Electromechanical control of cell adhesion and motility

简介(申请人提供)：组织工程学是一门相对较新的学科，但 迅速拓展的生物医学工程研究领域。因为细胞 粘附性和运动性是决定组织完整性的两个关键因素 以及细胞和分子的功能、阐明和调节 涉及细胞黏附和运动的机制是根本重要的 用于组织工程学。组织工程技术在临床中的应用 在软组织置换或再生方面的应用取得了成功。 然而，人造组织的普遍使用是有限的。详细 需要了解调节细胞黏附和运动的机制。 为受控细胞接种、改进的设计和 人造组织工程学。 无创电刺激(ES)的使用提供了一种新的非机械的 调节细胞黏附和运动的技术。涉及使用的项目 成纤维细胞、肝细胞和白细胞中的ES及其侧方和侧方的研究 膜蛋白的旋转动力学为进一步研究提供了良好的基础 本提案中提出的假设。作为对ES的响应，机械式传感器 整合素可能介导细胞的黏附和运动。整合素 重新分布在细胞表面，与细胞骨架相互作用，并积极 参与焦点粘合接触的动态形成。优化使用 ES已被证明可以重新分配整合素，重组细胞骨架，改变 钙稳态，并诱导引导细胞迁移而不产生不利影响 影响细胞活力的。该方案使用独特的非侵入性光学 技术，包括单粒子跟踪和激光光学陷阱，至1) 在单分子水平跟踪ES诱导的整合素运动的变化 在人成纤维细胞表面；2)测量强度的变化 ES在受控二维上诱导的整合素-细胞骨架相互作用 细胞外基质；以及3)表征机电诱导和 重组三维凝胶模型中整合素依赖的细胞运动。这个 拟议研究的长期目标是操纵和控制 通过最佳使用ES来提高细胞的粘附性和运动性，从而增强 工程组织的组织完整性和功能。

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.