Inorganic Chemistry; Organic Chemistry; Physical and Theoretical Chemistry; Computer Science Applications; Spectroscopy; Molecular Biology; General Medicine; Catalysis
Abstract :
[en] The castration of stallions is traditionally performed after puberty, at around the age of 2 years old. No studies have focused on the effects of early castration on osteoarticular metabolism. Thus, we aimed to compare early castration (3 days after birth) with traditional castration (18 months of age) in horses. Testosterone and estradiol levels were monitored from birth to 33 months in both groups. We quantified the levels of biomarkers of cartilage and bone anabolism (CPII and N-MID) and catabolism (CTX-I and CTX-II), as well as of osteoarthritis (HA and COMP) and inflammation (IL-6 and PGE2). We observed a lack of parallelism between testosterone and estradiol synthesis after birth and during puberty in both groups. The extra-gonadal synthesis of steroids was observed around the 28-month mark, regardless of the castration age. We found the expression of estrogen receptor (ESR1) in cartilage and bone, whereas androgen receptor (AR) expression appeared to be restricted to bone. Nevertheless, with respect to osteoarticular metabolism, steroid hormone deprivation resulting from early castration had no discernable impact on the levels of biomarkers related to bone and cartilage metabolism, nor on those associated with OA and inflammation. Consequently, our research demonstrated that early castration does not disrupt bone and cartilage homeostasis.
Disciplines :
Veterinary medicine & animal health Animal production & animal husbandry Anatomy (cytology, histology, embryology...) & physiology
Author, co-author :
Rouge, Marion ✱; Université de Caen-Normandie, OeReCa, 14000 Caen, France
Legendre, Florence ✱; Université de Caen Normandie BIOTARGEN, 14000 Caen, France
Elkhatib, Razan; Université de Caen-Normandie, OeReCa, 14000 Caen, France
Delalande, Christelle; Université de Caen-Normandie, OeReCa, 14000 Caen, France
Cognié, Juliette; INRAE, Université de Tours, Centre de Recherche de Tours, UMR PRC, 37380 Nouzilly, France
Reigner, Fabrice; INRAE, Université de Tours, Centre de Recherche de Tours, UEPAO, 37380 Nouzilly, France
Barrière, Philippe; INRAE, Université de Tours, Centre de Recherche de Tours, UEPAO, 37380 Nouzilly, France
Deleuze, Stefan ; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH) > FARAH: Médecine vétérinaire comparée
Hanoux, Vincent; Université de Caen-Normandie, OeReCa, 14000 Caen, France
Galéra, Philippe ; Université de Caen Normandie BIOTARGEN, 14000 Caen, France
Bouraïma-Lelong, Hélène; Université de Caen-Normandie, OeReCa, 14000 Caen, France
✱ These authors have contributed equally to this work.
Language :
English
Title :
Early Castration in Horses Does Not Impact Osteoarticular Metabolism
Région normandie [FR] MESR - France. Ministère de l'Enseignement supérieur et de la Recherche [FR] Fonds Eperon [FR]
Funding text :
This work was supported by the Région Normandie, Institut Français du Cheval et de l’Equitation (IFCE), by the Ministère de l’Enseignement Supérieur, de la Recherche et de l’Innovation (Ministry of Higher Education and Research), by Fonds Eperon (EQUISTEM, N80-2014, 917CB194), and by GIS CENTAURE equine research (EQUISTEM-G,014CJ061). These funding sources had no involvement in the study design, collection, analysis and interpretation of data, in the writing of the manuscript, or in the decision to submit the manuscript for publication.
