The interdependent relationship between the nitric oxide signaling pathway and primary cilia in pulse electromagnetic field‐stimulated osteoblastic differentiation

The FASEB JournalVolume 36, Issue 6 e22376 RESEARCH ARTICLE The interdependent relationship between the nitric oxide signaling pathway and primary cilia in pulse electromagnetic field-stimulated osteoblastic differentiation Wen-Fang He, Wen-Fang He orcid.org/0000-0002-0032-5978 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. China Department of Bioengineering, School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, P. R. China Key Laboratory of Stem Cells and Gene Drugs of Gansu Province, Lanzhou, P. R. ChinaSearch for more papers by this authorRong Qin, Rong Qin orcid.org/0000-0003-1848-1642 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorYu-Hai Gao, Yu-Hai Gao Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorJian Zhou, Jian Zhou orcid.org/0000-0001-7211-1680 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorJuan-Juan Wei, Juan-Juan Wei orcid.org/0000-0003-3013-6286 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorJing Liu, Jing Liu orcid.org/0000-0002-7005-611X Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorXue-Feng Hou, Xue-Feng Hou Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorHui-Ping Ma, Hui-Ping Ma orcid.org/0000-0003-4560-2155 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorCory J. Xian, Cory J. Xian UniSA Clinical and Health Sciences, University of South Australia, Adelaide, South Australia, AustraliaSearch for more papers by this authorXue-Yan Li, Corresponding Author Xue-Yan Li lixueyan@lut.cn Department of Bioengineering, School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, P. R. China Correspondence Xue-Yan Li, School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, P. R. China. Email: lixueyan@lut.cn Ke-Ming Chen, Institute of Orthopaedics, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou 730050, P. R. China. Email: chenkm@lut.cnSearch for more papers by this authorKe-Ming Chen, Corresponding Author Ke-Ming Chen chenkm@lut.cn orcid.org/0000-0002-9749-8356 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. China Key Laboratory of Stem Cells and Gene Drugs of Gansu Province, Lanzhou, P. R. China Correspondence Xue-Yan Li, School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, P. R. China. Email: lixueyan@lut.cn Ke-Ming Chen, Institute of Orthopaedics, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou 730050, P. R. China. Email: chenkm@lut.cnSearch for more papers by this author Wen-Fang He, Wen-Fang He orcid.org/0000-0002-0032-5978 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. China Department of Bioengineering, School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, P. R. China Key Laboratory of Stem Cells and Gene Drugs of Gansu Province, Lanzhou, P. R. ChinaSearch for more papers by this authorRong Qin, Rong Qin orcid.org/0000-0003-1848-1642 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorYu-Hai Gao, Yu-Hai Gao Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorJian Zhou, Jian Zhou orcid.org/0000-0001-7211-1680 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorJuan-Juan Wei, Juan-Juan Wei orcid.org/0000-0003-3013-6286 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorJing Liu, Jing Liu orcid.org/0000-0002-7005-611X Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorXue-Feng Hou, Xue-Feng Hou Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorHui-Ping Ma, Hui-Ping Ma orcid.org/0000-0003-4560-2155 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. ChinaSearch for more papers by this authorCory J. Xian, Cory J. Xian UniSA Clinical and Health Sciences, University of South Australia, Adelaide, South Australia, AustraliaSearch for more papers by this authorXue-Yan Li, Corresponding Author Xue-Yan Li lixueyan@lut.cn Department of Bioengineering, School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, P. R. China Correspondence Xue-Yan Li, School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, P. R. China. Email: lixueyan@lut.cn Ke-Ming Chen, Institute of Orthopaedics, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou 730050, P. R. China. Email: chenkm@lut.cnSearch for more papers by this authorKe-Ming Chen, Corresponding Author Ke-Ming Chen chenkm@lut.cn orcid.org/0000-0002-9749-8356 Fundamental Medical Science Research Laboratories, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou, P. R. China Key Laboratory of Stem Cells and Gene Drugs of Gansu Province, Lanzhou, P. R. China Correspondence Xue-Yan Li, School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, P. R. China. Email: lixueyan@lut.cn Ke-Ming Chen, Institute of Orthopaedics, The 940th Hospital of Joint Logistic Support Force, People's Liberation Army of China, Lanzhou 730050, P. R. China. Email: chenkm@lut.cnSearch for more papers by this author First published: 26 May 2022 https://doi.org/10.1096/fj.202101577RR Funding information National Natural Sciences Foundation of China, Grant/Award Numbers: 81770879; Youth Project of Gansu Science Foundation, Grant/Award Numbers: 20JR5RA589. Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Abstract Pulsed electromagnetic fields (PEMFs) have long been recognized being safe and effective in treating bone fracture nonunion and osteoporosis. However, the mechanism of osteogenic action of PEMFs is still unclear. While primary cilia are reported to be a sensory organelle for PEMFs, and nitric oxide (NO) plays an indispensable role in osteogenic effect of PEMFs, the relationship between NO and primary cilia is unknown. In this study, effects of treatment with 50 Hz 0.6 mT PEMFs on osteogenic differentiation and mineralization, NO secretion, and ciliary location of specific proteins were examined in rat calvarial osteoblasts (ROBs) with normal or abrogated primary cilia. It was found that PEMFs stimulated the osteogenic differentiation by activating the NOS/NO/sGC/cGMP/PKG signaling pathway, which need the existence of primary cilia. All components of the signaling pathway including iNOS, eNOS, sGC, PKG-1, and PKG-2 were localized to primary cilia, and eNOS was phosphorylated inside the primary cilia. Besides, primary cilia were elongated significantly by PEMF treatment and changed dynamically with the activation NO/cGMP pathway. When the pathway was blocked by L-NAME, PEMFs could no longer elongate the primary cilia and stimulate the osteoblastic differentiation. Thus, this study for the first time observed activation of the NO/cGMP signaling pathway in ciliary compartment of osteoblasts, and PEMFs could not stimulate the osteoblastic differentiation if the NO signaling pathway was blocked or the ciliogenesis was inhibited. Our findings indicate the interdependent relationship between NO and primary cilia in the PEMF-promoted osteogenesis. Open Research DATA AVAILABILITY STATEMENT The data that support the findings of this study are available from the corresponding author upon reasonable request. Volume36, Issue6June 2022e22376 RelatedInformation