上海交通大学学报（医学版）

上海交通大学学报(医学版) ›› 2020, Vol. 40 ›› Issue (09): 1184-1192.doi: 10.3969/j.issn.1674-8115.2020.09.004

• 论著·基础研究 • 上一篇    下一篇

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

刘立立1，崔文国1, 2   

  1. 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
  • 出版日期:2020-09-28 发布日期:2020-11-04
  • 通讯作者: 崔文国，电子信箱：wgcui80@hotmail.com。
  • 作者简介:刘立立（1993—），男，硕士生；电子信箱：13295152709@163.com。
  • 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

LIU Li-li1, CUI Wen-guo1, 2   

  1. 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:2020-09-28 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).

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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。

关键词: 微流控, 微球, 抗菌, 可注射水凝胶, 促成骨

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.

Key words: microfluidics, microsphere, bacteriostasis, injectable hydrogel, promoting osteogenesis

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