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建立一种由金黄色葡萄球菌引起的皮肤和软组织感染的猪模型,包括适用于测试局部抗菌剂的耐甲氧西林菌株。

Development of a porcine model of skin and soft-tissue infection caused by Staphylococcus aureus, including methicillin-resistant strains suitable for testing topical antimicrobial agents.

作者信息

Raška Filip, Lipový Břetislav, Kobzová Šárka, Vacek Lukáš, Jarošová Rea, Kleknerová Dominika, Matiašková Katarína, Makovický Peter, Vícenová Monika, Jeklová Edita, Pantůček Roman, Faldyna Martin, Janda Lubomír

机构信息

Department of Burns and Plastic Surgery, Faculty of Medicine, Institution Shared with University Hospital Brno, Masaryk University, Brno, Czech Republic.

Department of Burns Medicine, Third Faculty of Medicine, Charles University and University Hospital Kralovske Vinohrady, Vinohrady, Prague, Czech Republic.

出版信息

Animal Model Exp Med. 2025 Mar;8(3):544-557. doi: 10.1002/ame2.12495. Epub 2024 Oct 31.

Abstract

BACKGROUND

In view of the ever-increasing representation of Staphylococcus spp. strains resistant to various antibiotics, the development of in vivo models for evaluation of novel antimicrobials is of utmost importance.

METHODS

In this article, we describe the development of a fully immunocompetent porcine model of extensive skin and soft tissue damage suitable for testing topical antimicrobial agents that matches the real clinical situation. The model was developed in three consecutive stages with protocols for each stage amended based on the results of the previous one.

RESULTS

In the final model, 10 excisions of the skin and underlying soft tissue were created in each pig under general anesthesia, with additional incisions to the fascia performed at the base of the defects and immediately inoculated with Staphylococcus aureus suspension. One pig was not inoculated and used as the negative control. Subsequently, the bandages were changed on Days 4, 8, 11, and 15. At these time points, a filter paper imprint technique (FPIT) was made from each wound for semi-quantitative microbiological evaluation. Tissue samples from the base of the wound together with the adjacent intact tissue of three randomly selected defects of each pig were taken for microbiological, histopathological, and molecular-biological examination. The infection with the inoculated S. aureus strains was sufficient during the whole experiment as confirmed by both FPIT and from tissue samples. The dynamics of the inflammatory markers and clinical signs of infection are also described.

CONCLUSIONS

A successfully developed porcine model is suitable for in vivo testing of novel short-acting topical antimicrobial agents.

摘要

背景

鉴于对各种抗生素耐药的葡萄球菌属菌株的比例不断增加,开发用于评估新型抗菌药物的体内模型至关重要。

方法

在本文中,我们描述了一种完全具有免疫能力的猪模型的开发,该模型适用于测试与实际临床情况相匹配的局部抗菌剂,可造成广泛的皮肤和软组织损伤。该模型分三个连续阶段开发,每个阶段的方案根据上一阶段的结果进行修改。

结果

在最终模型中,每头猪在全身麻醉下进行10处皮肤及皮下软组织切除,并在缺损底部对筋膜进行额外切开,然后立即接种金黄色葡萄球菌悬液。有一头猪未接种,用作阴性对照。随后,在第4、8、11和15天更换绷带。在这些时间点,从每个伤口采用滤纸印迹技术(FPIT)进行半定量微生物学评估。从每头猪的三个随机选择的缺损伤口底部以及相邻完整组织采集组织样本,进行微生物学、组织病理学和分子生物学检查。通过FPIT和组织样本证实,在整个实验过程中,接种的金黄色葡萄球菌菌株感染充分。还描述了炎症标志物的动态变化和感染的临床症状。

结论

成功开发的猪模型适用于新型短效局部抗菌剂的体内测试。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d0f/11904108/a72d21cb0fbb/AME2-8-544-g006.jpg

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