Tissue Regeneration by Biophysical Signaling

描述（由申请人提供）：组织工程的传统方法侧重于生化鸡尾酒，以指导细胞走向组织特异性结果；在某些情况下，还利用了机械力。然而，也有大量文献详细介绍了生物物理信号在组织发育和组织再生过程中的作用，但迄今尚未纳入组织工程领域。发育生物学领域跟踪生物物理因素，如膜电压电位和离子通量，在组织再生，伤口愈合，胚胎模式和许多其他关键的组织相关事件中的作用。这些数据提供了膜电位和决定组织特异性结果的细胞行为之间的明确联系。然而，许多分子细节仍不清楚，这种新的细胞控制方式尚未被用于推进组织再生。本提案的重点是通过专门研究骨和脂肪组织再生，发育和模式的生物物理调节来填补这一空白。我们将利用三维人体组织系统的骨骼和脂肪组织。目的是确定生物物理因素，如膜电位，在Aim #1的组织再生、Aim #2的组织发育和Aim #3的组织模式的背景下对组织特异性结果的效用。我们将比较膜电位在组织再生和形成过程中的作用，以使用传统的生化鸡尾酒作为对照。在最后一个目标中，我们将专注于通过光激活离子运输调节组织结果的空间控制，通过3D光学可寻址支架系统介导，在体外产生组织模式，类似于肢体发育期间的形态控制。这项研究的结果将为体外组织形成的调控提供一种全新的方法，对再生医学的许多领域都有影响。理解和利用生物电信号在不可兴奋细胞组织结果中的作用，将为组织再生的基本控制提供新的见解，以及在体外和体内组织中产生复杂模式发育的新方法。

DESCRIPTION (provided by applicant): Traditional approaches to tissue engineering have focused on biochemical cocktails to direct cells toward tissue-specific outcomes; in some cases mechanical forces have also been utilized. However, there is also a significant literature that details the role of biophysical signaling during tissue development and tissue regeneration, which has not yet been incorporated into the field of tissue engineering to date. The field of developmental biology has tracked the role of biophysical factors, such as membrane voltage potential and ion fluxes, during tissue regeneration, in wound healing, in embryonic patterning, and in many other critical tissue- related events. These data provide a clear link between membrane potential and cell behavior that determine tissue-specific outcomes. However, many molecular details are still unclear and this novel cell control modality has not been capitalized upon to advance tissue regeneration. The focus of the present proposal is to fill this void by specifically studying biophysical regulation of bone and adipose tissue regeneration, development and patterning. We will utilize 3D human tissue systems for bone and adipose tissue. The goal is to determine the utility of biophysical factors, such as membrane potential, on tissue-specific outcomes in the context of tissue regeneration in Aim #1, tissue development in Aim #2, and tissue patterning in Aim #3. We will compare the role of membrane potential during tissue regeneration and formation to the use of traditional biochemical cocktails as the controls. In the last aim, we will focus on spatial control of tissue outcomes via light-activated regulation of ion transport, mediated via a 3D optically-addressable scaffold system, to generate tissue patterns in vitro, analogous to morphological control during limb development. The outcome of the proposed study will be an entirely new approach to the regulation of tissue formation in vitro, with implications in many areas of regenerative medicine. Understanding and exploiting the role of bioelectrical signals on tissue outcomes in non-excitable cells will provide new insight into fundamental control of tissue regeneration, as well as novel approaches toward generating complex pattern development in tissues both in vitro and in vivo. PUBLIC HEALTH RELEVANCE: Exploitation of biophysical control of tissue regeneration is virtually unexplored territory in the field of tissue engineering, despite extensive studies in developmental biology that have clearly shown the importance of changes in membrane potential and endogenous electric fields during tissue/organ development and regeneration. Thus, the goal of this program is to determine the impact of membrane potential-regulated signaling on bone and adipose tissue regeneration, formation, and patterning. The outcome of this program would be an entirely new approach to tissue formation and control in vitro, with major implications for regeneration in vivo. Rational modulation of these powerful biophysical controls, with or without more traditional biochemical controls, will allow greater control of tissue development and function. Building upon principles from developmental biology, progress from the proposed studies will have a profound impact on the field of tissue engineering. Understanding the role of biophysical factors on tissue behavior will yield insight into fundamental control mechanisms underlying tissue growth and regeneration, offering a new perspective to the current tissue engineering paradigm. Such an understanding will also define a set of well-characterized pharmacological and molecular-genetic tools to enable novel approaches to complex tissue patterning.