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体外冲击波对人骨骼肌细胞活力及基因表达的影响:一项体外对照研究

Exposure to radial extracorporeal shock waves modulates viability and gene expression of human skeletal muscle cells: a controlled in vitro study.

作者信息

Mattyasovszky Stefan G, Langendorf Eva K, Ritz Ulrike, Schmitz Christoph, Schmidtmann Irene, Nowak Tobias E, Wagner Daniel, Hofmann Alexander, Rommens Pol M, Drees Philipp

机构信息

Department of Orthopedics and Traumatology, University Medical Centre of the Johannes Gutenberg-University of Mainz, Langenbeckstraße 1, 55131, Mainz, Germany.

Extracorporeal Shock Wave Research Unit, Department of Anatomy II, Ludwig-Maximilians-University of Munich, Munich, Germany.

出版信息

J Orthop Surg Res. 2018 Apr 6;13(1):75. doi: 10.1186/s13018-018-0779-0.

Abstract

BACKGROUND

Recent clinical and animal studies have shown that extracorporeal shock wave therapy has a promoting influence on the healing process of musculoskeletal disorders. However, the underlying biological effects of extracorporeal shock wave therapy on human skeletal muscle cells have not yet been investigated.

METHODS

In this study, we investigated human skeletal muscle cells after exposure to radial extracorporeal shock waves in a standardized in vitro setup. Cells were isolated from muscle specimens taken from adult patients undergoing spine surgery. Primary muscle cells were exposed once or twice to radial extracorporeal shock waves in vitro with different energy flux densities. Cell viability and gene expression of the paired box protein 7 (Pax7), neural cell adhesion molecule (NCAM), and myogenic factor 5 (Myf5) and MyoD as muscle cell markers were compared to non-treated muscle cells that served as controls.

RESULTS

Isolated muscle cells were positive for the hallmark protein of satellite cells, Pax7, as well as for the muscle cell markers NCAM, MyoD, and Myf5. Exposure to radial extracorporeal shock waves at low energy flux densities enhanced cell viability, whereas higher energy flux densities had no further significant impact. Gene expression analyses of muscle specific genes (Pax7, NCAM, Myf5, and MyoD) demonstrated a significant increase after single exposure to the highest EFD (4 bar, 0.19 mJ/mm) and after double exposure with the medium EFDs (2 and 3 bar; 0.09 and 0.14 mJ/mm, respectively). Double exposure of the highest EFD, however, results in a significant down-regulation when compared to single exposure with this EFD.

CONCLUSIONS

This is the first study demonstrating that radial extracorporal shock wave therapy has the potential to modulate the biological function of human skeletal muscle cells. Based on our experimental findings, we hypothesize that radial extracorporal shock wave therapy could be a promising therapeutic modality to improve the healing process of sports-related structural muscle injuries.

摘要

背景

近期的临床和动物研究表明,体外冲击波疗法对肌肉骨骼疾病的愈合过程具有促进作用。然而,体外冲击波疗法对人骨骼肌细胞的潜在生物学效应尚未得到研究。

方法

在本研究中,我们在标准化的体外实验装置中,对暴露于径向体外冲击波后的人骨骼肌细胞进行了研究。细胞取自接受脊柱手术的成年患者的肌肉标本。原代肌肉细胞在体外以不同的能量通量密度接受一次或两次径向体外冲击波照射。将成对盒蛋白7(Pax7)、神经细胞黏附分子(NCAM)、生肌因子5(Myf5)和肌肉细胞标记物MyoD的细胞活力和基因表达与未处理的作为对照的肌肉细胞进行比较。

结果

分离出的肌肉细胞对卫星细胞的标志性蛋白Pax7以及肌肉细胞标记物NCAM、MyoD和Myf5呈阳性。在低能量通量密度下暴露于径向体外冲击波可提高细胞活力,而较高的能量通量密度则没有进一步的显著影响。对肌肉特异性基因(Pax7、NCAM、Myf5和MyoD)的基因表达分析表明,单次暴露于最高能量通量密度(4巴,0.19毫焦/平方毫米)后以及两次暴露于中等能量通量密度(分别为2和3巴;0.09和0.14毫焦/平方毫米)后,基因表达显著增加。然而,与单次暴露于该能量通量密度相比,最高能量通量密度的两次暴露导致显著下调。

结论

这是第一项证明径向体外冲击波疗法有可能调节人骨骼肌细胞生物学功能的研究。基于我们的实验结果,我们推测径向体外冲击波疗法可能是一种有前景的治疗方式,可改善与运动相关的结构性肌肉损伤的愈合过程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c19a/5889540/633bdc4ca0fd/13018_2018_779_Fig1_HTML.jpg

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