基于ε-聚赖氨酸和环糊精构建细菌生物膜抗生素递送系统治疗骨与关节感染的实验研究  

Role of antibiotic delivery system targeting bacterial biofilm based onε-poly-L-lysine and cyclodextrin in treatment of bone and joint infections

作　　者：刘铁鑫 林俊卿 郑宪友[1,2] LIU Tiexin;LIN Junqing;ZHENG Xianyou(Department of Orthopedics,Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai,200233,P.R.China;National Orthopedic Medical Center,Shanghai,200233,P.R.China)

机构地区：[1]上海交通大学医学院附属第六人民医院骨科,上海200233 [2]国家骨科医学中心,上海200233

出　　处：《中国修复重建外科杂志》2025年第3期362-369,共8页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：上海交通大学医工交叉研究基金资助项目(YG2022ZD017)。

摘　　要：目的探讨基于ε-聚赖氨酸(ε-poly-L-lysine,ε-PLL)和环糊精(cyclodextrin,CD)构建的细菌生物膜抗生素[利奈唑胺(linezolid,LZD)]递送系统(ε-PLL-CD-LZD)的作用机制,旨在通过提高抗生素的生物利用度,有效渗透并破坏生物膜结构,从而增强骨与关节感染治疗效果。方法采用化学合成法制备ε-PLL-CDLZD。通过核磁共振检测CD接枝率进行表征测定;与小鼠胚胎成骨细胞前体细胞MC3T3-E1、人脐静脉内皮细胞、小鼠胚胎成纤维细胞3T3-L1共培养后行活/死细胞染色,评估体外细胞相容性;采用金黄色葡萄球菌生物膜,通过富集能力研究评估ε-PLL-CD-LZD在生物膜中的富集作用,最低抑菌浓度(minimum inhibitory concentration,MIC)实验、扫描电镜和活/死细菌染色评估其清除生物膜的效果。构建雄性SD大鼠骨与关节感染模型,验证ε-PLL-CD-LZD抗菌效果。结果ε-PLL-CD-LZD中CD平均接枝率为9.88%,与3种细胞共培养后存活率均达90%以上,在生物膜中表现出较强富集能力,MIC为2 mg/L,扫描电镜观察示能有效破坏生物膜结构,具有高效生物膜清除能力。大鼠体内实验显示ε-PLL-CD-LZD能显著降低感染部位细菌计数及阳性率(P<0.05)。结论ε-PLL-CD通过提高抗生素生物利用度,并渗透、破坏生物膜,从而在动物体内表现出显著的抗感染效果。Objective To explore the mechanism of antibiotic delivery system targeting bacterial biofilm with linezolid(LZD)based onε-poly-L-lysine(ε-PLL)and cyclodextrin(CD)(ε-PLL-CD-LZD),aiming to enhance antibiotic bioavailability,effectively penetrate and disrupt biofilm structures,and thereby improve the treatment of bone and joint infections.Methodsε-PLL-CD-LZD was synthesized via chemical methods.The grafting rate of CD was characterized using nuclear magnetic resonance.In vitro biocompatibility was evaluated through live/dead cell staining after coculturing with mouse embryonic osteoblast precursor cells(MC3T3-E1),human umbilical vein endothelial cells,and mouse embryonic fibroblast cells(3T3-L1).The biofilm-enrichment capacity ofε-PLL-CD-LZD was assessed using Staphylococcus aureus biofilms through enrichment studies.Its biofilm eradication efficacy was investigated via minimum inhibitory concentration(MIC)determination,scanning electron microscopy,and live/dead bacterial staining.A bone and joint infection model in male Sprague-Dawley rats was established to validate the antibacterial effects ofε-PLL-CDLZD.Results Inε-PLL-CD-LZD,the average grafting rate of CD reached 9.88%.The cell viability exceeded 90%after co-culturing with three types cells.The strong biofilm enrichment capability was observed with a MIC of 2 mg/L.Scanning electron microscopy observations revealed the effective disruption of biofilm structure,indicating potent biofilm eradication capacity.In vivo rat experiments demonstrated thatε-PLL-CD-LZD significantly reduced bacterial load and infection positivity rate at the lesion site(P<0.05).Conclusion Theε-PLL-CD antibiotic delivery system provides a treatment strategy for bone and joint infections with high clinical translational significance.By effectively enhancing antibiotic bioavailability,penetrating,and disrupting biofilms,it demonstrated significant anti-infection effects in animal models.

关 键 词：骨和关节感染 Ε-聚赖氨酸 环糊精 抗生素递送 生物膜 大鼠 

分 类 号：R681.2[医药卫生—骨科学] R684.3[医药卫生—外科学]