摘要
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。
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-E1cells 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.
作者
杨勇
许顺恩
古先扬
华大炜
滕建祥
王桢
叶川
Yang Yong;Xu Shunen;Gu Xianyang;Hua Dawei;Teng Jianxiang;Wang Zhen;Ye Chuan(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)
出处
《中国组织工程研究》
CAS
北大核心
2023年第16期2534-2541,共8页
Chinese Journal of Tissue Engineering Research
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
electrospinning
biomaterial
scaffold
vancomycin
microsphere
infection
sodium alginate
bone
