Tissue Regeneration by Biophysical Signaling

描述（由申请人提供）：组织工程的传统方法集中于生物化学鸡尾酒，以引导细胞朝向组织特异性结果;在某些情况下，还利用了机械力。然而，也有重要的文献详细描述了生物物理信号在组织发育和组织再生过程中的作用，迄今为止尚未纳入组织工程领域。发育生物学领域已经追踪了生物物理因素（例如膜电压电位和离子通量）在组织再生、伤口愈合、胚胎图案化和许多其他关键组织相关事件中的作用。这些数据提供了膜电位和细胞行为之间的明确联系，决定组织特异性结果。然而，许多分子细节仍然不清楚，这种新的细胞控制模式尚未被利用来促进组织再生。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3D人体组织系统的骨骼和脂肪组织。目标是确定生物物理因素（如膜电位）在目标#1中的组织再生、目标#2中的组织发育和目标#3中的组织图案化背景下对组织特异性结局的效用。我们将比较膜电位在组织再生和形成过程中的作用，并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调节离子转运，通过3D光学可寻址支架系统介导的组织结果的空间控制，以在体外生成组织模式，类似于肢体发育过程中的形态控制。这项研究的结果将是一种全新的体外组织形成调控方法，对再生医学的许多领域都有影响。了解和利用生物电信号在非兴奋细胞中对组织结果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。 公共卫生相关性：开发组织再生的生物物理控制实际上是组织工程领域中未开发的领域，尽管发育生物学中的广泛研究已经清楚地表明了组织/器官发育和再生期间膜电位和内源性电场变化的重要性。因此，该计划的目标是确定膜电位调节信号对骨和脂肪组织再生，形成和模式的影响。该计划的结果将是一种全新的体外组织形成和控制方法，对体内再生具有重要意义。合理调节这些强大的生物物理控制，有或没有更传统的生化控制，将允许更大的控制组织的发展和功能。基于发育生物学原理，拟议研究的进展将对组织工程领域产生深远影响。了解生物物理因素对组织行为的作用将有助于深入了解组织生长和再生的基本控制机制，为当前的组织工程范式提供新的视角。这样的理解也将定义一套良好的表征药理学和分子遗传学工具，使新的方法来复杂的组织图案。

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.