引用本文:
吴道澄, 万明习, 张胜, 吴红, 胡蕴玉. 注射用人骨形成蛋白纳米微粒的制备与性质[J]. 应用化学,
2002, 19(7): 613-616.
Citation: WU Dao-Cheng, WAN Ming-Xi, ZHANG Sheng, WU Hong, HU Yun-Yu. Preparation and Property of Bone Morphogenetic Protein-2 Polycyanoacrylate Nanoparticle for Injection[J]. Chinese Journal of Applied Chemistry, 2002, 19(7): 613-616.
Citation: WU Dao-Cheng, WAN Ming-Xi, ZHANG Sheng, WU Hong, HU Yun-Yu. Preparation and Property of Bone Morphogenetic Protein-2 Polycyanoacrylate Nanoparticle for Injection[J]. Chinese Journal of Applied Chemistry, 2002, 19(7): 613-616.
注射用人骨形成蛋白纳米微粒的制备与性质
摘要:
用乳液聚合法制得注射用人骨形成蛋白-2(rhBMP-2)聚氰基丙烯酸正丁酯纳米微粒,考察其体内外生物活性.采用正交设计法优化制备工艺;用电子显微镜测定纳米微粒的形态和大小;紫外分光光度法测定rhBMP-2含量与包裹率;四唑盐比色法(MTT法)测定细胞增殖情况,用碱性磷酸酶(ALP)染色法测定rhBMP-2纳米微粒的生物活性.结果表明rhBMP-2纳米微粒形态规整,平均粒径(107±12)nm,rhBMP-2的包裹率为78%.4℃存放6个月生物活性无变化.体外缓释试验表明纳米微粒中80%的rhBMP-2在前10d释放,向培养基中加入rhBMP-2纳米微粒可以明显提高骨髓基质细胞(MSC)中的碱性磷酸酶水平且与rhBMP-2及空白对照有显著性差异(P<0.01).说明注射用rhBMP-2聚氰基丙烯酸正丁酯纳米微粒生物活性好,具有确定的缓释作用,可作为注射性骨诱导制剂.
English
Preparation and Property of Bone Morphogenetic Protein-2 Polycyanoacrylate Nanoparticle for Injection
Abstract:
The bone morphogenetic protein-2(rhBMP-2) was combined with polycyanoacrylate by improved emulsion polymerization method to give nanoparticles for injection use. Their biological activities were tested both in vivo and in vitro. The process of the recombination preparation was optimized by orthogonal factorization method. The size of the nanoparticle was determined by electronic microscopy. The content of rhBMP-2 and its encapsulation rate were determined by UV spectrophotometry. The proliferation of the cell was measured by MTT assay. The biological activity of the rhBMP-2 polycyanoacrylate nanoparticle was examined by alkaline phosphatase(ALP) coloration method. The results showed that the rhBMP-2 polycyanoacrylate nanoparticle was homogenous and stable with the average diameter (107±12) nm and rhBMP encapsulation rate of 78%. The in vivo experiment showed that 80% of rhBMP could be released in 10 days. The ALP level in the marrow stromal cell(MSC) cultural system increased significantly(P<0.01) by adding rhBMP nanoparticle to MSC system.
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