Probing the Bioeffects of Cavitation at Single-Cell Level

在单细胞水平上探讨空化的生物效应

• 批准号: 8770631
• 负责人: PEI ZHONG
• 金额: $ 7.85万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8770631
• 关键词: Address; Adverse effects; Animal Experiments; Antineoplastic Agents; Binding; Biological; Blast Cell; Blood - brain barrier anatomy; Cell Communication; Cell Culture Techniques; Cell Line; Cell surface; Cells; Chondrocytes; Clinic; Contrast Media; Cytolysis; Cytoskeleton; Development; Devices; Differentiation and Growth; Drug Delivery Systems; Epithelial Cells; Focused Ultrasound Therapy; Foundations; Fractionation; Fracture; Fracture Healing; Future; Goals; Healed; Hela Cells; In Vitro; Individual; Interdisciplinary Study; Investigation; Knowledge; Lasers; Location; Mechanics; Medical; Medical Technology; Membrane; Methods; Microfabrication; Microfluidics; Nature; Orthopedics; Pain management; Pattern; Phase; Pilot Projects; Process; Production; Regenerative Medicine; Research; Research Personnel; Resolution; Shock; Site; Sonication; Stem cells; Surface; System; Techniques; Technology; Therapeutic; Therapeutic Effect; Tissue Engineering; Tissues; Translational Research; Traumatic Brain Injury; Treatment Protocols; Ultrasonic Transducer; Ultrasonics; Ultrasonography; Validation; base; cancer therapy; cell type; clinical application; clinically relevant; design; follow-up; healing; improved; in vivo; injury and repair; insight; new technology; novel; public health relevance; soft tissue; treatment strategy

DESCRIPTION (provided by applicant): Because of their non-invasiveness and non-ionization nature, therapeutic ultrasound and focused shock waves have been used extensively in clinic for the treatment of cancers, targeted drug delivery (such as opening of blood-brain-barriers), disintegration of concretions, fractionation of soft tissues, and healing of non-union fractures, a well as pain therapy. Although in all these applications cavitation has often been implicated as a key mechanism whereby the desirable therapeutic effects and, in some cases, the undesirable adverse effects are produced, the dynamic processes of bubble(s)-tissue interaction and, in particular, bubble(s)-cell interaction, are largely unknown. This is primarily due to the lack of viable experimental systems that can be used to investigate such dynamic interactions with sufficient spatial and temporal resolutions. To overcome this primary challenge, we propose to develop a new experimental system and associated technologies to investigate the bioeffects produced by cavitation bubbles at the single cell level. This project will be carried out by a multidisciplinary research team with recognized expertise in therapeutic ultrasound and cavitation (Zhong Lab) and cell mechanics (Guilak Lab). Two specific aims are proposed: 1) development of a microfluidic based system that allows for precise control of the location and orientation of individual cells grown in each channel, as well as their spatial alignment with laser- and ultrasound-generated cavitation bubbles, 2) investigation of the bioeffects (cell lysis, membrane poration, Ca2+ influx, cytoskeleton re-arrangement, injury repair, viability, and proliferation) and mechanical deformation of the cell produced by the bubble(s)-cell interaction using representative cell lines (i.e., HeLa cells, MDCK epithelial cells, and chondrocytes) relevant to therapeutic ultrasound and shock wave therapy. With a better understanding of the bubble(s)-cell interaction and the resultant mechanical and biological consequences elicited in different cell types, we hope to gain insights that may be used in the future to improve the design of therapeutic ultrasound and shock wave devices, as well as treatment protocols for safe and more effective medical applications. Furthermore, the new knowledge acquired from this project may be used to develop novel ultrasonic techniques that can be applied to stimulate stem cells under precise mechanical loading conditions to impact their differentiation, growth and phenotypic expression with tissue engineering and regenerative medicine applications. The goal of this R03 application is to demonstrate the feasibility in constructing such a novel experimental system and developing associated technologies for assessing the bioeffects produced by cavitation bubbles at the single-cell level, while the comprehensive utilization of this novel system will be exploited in a follow-up RO1 application.

描述（由申请人提供）：由于其非侵入性和非电离性质，治疗超声和聚焦冲击波已广泛应用于临床治疗癌症、靶向药物输送（如打开血脑屏障） ）、结石崩解、软组织分割、不愈合骨折愈合以及疼痛治疗。尽管在所有这些应用中，空化通常被认为是产生理想治疗效果以及在某些情况下产生不良副作用的关键机制，但气泡与组织相互作用的动态过程，特别是气泡（ s)-细胞相互作用，很大程度上是未知的。这主要是由于缺乏可行的实验系统来研究具有足够空间和时间分辨率的动态相互作用。为了克服这一主要挑战，我们建议开发一种新的实验系统和相关技术来研究单细胞水平上空化气泡产生的生物效应。该项目将由多学科研究团队进行，该团队在治疗性超声和空化（Zhong 实验室）以及细胞力学（Guilak 实验室）方面拥有公认的专业知识。提出了两个具体目标：1）开发基于微流体的系统，可以精确控制每个通道中生长的单个细胞的位置和方向，以及它们与激光和超声产生的空化气泡的空间对准，2）生物效应研究（细胞裂解， 使用代表性细胞系（即 HeLa 细胞、MDCK 上皮细胞和软骨细胞）与治疗性超声和冲击波疗法相关。通过更好地了解气泡与细胞的相互作用以及在不同细胞类型中引起的机械和生物后果，我们希望获得可在未来用于改进治疗性超声和冲击波设备的设计的见解，以及安全和更有效的医疗应用的治疗方案。此外，从该项目中获得的新知识可用于开发新型超声波技术，该技术可用于在精确的机械负载条件下刺激干细胞，以通过组织工程和再生医学应用影响其分化、生长和表型表达。 R03应用的目标是证明构建这样一个新颖的实验系统和开发相关技术以评估单细胞水平空化气泡产生的生物效应的可行性，同时该新颖系统的综合利用将在以下领域得到开发：后续RO1申请。