Electromechanical control of cell adhesion and motility

描述（由申请人提供）：组织工程是一种相对较新的，但 迅速扩大的生物医学工程研究领域。因为蜂窝小区 粘附性和运动性是决定组织完整性的两个关键因素 细胞和分子的功能、阐明和调节 参与细胞粘附和运动的机制是非常重要的 用于组织工程。工程化组织在临床上的应用 在软组织替换或再生中的应用已经取得成功。 然而，人造组织的一般用途是有限的。详细 对调节细胞粘附和运动的机制的理解是期望的 为控制细胞接种、改进设计和 人工组织工程 非侵入性电刺激（ES）的使用提供了一种新的非机械性刺激。 调节细胞粘附和运动性技术。涉及使用 ES在成纤维细胞、肝细胞和白色细胞中的表达，以及侧和 膜蛋白质的旋转动力学提供了很好的基础， 在这个提案中提出了一些假设。为了响应ES，机械换能器 整联蛋白可能介导细胞粘附和运动。整合素 在细胞表面重新分布，与细胞骨架相互作用， 参与粘着斑接触的动态形成。的优化使用 ES已被证明可以重新分布整合素，重组细胞骨架，改变 钙稳态，并诱导引导细胞迁移，而不会产生不利影响 影响细胞活力。该提案使用独特的非侵入式光学 技术，包括单粒子跟踪和激光光学陷阱，以1） 在单分子水平上跟踪ES诱导的整合素运动的变化 在人成纤维细胞的表面上; 2）测量 ES诱导的整合素-细胞骨架相互作用 细胞外基质;和3）表征机电诱导的和 重组三维凝胶模型中整合素依赖性细胞运动性。的 拟议研究的长期目标是操纵和控制 细胞粘附和运动性的最佳使用ES，从而提高 组织完整性和工程组织的功能。

DESCRIPTION (provided by applicant): Tissue engineering is a relatively new but rapidly expanding field of biomedical engineering research. Because cell adhesion and motility are two critical factors in determining tissue integrity and function, elucidation and regulation of the cellular and molecular mechanisms involved in cell adhesion and motility is fundamentally important for tissue engineering. Implementation of engineered tissues in clinical applications has been successful in soft-tissue replacement or regeneration. The general use of artificial tissues is limited, however. Detailed understanding of mechanisms regulating cell adhesion and motility is expected to provide insights for controlled cell seeding, improved designing and engineering of artificial tissues. Use of non-invasive electrical stimulation (ES) offers a novel non-mechanical technique to regulate cell adhesion and motility. Projects involving the use of ES in fibroblasts, hepatocytes, and white cells and studies of lateral and rotational dynamics of membrane proteins provided an excellent basis for the proposed hypotheses in this proposal. In response to ES, the mechanotransducer integrin is likely to mediate cell adhesion and motility. Integrins redistribute on the cell surface, interact with cytoskeleton, and actively participate in dynamic formation of focal adhesion contacts. Optimized use of ES has been shown to redistribute integrins, reorganize cytoskeleton, alter calcium homeostasis, and induce guided cell migration without adversely affecting cell viability. This proposal uses unique non-invasive optical techniques, including single particle tracking and laser optical trap, to 1) track, at the single molecule level, changes in integrin motion induced by ES on the surface of human fibroblasts; 2) measure changes in the strength of integrin-cytoskeleton interactions induced by ES on controlled 2 dimensional extracellular matrices; and 3) characterize electromechanically induced and integrin-dependent cell motility in reconstituted 3 dimensional gel model. The long-term objectives of the proposed research are to manipulate and control cell adhesion and motility by the optimal use of ES and, thereby, to enhance tissue integrity and function of engineered tissues.