Effect of N,N-dimethylimidedicarboimide diamide (metformin hydrochloride) on osteogenic differentiation of mesenchymal stem cells of epicardial adipose tissue

Background. To date, no specific methods for the prevention and treatment of coronary artery calcification have been developed. A literature analysis demonstrates that the use of certain pharmacological agents in patients with coronary artery disease significantly reduces the risk of developing this pathological condition. Drugs whose mechanism of action involves regulating cellular metabolism may serve as a promising tool for managing osteogenic differentiation of multipotent cells in the cardiovascular system and, consequently, preventing the development of massive calcification of cardiac vessels.

The aim. To evaluate the effect of different concentrations of N,N-dimethylimidicarboimide diamide (DMDC) on the osteogenic differentiation of mesenchymal stem cells (MSCs) derived from epicardial adipose tissue (EAT).

Methods. MSCs were isolated from EAT biopsies collected during coronary artery bypass grafting. Cell cultures were passaged to the third passage, after which immunophenotyping and osteogenic differentiation were performed. For MSC differentiation, osteogenic medium was used, either native or supplemented with 10, 50, and 100 μmol DMDC. Osteogenesis was assessed by measuring protein levels (RUNX2, osteopontin, alkaline phosphatase) in cell culture supernatants and staining the cell cultures with subsequent determination of staining intensity.

Results. Throughout differentiation, cells incubated in medium supplemented with 50 μmol DMDC demonstrated high levels of osteogenic protein synthesis. Osteoblasts differentiated with 50 μmol DMDC synthesized 1.1 times more calcium than MSCs incubated with other DMDC concentrations, and 1.3 times more than in control cultures (p = 0.001).

Conclusion. Adding DMDC to the differentiation medium for EAT-MSCs from patients with coronary artery disease significantly enhances the synthesis of key markers of osteoblast differentiation. A particularly pronounced effect was observed at a concentration of 50 μmol of the drug.

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