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血管内动脉瘤修复 (EVAR) 移植物上的生物膜形成——体外模型的概念验证。

Biofilm formation on endovascular aneurysm repair (EVAR) grafts-a proof of concept in vitro model.

机构信息

Department of Clinical Microbiology, Rigshospitalet Copenhagen University Hospital, Copenhagen, Denmark; Division for Infection Medicine, Department for Clinical Sciences Lund, Faculty of Medicine, Lund University, Lund, Sweden; Clinical Microbiology and Infection Control, Office for Medical Services, Region Skåne, Lund, Sweden.

Department of Clinical Microbiology, Rigshospitalet Copenhagen University Hospital, Copenhagen, Denmark.

出版信息

Clin Microbiol Infect. 2023 Dec;29(12):1600.e1-1600.e6. doi: 10.1016/j.cmi.2023.09.012. Epub 2023 Sep 20.

Abstract

OBJECTIVES

An endovascular aneurysm repair (EVAR) graft is a catheter-implanted vascular prosthesis and is the preferred treatment for patients with aortic aneurysm. If an EVAR graft becomes the focus of infection, the treatment possibilities are limited because it is technically difficult to remove the graft to obtain source control. This study examines whether Pseudomonas aeruginosa and Staphylococcus aureus form biofilm on EVAR prostheses.

METHODS

EVAR graft sections were exposed to bacteria at 10 or 10 colony forming units (CFU)/mL in lysogeny broth and Krebs-Ringer at 37°C, bacterial biofilm formation was evaluated by scanning electron microscopy and counting CFU on the graft sections after antibiotic exposure at × 10 minimal inhibitory concentration. Bacteria were tested for tolerance to benzylpenicillin, tobramycin, and ciprofloxacin.

RESULTS

Bacterial exposure for 15 minutes established biofilms on all prosthesis fragments (6/6 replicates). After 4 hours, bacteria were firmly attached to the EVAR prostheses and resisted washing. After 18-24 hours, the median CFU/g of EVAR graft reached 5.2 × 10 (1.15 × 10-1.1 × 10) for S. aureus and 9.1 × 10 (3.5 × 10-6.25 × 10) for P. aeruginosa. Scanning electron microscopy showed bacterial attachment to the graft pieces. There was a time-dependent development of tolerance with approximately 20 (tobramycin), 560 (benzylpenicillin), and 600 (ciprofloxacin) times more S. aureus surviving antibiotic exposure in 24- compared with 0-hour-old biofilm. Five (tobramycin) and 170 times (ciprofloxacin) more P. aeruginosa survived antibiotic exposure in 24- compared with 0-hour-old biofilms.

DISCUSSION

Our results show that bacteria can rapidly adhere to and subsequently form antibiotic-tolerant biofilms on EVAR graft material in concentrations equivalent to levels seen in transient bacteraemia in vivo. Potentially, the system can be used for identifying optimal treatment combinations for infected EVAR prosthesis.

摘要

目的

血管内动脉瘤修复(EVAR)移植物是一种经导管植入的血管假体,是治疗主动脉瘤患者的首选方法。如果 EVAR 移植物成为感染的焦点,那么治疗的可能性就会受到限制,因为从技术上讲,很难移除移植物以获得源控制。本研究检查了铜绿假单胞菌和金黄色葡萄球菌是否在 EVAR 移植物上形成生物膜。

方法

将 EVAR 移植物切片在 10 或 10 个菌落形成单位(CFU)/mL 噬菌体肉汤和 Krebs-Ringer 中于 37°C 下暴露于细菌,通过扫描电子显微镜评估细菌生物膜形成情况,并在抗生素暴露后在移植物切片上计数 CFU × 10 最小抑菌浓度。测试细菌对苯唑西林、妥布霉素和环丙沙星的耐受性。

结果

细菌暴露 15 分钟即可在所有移植物碎片上形成生物膜(6/6 个重复)。4 小时后,细菌牢固地附着在 EVAR 移植物上,并且难以清洗。18-24 小时后,EVAR 移植物的 CFU/g 中位数达到金黄色葡萄球菌 5.2×10(1.15×10-1.1×10)和铜绿假单胞菌 9.1×10(3.5×10-6.25×10)。扫描电子显微镜显示细菌附着在移植物碎片上。随着时间的推移,细菌对生物膜的耐受性逐渐增强,与 0 小时相比,24 小时的生物膜中金黄色葡萄球菌的存活数量增加了约 20(妥布霉素)、560(苯唑西林)和 600 倍(环丙沙星),铜绿假单胞菌的存活数量增加了 5(妥布霉素)和 170 倍(环丙沙星)。

讨论

我们的结果表明,细菌可以在 EVAR 移植物材料上迅速附着,并随后形成抗生素耐药性生物膜,其浓度相当于体内短暂菌血症中观察到的水平。该系统有可能用于确定感染的 EVAR 移植物的最佳治疗组合。

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