Sporocidal Mechanism and Inline Monitoring of Peracetic Acid in Supercritical CO2

 描述（由申请人提供）：由于医院获得性感染（HAI）中抗生素耐药性的快速上升，对生物医学器械、异种移植物和同种异体移植物材料进行灭菌至关重要。对于一种主要的HAI病原体艰难梭菌，抗生素的使用促进了更高的感染率和更差的结果，因为天然肠道植物群被所使用的强效抗生素消除（Songer和安德森，2006）。C.艰难梭菌、金黄色葡萄球菌和铜绿假单胞菌是导致2002年超过100，000人死亡的估计的200万医院感染中的许多感染的原因，并且估计每二十名医院患者中就有一名将获得HAI（www.cdc.gov/HAI/burden.html，www.cdc.gov/HAI/pdfs/hai/infections_deaths.pdf）。HAI的估计费用远远超过250 - 350亿美元，而预防费用估计不到70亿美元（www.cdc.gov/HAI/pdfs/hai/Scott_CostPaper.pdf）。在使用任何灭菌系统或抗生素时，需要确定所涉及的分子或因子的杀微生物作用机制。从抗生素耐药性、抗生素促进C.艰难梭菌感染和杀生物剂抗性驱动了对表征新的和长期使用的杀生物剂的杀死机制的需求。了解杀生物剂的关键细胞靶点有助于开发创新、安全的方法来控制或消除HAI的发病率。[For一种新的杀菌方法，被接受用于内部和外部使用的医疗装置以及用于药物，美国食品和药物管理局（FDA）现在要求鉴定杀生物剂作用的机制。NovaSterilis，Inc.，已经完成了两项NIH第二阶段的赠款，从而在市场上推出了一种经验证的用于软组织和硬组织的超临界二氧化碳-过氧乙酸（scCO 2-PAA）灭菌系统，并且正在开发可吸收缝线的灭菌。本提案阐述了FDA对scCO 2中PAA杀灭微生物机制表征的要求。NovaSterilis的商业Nova 2200系统监测超临界CO2形成和维持所需的条件，但目前不监测溶解在scCO 2中的过氧乙酸（和稳定剂过氧化氢）的浓度。[The NovaSterilis第一阶段计划的目标是确定PAA和scCO 2中的氢气对枯草芽孢杆菌（最具抗性的生命形式之一）孢子的杀菌作用机制，并开发和测试在线仪表，以监测系统中PAA的杀菌水平。]这项工作将显著推进FDA监管机构对使用PAA和过氧化氢作为医疗器械和生物制品的共灭菌剂进行scCO 2灭菌的认可，以及使用PAA作为灭菌剂或共灭菌剂的食品和环境系统。

 DESCRIPTION (provided by applicant): It is critical to sterilize biomedical devices, xenograft and allograft materials, due to the rapid rise of antibiotic resistance in hospital-acquired infections (HAI). For one major HAI pathogen, Clostridium difficile, the use of antibiotics promotes greater infection rates and worse outcome, as the native intestinal flora is eliminated by the strong antibiotics used (Songer and Anderson, 2006). C. difficile, Staphylococcus aureus and Pseudomonas aeruginosa are responsible for many of the estimated 2 million nosocomial infections resulting in over 100,000 deaths in 2002 and with the estimate that one in twenty hospital patients will acquire an HAI (www.cdc.gov/HAI/burden.html, www.cdc.gov/HAI/pdfs/hai/infections_deaths.pdf). The estimated costs of HAI are well over $25-35 billion, with prevention costs estimated at below $7 billion (www.cdc.gov/HAI/pdfs/hai/Scott_CostPaper.pdf). With the use of any sterilization system or antibiotic, there is a need to identify the mechanism of microbicidal action of the molecule or factor involved. Lessons learned from antibiotic resistance, antibiotic promotion of C. difficile infection and biocide resistance drive the requirement for characterizing new and long-used biocide's mechanisms of kill. Understanding the critical cellular targets for biocides enables the development of innovative, safe methods to control or eliminate the incidence of HAI. [For a novel sterilization method to be accepted for medical devices for internal and external use and for pharmaceuticals, the U.S. Food and Drug Administration (FDA) now require identification of the mechanism of biocide action.] NovaSterilis, Inc., has completed two NIH Phase II grants, resulting in a validated supercritical Carbon dioxide- peracetic acid (scCO2-PAA) sterilization system on the market for soft and hard tissue and is developing sterilization of absorbable sutures. This proposal addresses the FDA requirement for characterization of the mechanism of microbial kill by PAA in scCO2. NovaSterilis' commercial Nova 2200 system monitors the conditions required for formation and maintenance of supercritical CO2, but does not currently monitor concentrations of peracetic acid (and stabilizer hydrogen peroxide) solubilized in scCO2. [The goals of the NovaSterilis Phase I program are to identify the mechanism of biocidal action of PAA and hydrogen in scCO2 on spores of the bacterium Bacillus subtilis, one of the most resistant forms of life, and to develop and test an inline meter to monitor microbicidal levels of PAA in the system.] This work will significantly advance FDA regulatory acceptance of scCO2 sterilization with PAA and hydrogen peroxide as co-sterilants for medical devices and biologicals, as well as for food and environmental systems utilizing PAA as a sterilant or co- sterilant.