Abstract
The use of in vitro systems to investigate the process of corneal wound healing offers the opportunity to reduce animal pain inflicted during in vivo experimentation. This study aimed to establish an easy-to-handle ex vivo organ culture model with porcine corneas for the evaluation and modulation of epithelial wound healing. Cultured free-floating cornea disks with a punch defect were observed by stereomicroscopic photo documentation. We analysed the effects of different cell culture media and investigated the impact of different wound sizes as well as the role of the limbus. Modulation of the wound healing process was carried out with the cytostatic agent Mitomycin C. The wound area calculation revealed that after three days over 90% of the lesion was healed. As analysed with TUNEL and lactate dehydrogenase assay, the culture conditions were cell protecting and preserved the viability of the corneal tissue. Wound healing rates differ dependent on the culture medium used. Mitomycin C hampered wound healing in a concentration-dependent manner. The porcine cornea ex vivo culture ideally mimics the in vivo situation and allows investigations of cellular behaviour in the course of wound healing. The effect of substances can be studied, as we have documented for a mitosis inhibitor. This model might aid in toxicological studies as well as in the evaluation of drug efficacy and could offer a platform for therapeutic approaches based on regenerative medicine.
Highlights
The cornea is a transparent, avascular tissue which forms an important barrier through tight junctions present at its superficial layer [1]
This highly specialized tissue has been used extensively as a model system to study wound healing in the human body as its anatomic location causes the cornea to be subject to abrasive forces and physical damage [2]
Wound healing of the cornea involves the removal of necrotic cells, infiltration of neutrophils, migration of cells from the wound edge covering the wound, mitosis as well as migration of limbal epithelial stem cells, and differentiation of newly formed epithelial cells to more mature, anterior, stratified epithelial cells to restore a smooth multilayered formation [4]
Summary
The cornea is a transparent, avascular tissue which forms an important barrier through tight junctions present at its superficial layer [1]. This highly specialized tissue has been used extensively as a model system to study wound healing in the human body as its anatomic location causes the cornea to be subject to abrasive forces and physical damage [2]. Wound healing speed might be impacted by various diseases such as diabetes, autoimmune disorders or infection of the tissue During this period, various eye drops are used to support healing and prevent infection. These agents should not adversely affect re-epithelization and reduce scar tissue formation [3]
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