摘要
目的制备重组人骨形态发生蛋白/壳聚糖/硫酸葡聚糖(CS/DS/rhBMP-2)复合微球,评估其体外生物相容性。方法采用离子交联法制备CS/DS/rhBMP-2复合微球及CS/DS空白微球,电镜观察CS/DS/rhBMP-2复合微球的形态。采用MTT法检测CS/DS/rhBMP-2微球组、CS/DS空白微球组、rhBMP-2组、对照组各组对L929小鼠成纤维细胞的细胞毒性。结果成功制备CS/DS/rhBMP-2复合微球,电镜检查结果示微球成球满意,分散度好。体外实验结果提示复合微球与L929细胞共培养1、3、5、7 d,其光密度(OD)值与对照组比较差异无统计学意义(P>0.05);CS/DS/rhBMP-2微球组、CS/DS空白微球组、rhBMP-2组、对照组浸提液对L929细胞作用2 d后的细胞相对增殖率(RGR)分别为102.41%、102.85%、99.34%、100%,毒性分级分别为0、0、1、0级。结论成功制备的CS/DS/rhBMP-2复合微球体外生物相容性好,无细胞毒性,在骨组织工程领域具有潜在的应用价值。
Objective To prepare recombinant human bone morphogenetic protein-2/chitosan/dextran sulfate （CS/DS/rhBMP-2） composite microspheres, and to evaluate their biocompatibility in vitro. Methods CS/DS/rhBMP-2 and blank CS/DS microspheres were prepared with ionic crosslinking method, and the morphology of CS/DS/rhBMP2 microspheres was observed by scanning electron microscope （SEM）. For biocompatibility examination in vitro, they were divided into CS/DS/rhBMP-2 group, CS/DS group, rhBMP-2 group and control group, cell cytotoxicity to L929 cell line of mouse fibroblasts in all groups were detected by MTT assay. Results SEM results showed CS/DS/rhBMP2 microspheres were well balling and well distributed, showing that the＆nbsp;composite microspheres were prepared successfully. Biocompatibility examinations revealed that there were no statistical significance of optical density （OD） among CS/DS/rhBMP-2 group and other control groups after 1, 3, 5, 7 day＇s co-culture with L929 cells. In CS/DS/rhBMP-2 group, CS/DS group, rhBMP-2 group and control group after 2 day＇s co-culture with L929, cell relative growth rate （RGR） were 102.41%, 102.85%, 99.34% and 100%, and the result indicated 0, 0, 1, 0 grade according to the toxicity classification respectively. Conclusion The prepared CS/DS/rhBMP-2 microspheres shown good dispersion and cytocompatibility in vitro, may be applied in bone tissue engineering field in the future due to its potential value.
出处
《中国骨科临床与基础研究杂志》
2013年第6期345-349,共5页
Chinese Orthopaedic Journal of Clinical and Basic Research
基金
国家自然科学基金青年基金(513031)
广东省自然科学基金项目(S2012010009743)
广州市科技计划项目(2013J4100120)
关键词
骨形态发生蛋白类
壳聚糖
硫酸葡聚糖
微球体
迟效制剂
生物相容性材料
细胞毒性
Bone morphogenetic proteins
Chitosan
Dextran sulfate
Microspheres
Delayed-action preparations
Biocompatible materials
Cytotoxicity
