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目的:观察rhBMP-2微球温敏型凝胶的药剂学特征及对人牙周膜细胞(PDLCs)增殖的影响。方法:采用W/O/W型乳化溶剂挥发法以聚乳酸-乙醇酸共聚物(PLGA)为材料制备空白微球,通过吸附法制备rhBMP-2载药微球。以单甲基聚乙二醇-羟基乙酸共聚物(MPEG-PLGA)为材料制备凝胶,采用夹心酶联免疫吸附法测定释药速度。采用四唑盐比色法(MTT)及酶动力学法测定细胞增殖和碱性磷酸酶活性。结果:rhBMP-2微球呈规则球形,表面多孔,粒径为(19.6±2.3)μm,微球凝胶剂相转变温度为36℃左右,微球凝胶剂体外释药曲线符合Higuchi方程,28d的累积释放度为80%。rhBMP-2微球凝胶剂能明显促进牙周膜细胞的增殖和提高ALP活性,随时间推移在第10d时rhBMP微球凝胶促PDLCs生长的作用明显强于rhBMP溶液。结论:rhBMP-2微球温敏型凝胶剂具有明显的缓释特征,与溶液剂相比促进PDLCs增殖作用更持久。
OBJECTIVE: To observe the pharmacological characteristics of rhBMP-2 microspheres and its effect on the proliferation of human periodontal ligament cells (PDLCs). Methods: Blank microspheres were prepared by W / O / W emulsion solvent evaporation method using polylactic-co-glycolic acid (PLGA) as the material and rhBMP-2 loaded microspheres were prepared by adsorption method. Gel was prepared from monomethyl polyethylene glycol-co-glycolic acid (MPEG-PLGA). The drug release rate was determined by sandwich enzyme-linked immunosorbent assay. Cell proliferation and alkaline phosphatase activity were determined by tetrazolium salt colorimetry (MTT) and enzyme kinetic method. Results: The rhBMP-2 microspheres were spherical in shape with porous surface, the particle size was (19.6 ± 2.3) μm and the phase transition temperature of the microsphere gel was about 36 ℃. The in vitro drug release curve of the microsphere gel was consistent with the Higuchi equation, The cumulative release of 28d is 80%. rhBMP-2 microspheres gel can significantly promote the proliferation of periodontal ligament cells and increase the activity of ALP. With the passage of time, the effect of rhBMP microspheres on the growth of PDLCs was stronger than that of rhBMP solution on the 10th day. CONCLUSION: The rhBMP-2 microspheres thermo-sensitive gelling agent has obvious sustained-release characteristics. Compared with the solution, rhBMP-2 gels can prolong the proliferation of PDLCs.