There is a clinical need for improved wound treatments that prevent both infection and excessive inflammation. TCP-25, a thrombin-derived peptide, is antibacterial and scavenges pathogen-associated molecular patterns (PAMPs), such as lipopolysaccharide, thereby preventing CD14 interaction and Toll-like receptor dimerization, leading to reduced downstream immune activation. Here, we describe the development of a hydrogel formulation that was functionalized with TCP-25 to target bacteria and associated PAMP-induced inflammation. In vitro studies determined the polymer prerequisites for such TCP-25-mediated dual action, favoring the use of noncharged hydrophilic hydrogels, which enabled peptide conformational changes and LPS binding. The TCP-25-functionalized hydrogels killed Gram-positive Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa bacteria in vitro, as well as in experimental mouse models of subcutaneous infection. The TCP-25 hydrogel also mediated reduction of LPS-induced local inflammatory responses, as demonstrated by analysis of local cytokine production and in vivo bioimaging using nuclear factor κB (NF-κB) reporter mice. In porcine partial thickness wound models, TCP-25 prevented infection with S. aureus and reduced concentrations of proinflammatory cytokines. Proteolytic fragmentation of TCP-25 in vitro yielded a series of bioactive TCP fragments that were identical or similar to those present in wounds in vivo. Together, the results demonstrate the therapeutic potential of TCP-25 hydrogel, a wound treatment based on the body's peptide defense, for prevention of both bacterial infection and the accompanying inflammation.

中文翻译：

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含有双作用肽的水凝胶靶向伤口感染和炎症。

临床上需要改善的伤口治疗，以防止感染和过度炎症。TCP-25是一种凝血酶衍生肽，具有抗菌作用，可清除病原体相关的分子模式（PAMP），例如脂多糖，从而防止CD14相互作用和Toll样受体二聚化，从而导致下游免疫活化降低。在这里，我们描述了一种水凝胶制剂的开发过程，该制剂可通过TCP-25进行功能化，以靶向细菌和相关的PAMP诱导的炎症。体外研究确定了此类TCP-25介导的双重作用的聚合物先决条件，这有利于使用不带电荷的亲水性水凝胶，该凝胶可实现肽的构象变化和LPS结合。TCP-25功能化的水凝胶可在体外以及皮下感染的实验小鼠模型中杀死革兰氏阳性金黄色葡萄球菌和革兰氏阴性铜绿假单胞菌细菌。TCP-25水凝胶还介导了LPS诱导的局部炎症反应的减少，如通过使用核因子κB（NF-κB）报告基因小鼠对局部细胞因子产生和体内生物成像的分析所证实的。在猪的部分厚度伤口模型中，TCP-25预防了金黄色葡萄球菌的感染并降低了促炎细胞因子的浓度。体外TCP-25的蛋白水解片段化产生了一系列生物活性TCP片段，这些片段与体内伤口中存在的片段相同或相似。这些结果共同证明了TCP-25水凝胶的治疗潜力，这是一种基于人体肽段防御能力的伤口治疗，