Abstract
The repair of wounds through collective movement of epithelial cells is a fundamental process in multicellular organisms. In stratified epithelia such as the cornea and skin, healing occurs in three steps that include a latent, migratory, and reconstruction phases. Several simple and inexpensive assays have been developed to study the biology of cell migration in vitro. However, these assays are mostly based on monolayer systems that fail to reproduce the differentiation processes associated to multilayered systems. Here, we describe a straightforward in vitro wound assay to evaluate the healing and restoration of barrier function in stratified human corneal epithelial cells. In this assay, circular punch injuries lead to the collective migration of the epithelium as coherent sheets. The closure of the wound was associated with the restoration of the transcellular barrier and the re-establishment of apical intercellular junctions. Altogether, this new model of wound healing provides an important research tool to study the mechanisms leading to barrier function in stratified epithelia and may facilitate the development of future therapeutic applications.
Highlights
Cells mostly change morphology and become flattened[10]
Understanding how re-epithelialization and induction of barrier function are orchestrated in multilayered wound healing models requires the development of injury assays that can be systematically and quantitatively reproduced[12]
Understanding the mechanisms that lead to restoration of barrier function in stratified epithelia during wound healing is essential to maintain tissue integrity and reduce complications such as infection and corneal melting
Summary
Cells mostly change morphology and become flattened[10]. This phase of wound repair is essential for the reestablishment of the normal appearance of the epithelia and the restoration of barrier function[11]. Most culture models evaluating migration of cells from stratified tissues are based on monolayer cultures, which fail to reproduce the three phases of epithelial healing. The aim of this study was to develop and characterize a simple and reproducible in vitro model of wound healing that could potentially be used to study the different phases of healing in stratified epithelium
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