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基于光学相干断层扫描技术检测耐甲氧西林金黄色葡萄球菌(MRSA)形成的骨科植入物生物膜。

Optical coherence tomography-based detection of orthopaedic implant biofilms formed by methicillin-resistant S.aureus (MRSA).

作者信息

Demidov V, Demidova N, Hazem D, Craig P L, Gunn J R, Jackson O P, Gitajn I L, Vitkin I A, Elliott J T

机构信息

Dept. of Orthopaedics, Dartmouth Health, 1 Medical Center Dr, Lebanon, NH USA 03766.

Geisel School of Medicine at Dartmouth, 1 Rope Ferry Rd, Hanover, NH USA 03755.

出版信息

Proc SPIE Int Soc Opt Eng. 2024 Jan-Feb;12822. doi: 10.1117/12.3001786. Epub 2024 Mar 12.

Abstract

Orthopaedic implant-associated infections cause serious complications primarily attributed to bacterial biofilm formation and often characterized by increased antibiotic resistance and diminished treatment response. There is currently a lack of imaging modalities that can directly visualize biofilms to determine the location and extent of contamination. Optical coherence tomography (OCT) is a portable, non-invasive, high-resolution imaging modality with the potential to fulfill this unmet need. In this study, we aim to evaluate the efficacy of OCT in detecting biofilms formed by life- and limb-threatening bacteria on orthopaedic implants. Bioluminescent strain SAP231 of methicillin-resistant S.aureus (MRSA) was used to grow biofilms on the surfaces of titanium and stainless-steel orthopaedic hardware situated inside custom-designed macrofluidic devices, allowing continuous nutrient broth supply and waste removal. Three-dimensional OCT images of each piece of hardware were obtained every 24 hours with subsequent bioluminescence imaging using the PerkinElmer IVIS Spectrum. OCT texture analysis based on multi-parametric fitting approach was developed and validated against IVIS quantification for accurate identification of live MRSA signatures. The monitoring of biofilm formation and measurement of film thicknesses starting at 12 micrometers and reaching 180 micrometers in 72 hours on metal hardware is demonstrated. This proof-of-concept study highlights the ability of OCT to detect and quantify the formation of MRSA bacterial biofilms in a high fidelity orthopaedic implant biofilm model , opening avenues for translation of this technique to preclinical models of contaminated orthopaedic trauma surgery and further clinical translation.

摘要

骨科植入物相关感染会引发严重并发症,主要归因于细菌生物膜的形成,其特征通常是抗生素耐药性增加且治疗反应减弱。目前缺乏能够直接可视化生物膜以确定污染位置和范围的成像方式。光学相干断层扫描(OCT)是一种便携式、非侵入性的高分辨率成像方式,有潜力满足这一未被满足的需求。在本研究中,我们旨在评估OCT检测由对生命和肢体构成威胁的细菌在骨科植入物上形成的生物膜的功效。耐甲氧西林金黄色葡萄球菌(MRSA)的生物发光菌株SAP231被用于在定制设计的宏观流体装置内的钛和不锈钢骨科硬件表面生长生物膜,该装置可实现营养液的持续供应和废物清除。每24小时获取每件硬件的三维OCT图像,随后使用珀金埃尔默IVIS Spectrum进行生物发光成像。基于多参数拟合方法开发了OCT纹理分析,并针对IVIS定量进行了验证,以准确识别活的MRSA特征。结果表明,在金属硬件上,生物膜形成的监测以及膜厚度的测量在72小时内从12微米开始并达到180微米。这项概念验证研究突出了OCT在高保真骨科植入物生物膜模型中检测和量化MRSA细菌生物膜形成的能力,为将该技术转化为受污染骨科创伤手术的临床前模型以及进一步的临床转化开辟了道路。

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