Volume 8, Issue 3 (12-2020)
2020, 8(3): 23-37 |
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1- Damghan University, Faculty of Biology, Cellular & Molecular Department
Abstract: (3386 Views)
Background & Aim: Extremely low-frequency electromagnetic field (ELF-EMF) and betaine are safe factors in bone fracture repair. This study aimed to compare the effects of these two stimuli on osteogenic differentiation of human adipose stem cells (hADSCs).
Methods: After obtaining written informed consent, cells were extracted from abdominal adipose tissue and then cultured in vitro until the third passage. Experimental groups were as follows: cells cultured in α-MEM and serum (control), in osteogenesis differentiation medium containing 10 mM Betaine (BET + OD), in α-MEM containing Betaine (BET), in OD and wave (50HZ and 1mT) (OD +EMF) and in the wave alone (EMF). Then, osteogenic differentiation was assessed by morphology, evaluating the qualitative and quantitative Alizarin red staining, alkaline phosphatase activity and real time PCR of RUNX2 and OCN genes.
Results: There was a significant increase in the formation of calcium deposits, alkaline phosphatase activity and also the expression of RUNX2 and OCN genes in OD + BET group compared to BET and OD + EMF groups. At the end of day 7, a significant decrease in alkaline phosphatase activity was observed in the EMF group compared to the OD + EMF and the BET groups. There was also a significant decrease in the expression of OCN gene in the EMF compared to the BET group.
Conclusion: In this study, synergistic effect of betaine and EMF on osteogenesis of hADSCs was demonstrated and this could be used for fracture repair and tissue engineering.
Methods: After obtaining written informed consent, cells were extracted from abdominal adipose tissue and then cultured in vitro until the third passage. Experimental groups were as follows: cells cultured in α-MEM and serum (control), in osteogenesis differentiation medium containing 10 mM Betaine (BET + OD), in α-MEM containing Betaine (BET), in OD and wave (50HZ and 1mT) (OD +EMF) and in the wave alone (EMF). Then, osteogenic differentiation was assessed by morphology, evaluating the qualitative and quantitative Alizarin red staining, alkaline phosphatase activity and real time PCR of RUNX2 and OCN genes.
Results: There was a significant increase in the formation of calcium deposits, alkaline phosphatase activity and also the expression of RUNX2 and OCN genes in OD + BET group compared to BET and OD + EMF groups. At the end of day 7, a significant decrease in alkaline phosphatase activity was observed in the EMF group compared to the OD + EMF and the BET groups. There was also a significant decrease in the expression of OCN gene in the EMF compared to the BET group.
Conclusion: In this study, synergistic effect of betaine and EMF on osteogenesis of hADSCs was demonstrated and this could be used for fracture repair and tissue engineering.
Type of Study: Research |
Subject:
Special
Received: 2020/09/22 | Accepted: 2020/12/13 | Published: 2021/01/3
Received: 2020/09/22 | Accepted: 2020/12/13 | Published: 2021/01/3
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