Biophysical Cues and Corneal Wound Healing

描述(申请人提供)：角膜伤口愈合失调可导致角膜疤痕和角膜混浊，导致视力丧失。尽管角膜创伤愈合的许多因素已经被很好地描述，但创伤空间的生物物理属性以及这些因素对伤口愈合至关重要的细胞活动的影响还没有得到充分的研究。与细胞外环境中存在的可溶信号分子一起，生物物理线索正在成为决定细胞行为的关键因素。我们建议对创面愈合过程中基质形态和顺应性的变化进行定量分析。我们的假设是，伤口间隙中的生物物理线索在伤口愈合过程中发生变化，这些线索直接影响角膜基质层角质形成细胞向肌成纤维细胞的转变。肌成纤维细胞反过来通过主动收缩改变伤口微环境的顺应性。最终，伤口床中生物物理线索的变化将激活细胞凋亡途径，促进肌成纤维细胞的去除，光学透明的角膜将回归。以生物物理环境中测量的变化为指导，我们将制备具有相似生物物理性质(顺应性和地形)的水凝胶基质，以确定这些变化对体外培养的肌成纤维细胞行为的细胞后果。我们获得的关于细胞行为和基因和蛋白质变化的信息可以使伤口愈合过程中的生物物理线索成为治疗性靶点。可以直接调节伤口床的属性，例如角膜基质细胞的收缩元素，以改善伤口愈合结果。 与公共卫生相关：角膜的伤口愈合可能导致角膜疤痕，并可能导致视力受损或丧失。尽管角膜伤口愈合的许多因素已经被很好地描述，但伤口空间的生物物理属性还没有得到充分的研究。随着细胞外环境中存在的可溶信号分子，生物物理线索正在成为细胞行为的关键调节因素。这一建议将决定整个角膜创面愈合过程中存在的生物物理线索(特别是地形图和顺应性)。我们的假设是，伤口间隙的生物物理属性在整个愈合过程中都会发生变化，角膜基质层细胞表型的转变受到伤口间隙生物物理线索的影响。此外，肌成纤维细胞是正常和失调伤口愈合的中心角色，它将改变伤口微环境的顺应性，从而帮助伤口修复，但最终也会触发防止角膜模糊所需的信号。如果被证实是真的，我们的假设将表明，直接调节角膜基质细胞的生物物理特性和收缩元素可以改善伤口愈合结果。

DESCRIPTION (provided by applicant): Dysregulated wound healing in the cornea can lead to corneal scars and corneal haze resulting in a loss of visual acuity. While many elements of corneal wound healing are well characterized, the biophysical attributes of the wound space and the impact of these on cellular activities critical to wound healing have not been adequately investigated. Along with soluble signaling molecules present in the extracellular milieu, biophysical cues are emerging as key factors in determining cell behaviors. We propose to quantitatively analyze the alterations in stromal topography and compliance throughout the wound healing process. Our hypothesis is that the biophysical cues in the wound space alter throughout wound healing and that these directly influence the transition of keratocytes to myofibroblasts in the corneal stromal layer. The myofibroblasts, in turn, would alter the compliance of the wound microenvironment through active contraction. Ultimately, the change in the biophysical cues in the wound bed would activate apoptotic pathways, promoting removal of the myofibroblasts and an optically clear cornea would return. Using the measured alterations in the biophysical environment as a guide, we will fabricate hydrogel substrates with similar biophysical properties (of compliance and topography) to determine the cellular consequences of these alterations on myofibroblast behavior in vitro. The information that we obtain about the cellular behaviors and gene and protein alterations could enable the therapeutic targeting of biophysical cues during wound healing. Attributes of the wound bed could be directly modulated, such as the contractile elements of the corneal stromal cells, to improve wound healing outcomes. PUBLIC HEALTH RELEVANCE: Wound healing in the cornea can lead to corneal scars and can lead to impairment or loss of vision. While many elements of corneal wound healing are well characterized, biophysical attributes of the wound space have not been adequately investigated. Along with soluble signaling molecules present in the extracellular milieu, biophysical cues are emerging as key regulators of cell behaviors. This proposal would determine the biophysical cues (specifically topography and compliance) present throughout corneal sromal wound healing. Our hypothesis is that the biophysical attributes of the wound space changes throughout the healing process and that the transition of phenotypes of the cells in the corneal stromal layer is modulated by alterations in the biophysical cues of the wound space. In addition, the myofibroblast, a central player in normal and dysregulated wound healing will modify the compliance of the wound microenvironment and thereby assist in wound repair, but also in ultimately triggering signals necessary to prevent corneal haziness. If proven true, our hypothesis would indicate that direct modulation of the biophysical properties and contractile elements of the corneal stromal cells could improve wound healing outcomes.