Day Anna, Alkhalil Abdulnaser, Carney Bonnie C, Hoffman Hilary N, Moffatt Lauren T, Shupp Jeffrey W
Anna Day, BS, is a Research Associate; Abdulnaser Alkhalil, PhD, is a Senior Research Scientist; Bonnie C. Carney, BS, is a Research Associate; Hilary N. Hoffman, BS, is a Research Associate; Lauren T. Moffatt, PhD, is the Laboratory Director; and Jeffrey W. Shupp, MD, is the Director, Firefighters' Burn and Surgical Research Laboratory, MedStar Health Research Institute, Washington, DC.
Adv Skin Wound Care. 2017 Dec;30(12):543-551. doi: 10.1097/01.ASW.0000526607.80113.66.
The aims of this study were to assess the effectiveness of a hypochlorous acid-based wound cleanser (Vashe Wound Solution [VWS], SteadMed Medical, Fort Worth, Texas) in disrupting methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa biofilms relative to other cleansers using an in vitro collagen biofilm model and to evaluate cleansers' cytotoxicity. The bioburden reduction of venous stasis wounds by VWS and another cleanser was evaluated.
Plates coated with collagen films incubated with active bacteria cultures to yield biofilm mimics were treated with VWS, 1% and 10% povidone-iodine (PI), 0.05% chlorhexidine wound solution (CWS), or normal saline for 3 or 10 minutes. Biofilms were then analyzed for biomass density using a crystal violet assay, quantitative cultures, and fluorescent microscopy. Cytotoxicity was measured using neutral red uptake by primary human dermal fibroblasts. Pre- and postcleansing exudates and swab samples obtained from venous stasis wounds of patients were processed and plated on a series of selective agar plates for bacteria typing and quantification.
All agents tested significantly neutralized methicillin-resistant S aureus and P aeruginosa biofilms compared with saline control as assessed by crystal violet assay and fluorescent microscopy assays. Undiluted VWS was significantly less cytotoxic compared with 1% PI, CWS, and 10% PI (in increasing order of cytotoxicity). There was no significant difference in bacterial reduction in wounds after treatment with VWS or CWS for any type of bacteria examined using selective media. In wounds that were treated with VWS or CWS, there was a similar percentage reduction in bacterial colony-forming units from precleansing levels when plated on tryptic soy agar, MacConkey, streptococcal, and mannitol salt agar plates. Plates treated with CWS trended toward higher bacterial reduction on nonselective and gram-negative agars, whereas VWS trended toward higher bacterial reduction in Streptococcus-selective agars.
These findings support the use of VWS in the treatment of wounds with biofilms and to reduce the bioburden of venous stasis ulcers. While VWS-treated biofilms had higher biomass than CWS- and saline-treated biofilms, most of the cellular component was not viable. Ultimately, VWS had a similar effectiveness to CWS in eliminating bacteria but with lower cytotoxicity.
本研究旨在使用体外胶原蛋白生物膜模型，评估一种基于次氯酸的伤口清洁剂（Vashe伤口溶液[VWS]，SteadMed Medical，得克萨斯州沃思堡）相对于其他清洁剂破坏耐甲氧西林金黄色葡萄球菌和铜绿假单胞菌生物膜的有效性，并评估清洁剂的细胞毒性。评估了VWS和另一种清洁剂对静脉淤滞性伤口生物负荷的降低情况。
用活性细菌培养物孵育涂有胶原蛋白膜的平板以产生生物膜模拟物，然后用VWS、1%和10%聚维酮碘（PI）、0.05%氯己定伤口溶液（CWS）或生理盐水处理3或10分钟。然后使用结晶紫测定法、定量培养和荧光显微镜分析生物膜的生物量密度。使用原代人皮肤成纤维细胞摄取中性红来测量细胞毒性。对从患者静脉淤滞性伤口获得的清洁前和清洁后渗出液及拭子样本进行处理，并接种在一系列选择性琼脂平板上进行细菌分型和定量。
通过结晶紫测定法和荧光显微镜测定法评估，与生理盐水对照相比，所有测试试剂均能显著中和耐甲氧西林金黄色葡萄球菌和铜绿假单胞菌生物膜。与1% PI、CWS和10% PI（细胞毒性由低到高）相比，未稀释的VWS细胞毒性显著更低。使用选择性培养基检查的任何类型细菌，用VWS或CWS处理后伤口中的细菌减少情况均无显著差异。在用VWS或CWS处理的伤口中，接种在胰蛋白胨大豆琼脂、麦康凯、链球菌和甘露醇盐琼脂平板上时，细菌菌落形成单位相对于清洁前水平的减少百分比相似。用CWS处理的平板在非选择性和革兰氏阴性琼脂上细菌减少趋势更高，而VWS在链球菌选择性琼脂上细菌减少趋势更高。
这些发现支持使用VWS治疗有生物膜的伤口并降低静脉淤滞性溃疡的生物负荷。虽然经VWS处理的生物膜比经CWS和生理盐水处理的生物膜具有更高的生物量，但大多数细胞成分无活力。最终，VWS在消除细菌方面与CWS具有相似的有效性，但细胞毒性更低。
