Comparing the effects of betaine and Pulsed Electromagnetic Field on the process of osteogenesis in the presence and absence of osteogenic differentiation medium
Comparing the effects of betaine and Pulsed Electromagnetic Field on the process of osteogenesis in the presence and absence of osteogenic differentiation medium
Tayebeh Sadat Tabatabai,1,*Tahereh Sadat Tabatabaie,2
1. Student Research Committee, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran 2. Student Research Committee, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran
Introduction: Human adipose mesenchymal stem cells (hAdMSCs) are frequently used in tissue engineering and regenerative medicine. Two secure treatments for bone fracture repair are betaine and pulsed electromagnetic fields (PEMF).
Methods: After receiving written permission, abdominal fat was used to remove mesenchymal stem cells, and flow cytometry was used to verify the stemness of the cells. Cells were cultured in -MEM medium with 10% serum as a negative control, in bone differentiation medium as a positive control, in bone differentiation medium and betaine (BET+OD), in betaine (BET), in the bone differentiation environment and waves (OD+ wave), and cells exposed to waves (wave). In this study, cells were cultured for 14 days (8 hours a day) with a 10 mM dosage of betaine and a sinusoidal electromagnetic field with a frequency of 50 HZ and 1 mT intensity. Alizarin red staining, alkaline phosphatase activity, and cell shape were used to assess osteogenic differentiation following treatment. Real-time PCR was used to assess bone gene expression.
Results: Both quantitative and qualitative alizarin red staining results demonstrated a significant reduction in calcium deposition in the OD+wave and wave groups compared to the positive control group, but no significant difference was seen between the other groups. Additionally, the positive control group demonstrated a much higher level of alkaline phosphatase activity than the negative control group. Comparing the BET, OD+wave, and wave groups to the positive control group, a significant reduction in the amount of calcium deposits was seen. In comparison to the negative control group, there was a considerable increase in the expression of the Runx2 and osteocalcin genes in the positive control group. In comparison to the positive control group, the expression level of these genes in the BET group was considerably lower. In comparison to the positive control group, a significant decrease was also seen in the OD+wave and wave groups.
Conclusion: Osteogenic effects of the osteogenic differentiation medium have decreased when betaine and waves are combined with it, whereas the osteogenic effects of the osteogenic differentiation medium alone have increased.
Keywords: human adipose mesenchymal stem cells, Pulsed Electromagnetic Field, betaine