水凝胶复合载辛伐他汀蛋白微球对成骨细胞增殖分化的影响  

Effect of hydrogel-loaded simvastatin microspheres on osteoblast proliferation and differentiation

作　　者：柳晓琳 穆新月 马子雨 刘树泰 王文龙 韩晓谦[1] 董志恒[6] Liu Xiaolin;Mu Xinyue;Ma Ziyu;Liu Shutai;Wang Wenlong;Han Xiaoqian;Dong Zhiheng(Department of Prosthodontics,Affiliated Hospital of Binzhou Medical College,Binzhou 256600,Shandong Province,China;Department of Stomatology,Affiliated Hospital of Jining Medical College,Jining 272007,Shandong Province,China;Binzhou Medical College,Yantai 264000,Shandong Province,China;Periodontal Department,Yantai Stomatological Hospital,Yantai 264000,Shandong Province,China;Department of Oral and Maxillofacial Surgery,Affiliated Hospital of Binzhou Medical College,Binzhou 256600,Shandong Province,China;Department of Children’s Stomatology,Affiliated Hospital of Binzhou Medical College,Binzhou 256600,Shandong Province,China)

机构地区：[1]滨州医学院附属医院口腔修复科,山东省滨州市256600 [2]济宁医学院附属医院口腔内科,山东省济宁市272007 [3]滨州医学院,山东省烟台市264000 [4]烟台市口腔医院牙周科,山东省烟台市264000 [5]滨州医学院附属医院口腔颌面外科,山东省滨州市256600 [6]滨州医学院附属医院儿童口腔科,山东省滨州市256600

出　　处：《中国组织工程研究》2023年第7期998-1003,共6页Chinese Journal of Tissue Engineering Research

基　　金：山东省医药卫生科技发展计划项目(ZR2019PH075),项目负责人:王文龙;滨州医学院科技计划项目(BY2019KJ15),项目负责人:韩晓谦。

摘　　要：背景:牙周炎患者逐年增加,采用传统的牙周治疗方法并不能恢复牙周软硬组织,因此需要制备出一种药物缓释材料辅助治疗牙周炎,恢复牙周破坏的软硬组织。目的:制备装载辛伐他汀的牛血清白蛋白微球复合水凝胶材料,检测其对辛伐他汀的双重缓释作用,并进一步研究此释药复合材料对成骨细胞黏附、增殖的影响。方法:①采用去溶剂法制备载辛伐他汀的牛血清白蛋白微球,采用透射电镜、扫描电镜观察微球的形态,测量其粒径,采用酶标仪检测载药微球的包封率、载药率及体外药物释放;②将载药微球负载到明胶水凝胶中,采用酶标仪检测水凝胶的体外药物释放,扫描电镜观察复合材料的形态;利用该水凝胶浸提液培养MC3T3-E1细胞,采用CCK-8法检测细胞增殖,碱性磷酸酶试剂盒检测细胞碱性磷酸酶的表达。结果与结论:①透射电镜显示,载药微球呈光滑的圆球形,较为分散,未见明显聚集,微球粒径也较为均匀,80%的微球粒径在0.2-0.8μm之间;扫描电镜显示,载药微球光滑,呈圆球形,分散性较好,粒径分布较为均匀,粒径在0.2-0.8μm之间;②载药微球的包封率为68.9%-87.5%,载药率为0.95%-1.21%;③载药微球中辛伐他汀的释放曲线是温和的持续释放过程,载药水凝胶中辛伐他汀的释放曲线为前期释放速度快后期缓慢的持续释放过程,其中载药水凝胶比载药微球能更快释放并达到药物的作用浓度,在后期缓慢释放维持药物的作用浓度;④扫描电镜显示,载药水凝胶呈多孔条状结构,适合成骨细胞的黏附和生长,在水凝胶表面可以看到圆球形的载药微球;⑤载药水凝胶可促进MC3T3-E1细胞的增殖与碱性磷酸酶表达;⑥结果表明,缓释辛伐他汀微球水凝胶复合材料具有良好的生物相容性,能够促进成骨细胞的增殖与成骨分化。BACKGROUND: The number of patients with periodontitis increases year by year, and the use of traditional periodontal treatment cannot restore the periodontal soft and hard tissue. Therefore, it is necessary to prepare a drug sustained-release material to assist the treatment of periodontitis and restore the damaged soft and hard tissues of periodontitis.OBJECTIVE: To prepare bovine serum albumin microspheres composite hydrogel material, detect the dual sustained release effect on simvastatin, and further study the effect of the composite material on the adhesion and proliferation of osteoblasts.METHODS:(1) Bovine serum albumin microspheres loaded with simvastatin were prepared by solvent removal method. The morphology of microspheres was observed by transmission electron microscopy and scanning electron microscopy and the particle size of microspheres was measured. The encapsulation rate, drug loading rate and in vitro release of microspheres were detected by a microplate analyzer.(2) Simvastatin albumin microspheres were loaded into the prepared hydrogel, and in vitro drug release from hydrogels was detected using a microplate reader. The morphology of the composite material was observed by scanning electron microscopy. The hydrogel extract was used to culture MC3T3-E1 cells. The proliferation of osteoblasts was detected by CCK-8 assay. The alkaline phosphatase expression in osteoblasts was detected by alkaline phosphatase kit.RESULTS AND CONCLUSION:(1) Transmission electron microscopy showed that bovine serum albumin microspheres and blank microspheres loaded with simvastatin were smooth, spherical and dispersed, with no obvious aggregation. The particle size of bovine serum albumin microspheres was uniform, and 80%of the microspheres had a particle size between 0.2 μm and 0.8 μm. Scanning electron microscopy showed that the microspheres were smooth and spherical with good dispersion and uniform particle size distribution, ranging from 0.2 μm to 0.8 μm.(2) The encapsulation rate and drug loading rate of sim

关 键 词：辛伐他汀 药物缓释 微球 水凝胶 成骨细胞 增殖与分化 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R783.1