Understanding Plasma Assisted Biofilm Inactivation

描述（由申请人提供）：生物膜每年在设备损坏、产品污染、感染以及器械和假体植入物的定植方面花费数十亿美元。传统的微生物根除方法的有效性大大削弱了生物膜，因为它们表现出非典型的特点相比，他们的寄生虫对应物。因此，消灭这些生物体的能力要求开发替代技术。该项目的广泛、长期目标是填补缺乏有效生物膜灭活/灭菌方法的空白。等离子体的使用，即第四种状态，提供了一种新的替代方案，因为等离子体含有单独称为杀菌剂的反应剂的混合物。虽然有证据表明，等离子体是有用的，以消除细菌生物膜，仍然有许多未解决的问题之前，我们可以理解的机制，导致灭活/灭菌，并将其应用于医疗或环境的目的。本研究的主要目的是了解导致气体放电等离子体辅助生物膜灭活的机制。具体目的是：1）确定血浆对生物膜形成细胞的生理状态的影响。我们将检验以下假设：血浆影响生物膜形成细胞的生理状态（活力、可培养性、代谢状态），并且该影响取决于生物膜暴露于血浆的时间量和生物膜的年龄。2)以确定血浆对生物膜形成细胞结构的影响。我们将测试等离子体影响生物膜形成细胞的结构（细胞包膜的完整性和孔的最终存在）的假设，并且该效果取决于生物膜暴露于等离子体的时间量和生物膜的年龄;以及3）将气体-等离子体组成与生物膜灭活/灭菌相关联。我们将确定细胞对血浆的反应是一种普遍现象，还是取决于所用血浆的类型。我们将选择具有特定多样性或丰富的反应剂的等离子体环境，并将使用它来评估其对生物膜形成细胞结构和生理状态的影响。这项研究将提供一个全面的了解等离子体辅助生物膜灭活及其机制，实现NIH的目标#3（到。扩大医学和相关科学的知识基础，以提高国家的经济福祉，并确保公共研究投资持续获得高回报）。通过实现这一主要目标，我们希望为未来的技术发展奠定基础，作为一个长期目标，解决一个严重的医疗问题，履行NIH的使命（追求有关生命系统的性质和行为的基本知识的科学，以及这些知识的应用，以延长健康的生命，减少疾病和残疾的负担）;特别是目标1（促进基本的创造性发现、创新性研究战略及其应用，以此为基础，大大提高国家保护和改善健康的能力）。 公共卫生相关性：生物膜污染常见的家庭表面和许多医疗器械（例如，导管、隐形眼镜、机械心脏瓣膜、关节置换等），导致疾病和死亡。破坏生物膜所需的大剂量抗菌剂是环境不期望的，并且可能对患者有害。因此，要有能力消灭这些生物体，就需要开发替代技术。等离子体，物质的第四种状态，提供了一个有吸引力的选择。本研究将研究等离子体辅助生物膜灭活的机理，长期目标是开发一种易于使用，无危害的灭菌方法来破坏生物膜。

DESCRIPTION (provided by applicant): Biofilms cost this nation billions of dollars yearly in equipment damage, product contamination, infections, and colonization of devices and prosthetic implants. The effectiveness of conventional methods of microbial eradication is drastically impaired with biofilms since they exhibit atypical characteristics when compared to their planktonic counterparts. Therefore the ability to destroy these organisms demands the development of alternative techniques. The broad, long term goal of this project is to fill a gap regarding the lack of effective biofilm inactivation/sterilization methods. The use of plasmas, the fourth state of the matter, offers a novel alternative since plasmas contain a mixture of reactive agents individually known as bacteria- killing agents. Although evidence suggests that plasma is useful to inactivate bacterial biofilms, there are still many unresolved questions before we can understand the mechanism that causes inactivation/sterilization and apply it for medical or environmental purposes. The main goal of this research is to understand the mechanisms leading to gas discharge plasma-assisted biofilm inactivation. The specific aims are: 1) to determine the effect of plasma on the physiological status of biofilms-forming cells. We will test the hypothesis that plasma affects the physiological status (viability, culturability, metabolic status) of biofilm- forming cells and the effect depends on the amount of time that biofilms have been exposed to plasma and the age of the biofilm. 2) to determine the effect of plasma on the structure biofilm-forming cells. We will test he hypothesis that plasma affects the structure (integrity of cell envelopes and eventual presence of pores) of biofilm-forming cells, and the effect depends on the amount of time that biofilms have been exposed to plasma and the age of the biofilm; and 3) to relate gas-plasma composition with biofilm inactivation/sterilization. We will determine whether the response of cells to plasma is a general phenomenon or if it depends on the type of plasma used. We will select a plasma environment with a particular diversity or abundance of reactive agents and we will use it to evaluate its effect on biofilm-forming cells structure and physiological status. This study will provide a comprehensive understanding of plasma-assisted-biofilm inactivation and its mechanisms, fulfilling NIH's goal #3 (to. expand the knowledge base in medical and associated sciences in order to enhance the Nation's economic well-being and ensure a continued high return on the public investment in research). By achieving this main goal we expect to build the basis for the future development of a technology that will, as a long-term goal, solve a serious medical problem, fulfilling NIH's mission (Science in pursuit of fundamental knowledge about the nature and behavior of living systems and the application of that knowledge to extend healthy life and reduce the burdens of illness and disability); and particularly goal # 1 (to foster fundamental creative discoveries , innovative research strategies, and their applications as a basis to advance significantly the Nation's capacity to protect and improve health). PUBLIC HEALTH RELEVANCE: Biofilms contaminate common household surfaces and many medical devices, (e.g., catheters, contact lenses, mechanical heart valves, joint replacements, etc.), causing illness and death. The huge doses of antimicrobials required to destroy biofilms are environmentally undesirable and may be hazardous to a patient. Therefore, the ability to destroy these organisms demands the development of alternative techniques. Plasmas, the fourth state of matter, offer an attractive alternative. This research will study the mechanisms of plasma-assisted- biofilm inactivation with the long term goal of developing an easy-to-use, non-hazardous sterilization method to destroy biofilms.