Electromechanical Imaging of Live Vascular Smooth Muscle Cells

活血管平滑肌细胞的机电成像

• 批准号: 7777889
• 负责人: Alexey A Vertegel
• 金额: $ 18.3万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2012-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7777889
• 关键词: Address; Atherosclerosis; Atomic Force Microscopy; Behavior; Biological; Biological Models; Biological Process; Biosensing Techniques; Blood Vessels; Calcified; Cell Culture Techniques; Cells; Collagen Fibril; Computer software; Coupling; Culture Media; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Electrolytes; Electron Beam; Elements; Environment; Functional disorder; Future; Goals; Heart; Image; Individual; Injury; Investigation; Ion Channel; Length; Life; Liquid substance; Maps; Mechanics; Medical; Methodology; Methods; Microfilaments; Microtubules; Modeling; Molecular; Motor; Muscle Cells; Patch-Clamp Techniques; Pathologic Processes; Phenotype; Physiological; Play; Process; Property; Proteins; Protons; Reaction; Resolution; Role; Scanning Probe Microscopy; Smooth Muscle Myocytes; Solutions; Stimulus; Structure; System; Techniques; Testing; Tissue Engineering; Tissues; Vision; aqueous; base; biological systems; cell behavior; cell type; design; electric field; instrument; laminin-5; member; molecular shape; nanoscale; novel diagnostics; novel therapeutic intervention; operation; prevent; public health relevance; response; restenosis; submicron; success; tool; voltage

DESCRIPTION (provided by applicant): Coupling between electrical and mechanical phenomena is a universal feature of all biological systems. Yet, not much is known about origins of biological electromechanical phenomena on the cellular and subcellular scale. Understanding the underlying molecular mechanisms may have tremendous impact on general understanding of biological processes and specific biomedical applications. Electromechanical stimulation of cells can become a valuable tool for their characterization and can eventually result in the development of novel therapeutic interventions. Insufficient information about electromechanical phenomena in biological systems is a result of the lack of characterization techniques capable of providing such information on the nanometer scale and capable of operation in liquid environment. Recently, piezoresponse force microscopy (PFM) has demonstrated potential for imaging structure of connective and calcified tissues with sub-10 nm resolution. Members of this team have also demonstrated high resolution piezoresponse imaging of model systems in aqueous solutions. The ability to map electromechanical properties in aqueous media opens the way to characterization of biological systems in native-like conditions. Here we propose to expand PFM for characterization and stimulation of live cells in physiological environment. Our long-term vision is to use electromechanical imaging as a diagnostic tool and ultimately, utilize electromechanical stimulation for induction of a desirable change in cell behavior. More specifically, we will focus on electromechanical properties of vascular smooth muscle cells as a model system. To accomplish the goals of this project, we will use electron-beam induced etching to fabricate shielded probes needed for PFM imaging in electrolyte solutions with physiological concentrations. We will perform PFM imaging of live VSMCs in aqueous media and compare piezoresponce of their synthetic and contractile phenotypes. As a result of this project we expect to develop experimental technique needed for investigation of local electromechanical phenomena in live cells and create a methodology for predicting cell behavior upon electromechanical stimulation. Public Health Relevance Statement: Electromechanical imaging of live cells can become a unique method to provide information about electrophysiological response of cells and tissues on the nanoscale. It can also be used in the future for diagnostic purposes. Even more broadly, it is envisioned that the use of electromechanical stimulations of live cells can find applications in tissue engineering, medical diagnosis, and biosensing.

描述（由申请人提供）：电气和机械现象之间的耦合是所有生物系统的普遍特征。然而，人们对细胞和亚细胞尺度上生物机电现象的起源知之甚少。了解潜在的分子机制可能会对生物过程和特定生物医学应用的一般理解产生巨大影响。细胞的机电刺激可以成为表征细胞的有价值的工具，并最终导致新型治疗干预措施的发展。 关于生物系统中机电现象的信息不足是由于缺乏能够提供纳米级信息并能够在液体环境中操作的表征技术的结果。最近，压电响应力显微镜 (PFM) 已展现出以亚 10 nm 分辨率对结缔组织和钙化组织结构进行成像的潜力。该团队的成员还展示了水溶液中模型系统的高分辨率压电响应成像。绘制水介质中机电特性的能力为在类似天然条件下表征生物系统开辟了道路。 在这里，我们建议扩展 PFM 以表征和刺激生理环境中的活细胞。我们的长期愿景是使用机电成像作为诊断工具，并最终利用机电刺激来诱导细胞行为发生所需的变化。更具体地说，我们将重点关注作为模型系统的血管平滑肌细胞的机电特性。 为了实现该项目的目标，我们将使用电子束诱导蚀刻来制造在生理浓度的电解质溶液中进行 PFM 成像所需的屏蔽探针。我们将对水介质中的活 VSMC 进行 PFM 成像，并比较其合成表型和收缩表型的压电响应。作为该项目的结果，我们希望开发研究活细胞中局部机电现象所需的实验技术，并创建一种预测机电刺激下细胞行为的方法。 公共健康相关性声明：活细胞机电成像可以成为提供有关纳米级细胞和组织电生理反应信息的独特方法。它也可以在未来用于诊断目的。更广泛地说，预计活细胞机电刺激的使用可以在组织工程、医学诊断和生物传感中找到应用。