Reduction of mechanical loading in tendons induces heterotopic ossification and activation of the β-catenin signaling pathway.
Reduction of mechanical loading in tendons induces heterotopic ossification and activation of the β-catenin signaling pathway.
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肌腱机械负荷的减少诱导异位骨化和 β-连环蛋白信号通路的激活
DOI：
10.1016/j.jot.2021.03.004
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发表时间：
2021-07
影响因子：
6.6
通讯作者：
Zheng MH
中科院分区：
文献类型：
--
作者：
Wang T;Chen P;Chen L;Zhou Y;Wang A;Zheng Q;Mitchell CA;Leys T;Tuan RS;Zheng MH
肌腱是使关节运动的力传递组织。过度的机械负荷通常被认为是导致肌腱病的主要因素，然而，越来越多的证据支持这样的假设，即过载造成胶原纤维的微损伤，导致损伤部位内细胞群的局部负荷降低。异位骨化是晚期肌腱病的并发症，它会严重影响肌腱的力学性能和内环境稳定。在这里，我们研究机械欠载对肌腱骨化的影响，并探讨其潜在的分子机制。将兔跟腱分离后置于低负荷环境（3%循环拉伸染色，0.25Hz，8h/d）中培养10、15和20 d。使用分离的腱源性干细胞（TDSC）生成3D构建体，培养并经受欠载环境6天。进行组织学评估以评价3D构建体的结构;采用qPCR和免疫组织化学研究TDSC分化，并通过Western印迹研究β-连环蛋白信号通路。机械测试用于确定肌腱承受力产生的能力。在欠载环境中长时间培养的肌腱表现出进行性异位骨化和生物力学强度降低。qPCR显示，在欠载环境中培养的3D TDSC构建体表现出几种成骨基因的表达增加：这些基因包括RUNX 2、ALP和骨钙素，与生腱分化标志物（巩膜轴和腱调节蛋白）相比。免疫组织化学分析进一步证实了在负荷不足的3D TDSC构建体中高的骨钙素产生。TDSC构建体的蛋白质印迹显示β-连环蛋白积累和易位与Ser 552磷酸化的增加和Ser 33磷酸化的减少相关。这些发现揭示了由于损伤部位TDSC负荷不足导致肌腱病异位骨化的潜在机制，并且β-连环蛋白可能是治疗肌腱异位骨化的潜在靶点。肌腱异位骨化严重影响患者的生活质量，尤其是有运动史的患者。本研究揭示了与机械负荷有关的肌腱异位骨化的可能机制。本研究为制定预防和治疗肌腱异位骨化的机械刺激方案提供了可能。
Tendons are the force transferring tissue that enable joint movement. Excessive mechanical loading is commonly considered as a primary factor causing tendinopathy, however, an increasing body of evidence supports the hypothesis that overloading creates microdamage of collagen fibers resulting in a localized decreased loading on the cell population within the damaged site. Heterotopic ossification is a complication of late stage tendinopathy, which can significantly affect the mechanical properties and homeostasis of the tendon. Here, we the examine the effect of mechanical underloading on tendon ossification and investigate its underlying molecular mechanism. Rabbit Achilles tendons were dissected and cultured in an underloading environment (3% cyclic tensile stain,0.25 Hz, 8 h/day) for either 10, 15 or 20 days. Using isolated tendon-derived stem cells (TDSCs) 3D constructs were generated, cultured and subjected to an underloading environment for 6 days. Histological assessments were performed to evaluate the structure of the 3D constructs; qPCR and immunohistochemistry were employed to study TDSC differentiation and the β-catenin signal pathway was investigated by Western blotting. Mechanical testing was used to determine ability of the tendon to withstand force generation. Tendons cultured for extended times in an environment of underloading showed progressive heterotopic ossification and a reduction in biomechanical strength. qPCR revealed that 3D TDSCs constructs cultured in an underloading environment exhibited increased expression of several osteogenic genes: these include RUNX2, ALP and osteocalcin in comparison to tenogenic differentiation markers (scleraxis and tenomodulin). Immunohistochemical analysis further confirmed high osteocalcin production in 3D TDSCs constructs subject to underloading. Western blotting of TDSC constructs revealed that β-catenin accumulation and translocation were associated with an increase in phosphorylation at Ser552 and decrease phosphorylation at Ser33. These findings unveil a potential mechanism for heterotopic ossification in tendinopathy due to the underloading of TDSCs at the damage sites, and also that β-catenin could be a potential target for treating heterotopic ossification in tendons. Tendon heterotopic ossification detrimentally affect quality of life especially for those who has atheletic career. This study reveals the possible mechanism of heterotpic ossification in tendon related to mechanical loading. This study provided the possible to develop a mechanical stimulation protocol for preventive and therapeutic purpose for tendon heterotopic ossification.
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影响因子：
2.8
作者：
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影响因子：
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作者：
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通讯作者：
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