Carreau S. Genissel C. Bilinska B. Levallet J. Sources of Oestrogen in the Testis and Reproductive Tract of the Male Int. J. Androl. 1999 22 211 223 10.1046/j.1365-2605.1999.00172.x 10442293
Payne A.H. Hales D.B. Overview of Steroidogenic Enzymes in the Pathway from Cholesterol to Active Steroid Hormones Endocr. Rev. 2004 25 947 970 10.1210/er.2003-0030
Andersson A.-M. Toppari J. Haavisto A.-M. Petersen J.H. Simell T. Simell O. Skakkeb N.E. Longitudinal Reproductive Hormone Profiles in Infants: Peak of Inhibin B Levels in Infant Boys Exceeds Levels in Adult Men J. Clin. Endocrinol. Metab. 1998 83 675 681 10.1210/jc.83.2.675
Zirkin B.R. Papadopoulos V. Leydig Cells: Formation, Function, and Regulation Biol. Reprod. 2018 99 101 111 10.1093/biolre/ioy059
Vermeulen A. Kaufman J.M. Goemaere S. Van Pottelberg I. Estradiol in Elderly Men Aging Male 2002 5 98 102 10.1080/tam.5.2.98.102
Fagerberg L. Hallström B.M. Oksvold P. Kampf C. Djureinovic D. Odeberg J. Habuka M. Tahmasebpoor S. Danielsson A. Edlund K. et al. Analysis of the Human Tissue-Specific Expression by Genome-Wide Integration of Transcriptomics and Antibody-Based Proteomics Mol. Cell. Proteom. 2014 13 397 406 10.1074/mcp.M113.035600
Cauley J.A. Estrogen and Bone Health in Men and Women Steroids 2015 99 11 15 10.1016/j.steroids.2014.12.010
Gokhale J.A. Frenkel S.R. Dicesare P.E. Estrogen and Osteoarthritis Am. J. Orthop. 2004 33 71 80
Lorentzon M. Swanson C. Andersson N. Mellström D. Ohlsson C. Free Testosterone Is a Positive, Whereas Free Estradiol Is a Negative, Predictor of Cortical Bone Size in Young Swedish Men: The GOOD Study: Estradiol predicts cortical bone size in men J. Bone Miner. Res. 2005 20 1334 1341 10.1359/JBMR.050404 16007330
Roman-Blas J.A. Castañeda S. Largo R. Herrero-Beaumont G. Osteoarthritis Associated with Estrogen Deficiency Arthritis Res. Ther. 2009 11 241 10.1186/ar2791 19804619
Bord S. Estrogen Receptors and Are Differentially Expressed in Developing Human Bone J. Clin. Endocrinol. Metab. 2001 86 2309 2314 10.1210/jc.86.5.2309 11344243
Khalid A.B. Krum S.A. Estrogen Receptors Alpha and Beta in Bone Bone 2016 87 130 135 10.1016/j.bone.2016.03.016 27072516
Nasatzky E. Schwartz Z. Soskolne W.A. Brooks B.P. Dean D.D. Boyan B.D. Ornoy A. Evidence for Receptors Specific for 17 Beta-Estradiol and Testosterone in Chondrocyte Cultures Connect. Tissue Res. 1994 30 277 294 10.3109/03008209409015043 7956206
Nilsson O. Chrysis D. Pajulo O. Boman A. Holst M. Rubinstein J. Martin Ritzén E. Sävendahl L. Localization of Estrogen Receptors-Alpha and -Beta and Androgen Receptor in the Human Growth Plate at Different Pubertal Stages J. Endocrinol. 2003 177 319 326 10.1677/joe.0.1770319 12740020
Vanderschueren D. Vandenput L. Boonen S. Lindberg M.K. Bouillon R. Ohlsson C. Androgens and Bone Endocr. Rev. 2004 25 389 425 10.1210/er.2003-0003 15180950
Khosla S. Monroe D.G. Regulation of Bone Metabolism by Sex Steroids Cold Spring Harb. Perspect. Med. 2018 8 a031211 10.1101/cshperspect.a031211 28710257
Nakamura T. Imai Y. Matsumoto T. Sato S. Takeuchi K. Igarashi K. Harada Y. Azuma Y. Krust A. Yamamoto Y. et al. Estrogen Prevents Bone Loss via Estrogen Receptor α and Induction of Fas Ligand in Osteoclasts Cell 2007 130 811 823 10.1016/j.cell.2007.07.025
