F-actin microfilaments affect the LIPUS-promoted osteogenic differentiation of BMSCs through TRPM7

作者全名："Yao, Huan; Tang, Li; Wang, Dong; Pang, Hua; Yang, Ke"

作者地址："[Yao, Huan; Tang, Li] Chongqing Med Univ, Affiliated Hosp 1, Dept Ultrasound, Chongqing, Peoples R China; [Wang, Dong] Univ Elect Sci & Technol China, Sichuan Prov Peoples Hosp, Dept Ultrasound, Chengdu, Peoples R China; [Pang, Hua] Chongqing Med Univ, Dept Nucl Med, Affiliated Hosp 1, Chongqing, Peoples R China; [Yang, Ke] Childrens Hosp Chongqing Med Univ, Pediat Res Inst, Natl Clin Res Ctr Child Hlth & Disorders, Minist Educ Key Lab Child Dev & Disorders, Chongqing 400014, Peoples R China"

通信作者："Yang, K (通讯作者)，Childrens Hosp Chongqing Med Univ, Pediat Res Inst, Natl Clin Res Ctr Child Hlth & Disorders, Minist Educ Key Lab Child Dev & Disorders, Chongqing 400014, Peoples R China."

来源：BIOTECHNOLOGY JOURNAL

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:001303104600001

JCR分区：Q2

影响因子：4.7

年份：2024

卷号：19

期号：8

开始页：　

结束页：　

文献类型：Article

关键词：BMSCs; LIPUS; microfilaments; osteogenic differentiation; TRPM7

摘要："The differentiation of bone marrow mesenchymal stem cells (BMSCs) toward osteogenesis can be induced by low-intensity pulsed ultrasound (LIPUS). However, the molecular mechanisms responsible for LIPUS stimulation are unclear. The possible molecular mechanisms by which LIPUS promotes osteogenic differentiation of BMSCs were investigated in this study. The quantification of alkaline phosphatase (ALP) activity, Alizarin Red S staining, ALP staining, and the establishment of a calvarial defect model were used to evaluate osteogenic effects. Immunofluorescence was performed to observe the expression of microfilaments and transient receptor potential melastatin 7 (TRPM7). The levels of F-actin/G-actin and osteogenesis-related proteins under LIPUS alone or LIPUS combined with cytoskeleton interfering drugs (Cytochalasin D [CytoD] or Jasplakinolide [JA]) were assayed by western blot. Quantitative real-time reverse transcription polymerase chain reaction was utilized to measure the expression of Trpm7 mRNA. Moreover, adenoviral Trpm7 knockdown was verified using western blot. The results demonstrated that LIPUS promoted bone formation in vivo. Under osteogenic induction in vitro, the osteogenesis of BMSCs induced by LIPUS was accompanied by the depolymerization and rearrangement of microfilaments and increased levels of TRPM7. By perturbing intracellular actin dynamics, CytoD enhanced the pro-osteogenicity of LIPUS and increased TRPM7 level, while JA inhibited the pro-osteogenicity of LIPUS and reduced TRPM7 level. Additionally, the knockdown of Trpm7 suppressed the osteogenic promotion of BMSCs induced by LIPUS. The transient depolymerization and rearrangement of the cytoskeleton microfilaments mediated by LIPUS can affect TRPM7 expression and subsequently promote the osteogenesis of BMSCs. This study provides further direction for exploring the molecular mechanism of LIPUS, as a mechanical stress, in facilitating the osteogenic differentiation of BMSCs. LIPUS has the potential to enhance the differentiation of BMSCs toward osteogenesis. The authors elucidate that LIPUS-induced transient depolymerization and rearrangement of cytoskeletal microfilaments can modulate TRPM7 expression, thereby promoting the osteogenic differentiation of BMSCs. This work provides a theoretical basis for the more effective clinical application of LIPUS in addressing impaired fracture healing. image"

基金机构：Chongqing Sciences and Technology Commission of China

基金资助正文：Chongqing Sciences and Technology Commission of China