Synergistic effect of metformin and vitamin D3 on osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells under high d-glucose conditions

Nhi Nguyen-Yen Ha; Thi Kim Tan Huynh; Ngoc Uyen Phuong Phan; Thi-Hiep Nguyen; Long Binh Vong; Nhu-Thuy Trinh

doi:10.1016/j.reth.2023.12.003

Synergistic effect of metformin and vitamin D₃ on osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells under high d-glucose conditions

Regen Ther. 2023 Dec 26:25:147-156. doi: 10.1016/j.reth.2023.12.003. eCollection 2024 Mar.

Authors

Nhi Nguyen-Yen Ha^{1

2

3}, Thi Kim Tan Huynh^{2

3}, Ngoc Uyen Phuong Phan⁴, Thi-Hiep Nguyen^{1

3}, Long Binh Vong^{1

3}, Nhu-Thuy Trinh^{1

3}

Affiliations

¹ School of Biomedical Engineering, International University, 700000 HCMC, Viet Nam.
² Faculty of Biology and Biotechnology, University of Science, 700000 HCMC, Viet Nam.
³ Vietnam National University-Ho Chi Minh City (VNU-HCMC), 700000 HCMC, Viet Nam.
⁴ Biotechnology Center of Ho Chi Minh City, 700000 HCMC, Viet Nam.

Abstract

Introduction: Vitamin D₃ plays a vital role in bone health, with low levels of vitamin D₃ being related to skeletal fragility, fractures, and metabolic disorders such as diabetes. Metformin is known as an antihyperglycemic agent for regulating blood sugar. A correlation between diabetes mellitus and osteoporosis is attracting considerable interest, and research to find the prevention and treatment is gradually being studied. In this study, we investigated the effect of metformin and vitamin D₃ on osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells (AT-MSCs) under high d-glucose concentrations and optimized by combining vitamin D₃ and metformin in the process.

Methods: ROS production of AT-MSCs under high d-glucose conditions was measured by DCFH-DA assay. The differentiated AT-MSCs were analyzed by Alizarin Red S staining and optical density measurement. The investigation involved the examination of osteogenic master genes' expressions using quantitative reverse transcription polymerase chain reaction (qRT-PCR) techniques.

Results: Interestingly, the results have shown that human AT-MSCs will exhibit high ROS accumulation and low osteogenic differentiation capabilities, indicated by low calcium deposition, as well as low expression of indicative genes such as ALP, Runx-2 under high d-glucose conditions. The combination of vitamin D₃ and metformin remarkedly accelerated the osteogenic differentiation of AT-MSCs under high d-glucose concentrations more effectively than the administration of either agent.

Conclusions: This study partially explains an aspect of an in vitro model for pre-clinical drug screening for osteoporosis-related diabetic pathological mechanisms, which can be applied for further research on the prevention or treatment of osteoporosis in diabetic patients.

Keywords: AT-MSCs; Diabetes mellitus; Mesenchymal stem cells; Metformin; Osteogenic differentiation; Osteoporosis; Vitamin D.