Poster Topical Area: Aging and Chronic Disease
Poster Board Number: 113
Objectives: Current treatments for Inflammatory Bowel Disease (IBD) are insufficient, commonly based on inhibiting pro-inflammatory cytokines and using immunomodulators. Not only is there concern about the possible negative long term effects of these treatments, but they also do not address the structural damage to the digestive tract. By increasing the rate of wound healing in the intestinal epithelium we can reduce intestinal damage, including the occurrences of fistulas. The aim of this study is to examine the effects of various micronutrients and peptides with antimicrobial and proliferation-inducing activities individually and in combination. These molecules have been shown to accelerate wound healing in model organisms, but their effect on intestinal epithelial cells requires further examination.
Methods: Using HT-29 colorectal cancer intestinal epithelial cells we are examining the effect of each micronutrient and peptide on wound closure for selection of the best combinations to determine the optimal conditions for micronutrient- and peptide-induced wound healing. The wound was created using silicone culture inserts for consistent wound size. HT-29 cells were allowed to grow to confluency in a 6-well plate, the insert was removed, and each well was treated with a different peptide or micronutrient. Pictures and measurements (in μm) of each well were taken every 24hrs for 5 days. Complete wound closure in the control occurs in approximately 120hrs and both peptides and micronutrients have shown to shorten that time.
Results: Curcumin is exhibiting concentration-dependent action; lower concentrations (1μM) are stimulating wound healing more strongly than higher concentrations (10μM). The antimicrobial peptide LL-37 (10 - 100μg/mL) had a stimulatory effect at all concentrations used. In HT-29 cells both Curcumin and Vitamin D stimulate Vitamin D receptor activity as shown through a Vitamin D response element luciferase reporter assay.
Conclusions: Antimicrobial peptides and micronutrients stimulate wound healing in HT-29 cells. In future experiments we will package the chosen peptide(s) and nutrient(s) in nanoparticles for targeted delivery. The results of this study can help address the issue of structural damage to the intestinal epithelium that occurs in IBD.
University of Miami Miller School of Medicine