论著·基础研究

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

  • LIU Li-li1 ,
  • CUI Wen-guo1 ,
  • 2
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  • 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
刘立立(1993—),男,硕士生;电子信箱:13295152709@163.com。

网络出版日期: 2020-11-04

基金资助

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)。

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

  • 刘立立1,崔文国1 ,
  • 2
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  • 1. College of Pharmaceutical Sciences, Soochow University, Suzhou 215000, China; 2. 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

Online published: 2020-11-04

Supported by

National Natural Science Foundation of China (51873107); Shanghai Talent Development Fund (2018099); Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant Support (20171906); Shanghai Jiao Tong University “Medical and Research” Program (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 thiol-terminated 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(~350 μm);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。

本文引用格式

LIU Li-li1 , CUI Wen-guo1 , 2 . 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[J]. 上海交通大学学报(医学版), 2020 , 40(9) : 1184 -1192 . DOI: 10.3969/j.issn.1674-8115.2020.09.004

Abstract

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 (~ 350 μm). 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.
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