{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T07:05:20Z","timestamp":1790838320577,"version":"4.1.0"},"reference-count":48,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2023,3,16]],"date-time":"2023-03-16T00:00:00Z","timestamp":1678924800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Japan Sports Medicine Foundation 2020"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>To date, it remains unclear how overuse affects the tendons and entheses at different stages of maturation. Therefore, we evaluated histological and morphological changes in the tendons and entheses in adolescent (4-week-old) and adult mice (8-week-old) by performing flat-land and downhill running exercises. The mice were divided into the Sedentary, High Flat (flat-land high-speed running; concentric-contraction exercise), Low Down (downhill low-speed running; eccentric-contraction exercise), and High Down (downhill high-speed running; eccentric-contraction exercise) groups. Histological changes and inflammatory factor expressions were compared in the entheses and tendons after 4 weeks of exercise. Downhill, but not flat-land high-speed running, induced muscle\u2013tendon complex hypertrophy in both adolescent and adult mice. Histological enthesis changes were induced in both groups during downhill running but were less pronounced in adult mice. Conversely, no significant cell aggregation or fiber orientation changes were observed in the tendon, but increased inflammatory factors were observed in both groups, with significantly higher expression in the tendons of adult mice. Downhill running induced histological and morphological enthesis changes and inflammatory factor increase in the tendons, regardless of running speed variations. These results may help elucidate the pathogenesis of enthesopathy and tendinopathy, which have different pathophysiologies despite having the same pathogenetic factors.<\/jats:p>","DOI":"10.3390\/biology12030456","type":"journal-article","created":{"date-parts":[[2023,3,16]],"date-time":"2023-03-16T03:14:35Z","timestamp":1678936475000},"page":"456","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["The Effects of Downhill Running and Maturation on Histological and Morphological Properties of Tendon and Enthesis in Mice"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2231-7183","authenticated-orcid":false,"given":"Kaichi","family":"Ozone","sequence":"first","affiliation":[{"name":"Department of Health and Social Services, Health and Social Services, Graduate School of Saitama Prefectural University, Koshigaya 343-8540, Japan"},{"name":"Department of Rehabilitation, University of Tsukuba Hospital, Tsukuba 305-8576, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0463-6124","authenticated-orcid":false,"given":"Yuki","family":"Minegishi","sequence":"additional","affiliation":[{"name":"Department of Health and Social Services, Health and Social Services, Graduate School of Saitama Prefectural University, Koshigaya 343-8540, Japan"},{"name":"Japan Society for the Promotion of Science, Tokyo 102-0083, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9107-6320","authenticated-orcid":false,"given":"Yuichiro","family":"Oka","sequence":"additional","affiliation":[{"name":"Department of Health and Social Services, Health and Social Services, Graduate School of Saitama Prefectural University, Koshigaya 343-8540, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michiaki","family":"Sato","sequence":"additional","affiliation":[{"name":"Department of Health and Social Services, Health and Social Services, Graduate School of Saitama Prefectural University, Koshigaya 343-8540, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3313-9717","authenticated-orcid":false,"given":"Naohiko","family":"Kanemura","sequence":"additional","affiliation":[{"name":"Department of Physical Therapy, Health and Social Services, Saitama Prefectural University, Koshigaya 343-8540, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/j.fcl.2005.01.013","article-title":"Achilles Tendon Disorders: Etiology and Epidemiology","volume":"10","author":"Kannus","year":"2005","journal-title":"Foot Ankle Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1007\/s004020100285","article-title":"Ultrasonography as a diagnostic modality in Osgood-Schlatter disease. A clinical study and review of the literature","volume":"121","author":"Blankstein","year":"2001","journal-title":"Arch. Orthop. Trauma Surg."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2325967118792192","DOI":"10.1177\/2325967118792192","article-title":"Pathogenic factors associated with Osgood-Schlatter disease in adolescent male soccer players: A prospective cohort study","volume":"6","author":"Watanabe","year":"2018","journal-title":"Orthop. J. Sport. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.jus.2008.09.001","article-title":"Overload syndromes of the knee in adolescents: Sonographic findings","volume":"11","author":"Draghi","year":"2008","journal-title":"J. Ultrasound"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1034\/j.1600-0838.2000.010004216.x","article-title":"Patellar tendinopathy in junior basketball players: A controlled clinical and ultrasonographic study of 268 patellar tendons in players aged 14\u201318 years","volume":"10","author":"Cook","year":"2000","journal-title":"Scand. J. Med. Sci. Sport."