Juul A. The Effects of Oestrogens on Linear Bone Growth Hum. Reprod. Update 2001 7 303 313 10.1093/humupd/7.3.303
Tanner J.M. Davies P.S.W. Clinical Longitudinal Standards for Height and Height Velocity for North American Children J. Pediatr. 1985 107 317 329 10.1016/S0022-3476(85)80501-1
Cadogan J. Blumsohn A. Barker M.E. Eastell R. A Longitudinal Study of Bone Gain in Pubertal Girls: Anthropometric and Biochemical Correlates J. Bone Miner. Res. 1998 13 1602 1612 10.1359/jbmr.1998.13.10.1602
Lu W.P. Cowell T. Lloyd-Jones A. Briody N. Howman-Giles R. Volumetric Bone Mineral Density in Normal Subjects, Aged 5–27 Years J. Clin. Endocrinol. Metab. 1996 81 1586 1590 8636372
Boot A.M. de Ridder M.A.J. van der Sluis I.M. van Slobbe I. Krenning E.P. de Muinck Keizer-Schrama S.M.P.F. Peak Bone Mineral Density, Lean Body Mass and Fractures Bone 2010 46 336 341 10.1016/j.bone.2009.10.003 19833245
Behre H.M. Kliesch S. Leifke E. Link T.M. Nieschlag E. Long-Term Effect of Testosterone Therapy on Bone Mineral Density in Hypogonadal Men J. Clin. Endocrinol. Metab. 1997 82 2386 2390 10.1210/jcem.82.8.4163 9253305
Bhasin S. Cunningham G.R. Hayes F.J. Matsumoto A.M. Snyder P.J. Swerdloff R.S. Montori V.M. Testosterone Therapy in Men with Androgen Deficiency Syndromes: An Endocrine Society Clinical Practice Guideline J. Clin. Endocrinol. Metab. 2010 95 2536 2559 10.1210/jc.2009-2354 20525905
Smith E. Korach K. Oestrogen Receptor Deficiency: Consequences for Growth Acta Paediatr. 1996 85 39 43 10.1111/j.1651-2227.1996.tb14292.x
Smith E.P. Boyd J. Frank G.R. Takahashi H. Cohen R.M. Specker B. Williams T.C. Lubahn D.B. Korach K.S. Estrogen Resistance Caused by a Mutation in the Estrogen-Receptor Gene in a Man N. Engl. J. Med. 1994 331 1056 1061 10.1056/NEJM199410203311604 8090165
Carani C. Qin K. Simoni M. Faustini-Fustini M. Serpente S. Boyd J. Korach K.S. Simpson E.R. Effect of Testosterone and Estradiol in a Man with Aromatase Deficiency N. Engl. J. Med. 1997 337 91 95 10.1056/NEJM199707103370204
Moverare S. Venken K. Eriksson A.-L. Andersson N. Skrtic S. Wergedal J. Mohan S. Salmon P. Bouillon R. Gustafsson J.-A. et al. Differential Effects on Bone of Estrogen Receptor and Androgen Receptor Activation in Orchidectomized Adult Male Mice Proc. Natl. Acad. Sci. USA 2003 100 13573 13578 10.1073/pnas.2233084100
Notini A.J. McManus J.F. Moore A. Bouxsein M. Jimenez M. Chiu W.M. Glatt V. Kream B.E. Handelsman D.J. Morris H.A. et al. Osteoblast Deletion of Exon 3 of the Androgen Receptor Gene Results in Trabecular Bone Loss in Adult Male Mice J. Bone Miner. Res. 2007 22 347 356 10.1359/jbmr.061117
Peng Z. Li X. Mäkelä S. Väänänen H.K. Poutanen M. Skeletal Changes in Transgenic Male Mice Expressing Human Cytochrome P450 Aromatase J. Bone Miner. Res. 2004 19 1320 1328 10.1359/JBMR.040510
Vico L. Vanacker J.-M. Sex Hormones and Their Receptors in Bone Homeostasis: Insights from Genetically Modified Mouse Models Osteoporos. Int. 2010 21 365 372 10.1007/s00198-009-0963-5
Khosla S. Amin S. Orwoll E. Osteoporosis in Men Endocr. Rev. 2008 29 441 464 10.1210/er.2008-0002 18451258
Nevitt M.C. Cummings S.R. Lane N.E. Hochberg M.C. Scott J.C. Pressman A.R. Genant H.K. Cauley J.A. Association of Estrogen Replacement Therapy with the Risk of Osteoarthritis of the Hip in Elderly White Women. Study of Osteoporotic Fractures Research Group Arch. Intern. Med. 1996 156 2073 2080 10.1001/archinte.1996.00440170081009 8862099
