聚乳酸羟基乙酸-甲状旁腺激素相关蛋白复合微球的制备形态表征及生物相容性研究  

A study on the preparation,morphological characterization and biocompatibility of polylactide-co-glycolide microspheres loaded with parathyroid hormone-related protein

作　　者：刘金晶 周强强 龙桂月 黄臻 廖红兵[1] LIU Jin-jing;ZHOU Qiang-qiang;LONG Gui-yue(College&Hospital of Stomatology,Guangxi Medical University,Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction,Guangxi Clinical Research Center for Craniofacial Deformity,Guangxi Health Commission Key Laboratory of Prevention and Treatment for Oral Infectious Diseases,Nanning 530021,China)

机构地区：[1]广西医科大学口腔医学院/附属口腔医院,广西口腔颌面修复与重建研究自治区级重点实验室,广西颅颌面畸形临床医学研究中心,广西壮族自治区卫生健康委员会口腔感染性疾病防治重点实验室,南宁530021

出　　处：《中国临床新医学》2022年第6期507-511,共5页CHINESE JOURNAL OF NEW CLINICAL MEDICINE

基　　金：国家自然科学基金项目(编号:81560190);广西自然科学基金项目(编号:2018GXNSFAA138032);广西医学高层次人才培养计划资助项目(编号:G201902004)。

摘　　要：目的制备聚乳酸羟基乙酸-甲状旁腺激素相关蛋白复合微球(PLGA-PTHrP)并观察其形态表征及其生物相容性。方法采用复乳溶剂挥发法制备空白聚乳酸羟基乙酸(PLGA)微球,通过表面浸提法将不同浓度的甲状旁腺激素相关蛋白(PTHrP)装载到PLGA微球。采用冷冻干燥法制备负载有效浓度PTHrP的PLGA微球。通过扫描电镜观察PLGA微球形态并计算微球粒径,检测PLGA-PTHrP的蛋白释放量,并绘制释放曲线。将PLGA-PTHrP与大鼠全骨髓细胞共培养,分析复合微球对细胞增殖分化的影响。结果采用复乳溶剂挥发法制备得到的空载PLGA微球呈球形或类球形,表面较平整,平均粒径为(56.73±7.22)μm。载蛋白微球的体外蛋白质释放实验结果显示,蛋白质的释放速率在前100 h较快,释放时间可持续达300 h。载蛋白微球释放的蛋白质占蛋白质总量的87.3%。实验结果显示,PLGA-PTHrP对大鼠全骨髓细胞增殖活力无明显影响。结论通过复乳溶剂挥发法可制备出表面较平整,粒径较均匀的PLGA空白微球,使用表面浸提法及冷冻干燥法能制备出有效的PLGA-PTHrP缓释体系,该缓释体系能在一定时间范围内实现PTHrP的缓释作用。Objective To prepare polylactide-co-glycolide microspheres loaded with parathyroid hormone-related protein(PLGA-PTHrP)and to observe their morphological characterization and biocompatibility.Methods The empty polylactide-co-glycolide(PLGA)microspheres were prepared by double emulsion-solvent evaporation method.Parathyroid hormone-related protein(PTHrP)was loaded into the empty PLGA microspheres by soaking empty PLGA microspheres in the different concentrations of PTHrP solution.PLGA microspheres loaded with effective concentration of PTHrP were prepared by freeze-drying method.The morphology of PLGA microspheres was observed by scanning electron microscope and the particle size of microspheres was calculated.The protein release amount of PLGA-PTHrP was detected,and the release curve was drawn.PLGA-PTHrP was co-cultured with rat whole bone marrow cells,and the effect of composite microspheres on cell proliferation and differentiation was analyzed.Results The empty PLGA microspheres prepared by the double emulsion-solvent evaporation method were spherical or quasi-spherical,with flat surface and average particle size of(56.73±7.22)μm.The results of in vitro protein release experiment of protein loaded microspheres showed that the release rate of protein was faster within the first 100 hours,and the total release time lasted up to 300 hours.The protein released by protein loaded microspheres accounted for 87.3%of the total protein.The experimental results showed that PLGA-PTHrP had no significant effect on the proliferation of activity rat whole bone marrow cells.Conclusion The PLGA empty microspheres with flatter surface and more uniform particle size can be successfully prepared by double emulsion-solvent evaporation method.The effective PLGA-PTHrP sustained-release system can be prepared by surface soaking and extraction and freeze-drying method and the sustained-release system can realize the sustained-release effect of PTHrP in a certain time range.

关 键 词：聚乳酸羟基乙酸-甲状旁腺激素相关蛋白复合微球 复乳溶剂挥发法 缓释体系 大鼠全骨髓细胞 细胞增殖 

分 类 号：R915[医药卫生—微生物与生化药学]