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
{"title":"The effects of primary cilia-mediated mechanical stimulation on nestin<sup>+</sup>-BMSCs during bone-tendon healing.","authors":"Huabin Chen, Han Xiao, Bing Wu, Xin Shi, Changbiao Guan, Jianzhong Hu, Tao Zhang, Hongbin Lu","doi":"10.1016/j.jare.2024.09.012","DOIUrl":null,"url":null,"abstract":"<p><strong>Introduction: </strong>Mechanical stimulation has been proven to promote bone-tendon interface (BTI) healing, but the mechanism remains unclear.</p><p><strong>Objective: </strong>To investigate the effects of mechanical stimulation on the biological behavior of nestin<sup>+</sup>-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<sup>+</sup>-BMSCs.</p><p><strong>Methods: </strong>Transgenic tracing mice (nestin cre<sup>ERT2</sup>:: IFT88<sup>fl/fl</sup>/ROSA26 YFP) with primary cilia on nestin<sup>+</sup>-BMSCs conditioned knocked out were constructed, and the littermates (nestin cre<sup>ERT2</sup>:: ROSA26 YFP) with normal cilia on nestin<sup>+</sup>-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<sup>+</sup>-BMSCs, detected by immunofluorescence staining. In vitro, nestin<sup>+</sup> BMSCs were sorted and stimulated by tensile force to study the mechanisms of primary cilium-mediated mechanosensitive basis.</p><p><strong>Results: </strong>Mechanical stimulation (MS) accelerated the recruitment of nestin<sup>+</sup>-BMSCs and promoted osteogenic and chondrogenic capacity. Histological, imaging and biomechanical results showed that the BTI healing quality of the IFT88<sup>+/+</sup>, MS group was better than that of the other groups. After the conditionally knockout IFT88 in nestin<sup>+</sup>-BMSCs, the repair ability of the BTI was obviously deteriorated, even though mechanical stimulation did not increase significantly (IFT88<sup>-/-</sup>, MS group). In vitro results showed the tensile loading enhanced the proliferation, migration and osteogenic or chondrogenic gene expression of nestin<sup>+</sup>-BMSCs with normal cilia. On the other hand, osteogenesis and chondrogenic expression were significantly decreased after inhibiting actin- Hippo/YAP pathway components.</p><p><strong>Conclusion: </strong>The primary cilia mediated mechanical stimulation regulated osteogenic and chondrogenic differentiation potential of nestin<sup>+</sup>-BMSCs through the actin- Hippo/YAP pathway, and then promoted the BTI healing process.</p>","PeriodicalId":94063,"journal":{"name":"Journal of advanced research","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of advanced research","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.jare.2024.09.012","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

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。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信