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Minoxidil decreases collagen I deposition and tissue-like contraction in clubfoot-derived cells: a way to improve conservative treatment of relapsed clubfoot?
Connective Tissue Research ( IF 2.9 ) Pub Date : 2020-09-20 , DOI: 10.1080/03008207.2020.1816992
Jarmila Knitlova 1 , Martina Doubkova 1, 2 , Martin Plencner 1 , David Vondrasek 1, 3 , Adam Eckhardt 1 , Martin Ostadal 4 , Jana Musilkova 1 , Lucie Bacakova 1 , Tomas Novotny 2, 5
Affiliation  

ABSTRACT

Aim

Clubfoot is a congenital deformity affecting the musculoskeletal system, resulting in contracted and stiff tissue in the medial part of the foot. Minoxidil (MXD) has an inhibitory effect on lysyl hydroxylase, which influences the quality of extracellular matrix crosslinking, and could therefore be used to reduce the stiffness and to improve the flexibility of the tissue. We assessed the in vitro antifibrotic effects of minoxidil on clubfoot-derived cells.

Methods

Cell viability and proliferation were quantified by xCELLigence, MTS, and LIVE/DEAD assays. The amount of collagen I deposited into the extracellular matrix was quantified using immunofluorescence with subsequent image segmentation analysis, hydroxyproline assay, and Second Harmonic Generation imaging. Extracellular matrix contraction was studied in a 3D model of cell-populated collagen gel lattices.

Results

MXD concentrations of 0.25, 0.5, and 0.75 mM inhibited the cell proliferation in a concentration-dependent manner without causing a cytotoxic effect. Exposure to ≥0.5 mM MXD resulted in a decrease in collagen type I accumulation after 8 and 21 days in culture. Changes in collagen fiber assembly were observed by immunofluorescence microscopy and nonlinear optical microscopy (second harmonic generation). MXD also inhibited the contraction of cell-populated collagen lattices (0.5 mM by 22%; 0.75 mM by 28%).

Conclusions

Minoxidil exerts an in vitro inhibitory effect on the cell proliferation, collagen accumulation, and extracellular matrix contraction processes that are associated with clubfoot fibrosis. This study provides important preliminary results demonstrating the potential relevance of MXD for adjuvant pharmacological therapy in standard treatment of relapsed clubfoot.



中文翻译:

米诺地尔减少马蹄足衍生细胞中胶原蛋白 I 的沉积和组织样收缩:改善复发性马蹄足保守治疗的方法?

摘要

目标

马蹄足是一种影响肌肉骨骼系统的先天性畸形,导致足部内侧组织收缩和僵硬。米诺地尔(MXD)对赖氨酰羟化酶有抑制作用,影响细胞外基质交联的质量,因此可用于降低组织的刚度和提高组织的柔韧性。我们评估了米诺地尔对马蹄足衍生细胞的体外抗纤维化作用。

方法

通过 xCELLigence、MTS 和 LIVE/DEAD 测定对细胞活力和增殖进行量化。使用免疫荧光以及随后的图像分割分析、羟脯氨酸测定和二次谐波成像来量化沉积到细胞外基质中的胶原蛋白 I 的量。在细胞填充胶原凝胶晶格的 3D 模型中研究了细胞外基质收缩。

结果

0.25、0.5 和 0.75 mM 的 MXD 浓度以浓度依赖性方式抑制细胞增殖,而不会引起细胞毒性作用。在培养 8 天和 21 天后,暴露于 ≥0.5 mM MXD 导致 I 型胶原蛋白积累减少。通过免疫荧光显微镜和非线性光学显微镜(二次谐波产生)观察胶原纤维组装的变化。MXD 还抑制了细胞填充的胶原蛋白晶格的收缩(0.5 mM 降低 22%;0.75 mM 降低 28%)。

结论

米诺地尔对与马蹄足纤维化相关的细胞增殖、胶原蛋白积累和细胞外基质收缩过程具有体外抑制作用。这项研究提供了重要的初步结果,证明了 MXD 在复发性马蹄足标准治疗中辅助药物治疗的潜在相关性。

更新日期:2020-09-20
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