{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T11:28:56Z","timestamp":1787138936980,"version":"build-2736575974"},"reference-count":48,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2026,8,16]],"date-time":"2026-08-16T00:00:00Z","timestamp":1786838400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100009103","name":"Special Project for Serving Local Development of Shaanxi Provincial Department of Education","doi-asserted-by":"publisher","award":["25JC090"],"award-info":[{"award-number":["25JC090"]}],"id":[{"id":"10.13039\/501100009103","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Doctoral Scientific Research Initiation Special Project","award":["2025BS13"],"award-info":[{"award-number":["2025BS13"]}]},{"name":"\u201cSupport Plan for Sanqin Scholars Innovation Team\u201d of Shaanxi Province","award":["[2020]45"],"award-info":[{"award-number":["[2020]45"]}]},{"name":"Science and Technology Planning Project of Xi\u2019an of China","award":["24GXFW0090"],"award-info":[{"award-number":["24GXFW0090"]}]},{"name":"Natural Science Basic Research Plan in Shaanxi Province of China","award":["2026JC-YBMS-0196"],"award-info":[{"award-number":["2026JC-YBMS-0196"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32070442"],"award-info":[{"award-number":["32070442"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["32070442"],"award-info":[{"award-number":["32070442"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Prolonged skeletal muscle disuse, such as extended inactivity and mechanical unloading, typically elicits severe muscle atrophy and progressive bone loss, yet hibernating mammals evade this pathological cascade via poorly defined adaptive mechanisms. Using the Daurian ground squirrel (Spermophilus dauricus) as a unique natural model of prolonged torpor, we demonstrate that skeletal muscle-derived small extracellular vesicles (Mu-EVs) orchestrate protective muscle\u2013bone crosstalk to maintain bone homeostasis during extended disuse. Morphological and microstructural analyses revealed no significant deficits in skeletal muscle and tibial bone between pre-hibernation (PRE) and torpor (TOR) states. Compared with PRE-Mu-EVs, TOR-Mu-EVs significantly enhanced osteogenic differentiation in MC3T3-E1 osteoblasts, markedly upregulating mRNA expression of the key osteogenic markers OCN and COL1A1 (p &lt; 0.05, p &lt; 0.01). Small RNA sequencing identified a novel unannotated miRNA (mature sequence: GCAGCAGCCCGGCTCTCCTAAT) sharply downregulated in TOR-Mu-EVs (p &lt; 0.01); this miRNA exhibits binding potential toward the transcript of Bmp7, a pivotal regulator of osteogenesis. In vitro functional assays confirmed that this miRNA suppresses osteoblast maturation; in a mouse hindlimb unloading (HLU) disuse osteoporosis model, miRNA antagomir partially alleviated bone loss, boosting Masson staining area by 27.13% (p &lt; 0.05) and bone volume fraction by 15.01% (n = 5, 0.05 &lt; p &lt; 0.1, Cohen\u2019s d = 0.71, 95% CI [\u22120.16, 1.38]). Collectively, hibernating Mu-EVs mitigate this BMP7-inhibiting miRNA to sustain osteogenic activity, hinting at a conserved regulatory cascade that could offer tentative translational clues for managing disuse osteoporosis.<\/jats:p>","DOI":"10.3390\/cells15161468","type":"journal-article","created":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T08:05:41Z","timestamp":1786953941000},"page":"1468","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Muscle-Derived Small Extracellular Vesicles Regulate Bone Maintenance During Hibernation Through miRNA-Mediated Signaling"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6859-8270","authenticated-orcid":false,"given":"Yue","family":"He","sequence":"first","affiliation":[{"name":"School of Nursing and Rehabilitation, Xi\u2019an Medical University, Xi\u2019an 710021, China"},{"name":"Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi\u2019an 710069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fangyang","family":"Pan","sequence":"additional","affiliation":[{"name":"Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi\u2019an 710069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Kong","sequence":"additional","affiliation":[{"name":"Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi\u2019an 710069, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyi","family":"Zhang","sequence":"additional","affiliation":[{"name":"Research Center for Special Medicine and Health Systems Engineering, School of Life Sciences, Northwestern Polytechnical University, Xi\u2019an 710129, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anni","family":"Wang","sequence":"additional","affiliation":[{"name":"Research Center for Special Medicine and Health Systems Engineering, School of Life Sciences, Northwestern Polytechnical University, Xi\u2019an 