Blockade of receptors of advanced glycation end products ameliorates diabetic osteogenesis of adipose‐derived stem cells through <scp>DNA</scp> methylation and Wnt signalling pathway

Abstract

Abstract Objectives Diabetes mellitus‐related osteoporosis is caused by the imbalance between bone absorption and bone formation. Advanced glycation end products ( AGE s) are considered a cause of diabetic osteoporosis. Although adipose‐derived stem cells ( ASC s) are promising adult stem cells in bone tissue regeneration, the ability of osteogenesis of ASC s in diabetic environment needs to explore. This study aimed to investigate the influence of AGE s on the osteogenic potential of ASC s and to explore the signalling pathways involved in its effect. Materials and methods ASC s were isolated from inguinal fat and cultured in osteogenic media with or without AGE s and FPS ‐ ZM 1, an inhibitor of receptor for AGE s ( RAGE ). Alizarin red‐S, Oil Red‐O and Alcian blue staining were used to confirm osteogenic, adipogenic and chondrogenic potential of ASC s, respectively. Immunofluorescence, western blotting and real‐time PCR were used to measure changes in markers of osteogenic differentiation, DNA methylation and Wnt signalling. Results The multipotentiality of ASC s was confirmed. Treated with AGE s, OPN and RUNX 2 expressions of ASC s were reduced and there was a noticeable loss of mineralization, concomitant with an increase in the expression of RAGE , 5‐ MC , DNMT 1 and DNMT 3a. AGE s treatment also led to a loss of Wnt signalling pathway markers, including β‐Catenin and LEF 1, with an increase in GSK ‐3β. Treatment with the RAGE inhibitor, FPS ‐ ZM 1, rescued AGE s‐induced loss of osteogenic potential, modulated DNA methylation and upregulated Wnt signalling in ASC s. Conclusions Our results demonstrate that AGE s‐ RAGE signalling inhibits the osteogenic potential of ASC s under osteoinductive conditions by modulating DNA methylation and Wnt signalling. FPS ‐ ZM 1 can rescue the negative effects of AGE s and provide a possible treatment for bone tissue regeneration in patients with diabetic osteoporosis.