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目的制备载血小板复合生长因子明胶缓释微球,观察其一般特性,检测其体外缓释效果。方法以明胶为原料,戊二醛为交联剂,采用改良的乳化冷凝聚合法制备明胶微球,以微球粒径为参考指标,通过单因素实验对明胶微球的制备条件进行优化。用光学显微镜、扫描电镜(SEM)对其大小结构形貌等一般特性进行表征;以血管内皮生长因子(vascular epithelial growth factor,VEGF)的含量为代表,酶联免疫吸附测定(ELISA)法检测其含量,计算微球吸附生长因子率、载生长因子量,测定体外缓释效果。结果当明胶溶液浓度为15%、Span8加入量为1.5 m L,搅拌速度为(750-855)r/min时可获得最优的明胶微球制备方案,获得的明胶微球平均粒径为15.95±3.79μm,其中7-20.5μm的微球占88.23%。制备的PG上清中VEGF的含量为(694.45±1.13)pg/m L,微球吸附生长因子率为87.7%,载VEGF量为6.09 pg/mg,体外缓释7 d累计释放量占总载药率的75.73%。结论制备的明胶缓释微球外观圆整,粒径均匀,分散度良好,吸附生长因子率、载生长因子量及体外缓释效果良好,初步实现了载血小板复合生长因子明胶缓释微球冻干制品的研制。
Objective To prepare sustained-release microspheres loaded with platelet-derived growth factor gelatin and observe its general characteristics. Methods Using gelatin as raw material and glutaraldehyde as crosslinking agent, gelatin microspheres were prepared by modified emulsification and condensation polymerization method. The microsphere diameter was taken as the reference index. The preparation conditions of gelatin microspheres were optimized by single factor experiment. The general characteristics such as size and structure were characterized by optical microscope and scanning electron microscope (SEM). The expression of vascular endothelial growth factor (VEGF) was detected by enzyme-linked immunosorbent assay (ELISA) Content, calculate the rate of microspheres adsorption of growth factors, containing the amount of growth factors, determination of sustained-release effect in vitro. Results The optimal gelatin microspheres preparation was obtained when the concentration of gelatin solution was 15%, the amount of Span8 was 1.5 mL and the stirring speed was (750-855) r / min. The average particle size of gelatin microspheres was 15.95 ± 3.79μm, of which 7-20.5μm microspheres accounted for 88.23%. The content of VEGF in the prepared supernatant of PG was (694.45 ± 1.13) pg / m L, the rate of microspheres adsorbing growth factor was 87.7%, the amount of VEGF loaded was 6.09 pg / mg and the cumulative release in vitro was 7 days Drug rate of 75.73%. Conclusion The prepared gelatin sustained-release microspheres have the advantages of round appearance, uniform particle size, good dispersion, good adsorption rate of growth factor, and good sustained release in vitro. The sustained release microspheres loaded with platelet growth factor gelatin Development of dry products.