Tracing Cellular Senescence in Bone: Time‐Dependent Changes in Osteocyte Cytoskeleton Mechanics and Morphology

Abstract Aging‐related bone loss significantly impacts the growing elderly population globally, leading to debilitating conditions such as osteoporosis. Senescent osteocytes play a crucial role in the aging process of bone. This longitudinal study examines the impact of continuous local and paracrine exposure to senescence‐associated secretory phenotype (SASP) factors on biophysical and biomolecular markers in osteocytes. Significant cytoskeletal stiffening in irradiated (IR) osteocytes are found, accompanied by expansion of F‐actin areas and a decline in dendritic integrity. These changes, correlating with alterations in pro‐inflammatory cytokine levels and osteocyte‐specific gene expression, support the reliability of biophysical markers for identifying senescent osteocytes. Notably, local accumulation of SASP factors have a more pronounced impact on osteocyte biophysical properties than paracrine effects, suggesting that the interplay between local and paracrine exposure can substantially influence cellular aging. This study underscores the importance of osteocyte mechanical and morphological properties as biophysical markers of senescence, highlighting their time dependence and differential effects of local and paracrine SASP exposure. Collectively, the investigation into biophysical senescence markers offers unique and reliable functional hallmarks for the non‐invasive identification of senescent osteocytes, providing insights that can inform therapeutic strategies to mitigate aging‐related bone loss.