骨科内植物念珠菌生物被膜体外模型的构建及药物敏感性  

Construction and drug sensitivity of an in vitro model of Candida biofilm in orthopedic implants

作　　者：陈美 石海燕 谭丽华 鲁厚根[2] 孙毅[3] CHEN Mei;SHI Hai-Yan;TAN Li-Hua;LU Hou-Gen;SUN Yi(The Second Clinical Medical College,Yangtze University,Jingzhou,Hubei 434020,China;Department of Orthopedics,Jingzhou Central Hospital,Jingzhou,Hubei 434020,China;Department of Dermatology,Jingzhou Central Hospital,Jingzhou,Hubei 434020,China)

机构地区：[1]长江大学第二临床医学院,湖北荆州434020 [2]荆州市中心医院骨科,湖北荆州434020 [3]荆州市中心医院皮肤科,湖北荆州434020

出　　处：《菌物学报》2021年第6期1380-1387,共8页Mycosystema

基　　金：湖北省卫计委科研项目(WJ2018H178);湖北省自然科学基金(2019CFB567)。

摘　　要：利用24孔板在骨科内植物表面构建骨科内植物念珠菌生物被膜模型,并使用荧光镜检和菌落计数法(colony forming unit,CFU)检测醋酸氯己定(以下简称氯己定)与氟康唑对骨科内植物念珠菌生物被膜的联合抗菌效应。本研究成功在体外构建出骨科内植物念珠菌生物被膜模型,并发现无论是白念珠菌(SC5314)、近平滑念珠菌(ATCC22019)还是克柔念珠菌(ATCC00279)氟康唑单药组组间没有显著性差异;氯己定对骨科内植物念珠菌生物被膜的80%最低抑菌浓度(SMIC80)均≥16μg/mL,氟康唑对念珠菌生物被膜的80%最低抑菌浓度(SMIC80)均>64μg/mL。读取SMIC80时,氯己定与氟康唑的协同率高达100%,而两种药物的联合能够使氟康唑和醋酸氯己定的用药剂量减少4-8倍。本研究还发现在体外白念珠菌(SC5314)、近平滑念珠菌(ATCC22019)、克柔念珠菌(ATCC00279)、氟康唑耐药白念珠菌(64550)以及耳念珠菌(0389)均可以在骨科内植物表面形成真菌生物被膜并对氟康唑产生了耐药性;氯己定与抗真菌药物氟康唑对骨科内植物念珠菌生物被膜的杀伤具有明显的协同作用,且明显减少单药用药剂量。An in vitro model of Candida biofilm in orthopedic implants was constructed by using 24-well plates, and the combined antibiotic effects of chlorhexidine acetate(hereinafter ’chlorhexidine’ for short) and fluconazole on the biofilm were detected by fluorescence microscopy and colony forming unit. It was found that there was no significant antibiotic difference against Candida albicans(SC5314), Candida parapsilosis(ATCC22019) or Candida krusei(ATCC00279) under fluconazole monotherapy. The 80% minimum inhibitory concentration(SMIC80) of chlorhexidine against Candida biofilms in orthopedic implants was ≥16μg/mL, and that of fluconazole against Candida biofilms were all >64μg/mL. When SMIC80 was read, the synergism of chlorhexidine and fluconazole was up to 100%, and the combination of the two drugs could reduce the dose of fluconazole and chlorhexidine by 4–8 times. This study also found that in vitro Candida albicans(SC5314), Candida parapsilosis(ATCC22019), Candida krusei(ATCC00279), fluconazole-resistant Candida albicans(64550) and Candida auris(0389) can form fungal biofilms on the surface of orthopedic implants and develop resistance to fluconazole. Chlorhexidine and fluconazole have obvious synergistic effect on killing Candida biofilms in orthopedic implants and significantly reduce the dose of single drug.

关 键 词：念珠菌 生物被膜 骨科内植物 抗真菌药物 氯己定 

分 类 号：R446.5[医药卫生—诊断学]