PROJECT SUMMARY An individuals’ biological sex significantly affects their ability to repair and regenerate tissue. A clear example of this is the reduced ability for women to heal and regenerate new, healthy tissue after menopause, which results from a significant loss of sex hormone signaling. This reduction in hormone levels disproportionately enhances the risk for many degenerative diseases including osteoporosis, osteoarthritis, cardiovascular disease, and degenerative brain diseases in which the rate of tissue breakdown exceeds the rate of tissue repair. While it is known that several factors contribute to sex differences in tissue repair including biomechanics, nutrition, physical activity level and sex hormones, the interplay between these parameters is not well understood. Specifically, it is unknown how the native sex differences in tissue structure and the resulting differences in mechanical function dictate cell phenotype and behavior and how this effect interacts with estrogen signaling to overall control tissue repair. Thus, a fundamental, mechanistic understanding of how a cell responds to the spatial and mechanical cues of its environment while mediating estrogen signaling is critical to understand why sex differences occur in tissue repair and homeostasis and for future patient-centered repair and regeneration strategies. The overall goal of our research program aims to develop biomaterial tools to interrogate sex differences in tissue repair and homeostasis. Theme 1: Do male and female MSCs respond to spatial and mechanical properties of the cell microenvironment differently? There is evidence in many tissues that extracellular matrix structure, organization, and resulting function differs between age-matched males and females. However, there are no studies showing how this affects cell response. Biomaterials engineered to mimic both the fibrous properties of structural collagens and the viscoelastic properties of proteoglycans in the native extracellular matrix will be used to assess sex differences in cell response to controlled changes in matrix properties. Theme 2: How does estrogen presentation to the cell affect downstream transcription and behavior? While estrogen is known to play a role on cell processes, these results are dependent on the concentration and the temporal presentation of estrogen to the cell. To address this limitation, we will use concentration gradient generator microchips to quickly and accurately determine the effect of estrogen concentration and timing on cell transcriptional activity. Theme 3: Can we engineer biomaterial systems to control release and presentation of estrogen to the cells? Release rates in a range of hours to months will be controlled by modulating diffusion out of the biomaterials via material chemistry and architecture. The ability to control the rate of release and localize to a specific tissue in the body is critical to promote the estrogen effects at the site while reducing the ...