Mhaouty-Kodja S. Naulé L. Capela D. Sexual Behavior: From Hormonal Regulation to Endocrine Disruption Neuroendocrinology 2018 107 400 416 10.1159/000494558 30326485
Eisenhauer K.M. McCue P.M. Nayden D.K. Osawa Y. Roser J.F. Localization of Aromatase in Equine Leydig Cells Domest. Anim. Endocrinol. 1994 11 291 298 10.1016/0739-7240(94)90020-5 7956176
Raeside J.I. The Isolation of Estrone Sulfate and Estradiol-17 Beta Sulfate from Stallion Testes Can. J. Biochem. 1969 47 811 815 10.1139/o69-124 4309376
Line S.W. Hart B.L. Sanders L. Effect of Prepubertal versus Postpubertal Castration on Sexual and Aggressive Behavior in Male Horses J. Am. Vet. Med. Assoc. 1985 186 249 251
Moll H.D. Pelzer K.D. Pleasant R.S. Modransky P.D. May K.A. A Survey of Equine Castration Complications J. Equine Vet. Sci. 1995 15 522 526 10.1016/S0737-0806(07)80421-7
Cognie J. Freret S. Lansade L. Parias C. Barriere P. Gesbert A. Reigner F. Deleuze S. Early Castration in Foals: Consequences on Physical and Behavioural Development Equine Vet. J. 2023 55 214 221 10.1111/evj.13580
Sims N.A. Clément-Lacroix P. Minet D. Fraslon-Vanhulle C. Gaillard-Kelly M. Resche-Rigon M. Baron R. A Functional Androgen Receptor Is Not Sufficient to Allow Estradiol to Protect Bone after Gonadectomy in Estradiol Receptor–Deficient Mice J. Clin. Investig. 2003 111 1319 1327 10.1172/JCI200317246
Rouge M. Elkhatib R. Delalande C. Cognié J. Reigner F. Barriere P. Deleuze S. Cousty M. Legendre F. Galera P. et al. Investigation of Equine Testis Contribution to Vitamin D Bioactivation Domest. Anim. Endocrinol. 2022 79 106691 10.1016/j.domaniend.2021.106691 34844012
Chen L. Wang R. Wang W. Lu W. Xiao Y. Wang D. Dong Z. Hormone Inhibition during Mini-Puberty and Testicular Function in Male Rats Int. J. Endocrinol. Metab. 2015 13 e25465 10.5812/ijem.25465 26587029
Winter J.S. Hughes I.A. Reyes F.I. Faiman C. Pituitary-Gonadal Relations in Infancy: 2. Patterns of Serum Gonadal Steroid Concentrations in Man from Birth to Two Years of Age J. Clin. Endocrinol. Metab. 1976 42 679 686 10.1210/jcem-42-4-679 1262442
Jost A. Vigier B. Prépin J. Perchellet J.P. Studies on Sex Differentiation in Mammals Recent Progress in Hormone Research, Proceedings of the 1972 Laurentian Hormone Conference Elsevier Amsterdam, The Netherlands 1973 1 41 978-0-12-571129-6
Wilson C.A. Davies D.C. The Control of Sexual Differentiation of the Reproductive System and Brain Reproduction 2007 133 331 359 10.1530/REP-06-0078
Gao W. Alcauter S. Elton A. Hernandez-Castillo C.R. Smith J.K. Ramirez J. Lin W. Functional Network Development During the First Year: Relative Sequence and Socioeconomic Correlations Cereb. Cortex 2015 25 2919 2928 10.1093/cercor/bhu088 24812084
Knickmeyer R.C. Gouttard S. Kang C. Evans D. Wilber K. Smith J.K. Hamer R.M. Lin W. Gerig G. Gilmore J.H. A Structural MRI Study of Human Brain Development from Birth to 2 Years J. Neurosci. 2008 28 12176 12182 10.1523/JNEUROSCI.3479-08.2008
Lyall A.E. Shi F. Geng X. Woolson S. Li G. Wang L. Hamer R.M. Shen D. Gilmore J.H. Dynamic Development of Regional Cortical Thickness and Surface Area in Early Childhood Cereb. Cortex 2015 25 2204 2212 10.1093/cercor/bhu027
