The effects of primary cilia-mediated mechanical stimulation on nestin+-BMSCs during bone-tendon healing.

Huabin Chen, Han Xiao, Bing Wu, Xin Shi, Changbiao Guan, Jianzhong Hu, Tao Zhang, Hongbin Lu
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Abstract

Introduction: Mechanical stimulation has been proven to promote bone-tendon interface (BTI) healing, but the mechanism remains unclear.

Objective: To investigate the effects of mechanical stimulation on the biological behavior of nestin+-bone mesenchymal stem cells (BMSCs) during the BTI healing, and to reveal the mechanisms of mechanical stimulation affecting BTI healing by primary cilia on the nestin+-BMSCs.

Methods: Transgenic tracing mice (nestin creERT2:: IFT88fl/fl/ROSA26 YFP) with primary cilia on nestin+-BMSCs conditioned knocked out were constructed, and the littermates (nestin creERT2:: ROSA26 YFP) with normal cilia on nestin+-BMSCs were the control. After establishing mouse supraspinatus insertion injury models, samples were collected at week-2 (n = 5 per group), 4 and 8 (n = 15 per group, respectively). In vivo, the repair efficiency was evaluated by histology, imaging, biomechanics, and the migration of nestin+-BMSCs, detected by immunofluorescence staining. In vitro, nestin+ BMSCs were sorted and stimulated by tensile force to study the mechanisms of primary cilium-mediated mechanosensitive basis.

Results: Mechanical stimulation (MS) accelerated the recruitment of nestin+-BMSCs and promoted osteogenic and chondrogenic capacity. Histological, imaging and biomechanical results showed that the BTI healing quality of the IFT88+/+, MS group was better than that of the other groups. After the conditionally knockout IFT88 in nestin+-BMSCs, the repair ability of the BTI was obviously deteriorated, even though mechanical stimulation did not increase significantly (IFT88-/-, MS group). In vitro results showed the tensile loading enhanced the proliferation, migration and osteogenic or chondrogenic gene expression of nestin+-BMSCs with normal cilia. On the other hand, osteogenesis and chondrogenic expression were significantly decreased after inhibiting actin- Hippo/YAP pathway components.

Conclusion: The primary cilia mediated mechanical stimulation regulated osteogenic and chondrogenic differentiation potential of nestin+-BMSCs through the actin- Hippo/YAP pathway, and then promoted the BTI healing process.

原始纤毛介导的机械刺激对骨肌腱愈合过程中 nestin+-BMSCs 的影响。
导言:机械刺激已被证实可促进骨-肌腱界面(BTI)愈合,但其机制仍不清楚:研究机械刺激对 BTI 愈合过程中 nestin+ 骨间充质干细胞(BMSCs)生物学行为的影响,并通过 nestin+-BMSCs 上的初级纤毛揭示机械刺激影响 BTI 愈合的机制:方法:构建nestin+-BMSCs上原始纤毛被条件性敲除的转基因追踪小鼠(nestin creERT2:: IFT88fl/fl/ROSA26 YFP),并以nestin+-BMSCs上正常纤毛的同窝小鼠(nestin creERT2:: ROSA26 YFP)为对照。建立小鼠冈上肌插入损伤模型后,分别在第2周(每组5个)、第4周和第8周(每组15个)采集样本。在体内,通过组织学、成像、生物力学和免疫荧光染色检测的 nestin+ BMSCs 迁移来评估修复效率。在体外,对 nestin+ BMSCs 进行分拣并用拉力刺激,以研究原发性纤毛介导的机械敏感性基础的机制:结果:机械刺激(MS)加速了nestin+-BMSCs的募集,促进了成骨和软骨能力。组织学、影像学和生物力学结果显示,IFT88+/+、MS 组的 BTI 愈合质量优于其他组。有条件敲除 nestin+-BMSCs 中的 IFT88 后,虽然机械刺激没有明显增加,但 BTI 的修复能力明显下降(IFT88-/-, MS 组)。体外实验结果表明,拉伸载荷增强了具有正常纤毛的 nestin+-BMSCs 的增殖、迁移和成骨或软骨基因表达。另一方面,抑制肌动蛋白-Hippo/YAP通路成分后,成骨和软骨基因表达明显降低:结论:原发性纤毛介导的机械刺激可通过肌动蛋白-Hippo/YAP通路调控nestin+-BMSCs的成骨和软骨分化潜能,进而促进BTI愈合过程。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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