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目的:制备重组人胰岛素样生长因子缓释明胶微球(rhIGF-Ⅰ-GMs),观察其一般性质、体外释药特性、对人牙周膜成纤维细胞(HPDLFs)的增殖及Ⅰ型胶原合成的影响。方法:改良乳化冷凝聚合法制备rhIGF-Ⅰ-GMs,扫描电镜(SME)观察其一般性质;ELISA法测定rhIGF-Ⅰ的含量,计算微球包封率和载药率及绘制微球体外释药曲线。体外培养HPDLFs,四唑盐比色法(MTT)和羟脯氨酸试剂盒消化法动态观察rhIGF-Ⅰ和rhIGF-Ⅰ-GMs梯度含量分别对HPDLFs增殖及Ⅰ型胶原合成的影响。结果:微球表面光滑圆整,球体均匀度好,平均粒径为(12.51±3.53)μm,冻干粉剂为白色粉末状,再分散性良好,微球载药量和包封率分别为920ng/g和92.0%,体外7d内药物缓释72.5%;rhIGF-Ⅰ和rhIGF-Ⅰ-GMs均明显促进HPDLFs增殖及Ⅰ型胶原分泌(P<0.05);rhIGF-Ⅰ-GMs较单独应用rhIGF-Ⅰ的效果更加显著(P<0.01),呈浓度及时间依赖性。结论:rhIGF-Ⅰ-GMs及其冻干粉剂制备良好;rhIGF-Ⅰ-GMs通过对rhIGF-Ⅰ的缓释作用促进HPDLFs增殖及Ⅰ型胶原分泌的效果明显优于单独使用rhIGF-Ⅰ。
OBJECTIVE: To prepare rhIGF-Ⅰ-GMs microspheres (rhIGF-Ⅰ-GMs) and observe its general properties, in vitro release characteristics, proliferation of human periodontal ligament fibroblasts (HPDLFs) and collagen type Ⅰ synthesis Impact. Methods: rhIGF-Ⅰ-GMs were prepared by modified emulsion polymerization. The general properties of rhIGF-Ⅰ-GMs were observed by scanning electron microscopy (SME). The content of rhIGF-Ⅰ was measured by ELISA. The microsphere encapsulation efficiency and drug loading rate curve. HPDLFs, tetrazolium assay (MTT) and hydroxyproline kit digestion method were used to observe the effect of gradient of rhIGF-Ⅰ and rhIGF-Ⅰ-GMs on the proliferation of HPDLFs and the synthesis of type Ⅰ collagen. Results: The surface of the microspheres was smooth and round, and the average particle size was (12.51 ± 3.53) μm. The lyophilized powder was white powder with good redispersibility. The drug loading and entrapment efficiency of the microspheres were 920 ng / g and 92.0%, respectively. The drug release was 72.5% in vitro for 7 days. Both rhIGF-Ⅰand rhIGF-Ⅰ-GMs significantly promoted the proliferation and typeⅠcollagen secretion of HPDLFs (P <0.05). Compared with rhIGF- Ⅰ effect was more significant (P <0.01), in a concentration and time-dependent manner. Conclusion: rhIGF-Ⅰ-GMs and its lyophilized powder are well prepared. The effect of rhIGF-Ⅰ-GMs on the proliferation and type I collagen secretion of HPDLFs is better than that of rhIGF-Ⅰ alone.