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疗问题。在伤口上定植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耐药性菌株的快速进化，这进一步使治疗复杂化。伤口感染对卫生资源和成本有重大影响。在伤口上定植的细菌形成粘附的多细胞群落，称为生物膜。生物膜中的细胞产生细胞外聚合物基质，其将细胞聚集在一起形成团块，并将细菌团块牢固地附着在受伤的组织上。生物膜基质固有的保护性质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经显示出抑制生物膜形成，分离预形成的生物膜，并使预形成的生物膜对体外抗生素和局部杀生物剂的杀伤敏感。我们的中心假设是，基质降解酶将是有用的局部药物用于预防和治疗伤口感染。本发明的目的是测量两种生物膜基质降解酶，DispersinB（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局部杀生物剂根除非创伤皮肤细菌定植的能力？(2)基质降解酶是否能增强局部杀生物剂对新鲜污染的全层伤口的去污能力？(3)基质降解酶是否能增强脉冲冲洗治疗对新鲜污染的全层伤口的去污能力？将使用猪皮肤定植和伤口污染模型进行研究。所有实验将使用常见的伤口病原体金黄色葡萄球菌进行。局部杀菌剂包括聚维酮碘和葡萄糖酸氯己定。皮肤定植模型的主要结果为CFU/cm 2，伤口污染模型的主要结果为CFU/伤口。预计拟议的实验将导致开发基于酶的局部伤口护理制剂，用于人体临床试验。使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常规抗生素的需求。细菌感染是伤口损伤的严重并发症。研究表明，在感染伤口中生长的细菌被一层粘性物质包围，称为粘液层。在这个项目中，我们计划测试粘液降解酶是否可以用于治疗和预防猪的伤口感染。如果成功的话，这些粘液降解酶将被掺入局部软膏中，可用于治疗和预防人们的伤口感染。 公共卫生相关性：细菌感染是伤口损伤的严重并发症。研究表明，在感染伤口中生长的细菌被一层粘性物质包围，称为粘液层。在这个项目中，我们计划测试粘液降解酶是否可以用于治疗和预防猪的伤口感染。如果成功的话，这些粘液降解酶将被掺入局部软膏中，可用于治疗和预防人们的伤口感染。

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. Project Narrative Bacterial infections are a serious complication of wound injuries. Studies have shown that bacteria growing in infected wounds are surrounded by a layer of sticky material that is referred to as the slime layer. In this project we plan to test whether slime-degrading enzymes can be used to treat and prevent wound infections in pigs. If successful, these slime-degrading enzymes will be incorporated into a topical ointment that can be used to treat and prevent wound infections in people. PUBLIC HEALTH RELEVANCE: Bacterial infections are a serious complication of wound injuries. Studies have shown that bacteria growing in infected wounds are surrounded by a layer of sticky material that is referred to as the slime layer. In this project we plan to test whether slime-degrading enzymes can be used to treat and prevent wound infections in pigs. If successful, these slime-degrading enzymes will be incorporated into a topical ointment that can be used to treat and prevent wound infections in people.