Corneal and Limbal Alkali Injury Induction Using a Punch-Trephine Technique in a Mouse Model

Our aim is to induce a precise and localized alkaline injury into the cornea and limbal area in a mouse model. We develop a novel punch-trephine method, and then studied how effective this method is in creating a corneal and limbal stem cell deficiency. Advanced techniques in the field of corneal opacity treatment and corneal regeneration, including limbal stem cell transplantation, exosome transplantation, and mitochondria transplantation are based on studies that investigated corneal responses to various injuries as well as treatments.

Therefore, it is critical to have a standard, accurate, and reliable injury model among researchers. Several techniques and animal models have been employed to induce various corneal injuries and investigate various aspects of the corneal wound healing process. The alkali burn model is well-established injury model, which is performed by applying sodium hydroxide directly to the corneal surface or by using flat filter paper.

We develop a novel punch-trephine method to create a mouse model of limbal stem cell deficiency accurately and reliably, overcome the limitations of previous techniques, and minimizing the risk of insufficient injury to the limbal area, or additional injury to the fornix and conjunctiva. This technique has several advantages. Effective chemical injury to the entire corneal surface and limbus in mice, localized and well-circumscribed injury to the cornea, application of any liquid for a predetermined duration, induction of various corneal injury sizes with suitable biopsy punches.

It is also feasible for rat and rabbit injury models.