Matrix viscoelasticity drives cell cluster formation to counteract cellular senescence†

Abstract

During tissue repair, stress-induced cellular senescence represents a critical factor that impedes the regenerative potential of tissues. While the regulatory effects of matrix viscoelasticity on cellular behavior have been documented, their role and correlated mechanisms underlying cellular senescence remain unclear. In this study, we engineered a viscoelastic gel matrix exhibiting a storage modulus of approximately 3 kPa, with a tunable loss modulus ranging from 0 to 300 Pa by incorporating linear alginate and modulating the compactness of a polyacrylamide-based covalent network. Utilizing a UV-induced senescence model, we observed that increasing the matrix's viscoelasticity from 0 Pa to 300 Pa led to a significant reduction in the proportion of senescent cells, from 90.5% to 22.7%. Furthermore, cells cultured in these matrices exhibited a tendency to form cell aggregation, with the cell populations demonstrating a collective resistance to stresses. This indicated that viscoelastic materials would promote enhanced cellular interactions, thereby strengthening cellular resilience against UV-induced stresses. Furthermore, combined with microarray analysis, it was concluded that the presence of viscoelastic components activated the connexin 43 (Cx43)-modulated gap junction for cluster formation, thereby suppressing the senescence-associated signaling pathways, including Wnt/β-catenin, MAPK, NF-κB, and TGF-β. Additionally, the integrin–cytoskeleton–Yes-associated protein (YAP) signaling axis played an active role in delaying cell aging. These results provide novel insights into the regulatory role of viscoelastic materials in cellular senescence and offer a compelling foundation for the development of advanced biomaterials for tissue repair.