负载骨形态发生蛋白2微球的可注射水凝胶的抑菌和成骨性能评价  

Injectable hydrogel loaded with bone morphogenetic protein-2 microspheres for bacteriostasis and osteogenesis

作　　者：刘立立 崔文国[1,2] LIU Li-li;CUI Wen-guo(College of Pharmaceutical Sciences,Soochow University,Suzhou 215000,China;Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint Disease,Shanghai Institute of Traumatology and Orthopaedics,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)

机构地区：[1]苏州大学药学院,苏州215000 [2]上海交通大学医学院附属瑞金医院,上海市伤骨科研究所,上海市中西医结合防治骨与关节病损重点实验室,上海200025

出　　处：《上海交通大学学报（医学版）》2020年第9期1184-1192,共9页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(51873107);上海市人才发展基金(2018099);上海市教育委员会高峰高原学科建设计划(20171906);上海交通大学医学转化交叉基金(ZH2018ZDA04)。

摘　　要：目的·构建一种负载骨形态发生蛋白2(bone morphogenetic protein-2,BMP-2)微球的新型抗菌和可注射水凝胶,检测其生物相容性、抗菌和促成骨性能。方法·通过微流控技术制备负载BMP-2的光交联明胶微球,将微球与巯基化的聚乙二醇[4-arm thiolterminated poly(ethylene glycol),4SH-PEG]混合,用Ag+交联4SH-PEG,制备含有微球的可注射4SH-PEG复合水凝胶(BMP/Gel/SH-Ag)。光学显微镜和扫描电子显微镜(scanning electron microscope,SEM)观察微球和水凝胶的微观形态;在100转/min、37℃的摇床中进行药物释放实验;使用针头直径为0.5 mm的注射器注射BMP/Gel/SH-Ag,评价其可注射性;通过琼脂扩散实验评价BMP/Gel/SH-Ag对金黄色葡萄球菌(Staphylococcus aureus,S.aureus)和大肠埃希菌(Escherichia coli,E.coli)的抗菌性能;通过CCK-8实验考察BMP/Gel/SH-Ag的生物相容性;通过骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)的碱性磷酸酶(alkaline phosphatase,ALP)染色和钙结节染色评价其促成骨活性。结果·明胶微球外表光滑,粒径分布均匀(~350μm);BMP/Gel/SH-Ag呈现孔洞状的微观结构;在摇床中温育8 d,BMP/Gel/SH-Ag中的BMP-2累积释放量为(81.8±3.6)%;BMP/Gel/SH-Ag可以用直径小至0.5 mm的注射器针头来注射,以产生水凝胶细丝;BMP/Gel/SH-Ag对S.aureus和E.coli有抑制作用;CCK8结果表明,BMP/Gel/SH-Ag具有良好的生物相容性;BMP/Gel/SH-Ag可提高BMSCs的ALP的表达和钙结节的含量。结论·该BMP/Gel/SH-Ag水凝胶在促进成骨和抗感染方面有良好的表现。Objective·To construct a novel antibacterial and injectable hydrogel(BMP/Gel/SH-Ag)loaded with bone morphogenetic protein-2(BMP-2)microspheres,and investigate its biocompatibility,antibacterial properties and bone-promoting properties.Methods·The photocrosslinked gelatin microspheres loaded with BMP-2 were prepared by microfluidics.The microspheres were mixed with 4-arm thiol-terminated poly(ethylene glycol)(4SH-PEG)and crosslinked with Ag+to prepare injectable sulfhydrylated PEG hydrogels(BMP/Gel/SH-Ag).The micromorphology of microspheres and hydrogels was observed by light microscope and scanning electron microscope.The drug release profile was investigated at 37℃in a shaker(100 r/min).The injectability of BMP/Gel/SH-Ag was evaluated by injecting hydrogel using a syringe with a tip diameter of 0.5 mm.The antibacterial activity of BMP/Gel/SH-Ag against Staphylococcus aureus(S.aureus)and Escherichia coli(E.coli)was evaluated by agar diffusion test.The biocompatibility of BMP/Gel/SH-Ag was verified by CCK-8,and the bone-promoting activity was evaluated by alkaline phosphatase(ALP)assay and calcium nodule staining in bone marrow mesenchymal stem cells(BMSCs).Results·Gelatin microspheres had smooth appearance and uniform particle size distribution(~350μm).BMP/Gel/SH-Ag had porous microstructure and can be injected with a syringe needle with a diameter of up to 0.5 mm in diameter to produce hydrogel filament.The cumulative release of BMP-2 from BMP/Gel/SH-Ag was(81.8±3.6)%after being incubated for 8 d.BMP/Gel/SH-Ag had obvious inhibitory effect on S.aureus and E.coli.CCK-8 results showed that BMP/Gel/SH-Ag had good biocompatibility.BMP/Gel/SH-Ag can increase the expression of ALP and the content of calcium nodules in rat BMSCs.Conclusion·The BMP/Gel/SH-Ag has good performance in promoting osteogenesis and anti-infection.

关 键 词：微流控 微球 抗菌 可注射水凝胶 促成骨 

分 类 号：R94[医药卫生—药剂学]