Osteoarthritis (OA), the leading cause of disability, is driven by articular cartilage degeneration and inflammation. Mechanosensitive ion channels PIEZO1 and PIEZO2 have been implicated in OA progression via calcium signaling in chondrocytes. This study investigates the role of conditional Piezo channel knockout (cKO) in modulating calcium signaling and senescence in OA, particularly under the supra-physiologic inflammatory condition of a high-fat diet (HFD). Using calcium imaging, we found that Piezo1 cKO and dual Piezo1/2 cKO significantly reduced Yoda1-induced calcium signaling, while Piezo2 cKO alone had no effect. Under HFD conditions, Piezo1 cKO showed a non-significant trend toward reduced calcium influx, potentially due to high inflammatory burden or altered Piezo1 expression. Immunohistochemical analysis revealed that Piezo1 cKO significantly attenuated expression of the senescence marker p16, even under HFD conditions, suggesting downstream protective effects. In contrast, p21 expression patterns were inconsistent and appeared influenced by metabolic or stress-related pathways outside of senescence. These findings identify Piezo1 as a key contributor to OArelated mechanotransduction and senescence, supporting its potential as a therapeutic target and warranting further investigation into combinatorial strategies for OA intervention.