Barnes R.J. Nathanielsz P.W. Rossdale P.D. Comline R.S. Silver M. Plasma Progestagens and Oestrogens in Fetus and Mother in Late Pregnancy J. Reprod. Fertil. Suppl. 1975 617 623
Schlinger B.A. Lane N.I. Grisham W. Thompson L. Androgen Synthesis in a Songbird: A Study of Cyp17 (17alpha-Hydroxylase/C17,20-Lyase) Activity in the Zebra Finch Gen. Comp. Endocrinol. 1999 113 46 58 10.1006/gcen.1998.7179
Witchel S.F. Pinto B. Burghard A.C. Oberfield S.E. Update on Adrenarche Curr. Opin. Pediatr. 2020 32 574 581 10.1097/MOP.0000000000000928 32692055
Turcu A.F. Auchus R.J. Adrenal Steroidogenesis and Congenital Adrenal Hyperplasia Endocrinol. Metab. Clin. N. Am. 2015 44 275 296 10.1016/j.ecl.2015.02.002
Labrie F. Bélanger A. Cusan L. Gomez J.L. Candas B. Marked Decline in Serum Concentrations of Adrenal C19 Sex Steroid Precursors and Conjugated Androgen Metabolites during Aging J. Clin. Endocrinol. Metab. 1997 82 2396 2402 10.1210/jcem.82.8.4160 9253307
Migeon C.J. Keller A.R. Lawrence B. Shepard T.H. Dehydroepiandrosterone and Androsterone Levels in Human Plasma: Effect of Age and Sex; Day-to-Day and Diurnal Variations J. Clin. Endocrinol. Metab. 1957 17 1051 1062 10.1210/jcem-17-9-1051
Remer T. Manz F. Hartmann M.F. Schoenau E. Wudy S.A. Prepubertal Healthy Children’s Urinary Androstenediol Predicts Diaphyseal Bone Strength in Late Puberty J. Clin. Endocrinol. Metab. 2009 94 575 578 10.1210/jc.2008-1375 19001521
Holló I. Fehér T. Szücs J. Serum Dehydroepiandrosterone, Androsterone and Cortisol Level in Primary Postmenopausal and Other Type Osteoporosis Acta Medica Acad. Sci. Hung. 1970 27 155 160
Zhao H. Tian Z. Hao J. Chen B. Extragonadal Aromatization Increases with Time after Ovariectomy in Rats Reprod. Biol. Endocrinol. 2005 3 6 10.1186/1477-7827-3-6
Juliand V. Martin-Rosset W. The Growing Horse: Nutrition and Prevention of Growth Disorders EAAP Scientific Series Wageningen Academic Publishers Wageningen, The Netherlands 2005 Volume 114 978-90-76998-62-6
Nelson L.R. Bulun S.E. Estrogen Production and Action J. Am. Acad. Dermatol. 2001 45 S116 S124 10.1067/mjd.2001.117432
Khosla S. Melton L.J. Atkinson E.J. O’Fallon W.M. Klee G.G. Riggs B.L. Relationship of Serum Sex Steroid Levels and Bone Turnover Markers with Bone Mineral Density in Men and Women: A Key Role for Bioavailable Estrogen J. Clin. Endocrinol. Metab. 1998 83 2266 2274 10.1210/jcem.83.7.4924
Falahati-Nini A. Riggs B.L. Atkinson E.J. O’Fallon W.M. Eastell R. Khosla S. Relative Contributions of Testosterone and Estrogen in Regulating Bone Resorption and Formation in Normal Elderly Men J. Clin. Investig. 2000 106 1553 1560 10.1172/JCI10942
Verhas M. Schoutens A. L’hermite-Baleriaux M. Dourov N. Verschaeren A. Mone M. Heilporn A. The Effect of Orchidectomy on Bone Metabolism in Aging Rats Calcif. Tissue Int. 1986 39 74 77 10.1007/BF02553294 3091224
Irie T. Aizawa T. Kokubun S. The Role of Sex Hormones in the Kinetics of Chondrocytes in the Growth Plate: A study in rabbit J. Bone Jt. Surg. Br. 2005 87-B 1278 1284 10.1302/0301-620X.87B9.15583
Sniekers Y.H. Weinans H. Bierma-Zeinstra S.M. van Leeuwen J.P.T.M. van Osch G.J.V.M. Animal Models for Osteoarthritis: The Effect of Ovariectomy and Estrogen Treatment—A Systematic Approach Osteoarthr. Cartil. 2008 16 533 541 10.1016/j.joca.2008.01.002