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1038\/nrrheum.2010.43","article-title":"The pathogenesis of tendinopathy: Balancing the response to loading","volume":"6","author":"Magnusson","year":"2010","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"75","DOI":"10.2165\/00007256-200333010-00006","article-title":"Overview of Injuries in the Young Athlete","volume":"33","author":"Adirim","year":"2003","journal-title":"Sport. Med."},{"key":"ref_8","first-page":"1539","article-title":"Tendinopathy","volume":"7","author":"Millar","year":"2021","journal-title":"Nat. Rev. Dis. Prim."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1642","DOI":"10.1111\/sms.13735","article-title":"Biomechanics of graded running: Part II\u2014Joint kinematics and kinetics","volume":"30","author":"Khassetarash","year":"2020","journal-title":"Scand. J. Med. Sci. Sport."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1097\/JES.0b013e3182a4e6d5","article-title":"How tendons buffer energy dissipation by muscle","volume":"41","author":"Roberts","year":"2013","journal-title":"Exerc. Sport Sci. Rev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2076","DOI":"10.1002\/jor.25233","article-title":"Structural and pathological changes in the enthesis are influenced by the muscle contraction type during exercise","volume":"40","author":"Ozone","year":"2022","journal-title":"J. Orthop. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1002\/jor.25399","article-title":"Eccentric contraction-dominant exercise leads to molecular biological changes in enthesis and enthesopathy-like morphological changes","volume":"41","author":"Ozone","year":"2022","journal-title":"J. Orthop. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1067\/mse.2000.101962","article-title":"Overuse activity injures the supraspinatus tendon in an animal model: A histologic and biomechanical study","volume":"9","author":"Soslowsky","year":"2000","journal-title":"J. Shoulder Elb. Surg."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1002\/jor.20347","article-title":"Rat supraspinatus tendon expresses cartilage markers with overuse","volume":"25","author":"Archambault","year":"2007","journal-title":"J. Orthop. Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1152\/jappl.1983.54.1.80","article-title":"Eccentric exercise-induced injury to rat skeletal muscle","volume":"54","author":"Armstrong","year":"1983","journal-title":"J. Appl. Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Portier, H., Benaitreau, D., and Pallu, S. (2020). Does physical exercise always improve bone quality in rats?. Life, 10.","DOI":"10.3390\/life10100217"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"182","DOI":"10.32098\/mltj.02.2014.17","article-title":"Novel characteristics of normal supraspinatus insertion in rats: An ultrastructural analysis using three-dimensional reconstruction using focused ion beam\/scanning electron microscope tomography","volume":"4","author":"Kanazawa","year":"2014","journal-title":"Muscles. Ligaments Tendons J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_20","unstructured":"(2022). JASP Team JASP, JASP Team. Version 0.16.3."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.18637\/jss.v088.i02","article-title":"JASP: Graphical statistical software for common statistical designs","volume":"88","author":"Love","year":"2019","journal-title":"J. Stat. Softw."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1089\/ten.tea.2016.0546","article-title":"Muscle-Secreted Factors Improve Anterior Cruciate Ligament Graft Healing: An In Vitro and In Vivo Analysis","volume":"24","author":"Ghebes","year":"2018","journal-title":"Tissue Eng. Part A"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4665","DOI":"10.1113\/JP270737","article-title":"The exercise-induced biochemical milieu enhances collagen content and tensile strength of engineered ligaments","volume":"593","author":"West","year":"2015","journal-title":"J. Physiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2823","DOI":"10.1002\/jor.24064","article-title":"Effect of micro-RNA on tenocytes and tendon-related gene expression: A systematic review","volume":"36","author":"Dubin","year":"2018","journal-title":"J. Orthop. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e353","DOI":"10.1111\/sms.12089","article-title":"Increased mast cell numbers in a calcaneal tendon overuse model","volume":"23","author":"Pingel","year":"2013","journal-title":"Scand. J. Med. Sci. Sport."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1242\/jeb.044487","article-title":"Effects of grade and mass distribution on the mechanics of trotting in dogs","volume":"214","author":"Lee","year":"2011","journal-title":"J. Exp. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1093\/icb\/icu015","article-title":"The scaling of uphill and downhill locomotion in legged animals","volume":"54","author":"Higham","year":"2014","journal-title":"Integr. Comp. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"924","DOI":"10.1016\/j.orthres.2004.02.016","article-title":"Inhomogeneous mechanical behavior of the human supraspinatus tendon under uniaxial loading","volume":"23","author":"Huang","year":"2005","journal-title":"J. Orthop. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1623","DOI":"10.1589\/jpts.26.1623","article-title":"Induction of muscle hypertrophy in rats through low intensity eccentric contraction","volume":"26","author":"Tsumiyama","year":"2014","journal-title":"J. Phys. Ther. