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.