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Glycerin is a widely used ingredient in cosmetics due to its cost-effectiveness and safety. While often used to enhance the texture of cosmetics, current research has demonstrated that it improves cutaneous properties such as enhanced skin hydration and moisturization. Due to its widespread use in cosmetics, enhancing the functional capacities of glycerin provides a promising method to improve the effectiveness of numerous cosmetics. Ozonized glycerin has emerged as a novel technology able to enhance glycerin’s effectiveness with reported anti-inflammatory, antibacterial, and antiviral effects. This approach leverages ozone stabilization in glycerin for improved stability and prolonged release to the skin. Clinical application of ozonized glycerin has exhibited lightening of aging spots through promoting skin turnover. The objective of this study was to evaluate the enhanced properties of glycerin when ozonized in terms of skin repair and inflammation. Through topical pretreatment of epidermal 3D wound healing models (13 days) and ex vivo human skin biopsies (4 days), ozonized glycerin was able to improve wound closure, enhance skin barrier and extracellular matrix protein expression, and reduce inflammation. Notably, ozonized glycerin enhanced wound closure by 6.8% compared to glycerin, as well as significantly protecting against LPS-induced elastin degradation (67.7% difference from LPS). These data provide evidence for the use of ozonized glycerin as a new technology to prevent and diminish skin inflammation and improve wound repair.