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1093\/gerona\/58.5.M417","article-title":"The positives of negatives: Clinical implications of eccentric resistance exercise in old adults","volume":"58","year":"2003","journal-title":"J. Gerontol.-Ser. A Biol. Sci. Med. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"674","DOI":"10.1242\/jeb.050112","article-title":"Muscle damage and muscle remodeling: No pain, no gain?","volume":"214","author":"Flann","year":"2011","journal-title":"J. Exp. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"447","DOI":"10.3389\/fphys.2017.00447","article-title":"V Skeletal muscle remodeling in response to eccentric vs. concentric loading: Morphological, molecular, and metabolic adaptations","volume":"8","author":"Franchi","year":"2017","journal-title":"Front. Physiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1111\/j.1365-2613.2007.00552.x","article-title":"Tendon matrix composition and turnover in relation to functional requirements","volume":"88","author":"Birch","year":"2007","journal-title":"Int. J. Exp. Pathol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2074","DOI":"10.1096\/fj.12-225599","article-title":"Lack of tissue renewal in human adult Achilles tendon is revealed by nuclear bomb 14 C","volume":"27","author":"Heinemeier","year":"2013","journal-title":"FASEB J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s40798-015-0009-9","article-title":"Human tendon adaptation in response to mechanical loading: A systematic review and meta-analysis of exercise intervention studies on healthy adults","volume":"1","author":"Bohm","year":"2015","journal-title":"Sport. Med.-Open"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1016\/S1095-6433(02)00138-1","article-title":"The skeletal attachment of tendons-tendon \u201centheses\u201d","volume":"133","author":"Benjamin","year":"2002","journal-title":"Comp. Biochem. Physiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1046\/j.1469-7580.1998.19340481.x","article-title":"Fibrocartilage in tendons and ligaments\u2014An adaptation to compressive load","volume":"193","author":"Benjamin","year":"1998","journal-title":"J. Anat."},{"key":"ref_38","first-page":"323","article-title":"Tendon structure and response to changing mechanical load","volume":"3","author":"Vogel","year":"2003","journal-title":"J. Musculoskelet. Neuronal Interact."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1006\/abbi.1997.0102","article-title":"Mechanical loading and TGF-\u03b2 regulate proteoglycan synthesis in tendon","volume":"342","author":"Robbins","year":"1997","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1006\/abbi.1993.1574","article-title":"Proteoglycan synthesis in fetal tendon is differentially regulated by cyclic compression in vitro","volume":"307","author":"Evanko","year":"1993","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"348","DOI":"10.1007\/s00776-008-1240-8","article-title":"Mechanical environment of the supraspinatus tendon: Three-dimensional finite element model analysis","volume":"13","author":"Seki","year":"2008","journal-title":"J. Orthop. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1111\/j.1469-7580.2006.00540.x","article-title":"Where tendons and ligaments meet bone: Attachment sites (\u2018entheses\u2019) in relation to exercise and\/or mechanical load","volume":"208","author":"Benjamin","year":"2006","journal-title":"J. Anat."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1038\/nrrheum.2017.188","article-title":"Enthesitis: From pathophysiology to treatment","volume":"13","author":"Schett","year":"2017","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1002\/art.39458","article-title":"Review: Enthesitis: New insights into pathogenesis, diagnostic modalities, and treatment","volume":"68","author":"Kehl","year":"2016","journal-title":"Arthritis Rheumatol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1007\/s11926-018-0751-3","article-title":"Enthesitis: Much more than focal insertion point inflammation","volume":"20","author":"Watad","year":"2018","journal-title":"Curr. Rheumatol. Rep."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2264","DOI":"10.1002\/jbmr.3844","article-title":"Cytokine-induced and stretch-induced sphingosine 1-phosphate production by enthesis cells could favor abnormal ossification in spondyloarthritis","volume":"34","author":"Briolay","year":"2019","journal-title":"J. Bone Miner. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/S1058-2746(99)90081-2","article-title":"Lateral tennis elbow: \u201cIs there any science out there?\u201d","volume":"8","author":"Boyer","year":"1999","journal-title":"J. Shoulder Elb. Surg."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"591","DOI":"10.2147\/DDDT.S154660","article-title":"Tendinopathy: Injury, repair, and current exploration","volume":"12","author":"Lipman","year":"2018","journal-title":"Drug Des. Devel. Ther."}],"container-title":["Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-7737\/12\/3\/456\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:56:25Z","timestamp":1760122585000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-7737\/12\/3\/456"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,16]]},"references-count":48,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["biology12030456"],"URL":"https:\/\/doi.org\/10.3390\/biology12030456","relation":{},"ISSN":["2079-7737"],"issn-type":[{"value":"2079-7737","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,16]]}}}