710129, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mu","family":"Cui","sequence":"additional","affiliation":[{"name":"School of Nursing and Rehabilitation, Xi\u2019an Medical University, Xi\u2019an 710021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuhong","family":"Niu","sequence":"additional","affiliation":[{"name":"School of Nursing and Rehabilitation, Xi\u2019an Medical University, Xi\u2019an 710021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuan","family":"Gao","sequence":"additional","affiliation":[{"name":"Research Center for Special Medicine and Health Systems Engineering, School of Life Sciences, Northwestern Polytechnical University, Xi\u2019an 710129, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Dang","sequence":"additional","affiliation":[{"name":"Research Center for Special Medicine and Health Systems Engineering, School of Life Sciences, Northwestern Polytechnical University, Xi\u2019an 710129, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongai","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Nursing and Rehabilitation, Xi\u2019an Medical University, Xi\u2019an 710021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1007\/s11914-010-0013-4","article-title":"Disuse osteopenia","volume":"8","author":"Bloomfield","year":"2010","journal-title":"Curr. Osteoporos. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1713","DOI":"10.1007\/s00198-018-4570-1","article-title":"Impact of muscle atrophy on bone metabolism and bone strength: Implications for muscle-bone crosstalk with aging and disuse","volume":"29","author":"Bettis","year":"2018","journal-title":"Osteoporos. Int."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1016\/j.cbpa.2011.11.009","article-title":"Skeletal muscle is protected from disuse in hibernating dauria ground squirrels","volume":"161","author":"Gao","year":"2012","journal-title":"Comp. Biochem Physiol. A Mol. Integr. Physiol."},{"key":"ref_4","first-page":"2587","article-title":"The effects of hibernation on the contractile and biochemical properties of skeletal muscles in the thirteen-lined ground squirrel, Ictidomys tridecemlineatus","volume":"216","author":"James","year":"2013","journal-title":"J. Exp. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1086\/684619","article-title":"Arctic Ground Squirrels Limit Bone Loss during the Prolonged Physical Inactivity Associated with Hibernation","volume":"89","author":"Wojda","year":"2016","journal-title":"Physiol. Biochem. Zool."},{"key":"ref_6","first-page":"1240","article-title":"Thirteen-lined ground squirrels (Ictidomys tridecemlineatus) show microstructural bone loss during hibernation but preserve bone macrostructural geometry and strength","volume":"15","author":"Stoll","year":"2011","journal-title":"J. Exp. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2582","DOI":"10.1242\/jeb.128348","article-title":"Prevention of muscle wasting and osteoporosis: The value of examining novel animal models","volume":"219","author":"Reilly","year":"2016","journal-title":"J. Exp. Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3247","DOI":"10.1242\/dev.063768","article-title":"Muscle force regulates bone shaping for optimal load-bearing capacity during embryogenesis","volume":"138","author":"Sharir","year":"2011","journal-title":"Development"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s11914-014-0244-x","article-title":"Biomechanical aspects of the muscle-bone interaction","volume":"13","author":"Avin","year":"2015","journal-title":"Curr. Osteoporos. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.bone.2015.02.010","article-title":"Bone and muscle: Interactions beyond mechanical","volume":"80","author":"Brotto","year":"2015","journal-title":"Bone"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1111\/nyas.13411","article-title":"Muscle-bone interactions: Movement in the field of mechano-humoral coupling of muscle and bone","volume":"1402","author":"Cardozo","year":"2017","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1242\/jeb.048074","article-title":"Muscles and their myokines","volume":"214","author":"Pedersen","year":"2011","journal-title":"J. Exp. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1337","DOI":"10.1002\/j.2040-4603.2013.tb00522.x","article-title":"Muscle as a secretory organ","volume":"3","author":"Pedersen","year":"2013","journal-title":"Compr. Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Kalra, H., Simpson, R.J., Ji, H., Aikawa, E., Altevogt, P., Askenase, P., Bond, V.C., Borr\u00e0s, F.E., Breakefield, X., and Budnik, V. (2012). Vesiclepedia: A compendium for extracellular vesicles with continuous community annotation. PLoS Biol., 10.","DOI":"10.1371\/journal.pbio.1001450"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1007\/s11914-019-00537-7","article-title":"Exosomes and Extracellular RNA in Muscle and Bone Aging and Crosstalk","volume":"17","author":"Qin","year":"2019","journal-title":"Curr. Osteoporos. