摘要
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-loaded gelatin microspheres（rhIGF-Ⅰ-GMS）and investigate their general properties,in vitro drug release characteristics and evaluate the biological effects of rhIGF-Ⅰ-GMs on the proliferation and collagen Ⅰ type in human periodontal ligament fibroblasts （HPDLFs）. Method：The rhIGF-Ⅰ-load-ed gelatin microspheres were prepared by improved emulsified cold-condensation method. Their general properties were observed by SEM. To measure the content of rhIGF-Ⅰ-GMs by ELISA method to study their release characteristics in tivro. The proliferation of the cultured fibroblast was measured with MTT method and collagen typeⅠwas measured with hydroxyprolline kit after rhIGF-Ⅰ-GMs and rhIGF-Ⅰ being added to the DMEM culture medium. Result：The rhIGF-Ⅰ-GMs were good,even and uniform spheres with mean particle size o（f12.51±3.53）μm. The lyophilized powder was pure white with good disparity. The drug loading amount and encapsulation efficiency of rhIGF-Ⅰ-GMs were 920 ng/g and 92.0 %. The drug release rate was 72.5 % at the 7th days after plate culturing. Both rhIGF-Ⅰand rhIGF-Ⅰ-GMs promoted the proliferation and collagenⅠ type of HPDLFs in a concentration and time related manner. Singnificantly greater proliferation and expression were found in rhIGF-Ⅰ-GMs group. Conclusion：The rhIGF-Ⅰ-GMs and their lyophilized powder have excellent pharmaceutical properties. The rhIGF-Ⅰ-GMs can significantly promote the proliferation and increase collagen typeⅠof HPDLFs by sustained release rhIGF-Ⅰ.
出处
《临床口腔医学杂志》
2010年第1期21-24,共4页
Journal of Clinical Stomatology
