Wound healing is a complex biological process requiring the coordinated function of multiple proteins. In chronic inflammatory bowel disease (IBD), particularly Crohn’s Colitis (CC), this process is dysregulated, leading to delayed tissue repair and increased disease burden. Prior studies indicate that Human Alpha Defensin 5 (HD5) is over-expressed in the colonic epithelium of CC patients, yet its impact on wound healing remains poorly understood.Our laboratory has identified that HD5 may impair wound healing by modulating key structural and regulatory proteins. Preliminary findings suggest that HD5 reduces the expression and function of desmoplakin, a critical desmosomal protein essential for maintaining epithelial integrity and promoting wound closure. Utilizing an assortment of biological techniques, including wound healing assays, quantitative polymerase chain reaction (qPCR), and Western blotting, we aim to elucidate the mechanistic relationship between HD5 and desmoplakin in epithelial wound healing using NCM460D cells as a model system. We hypothesize that increasing concentrations of HD5 down-regulate desmoplakin protein and mRNA expression, compromising its ability to mediate cellular adhesion and epithelial repair. This disruption is predicted to result in reduced wound closure and impaired epithelial barrier restoration in HD5-treated cells compared to untreated controls. Understanding the interaction between HD5 and desmoplakin provides a novel perspective on the molecular mechanism of IBD-associated epithelial cell damage. This research may lead to the development of targeted therapies aimed at targeting colonic HD5, thereby improving wound healing and reducing disease burden in CC patients.