Biofilm matrix-degrading enzymes for the treatment and prevention of S. aureus-as

描述（由申请人提供）：伤口感染是一个主要的医学问题。定植在伤口上的细菌会延迟伤口愈合，并成为继发感染的焦点。此外，使用抗生素治疗伤口感染导致抗生素耐药菌株的快速进化，这进一步使治疗复杂化。伤口感染对卫生资源和费用有重大影响。细菌在伤口上定植，形成附着的多细胞群落，称为生物膜。生物膜中的细胞产生细胞外聚合基质，将细胞聚集在一起，并将细菌团牢牢地附着在受伤的组织上。生物膜基质固有的保护性使得伤口感染难以治疗。大多数生物膜的细胞外基质含有聚合物，如多糖和DNA。降解生物膜基质聚合物的酶已被证明可以抑制生物膜的形成，分离预形成的生物膜，并使预形成的生物膜对抗生素和局部杀菌剂的体外杀伤敏感。我们的中心假设是，基质降解酶将是有用的局部剂预防和治疗伤口感染。本研究的目的是测定两种生物膜基质降解酶——分散酶b (R)（糖苷水解酶）和Pulmozyme(R) （dna酶I）的体内抗生物膜活性。本研究的目的是回答以下问题：(1)基质降解酶是否增强了局部杀菌剂根除非损伤皮肤细菌定植的能力？(2)基质降解酶是否增强了局部杀菌剂对新污染的全层伤口的去污能力？(3)基质降解酶是否增强了脉冲灌洗治疗对新污染全层创面的去污能力？研究将使用猪皮肤定植和伤口污染模型进行。所有实验将使用常见的伤口病原菌金黄色葡萄球菌进行。局部杀菌剂将包括聚维酮碘和葡萄糖酸氯己定。主要结果将是皮肤定植模型的CFU/cm2和伤口污染模型的CFU/伤口。拟议的实验预计将导致基于酶的局部伤口护理配方的发展，用于人体临床试验。利用酶破坏生物膜基质的物理完整性是一种有吸引力的伤口治疗策略，因为它可以减少或消除对传统抗生素的需求。

DESCRIPTION (provided by applicant): Wound infections are a major medical problem. Bacteria that colonize wounds can delay wound healing and act as a focus for secondary infections. In addition, the use of antibiotics to treat wound infections results in the rapid evolution of antibiotic-resistant strains, which further complicates treatment. Wound infections have a significant impact on health resources and costs. Bacteria that colonize wounds form adherent, multicellular communities known as biofilms. The cells in a biofilm produce an extracellular polymeric matrix that holds the cells together in a mass and firmly attaches the bacterial mass to the wounded tissue. The inherent protective nature of the biofilm matrix makes wound infections difficult to treat. The extracellular matrix of most biofilms contains polymers such as polysaccharides and DNA. Enzymes that degrade biofilm matrix polymers have been shown to inhibit biofilm formation, detach pre-formed biofilms, and sensitize pre-formed biofilms to killing by antibiotics and topical biocides in vitro. Our central hypothesis is that matrix-degrading enzymes will be useful topical agents for the prophylaxis and treatment of wound infections. The objective of the present proposal is to measure the in vivo anti-biofilm activities of two biofilm matrix-degrading enzymes, DispersinB(R) (a glycoside hydrolase) and Pulmozyme(R) (DNase I). The aims of this study are to answer the following questions: (1) Do matrix-degrading enzymes potentiate the ability of topical biocides to eradicate bacterial colonization of non-wounded skin? (2) Do matrix-degrading enzymes potentiate the ability of topical biocides to decontaminate freshly contaminated full-thickness wounds? (3) Do matrix-degrading enzymes potentiate the ability of pulse irrigation treatment to decontaminate freshly contaminated full-thickness wounds? Studies will be carried out using porcine skin colonization and wound contamination models. All experiments will be carried out using the common wound pathogen Staphylococcus aureus. Topical biocides will include povidone iodine and chlorhexidine gluconate. Major outcomes will be CFU/cm2 for skin colonization models and CFU/wound for wound contamination models. The proposed experiments are expected to lead to the development of enzyme-based topical wound care formulations for testing in human clinical trials. The use of enzymes to destroy the physical integrity of the biofilm matrix is an attractive wound treatment strategy because it may reduce or eliminate the need for conventional antibiotics.