Project Grant R03AA031535

INVESTIGATION OF COLD PLASMA FOR HEALING ALCOHOL-INDUCED TISSUE INJURY - ABSTRACT: BINGE DRINKING OF ALCOHOL BEVERAGES HINDERS TISSUE REPAIR AND AMPLIFIES THE SUSCEPTIBILITY TO INFECTION. THE PREVALENCE AS WELL AS THE RISKS ASSOCIATED WITH BINGE DRINKING POSES A SIGNIFICANT CONCERN IN THE UNITED STATES. BINGE DRINKING IMPAIRS THE THREE KEY PHASES OF WOUND HEALING: INFLAMMATION, PROLIFERATION, AND REMODELING. SPECIFICALLY, DURING THE INFLAMMATION PHASE, BINGE DRINKING DIMINISHES THE FUNCTION OF PRO-INFLAMMATORY CYTOKINES, NEUTROPHILS, AND MACROPHAGES, THUS COMPROMISING THE INNATE IMMUNE SYSTEM'S ABILITY TO COMBAT PATHOGENS. THE EFFICACY OF CURRENT WOUND HEALING METHODS IS CONSTRAINED BY THE VULNERABILITY TO WOUND INFECTION WHICH IS EXACERBATED BY CONCURRENT BINGE DRINKING. COLD PLASMA (CP) HAS RECENTLY GAINED SUBSTANTIAL ATTENTION IN CLINICAL PRACTICE DUE TO ITS REGENERATIVE PROPERTIES AND ITS ABILITY TO PREVENT INFECTION. PREVIOUS STUDIES HAVE SHOWN THAT CP TREATMENT IS INVOLVED IN THE WOUND HEALING MEDIATED BY THE NRF2 PATHWAY AND WNT/BCATENIN SIGNALING PATHWAY. TAKEN THESE SOLID PREMISES TOGETHER, WE HYPOTHESIZE THAT CP WILL IMPROVE IMPAIRED WOUND HEALING AND MITIGATE THE INCREASED VULNERABILITY TO INFECTION CAUSED BY BINGE DRINKING. TO SUBSTANTIATE THESE HYPOTHESES, WE HAVE PROPOSED TWO FOLLOWING SPECIFIC AIMS USING F344 RAT MODEL AND HUMAN SKIN ORGANOID MODEL, RESPECTIVELY. AIM 1 IS TO CHARACTERIZE WOUND HEALING PROPERTIES OF CP IN F344 RATS FOLLOWING BINGE EXPOSURE TO ALCOHOL. THIS AIM IS TO EXAMINE WOUND CLOSURE PROPERTIES OF CP AND TO REVEAL MOLECULAR MECHANISMS UNDERLYING CP MODULATION OF WOUND HEALING IN BINGE ETOH INTOXICATED F344 RATS. AIM 2 IS TO GENERATE AN IN-VITRO WOUND HEALING MODEL USING HIPSC DERIVED SKIN ORGANOIDS AND TO INVESTIGATE THE POTENTIAL WOUND HEALING PROPERTIES OF CP IN THESE SKIN ORGANOIDS GIVEN BINGE LEVEL OF ETOH. THIS AIM REPRESENTS THE TRANSLATIONAL ASPECT OF OUR STUDIES. WE FIRST WILL DEVELOP AND CHARACTERIZE 3D SKIN ORGANOIDS USING HUMAN INDUCED PLURIPOTENT STEM CELLS (HIPSC) AND THEN IDENTIFY THE POTENTIAL WOUND HEALING PROPERTIES OF CP IN ALCOHOL-TREATED HUMAN SKIN ORGANOIDS. IN ADDITION, WE WILL STUDY MOLECULAR MECHANISMS AND SIGNAL PATHWAYS UNDERLYING CP MODULATION OF WOUND HEALING IN ALCOHOL TREATED SKIN ORGANOIDS USING BOTH IMMUNOFLUORESCENCE AND GENE EXPRESSION ANALYSIS. THESE STUDIES ARE HIGHLY SIGNIFICANT BECAUSE 1) WE ARE TO DEMONSTRATE THE MEDICAL USE OF CP IN WOUND HEALING ASSOCIATED WITH ETOH INTOXICATION, AND 2) WE ARE TO ADDRESS THE TRANSLATIONAL VALUE OF THE PROPOSED STUDIES USING F344 RAT MODEL AND HUMAN CULTURE OF SKIN ORGANOIDS. SUCCESSFUL COMPLETION OF THIS SMALL R03 GRANT AWARD WILL PROVIDE VALUABLE THERAPEUTIC CLUES IN CP MODULATION OF THE WOUND HEALING DURING ETOH INTOXICATION. THE STUDY OUTCOMES SHALL SHIFT THE PARADIGMS REGARDING THE INTERPLAY BETWEEN WOUND HEALING PROPERTIES OF CP AND ALCOHOL- INDUCED IMPAIRMENT OF WOUND HEALING, EVENTUALLY LEADING TO CLINICAL APPLICATION OF CP IN WOUND HEALING DURING ETOH INTOXICATION. WE EXPECT A FULL R01 PROJECT TO BE DEVELOPED IN CONCLUSION OF THIS SMALL R03 PROJECT.