Vitamin C in the genic expression of oxidative stress markers in primary culture of keratinocytes of burned patients

Abstract

Background: burns are a major problem of public health in the world. The initial response to burns, mediated by activated neutrophils and production of reactive oxigen species (ROS) is frequently associated with secondary damage in other tissues, distant from the thermally injured site. This process, known as oxidative stress, aggravates the conditions of burned patients. In this situation, the antioxidant defenses are essential, however, they are exhausted, thus being necessary the supplementation of substances with such property. Vitamin C (ascorbic acid) is a powerful antioxidant on human plasma, contributing to ROS' elimination and protecting against lipidic peroxidation. The present study's aim is to evaluate the action of vitamin C on the expression of 84 genes associated with oxidative stress in primary keratinocytes cultivated from burned patients' skin. Methods: In this study, will be included 10 patients with severe burns and 5 healthy patients for control. Division into four experimental groups: I - Keratinocytes growth of burned patient, growth medium supplemented with ascorbic acid; II - Keratinocytes growth of unburned patient, growth medium supplemented with ascorbic acid; III - Keratinocytes growth of burned patient, without ascorbic acid supplementation; IV - Keratinocytes growth of unburned patient, without acid ascorbic supplementation. The growth will be initiated by enzimatic method using dispase and collagenase. In the experimental groups supplemented with vitamin C Sodium Ascorbate 100 ¼M will be used, after the first passage, at each growth medium exchange until it reaches the sub-confluence. 84 genes that mark oxidative stress will be analyzed (kit RT² Profiler" PCR Array Human Oxidative Stress.). The experiments will be done in triplicate.