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Development of a wound-on-chip model to study stromal-epithelial interactions during tissue repair

开发芯片伤口模型来研究组织修复过程中基质-上皮相互作用

基本信息

批准号：
10316239
负责人：
Jeroen Eyckmans
金额：
$ 20.63万
依托单位：
BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10316239
关键词：
3-Dimensional Address Adipose tissue Affect Aging Alpha Granule American Amputation Animal Model Animals Architecture Benchmarking Biological Assay Biological Models Biomimetics CRISPR-mediated transcriptional activation Cell Line Cells Cicatrix Coculture Techniques Collagen Communities Data Dependence Deposition Dermal Dermis Development Embryo Engineering Epidermis Epithelial Epithelial Cells Epithelial-Stromal Communication Exhibits Extracellular Matrix Failure Fibroblasts Fibrosis Gene Expression Genes Goals Granulation Tissue Hemostatic function Human Hydrogels In Vitro Inflammation Injury Link Measures Mediating Medical Methods Modeling Molecular Morphogenesis Natural regeneration Outcome Pathologic Phase Process Proxy Publishing Research Skin Tissue Source Speed System Techniques Technology Testing Thick Time Tissues Translations Wound models base chronic ulcer chronic wound cost estimate diabetic patient epithelial injury epithelial wound genome editing healing in vitro Model in vivo in vivo Model individual patient insight interest keratinocyte knock-down migration monolayer novel open wound overexpression regenerative repaired response restoration species difference tissue regeneration tissue repair wound wound bed wound closure wound healing

项目摘要

The goal of this proposal is to engineer a novel in vitro biomimetic wound healing model to study how human fibroblasts and epithelial cells coordinate tissue closure at the cellular and molecular level. In vivo wound healing is a dynamic morphogenetic process with the goal to close and restore the damaged tissue. A critical stage during tissue closure is re-epithelialization of wounds, a process by which epithelial cells migrate over the denuded wound bed, to restore the barrier. Failure of wounds to re-epithelialize results in chronic wound formation, a condition that affect 6 million Americans annually and carries an estimated cost of US $25 billion per year for the medical system. Hence, understanding the mechanisms that drive re-epithelialization has been a central focus in wound healing research. Due to limitations with animal models, in vitro models have been instrumental to study re-epithelialization by human epithelial cells. Traditional models, such as the scratch wound assay, involve scratching of a monolayer of epithelial cells adherent to a planar substrate, and the time for migrating cells to repopulate the scratch is measured as a proxy for healing. In more advanced co-culture models, the planar substrate is either replaced by a fibroblast-laded collagen hydrogel or by a dermal tissue explant. Whereas these models have a pre-defined substrate as a migration base for epithelial cells, in vivo studies have shown that for full-thickness wounds, the deeper fibrous layers must heal first through the formation of granulation tissue by fibroblasts, before epithelial cells can migrate over this provisional tissue to close the wound. Thus, in in vivo settings, re-epithelialization occurs as fibroblasts deposit a provisional template and reciprocal interactions between fibroblasts an epithelial cells coordinate closure of these two tissue layers. Current in vitro models don't capture this intricate tissue dynamics. Given the dependency of re-epithelialization on the underlying substrate, we hypothesize that fibroblasts mediate the rate of re-epithelialization during wound closure. To address this hypothesis, we propose in aim 1 to build a biomimetic in vitro wound closure model wherein re-epithelialization ensues fibrous tissue repair in wounded engineered microtissues to emulate healing of full thickness wounds. In aim 2, we will use state-of-the art genome editing techniques to elucidate fibroblast- epithelial interactions that regulate fibrous tissue closure and re-epithelialization. These studies will also validate and benchmark our 3D biomimetic model to other wound healing models. In aim 3, we will explore whether fibroblasts from different healthy and pathological tissue sources affect re-epithelialization in our biomimetic model. Ultimately, this project aims to establish a basis for optimizing a wound bed that enables rapid re- epithelialization as a paradigm for promoting tissue regeneration and minimizing scarring.

该提案的目标是设计一种新的体外仿生伤口愈合模型，以研究人类如何在伤口愈合过程中发挥作用。成纤维细胞和上皮细胞在细胞和分子水平上协调组织闭合。体内伤口愈合是一个动态的形态发生过程，目的是闭合和恢复受损组织。关键阶段在组织闭合过程中，伤口的再上皮化是上皮细胞在伤口上迁移的过程。裸露的伤口床恢复屏障伤口不能再上皮化导致慢性伤口这种疾病每年影响600万美国人，估计花费250亿美元每年，医疗系统。因此，了解驱动上皮再生的机制已经成为伤口愈合研究的中心焦点。由于动物模型的局限性，体外模型已经被广泛应用。有助于研究人上皮细胞的上皮再形成。传统的模型，比如抓伤测定，包括刮擦粘附于平面基底的单层上皮细胞，以及刮擦的时间。迁移细胞以重新填充划痕被测量为愈合的代表。在更先进的共培养模型中，平面基底或者被载有成纤维细胞的胶原水凝胶或者被真皮组织外植体替代。尽管这些模型具有预定义的底物作为上皮细胞的迁移基础，但体内研究已经证实了这些模型的有效性。表明对于全层伤口，较深的纤维层必须首先通过形成在上皮细胞可以在该临时组织上迁移以闭合肉芽组织之前，伤口因此，在体内环境中，当成纤维细胞存款临时模板时发生上皮再形成，成纤维细胞和上皮细胞之间的相互作用协调这两个组织层的闭合。目前的体外模型无法捕捉到这种复杂的组织动力学。考虑到上皮再生的依赖性在底层基质上，我们假设成纤维细胞介导了创伤过程中的再上皮化速率结束为了解决这一假设，我们在目标1中提出建立一个仿生体外伤口闭合模型其中再上皮化增强受伤的工程化微组织中的纤维组织修复以模拟愈合全层伤口在目标2中，我们将使用最先进的基因组编辑技术来阐明成纤维细胞- 调节纤维组织闭合和再上皮化的上皮相互作用。这些研究还将验证并将我们的3D仿生模型与其他伤口愈合模型进行比较。在目标3中，我们将探讨来自不同健康和病理组织来源的成纤维细胞影响我们的仿生模型最终，该项目旨在为优化伤口床奠定基础，使其能够快速重新定位。上皮形成作为促进组织再生和最小化瘢痕形成的范例。