔骨髓间充质干细胞联合植入兔局部骨缺损处,促进黏附增殖,诱导成骨分化填补骨缺损,BMP-4/SDF-1壳聚糖缓释体支架可以作为理想的骨缺损修复材料。
Objective To investigate effects of hydrogel release-controlled mierobody carrying bone morphogenetie protein-4 （ BMP-4 ）/strernal cell-derived faetar-1 （ SDF-1 ） on repairing bone marrow rnesenchyreal stem cells （ BMSCs ） osteogenie differentiation. Methods BMP-4/SDF-1 loaded ehitosan hydrogel release-controlled mierobody was prepared by freeze drying and emulsion erosslinking method. The morpho- logical characteristics of release-controlling microbody was observed by electron microscope scanning, and the effect was analyzed by enzyme linked immunosorbent assays （ELISA）. Rabbit BMSCs were euhured in vitro Transwell was used to detect BMSCs ehemotaxis of the release body, mononuclear cell direct eytotoxieity assay was used for the detection of effect of cytokine release on rabbit BMSCs proliferation, quantitative polymerase chain reaction assay was used to detect bone indices of alkaline phasphatase （ALP）/osteoealein（ OCN ）, and alizarin red staining was used to observe the release microbody effect on BMSCs mineralized nodule formation. Results The rabbit BMSCs proliferation of blank group, BMP-4 group, SDF-1 group, BMP4/SDF group gradually increased over time, and the rabbit BMSCs proliferation rate of BMP-4 group was significantly higher than other groups （ P 〈 0.05 ）. The co-cultured rabbit BMSCs small chamber crystal violet staining result showed that membrane penetrating cell number of BMP-4/SDF-lgroup was significantly more than the blank group, BMP-4 group and SDF-1 group （P 〈 0.05 ）. Sustained release composite body with After 7 days of co-culture of release-eantrolling micorbody with rabbit BMSCs, the ALP and OCN expression of BMP-4/ SDF-1 group were significantly higher than other groups, the difference was statistically significant （P 〈 0. 05 ）. BMP-4/SDF-1 mineralized nodules were significantly increased than the eantrol group, BMP-4 group and SDF-1 group. Conclusion Combining the SDF-1/BMP-2 chitosan with rabbit BMSCs to prepare the rabbit bone defect model c
出处
《医学综述》
2016年第4期807-810,F0003,共5页
Medical Recapitulate
基金
武汉市卫生局临床医学科研项目
关键词
成骨分化
壳聚糖
基质细胞衍生因子1
骨形成蛋白4
Osteoblast differentiation
Chitosan
Stromal cell-derived factor-I
Bone morphogenetic protein-4
