中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (16): 2534-2541.doi: 10.12307/2023.462

• 药物控释材料 drug delivery materials • 上一篇    下一篇

静电纺丝复合缓释抗菌微球的制备及性能分析

杨  勇1，许顺恩2，古先扬1，华大炜1，滕建祥1，王  桢1，叶  川1，2，3   

  1. 1贵州医科大学，贵州省贵阳市  550004；2贵州医科大学附属医院骨科，贵州省贵阳市  550004；3贵阳医学院附属医院骨科，贵州省贵阳市  550004
  • 收稿日期:2022-04-11 接受日期:2022-06-02 出版日期:2023-06-08 发布日期:2022-11-10
  • 通讯作者: 叶川，主任医师，贵州医科大学附属医院骨科，贵州省贵阳市 550004；贵州医科大学，细胞工程生物医药技术国家地方联合工程实验室，组织工程与干细胞实验中心，贵州省贵阳市 550004；贵阳医学院附属医院骨科，贵州省贵阳市 550004
  • 作者简介:杨勇，男，1993年生，贵州省贵阳市人，汉族，贵州医科大学在读硕士，医师，主要从事生物支架、生物材料、干细胞研究。

Preparation and property analysis of electrospinning composite sustained-release antibacterial microspheres

Yang Yong1, Xu Shunen2, Gu Xianyang1, Hua Dawei1, Teng Jianxiang1, Wang Zhen1, Ye Chuan1, 2, 3   

  1. 1Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 2Department of Orthopedics, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 3Department of Orthopedics, Affiliated Hospital of Guiyang Medical College, Guiyang 550004, Guizhou Province, China
  • Received:2022-04-11 Accepted:2022-06-02 Online:2023-06-08 Published:2022-11-10
  • Contact: Ye Chuan, Chief physician, Guizhou Medical University, Guiyang 550004, Guizhou Province, China; Department of Orthopedics, Affiliated Hospital of Guizhou Medical University, Guiyang 550004, Guizhou Province, China; Department of Orthopedics, Affiliated Hospital of Guiyang Medical College, Guiyang 550004, Guizhou Province, China
  • About author:Yang Yong, Master candidate, Physician, Guizhou Medical University, Guiyang 550004, Guizhou Province, China

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摘要：


文题释义：

静电纺丝：其本质上是高分子流体(黏度较高的非牛顿流体)在正极1-50 kV电压的推动下，形成静电雾化的特殊形式，最终在收集装置上固化成纤维。电纺纤维直径通常介于数十纳米至数微米之间，具有极大的比表面积、高孔隙率和相互连通的三维网状结构，构造与天然的细胞外基质接近。
静电喷雾：原理类似静电纺丝，作为电流体动力学射流技术的另一产物，其优势在于可将生长因子、多肽、药物等包封于其中。

背景：临床静脉注射抗生素治疗骨髓炎时，由于骨骼血供较差，药物难以在骨髓炎局部病灶处达到有效治疗浓度，治疗效果不佳一直是临床瓶颈。
目的：研制一种可局部缓释盐酸万古霉素、同时具备成骨作用的可降解生物材料植入物，在治疗骨髓炎的同时尽可能起到促进局部骨组织修复的作用。
方法：将6，12，18 g/L的盐酸万古霉素-海藻酸钠溶液以电喷的方式制成微球，进行真空冷冻干燥处理；制备聚3-羟基丁酸酯4-羟基丁酸酯电纺溶液，电纺同时，向接收装置上加入载盐酸万古霉素微球，使其包裹于电纺丝中，制备了含6，12，18 g/L盐酸万古霉素微球的静电纺丝支架，分别记为P@VancoMs6、P@VancoMs12、P@VancoMs18。①体外实验：将MC3T3-E1细胞分别接种于3种支架上，检测细胞黏附、增殖，筛选适宜额盐酸万古霉素质量浓度，用于后续实验。将MC3T3-E1细胞接种于P@VancoMs6、P@VancoMs12上，加入成骨诱导培养基，通过Ⅰ型胶原和骨钙蛋白免疫荧光染色评价支架体外成骨性能。将P@VancoMs6、P@VancoMs12分别与金黄色葡萄球菌共培养，采用抑菌圈实验分析复合物的抗菌性能。②将聚3-羟基丁酸酯4-羟基丁酸酯支架与P@VancoMs6、P@VancoMs12分别植入SD大鼠皮下，组织学切片观察支架的生物相容性。
结果与结论：①体外实验：细胞黏附与增殖实验显示，P@VancoMs6、P@VancoMs12对细胞增殖无影响，可用于后续实验。P@VancoMs6、P@VancoMs12可抑制金黄色葡萄球菌的生长，并且P@VancoMs12的抑制作用更明显。免疫荧光染色显示，P@VancoMs6、P@VancoMs12可促进细胞分泌Ⅰ型胶原和骨钙蛋白。②体内实验：皮下植入4 d后，3组植入物均可见炎细胞浸润，但未见坏死灶。③结果表明：含6，12 g/L载盐酸万古霉素微球的静电纺丝支架具有良好的抗菌作用与促成骨作用。
https://orcid.org/0000-0001-5670-6289(叶川)；https://orcid.org/0000-0002-0359-7696(杨勇)
中国组织工程研究杂志出版内容重点：生物材料；骨生物材料；口腔生物材料；纳米材料；缓释材料；材料相容性；组织工程

