缓释bFGF微球的制备及微球对成骨细胞的作用  

Preparation of sustained release microspheres of bFGF and its effect on osteoblasts

作　　者：段宏[1] 樊瑜波[2] 屠重棋[1] 刘钰[3] 裴福兴[1] 

机构地区：[1]四川大学华西医院骨科,四川成都610041 [2]北京航空航天大学生物工程系 [3]中国科学院成都有机化学研究所,四川成都610041

出　　处：《华西药学杂志》2007年第3期259-262,共4页West China Journal of Pharmaceutical Sciences

基　　金：国家自然科学基金资助项目(批准号:10402025)

摘　　要：目的应用W/O/W复乳-干燥法制备碱性成纤维细胞生长因子-聚乳酸-聚羟基乙酸共聚物（bFGF-PLGA）微球,考察其对成骨细胞的作用。方法应用正交设计试验进一步优化bFGF微球的制备工艺,采用优化处方制备bFGF-PLGA微球,并考察其外观形态和粒径分布,检测其载药量和包封率,研究其体外释药的性质。将bFGF和bFGF-PLGA微球加入成骨细胞培养液中,用MTT法、流式细胞仪观察细胞的增殖情况,并检测成骨细胞上清液中骨钙素（BGP）的含量。结果所制微球表面光滑圆整,球体均匀度好,无黏连现象。微球粒径的98%分布在1.22-10.24μm之间,平均粒径为5.52μm,径距1.43±0.18。载药量48×10^-3%±1×10^-3%,包封率为72.37%±1.16%。突释期内微球的体外释放度仅为19.26%,21 d后体外累积释放度高达85.46%,释出bFGF的平均浓度为72.47±6.26 ng·ml^-1,微球的体外释药规律符合H igu ich i方程（r=0.9978）。培养1 d后,3组吸光度的差异均无显著性意义;4、6、8 d时,PLGA微球组明显优于其余两组。培养2 d后,bFGF组的G2/M＋S期百分数最高,4、8 d后,PLGA微球组G2/M＋S百分数最高。成骨细胞上清液中PLGA微球组的BGP含量最高,其次为bFGF组。结论所制备的bFGF-PLGA微球表征良好,载药量和包封率高;微球通过较长时间地持续释放活性bFGF,可明显促进成骨细胞的增殖和分化。OBJECTIVE To prepare the basicfibroblast growth factor （bFGF） sustained release microspheres （MS） by W/O/W multiple emulsion evaporation method, and investigate their effects on the cultured osteoblasts. METHODS The formulation of bFGF- PLGA[ poly（ lactic -co- glycolic -acid）] -MS was optimized by orthogonal design. The morphology, size distribution, drug loading volume encapsulation efficiency and in vitro drug release behavior of bFGF - PLGA - MS being measured. After bFGF and bF- GF microspheres being added to the DMEM culture medium of osteoblast, the proliferation of cultured osteoblasts was measured with MTT method and flow cytometry. And the content of BGP（ bone gla protein） secreted by osteoblast was also measured with RIA method. RESULTS The bFGF MS prepared based on optimized formulation exhibited well - defind properties, with the even and uniform sphere in appearance, regular particles without adhesion, about 98% of particles with a size distribution between 1.22 to 10.24 pan, with a mean particle size of 5.22 μm and size span of 1.43 ±0. 18. The drug loading volume and encapsulation efficiency of bFGF MS reached 48 ×10^3% ± 1 ×10^3% and 72.37% ± 1.16%, respectively. In the burst release phase, the rate of in vitro drug release amounted to only 19.26%, but rose to 85.46% accumulatively 21 days later, with stable concentration of bFGF released from MS with the mean concentration of 72.47 ±6.26 ng. ml-1. The in vitro drug release of bFGF MS corresponded with Higuichi equation（ r = 0. 9978）. The in vitro cellular study showed no significant difference in the A values of three groups 1 day after plate culture. The A values in the bFGF MS group were higher than those in other two groups 4 and 6 days after plate culture. The flow cytometrical examination showed that the G2/M ＋ S percentage in the bFGF group reached the highest 2 days after plate culture and the G2/M ＋ S percentage in the bFGF MS group went the highest 4 and 8 days after plate culture. Among three groups, the c

关 键 词：碱性成纤维细胞生长因子 聚乳酸-聚羟基乙酸共聚物 微球 缓释 正交设计 成骨细胞 

分 类 号：R94[医药卫生—药剂学] R963[医药卫生—药学]