Project Summary Genitourinary syndrome of menopause (GSM) is an extremely prevalent condition consequent to the hypoestrogenic changes in the genitourinary tract that affects up to 85% of perimenopausal and menopausal women. Major symptoms of GSM include vaginal dryness, itching, discomfort, burning, and pain; thus, GSM significantly impacts quality of life during everyday activities and severely impairs sexual function. Despite its high prevalence and interference with healthy aging, current treatments for GSM are suboptimal, with many issues related to accessibility or long-term efficacy. Despite an astoundingly low satisfaction of only 35% with treatments for GSM, whether prescribed or over-the-counter, women continue to suffer from lack of better options. Thus, there remains a need for a safe and accessible therapy for this morbid condition that can both alleviate symptoms and restore a healthy vaginal phenotype. The proposed study will determine if a low-cost, acellular, tissue-specific, and minimally invasive regenerative therapy can repair the atrophic vaginal tissue, resulting in an accessible, high-impact intervention for GSM. Given our previous successes with tissue-specific pro-regenerative biomaterials, we opine that a novel vaginal tissue-derived ECM hydrogel (vECM) will reverse vaginal atrophy by inducing epithelial cell proliferation and differentiation as well as neovascularization when delivered as a topical treatment, and by improving tissue elasticity and smooth muscle phenotype when delivered via injection. Using a validated preclinical model of GSM and an array of diverse multi-scale tools, we will comprehensively characterize alterations in vaginal structural and functional properties due to menopause. We will then create a novel biomaterial designed specifically to women’s health and assess its efficacy in treating GSM – a chronic and understudied condition that negatively impact lives of millions of women world-wide.