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目的制备负载重组人骨形态发生蛋白-2(rhBMP-2)壳聚糖纳米微球,并检测其粒径、形态、降解及药理特性,以评估壳聚糖纳米微球作为rhBMP-2缓释载体的可行性。方法以壳聚糖为原料、三聚磷酸钠为交联剂,通过离子交联法制备负载rhBMP-2壳聚糖纳米微球,应用透视电镜观察微球的形态、激光粒径,分析其粒径分布、溶菌酶降解,了解降解特性。通过酶联免疫吸附实验(ELISA)检测rhBMP-2壳聚糖微球的载药率、包封率和释药规律。结果离子交联法制备的壳聚糖纳米微球,平均粒径大小为230nm,成球性较好,包封率和载药率分别为(66.867±4.575)%、(33.437±2.290)μg/mg;体外释药试验rhBMP-2可以从壳聚糖纳米微球中缓慢释放,释放行为符合双向动力学规律,整个释放过程可达30 d。结论离子交联法可成功制备壳聚糖纳米微球并具有缓释rhBMP-2的能力,为进一步应用于骨组织工程研究提供实验依据。
Objective To prepare recombinant human bone morphogenetic protein-2 (rhBMP-2) loaded chitosan nanospheres and investigate its particle size, morphology, degradation and pharmacological properties to evaluate the effect of chitosan nanospheres as a sustained-release carrier of rhBMP-2 Feasibility. Methods Chitosan as raw material and sodium tripolyphosphate as crosslinking agent were prepared by ion-crosslinking rhBMP-2 loaded chitosan nanospheres. The morphologies and laser particle size of microspheres were observed by transmission electron microscope. Path distribution, lysozyme degradation, to understand the degradation characteristics. The drug loading rate, entrapment efficiency and drug release of rhBMP-2 chitosan microspheres were detected by enzyme-linked immunosorbent assay (ELISA). Results The average diameter of chitosan nanospheres prepared by ion-crosslinking method was 230 nm with good sphericity. The encapsulation efficiency and drug-loading rate were (66.867 ± 4.575)% and (33.437 ± 2.290) μg / mg; In vitro release test rhBMP-2 can be slowly released from the chitosan nanospheres, the release behavior in line with the two-way kinetics, the release process up to 30 d. Conclusion Ion-crosslinking method can successfully prepare chitosan nanospheres and have the ability of sustained release of rhBMP-2, which can provide experimental evidence for further application in bone tissue engineering.