关键词: 静电纺丝, 生物材料, 支架, 盐酸万古霉素, 微球, 感染, 海藻酸钠, 骨

Abstract: BACKGROUND: Due to poor bone blood supply, intravenous antibiotics administration for treating osteomyelitis, it is difficult to achieve effective drug concentration in the focal area of osteomyelitis, poor treatment effect has been clinical bottlenecks. 
OBJECTIVE: To develop a biodegradable biomaterial implant with local sustained release of vancomycin and osteogenic effect, which can promote local bone tissue repair as much as possible while treating osteomyelitis.
METHODS: Vancomycin sodium alginate solutions with concentrations of 6, 12 and 18 g/L were prepared into microspheres by electrospraying; and then the microspheres were vacuum freeze-dried. Poly3-hydroxybutyrate 4-hydroxybutyrate was prepared into electrospinning solution. While electrospinning, vancomycin microspheres were added to the receiving device to wrap them in nanofibers. The electrospinning composites of vancomycin-loaded microspheres with concentrations of 6, 12, and 18 g/L were prepared respectively (denoted as P@VancoMs6, P@VancoMs12, P@VancoMs18). (1) In vitro test: MC3T3-E1 cells were seeded on three kinds of scaffolds, and cell adhesion and proliferation were detected, and the appropriate concentration of vancomycin hydrochloride was screened for subsequent experiments. MC3T3-E1 cells were seeded on P@VancoMs6 and P@VancoMs12, and osteogenic induction medium was added. The in vitro osteogenic properties of the scaffolds were evaluated by immunofluorescence staining of type I collagen and osteocalcin. P@VancoMs6 and P@VancoMs12 were co-cultured with Staphylococcus aureus, respectively, and the antibacterial properties of the complexes were analyzed by the inhibition zone test. (2) The poly-3-hydroxybutyrate 4-hydroxybutyrate scaffolds, P@VancoMs6 and P@VancoMs12 were implanted subcutaneously in SD rats separately, and the biocompatibility of the scaffolds was observed by histological sections.
RESULTS AND CONCLUSION: (1) In vitro experiments: cell adhesion and proliferation experiments showed that P@VancoMs6 and P@VancoMs12 had no effect on cell proliferation and could be used in subsequent experiments. P@VancoMs6 and P@VancoMs12 could inhibit the growth of Staphylococcus aureus, and the inhibitory effect of P@VancoMs12 was more obvious. Immunofluorescence staining showed that P@VancoMs6 and P@VancoMs12 could promote cells to secrete type I collagen and osteocalcin. (2) In vivo experiment: After 4 days of subcutaneous implantation, inflammatory cell infiltration was observed in the three groups of implants, but no necrosis was found. (3) The results showed that the electrospinning scaffolds containing 6 and 12 g/L vancomycin hydrochloride-loaded microspheres played good antibacterial and osteogenic effects.

Key words: electrospinning, biomaterial, scaffold, vancomycin, microsphere, infection, sodium alginate, bone

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