Inflammatory and Antimicrobial Responses to Methicillin-Resistant Staphylococcus aureus in an In Vitro Wound Infection Model
Inflammatory and Antimicrobial Responses to Methicillin-Resistant Staphylococcus aureus in an In Vitro Wound Infection Model
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DOI：
10.1371/journal.pone.0082800
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发表时间：
2013-12-10
期刊：
影响因子：
3.7
通讯作者：
Nibbering, Peter H.
中科院分区：
文献类型：
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作者：
Haisma, Elisabeth M.;Rietveld, Marion H.;Nibbering, Peter H.
烧伤创面感染患者的治疗可能会因为抗生素耐药细菌和生物膜的存在而变得复杂。在这里，我们展示了一种使用人类皮肤等效物(HSE)和生物被膜形成甲氧西林耐药金黄色葡萄球菌(MRSA)的体外热伤口感染模型，以测试对抗此类感染的试剂。在HSE上应用液氮冷却金属装置产生了可复制的创面，其特征是角质形成细胞死亡，表皮层从真皮脱落，并重新上皮化。热损伤伴随着角质形成细胞活化、炎症和抗菌反应标志物的上调。热损伤HSE暴露于MRSA后1小时可见大量粘附性MRSA/HSE，24-48小时后细菌大量增殖。与对照组相比，暴露于MRSA的烫伤大鼠的炎症介质如TLR2(但不是TLR3)、IL-6和IL-8以及抗菌蛋白人β-防御素-2、-3和RNAse7的表达增加。此外，局部应用莫匹罗星有效地在24小时内将(热损伤)HSE上的MRSA计数减少了99.9%以上。总之，这些数据表明，这种热伤口感染模型是一种很有前途的工具，可以用来研究伤口定植和感染的初始阶段，以及评估候选抗菌剂的局部效果。
Treatment of patients with burn wound infections may become complicated by the presence of antibiotic resistant bacteria and biofilms. Herein, we demonstrate an in vitro thermal wound infection model using human skin equivalents (HSE) and biofilm-forming methicillin-resistant Staphylococcus aureus (MRSA) for the testing of agents to combat such infections. Application of a liquid nitrogen-cooled metal device on HSE produced reproducible wounds characterized by keratinocyte death, detachment of the epidermal layer from the dermis, and re-epithelialization. Thermal wounding was accompanied by up-regulation of markers for keratinocyte activation, inflammation, and antimicrobial responses. Exposure of thermal wounded HSEs to MRSA resulted in significant numbers of adherent MRSA/HSE after 1 hour, and multiplication of these bacteria over 24-48 hours. Exposure to MRSA enhanced expression of inflammatory mediators such as TLR2 (but not TLR3), IL-6 and IL-8, and antimicrobial proteins human beta-defensin-2, -3 and RNAse7 by thermal wounded as compared to control HSEs. Moreover, locally applied mupirocin effectively reduced MRSA counts on (thermal wounded) HSEs by more than 99.9% within 24 hours. Together, these data indicate that this thermal wound infection model is a promising tool to study the initial phase of wound colonization and infection, and to assess local effects of candidate antimicrobial agents.