Abstract
Type 2 diabetes mellitus (T2D) is a globally prevalent metabolic disorder increasingly recognized for its detrimental effects on skeletal integrity. Patients with T2D exhibit increased bone fragility, which results in a heightened risk of bone traumas, yet the underlying mechanisms remain poorly understood. Chronic hyperglycemia and the production of advanced glycation end-products (AGEs) are thought to play a critical role in skeletal deterioration by activating the receptor for advanced glycation end-products (RAGE) pathway, promoting inflammation and bone resorption. Despite this association, the cellular responses of osteocytes to varying glycemic conditions have not been fully characterized in vitro. This study investigates the OCY454-12H cell line, an osteocyte-like, murine-derived conditionally immortalized model with enhanced SOST expression, as an in vitro model for T2D associated bone dysfunction. Cells were exposed to varying glucose concentrations for 10 days to model hyperglycemic conditions. Then, changes in gene and protein expression of key biomarkers, SOST/sclerostin and RAGE, were quantified using qPCR and ELISA analyses across glucose conditions. Significant glucose-dependent increases in gene and protein expression were identified, with 22 mM and 34 mM glucose most closely matching biomarker profiles reported in T2D-associated bone dysfunction in vivo. These findings provide preliminary support for the OCY454-12H cell line as a platform for investigating hyperglycemia-induced osteocyte dysfunction and establish candidate glucose concentrations for future in vitro T2D bone models.