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Short-wave enhances mesenchymal stem cell recruitment in fracture healing by increasing HIF-1 in callus.
Stem Cell Research & Therapy ( IF 7.5 ) Pub Date : 2020-09-07 , DOI: 10.1186/s13287-020-01888-0
Dongmei Ye 1 , Chen Chen 2 , Qiwen Wang 1, 3 , Qi Zhang 1 , Sha Li 1 , Hongwei Liu 1
Affiliation  

As a type of high-frequency electrotherapy, a short-wave can promote the fracture healing process; yet, its underlying therapeutic mechanisms remain unclear. To observe the effect of Short-Wave therapy on mesenchymal stem cell (MSC) homing and relative mechanisms associated with fracture healing. For in vivo study, the effect of Short-Wave therapy to fracture healing was examined in a stabilized femur fracture model of 40 SD rats. Radiography was used to analyze the morphology and microarchitecture of the callus. Additionally, fluorescence assays were used to analyze the GFP-labeled MSC homing after treatment in 20 nude mice with a femoral fracture. For in vitro study, osteoblast from newborn rats simulated fracture site was first irradiated by the Short-Wave; siRNA targeting HIF-1 was used to investigate the role of HIF-1. Osteoblast culture medium was then collected as chemotaxis content of MSC, and the migration of MSC from rats was evaluated using wound healing assay and trans-well chamber test. The expression of HIF-1 and its related factors were quantified by q RT-PCR, ELISA, and Western blot. Our in vivo experiment indicated that Short-Wave therapy could promote MSC migration, increase local and serum HIF-1 and SDF-1 levels, induce changes in callus formation, and improve callus microarchitecture and mechanical properties, thus speeding up the healing process of the fracture site. Moreover, the in vitro results further indicated that Short-Wave therapy upregulated HIF-1 and SDF-1 expression in osteoblast and its cultured medium, as well as the expression of CXCR-4, β-catenin, F-actin, and phosphorylation levels of FAK in MSC. On the other hand, the inhibition of HIF-1α was significantly restrained by the inhibition of HIF-1α in osteoblast, and it partially inhibited the migration of MSC. These results suggested that Short-Wave therapy could increase HIF-1 in callus, which is one of the crucial mechanisms of chemotaxis MSC homing in fracture healing.

中文翻译:

短波通过增加愈伤组织中的HIF-1增强骨折愈合中的间充质干细胞募集。

作为一种高频电疗,短波可以促进骨折愈合过程。然而,其潜在的治疗机制仍不清楚。观察短波疗法对间充质干细胞(MSC)归巢和骨折愈合相关机制的影响。对于体内研究,在稳定的40只SD大鼠股骨骨折模型中检查了短波疗法对骨折愈合的影响。射线照相术用于分析愈伤组织的形态和微结构。另外,在20只患有股骨骨折的裸鼠中治疗后,使用荧光测定法来分析GFP标记的MSC归巢。为了进行体外研究,首先用短波辐照来自新生大鼠模拟骨折部位的成骨细胞。靶向HIF-1的siRNA用于研究HIF-1的作用。然后收集成骨细胞培养基作为MSC的趋化成分,并使用伤口愈合试验和透孔室试验评估大鼠中MSC的迁移。通过q RT-PCR,ELISA和Western blot对HIF-1及其相关因子的表达进行定量。我们的体内实验表明,短波疗法可促进MSC迁移,增加局部和血清HIF-1和SDF-1的水平,诱导愈伤组织形成的变化,并改善愈伤组织的微结构和力学性能,从而加快愈合过程。骨折部位。而且,体外结果进一步表明,短波疗法上调了成骨细胞及其培养基中HIF-1和SDF-1的表达,以及CXCR-4,β-catenin,F-actin和磷酸化水平的表达。 MSC中FAK的名称。另一方面,HIF-1α的抑制作用明显受到成骨细胞中HIF-1α的抑制作用,部分抑制了MSC的迁移。这些结果表明,短波疗法可增加愈伤组织中的HIF-1,这是趋化性MSC归巢在骨折愈合中的关键机制之一。
更新日期:2020-09-08
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