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体外冲击波触发人脂肪来源干细胞的腱细胞分化。

Extracorporeal shock waves trigger tenogenic differentiation of human adipose-derived stem cells.

机构信息

a Department of Medical Sciences , University of Turin , Turin , Italy.

b Plastic Surgery Unit , University of Turin , Turin , Italy.

出版信息

Connect Tissue Res. 2018 Nov;59(6):561-573. doi: 10.1080/03008207.2018.1424147. Epub 2018 Jan 9.

Abstract

PURPOSES

Incomplete tendon healing impairs the outcome of tendon ruptures and tendinopathies. Human Adipose-derived Stem Cells (hASCs) are promising for tissue engineering applications. Extracorporeal Shock Waves (ESW) are a leading choice for the treatment of several tendinopathies. In this study, we investigated the effects of ESW treatment and tenogenic medium on the differentiation of hASCs into tenoblast-like cells.

MATERIALS AND METHODS

hASCs were treated with ESW generated by a piezoelectric device and tenogenic medium. Quantitative real-time PCR was used to check the mRNA expression levels of tenogenic transcription factors, extracellular matrix proteins, and integrins. Western blot and immunofluorescence were used to detect collagen 1 and fibronectin. Collagen fibers were evaluated by Masson staining. Calcium deposition was assessed by Alizarin Red staining.

RESULTS

The combined treatment improved the expression of the tendon transcription factors scleraxis and eyes absent 2, and of the extracellular matrix proteins fibronectin, collagen I, and tenomodulin. Cells acquired elongated and spindle shaped fibroblastic morphology; Masson staining revealed the appearance of collagen fibers. Finally, the combined treatment induced the expression of alpha 2, alpha 6, and beta 1 integrin subunits, suggesting a possible role in mediating ESW effects.

CONCLUSIONS

ESW in combination with tenogenic medium improved the differentiation of hASCs toward tenoblast-like cells, providing the basis for ESW and hASCs to be used in tendon tissue engineering.

摘要

目的

不完全的肌腱愈合会损害肌腱断裂和肌腱病的治疗效果。人脂肪来源干细胞(hASCs)是组织工程应用的有前途的选择。体外冲击波(ESW)是治疗多种肌腱病的首选方法。在这项研究中,我们研究了 ESW 治疗和肌腱形成培养基对 hASCs 向肌腱样细胞分化的影响。

材料和方法

用压电装置产生的 ESW 和肌腱形成培养基处理 hASCs。实时定量 PCR 用于检查肌腱转录因子、细胞外基质蛋白和整合素的 mRNA 表达水平。Western blot 和免疫荧光用于检测胶原蛋白 1 和纤维连接蛋白。用 Masson 染色评估胶原纤维。用茜素红染色评估钙沉积。

结果

联合治疗可提高肌腱转录因子 Scleraxis 和 Eyes Absent 2 的表达,以及细胞外基质蛋白纤维连接蛋白、胶原蛋白 I 和肌腱调蛋白的表达。细胞呈现出拉长的梭形纤维细胞形态;Masson 染色显示出胶原纤维的出现。最后,联合治疗诱导了 alpha 2、alpha 6 和 beta 1 整合素亚基的表达,表明其可能在介导 ESW 效应中发挥作用。

结论

ESW 联合肌腱形成培养基可促进 hASCs 向肌腱样细胞分化,为 ESW 和 hASCs 在肌腱组织工程中的应用提供了基础。

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