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Herrmann, M., Engelke, K., Ebert, R., M\u00fcller-Deubert, S., Rudert, M., Ziouti, F., Jundt, F., Felsenberg, D., and Jakob, F. (2020). Interactions between Muscle and Bone\u2014Where Physics Meets Biology. Biomolecules, 10.","DOI":"10.3390\/biom10030432"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"14262","DOI":"10.1002\/jcb.28946","article-title":"Muscle-bone crosstalk and potential therapies for sarco-osteoporosis","volume":"120","author":"Li","year":"2019","journal-title":"J. Cell. Biochem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.bbrc.2018.02.144","article-title":"Exosomes from C2C12 myoblasts enhance osteogenic differentiation of MC3T3-E1 pre-osteoblasts by delivering miR-27a-3p","volume":"498","author":"Xu","year":"2018","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1007\/s00223-020-00772-6","article-title":"MicroRNA-196a-5p in Extracellular Vesicles Secreted from Myoblasts Suppresses Osteoclast-like Cell Formation in Mouse Cells","volume":"108","author":"Takafuji","year":"2021","journal-title":"Calcif. Tissue Int."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1186\/s10020-023-00649-y","article-title":"Myoblast-derived exosomal Prrx2 attenuates osteoporosis via transcriptional regulation of lncRNA-MIR22HG to activate Hippo pathway","volume":"29","author":"Li","year":"2023","journal-title":"Mol. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2028","DOI":"10.1016\/j.cmet.2023.10.013","article-title":"Skeletal muscle-derived extracellular vesicles transport glycolytic enzymes to mediate muscle-to-bone crosstalk","volume":"35","author":"Ma","year":"2023","journal-title":"Cell Metab."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"R25","DOI":"10.1186\/gb-2009-10-3-r25","article-title":"Ultrafast and memory-efficient alignment of short DNA sequences to the human genome","volume":"10","author":"Langmead","year":"2009","journal-title":"Genome Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2933","DOI":"10.1093\/bioinformatics\/btt509","article-title":"Infernal 1.1: 100-fold faster RNA homology searches","volume":"29","author":"Nawrocki","year":"2013","journal-title":"Bioinformatics"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1093\/nar\/gkr688","article-title":"miRDeep2 accurately identifies known and hundreds of novel microRNA genes in seven animal clades","volume":"40","author":"Mackowiak","year":"2012","journal-title":"Nucleic Acids Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1007\/978-94-007-5590-1_3","article-title":"MicroRNA target prediction and validation","volume":"774","author":"Ritchie","year":"2013","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"John, B., Enright, A.J., Aravin, A., Tuschl, T., Sander, C., and Marks, D.S. (2004). Human MicroRNA targets. PLoS Biol., 2.","DOI":"10.1371\/journal.pbio.0020363"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1152\/japplphysiol.00969.2001","article-title":"Hindlimb unloading rodent model: Technical aspects","volume":"92","author":"Globus","year":"2002","journal-title":"J. Appl. Physiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"115826","DOI":"10.1016\/j.bone.2020.115826","article-title":"A temporal study on musculoskeletal morphology and metabolism in hibernating Daurian ground squirrels (Spermophilus dauricus)","volume":"144","author":"Zhang","year":"2021","journal-title":"Bone"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1242\/jeb.125401","article-title":"Skeletal muscle mass and composition during mammalian hibernation","volume":"219","author":"Cotton","year":"2016","journal-title":"J. Exp. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1007\/s00360-010-0505-7","article-title":"Muscle plasticity in hibernating ground squirrels (Spermophilus lateralis) is induced by seasonal, but not low-temperature, mechanisms","volume":"181","author":"Nowell","year":"2011","journal-title":"J. Comp. Physiol. B"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1002\/jmor.20007","article-title":"Hibernation does not reduce cortical bone density, area or second moments of inertia in woodchucks (Marmota monax)","volume":"273","author":"Doherty","year":"2012","journal-title":"J. Morphol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2746","DOI":"10.1242\/jeb.032854","article-title":"Bone strength is maintained after 8 months of inactivity in hibernating golden-mantled ground squirrels, Spermophilus lateralis","volume":"212","author":"Utz","year":"2009","journal-title":"J. Exp. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"51S","DOI":"10.1152\/jappl.1992.73.2.S51","article-title":"New Collective Author. Rat soleus muscle fiber responses to 14 days of spaceflight and hindlimb suspension","volume":"73","author":"Ohira","year":"1992","journal-title":"J. Appl. Physiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1016\/j.biopha.2017.10.012","article-title":"Identification of the optimal dose and calpain system regulation of tetramethylpyrazine on the prevention of skeletal muscle atrophy in hindlimb unloading rats","volume":"96","author":"Zhang","year":"2017","journal-title":"Biomed. Pharmacother."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Aswad, H., Jalabert, A., and Rome, S. (2016). Depleting extracellular vesicles from fetal bovine serum alters proliferation and differentiation of skeletal muscle cells in vitro. BMC Biotechnol., 16.","DOI":"10.1186\/s12896-016-0262-0"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2155","DOI":"10.1007\/s00125-014-3337-2","article-title":"Exosomes participate in the alteration of muscle homeostasis during lipid-induced insulin resistance in mice","volume":"57","author":"Aswad","year":"2014","journal-title":"Diabetologia"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.gpb.2015.02.001","article-title":"Exosome and exosomal microRNA: Trafficking, sorting, and function","volume":"13","author":"Zhang","year":"2015","journal-title":"Genom. Proteom. Bioinform."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"10849","DOI":"10.1074\/jbc.M112.446831","article-title":"Neutral sphingomyelinase 2 (nSMase2)-dependent exosomal transfer of angiogenic microRNAs regulate cancer cell metastasis","volume":"288","author":"Kosaka","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"C551","DOI":"10.1152\/ajpcell.00266.2013","article-title":"miR-23a is decreased during muscle atrophy by a mechanism that includes calcineurin signaling and exosome-mediated export","volume":"306","author":"Hudson","year":"2014","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e13107","DOI":"10.1111\/acel.13107","article-title":"Muscle atrophy-related myotube-derived exosomal microRNA in neuronal dysfunction: Targeting both coding and long noncoding RNAs","volume":"19","author":"Yang","year":"2020","journal-title":"Aging Cell"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"12888","DOI":"10.1038\/s41598-017-13105-9","article-title":"Denervation-related alterations and biological activity of miRNAs contained in exosomes released by skeletal muscle fibers","volume":"7","author":"Hamidi","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Rome, S., Forterre, A., Mizgier, M.L., and Bouzakri, K. (2019). Skeletal Muscle-Released Extracellular Vesicles: State of the Art. Front. Physiol., 10.","DOI":"10.3389\/fphys.2019.00929"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"103940","DOI":"10.1016\/j.jprot.2020.103940","article-title":"Quantitative proteomics and reverse engineer analysis identified plasma exosome derived protein markers related to osteoporosis","volume":"228","author":"Chen","year":"2020","journal-title":"J. Proteom."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"4635","DOI":"10.1016\/j.biomaterials.2007.07.009","article-title":"Combination of scaffold and adenovirus vectors expressing bone morphogenetic protein-7 for alveolar bone regeneration at dental implant defects","volume":"28","author":"Zhang","year":"2007","journal-title":"Biomaterials"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/j.molimm.2019.07.027","article-title":"miR-342-5p inhibits expression of Bmp7 to regulate proliferation, differentiation and migration of osteoblasts","volume":"114","author":"Li","year":"2019","journal-title":"Mol. Immunol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"116676","DOI":"10.1016\/j.lfs.2019.116676","article-title":"Regulation of Runx2 by MicroRNAs in osteoblast differentiation","volume":"232","author":"Narayanan","year":"2019","journal-title":"Life Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1002\/jcsm.12296","article-title":"miRNA-23a\/27a attenuates muscle atrophy and renal fibrosis through muscle-kidney crosstalk","volume":"9","author":"Zhang","year":"2018","journal-title":"J. Cachexia Sarcopenia Muscle"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1864","DOI":"10.7150\/thno.29579","article-title":"miR-26a Limits Muscle Wasting and Cardiac Fibrosis through Exosome-Mediated microRNA Transfer in Chronic Kidney Disease","volume":"9","author":"Wang","year":"2019","journal-title":"Theranostics"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/16\/1468\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,19]],"date-time":"2026-08-19T11:14:33Z","timestamp":1787138073000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/16\/1468"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,16]]},"references-count":48,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["cells15161468"],"URL":"https:\/\/doi.org\/10.3390\/cells15161468","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,16]]}}}