Metformin enhances osteogenic differentiation of stem cells from human exfoliated deciduous teeth through AMPK pathway

Abstract

Stem cells from human exfoliated deciduous teeth (SHEDs) are ideal seed cells in bone tissue engineering. As a first‐line antidiabetic drug, metformin has recently been found to promote bone formation. The purpose of this study was to investigate the effect of metformin on the osteogenic differentiation of SHEDs and its underlying mechanism. SHEDs were isolated from the dental pulp of deciduous teeth from healthy children aged 6 to 12, and their surface antigen markers of stem cells were detected by flow cytometry. The effect of metformin (10–200 μM) treatment on SHEDs cell viability, proliferation, and osteogenic differentiation was analyzed. The activation of adenosine 5′‐monophosphate‐activated protein kinase (AMPK) phosphorylation Thr172 (p‐AMPK) was determined by western blot assay. SHEDs were confirmed as mesenchymal stem cells (MSCs) on the basis of the expression of characteristic surface antigens. Metformin (10–200 μM) did not affect the viability and proliferation of SHEDs but significantly increased the expression of osteogenic genes, alkaline phosphatase activity, matrix mineralization, and p‐AMPK level expression in SHEDs. Compound C, a specific inhibitor of the AMPK pathway, abolished metformin‐induced osteogenic differentiation of SHEDs. Moreover, metformin treatment enhanced the expression of proangiogenic/osteogenic growth factors BMP2 and VEGF but reduced the osteoclastogenic factor RANKL/OPG expression in SHEDs. In conclusion, metformin could induce the osteogenic differentiation of SHEDs by activating the AMPK pathway and regulates the expression of proangiogenic/osteogenic growth factors and osteoclastogenic factors in SHEDs. Therefore, metformin‐pretreated SHEDs could be a potential source of seed cells during stem cell‐based bone tissue engineering.

Chitosan Hydrogel Supplemented with Metformin Promotes Neuron–like Cell Differentiation of Gingival Mesenchymal Stem Cells

“…After 21 days of induction, they were stained with Alizarin red, Oil Red O, and Alcian blue; the additional metformin had no effect on multidirectional differentiation, but it can still maintain the potential for multi–lineage differentiation. There is evidence that metformin can inhibit stem–cell aging through the AMPK pathway [ 36 ]; moreover, it can promote osteogenic differentiation of stem cells [ 37 ].…”

AMPK/mTOR Pathway Is Involved in Autophagy Induced by Magnesium-Incorporated TiO2 Surface to Promote BMSC Osteogenic Differentiation

“…AMPK has been reported to control the osteogenic differentiation of human MSCs through early mTOR inhibition-mediated autophagy [ 41 , 42 ]. The AMPK activator has been demonstrated to have a role in inducing osteogenic differentiation and mineralization of osteoblastic cell lines and bone marrow progenitor cells [ 43 , 44 , 45 , 46 ]. A recent study reported that magnesium released from biomaterials could activate the AMPK/mTOR signaling pathway, thus upregulating autophagy in osteosarcoma cells [ 35 ].…”

Therapeutic Potential of Stem Cells from Human Exfoliated Deciduous Teeth Infusion into Patients with Type 2 Diabetes Depends on Basal Lipid Levels and Islet Function

“…Considering that the types and doses of oral hypoglycemic drugs were not adjusted in the experimental period, there would have limited effects in terms of influencing the outcome of this study. Nevertheless, these drugs such as metformin may improve SHED function, as documented in vitro, 42,43 which might exert synergistic effects to alleviate T2DM. 44,45 Previous studies showed that MSC transplantation can significantly reduce the insulin dose needed by patients with diabetes, or even help them cease insulin injections.…”