Department of Cellular and Molecular Biology, School of Biology and Institute of Biological Sciences, Damghan University, Damghan, Iran.
Department of Medical Sciences, School of Paramedical, Shahroud University of Medical Sciences, Shahroud, Iran.
In Vitro Cell Dev Biol Anim. 2021 Apr;57(4):468-476. doi: 10.1007/s11626-021-00558-6. Epub 2021 Mar 26.
Human adipose tissue-derived mesenchymal stem cells (hADSCs) due to easy extraction, relative abundance, in vitro expansion and differentiation potential, frozen storage capability, and ability to secrete cytokines, compared to other stem cells, are appropriate candidate in regenerative medicine. Extremely low-frequency electromagnetic fields (ELF-EMF) and betaine are two safe factors in bone lesions repair. This study was designed to assess the osteogenic differentiation potential of these factors on hADSCs. The samples were collected from women undergoing liposuction after obtaining written consent. The hADSCs were extracted and treated with osteogenesis differentiation medium (OD) as the positive control, with OD and betaine (BET group), with OD and EMF (EMF group), and with OD and betaine and EMF (BET+EMF group) for 21 d; the negative control consisted of cells without treatment. Betaine 10 mM and EMF with 50-Hz frequency, 1-mT intensity (8 h daily), and in the form of sinus wave were used. Osteogenic differentiation was evaluated by Alizarin Red staining, alkaline phosphatase activity, calcium deposition, and real-time PCR. A significant increase in calcium deposition in the BET+EMF group was observed compared to the other groups. The activity of alkaline phosphatase in the positive control and BET groups was increased significantly compared to EMF and BET + EMF groups and a significant increase of this enzyme activity in the BET + EMF compared to EMF group was observed. The expression of RUNX2 and OCN genes in the EMF-treated groups were significantly reduced compared to the non-EMF-treated groups, and BET+EMF showed a significant increase of RUNX2 gene expression as compared the EMF group. The ELF-EMF leads to a decrease in the osteogenic differentiation and the expression RUNX2 and OCN genes in hADSCs. But osteogenic differentiation and RUNX2 gene expression were increased post-induction by betaine. The synergic effect of betaine and EMF on the osteogenic differentiation and related genes expression of hADSCs was higher than EMF.
人脂肪组织来源的间充质干细胞（hADSCs）由于易于提取、相对丰富、体外扩增和分化潜能、冷冻储存能力以及分泌细胞因子的能力，与其他干细胞相比，是再生医学的合适候选物。极低频率电磁场（ELF-EMF）和甜菜碱是骨病变修复的两种安全因素。本研究旨在评估这些因素对 hADSCs 成骨分化潜能的影响。样本取自接受吸脂术的女性，在获得书面同意后收集。将 hADSCs 提取并在成骨分化培养基（OD）中处理作为阳性对照，在 OD 和甜菜碱（BET 组）、OD 和 EMF（EMF 组）以及 OD 和甜菜碱和 EMF（BET+EMF 组）中处理 21 天；阴性对照组由未经处理的细胞组成。使用 10mM 甜菜碱和 50Hz 频率、1mT 强度（每天 8 小时）的正弦波形式的 EMF。通过茜素红染色、碱性磷酸酶活性、钙沉积和实时 PCR 评估成骨分化。与其他组相比，BET+EMF 组的钙沉积明显增加。与 EMF 和 BET+EMF 组相比，阳性对照和 BET 组的碱性磷酸酶活性显著增加，并且 BET+EMF 组的碱性磷酸酶活性显著高于 EMF 组。与未处理组相比，EMF 处理组的 RUNX2 和 OCN 基因表达显著降低，而 BET+EMF 组与 EMF 组相比，RUNX2 基因表达显著增加。ELF-EMF 导致 hADSCs 成骨分化和 RUNX2 和 OCN 基因表达减少。但甜菜碱诱导后，成骨分化和 RUNX2 基因表达增加。甜菜碱和 EMF 对 hADSCs 成骨分化和相关基因表达的协同作用高于 EMF。