Peshkova M. Lychagin A. Lipina M. Di Matteo B. Anzillotti G. Ronzoni F. Kosheleva N. Shpichka A. Royuk V. Fomin V. et al. Gender-Related Aspects in Osteoarthritis Development and Progression: A Review Int. J. Mol. Sci. 2022 23 2767 10.3390/ijms23052767 35269906
Eastell R. Hannon R.A. Biomarkers of Bone Health and Osteoporosis Risk Proc. Nutr. Soc. 2008 67 157 162 10.1017/S002966510800699X
Nagasue K. Inaba M. Okuno S. Kitatani K. Imanishi Y. Ishimura E. Miki T. Kim M. Nishizawa Y. Serum N-Terminal Midfragment vs. Intact Osteocalcin Immunoradiometric Assay as Markers for Bone Turnover and Bone Loss in Hemodialysis Patients Biomed. Pharmacother. Biomed. Pharmacother. 2003 57 98 104 10.1016/S0753-3322(02)00344-X 12842495
Herrmann D. Intemann T. Lauria F. Mårild S. Molnár D. Moreno L.A. Sioen I. Tornaritis M. Veidebaum T. Pigeot I. et al. Reference Values of Bone Stiffness Index and C-Terminal Telopeptide in Healthy European Children Int. J. Obes. 2014 38 S76 S85 10.1038/ijo.2014.138
Rauchenzauner M. Schmid A. Heinz-Erian P. Kapelari K. Falkensammer G. Griesmacher A. Finkenstedt G. Högler W. Sex- and Age-Specific Reference Curves for Serum Markers of Bone Turnover in Healthy Children from 2 Months to 18 Years J. Clin. Endocrinol. Metab. 2007 92 443 449 10.1210/jc.2006-1706
Ambroszkiewicz J. Gajewska J. Rowicka G. Klemarczyk W. Chelchowska M. Assessment of Biochemical Bone Turnover Markers and Bone Mineral Density in Thin and Normal-Weight Children Cartilage 2018 9 255 262 10.1177/1947603516686145
Bleasel J.F. Poole A.R. Heinegård D. Saxne T. Holderbaum D. Ionescu M. Jones P. Moskowitz R.W. Changes in Serum Cartilage Marker Levels Indicate Altered Cartilage Metabolism in Families with the Osteoarthritis-Related Type II Collagen Gene COL2A1 Mutation Arthritis Rheum. 1999 42 39 45 10.1002/1529-0131(199901)42:1<39::AID-ANR5>3.0.CO;2-Y
Mouritzen U. Christgau S. Lehmann H.-J. Tankó L.B. Christiansen C. Cartilage Turnover Assessed with a Newly Developed Assay Measuring Collagen Type II Degradation Products: Influence of Age, Sex, Menopause, Hormone Replacement Therapy, and Body Mass Index Ann. Rheum. Dis. 2003 62 332 336 10.1136/ard.62.4.332 12634232
Hoch J.M. Mattacola C.G. Medina McKeon J.M. Howard J.S. Lattermann C. Serum Cartilage Oligomeric Matrix Protein (SCOMP) Is Elevated in Patients with Knee Osteoarthritis: A Systematic Review and Meta-Analysis Osteoarthr. Cartil. 2011 19 1396 1404 10.1016/j.joca.2011.09.005 22001901
Saruga T. Sasaki E. Inoue R. Chiba D. Ota S. Iwasaki H. Uesato R. Nakaji S. Ishibashi Y. Usefulness of Serum Hyaluronic Acid Levels as a Predictor of Incidence of Hand Osteoarthritis Analyzed by Longitudinal Analysis from the Iwaki Cohort Sci. Rep. 2021 11 4074 10.1038/s41598-021-83693-0 33603120
Sasaki E. Tsuda E. Yamamoto Y. Maeda S. Inoue R. Chiba D. Fujita H. Takahashi I. Umeda T. Nakaji S. et al. Serum Hyaluronic Acid Concentration Predicts the Progression of Joint Space Narrowing in Normal Knees and Established Knee Osteoarthritis—A Five-Year Prospective Cohort Study Arthritis Res. Ther. 2015 17 283 10.1186/s13075-015-0793-0
Bertoni L. Jacquet-Guibon S. Branly T. Legendre F. Desancé M. Mespoulhes C. Melin M. Hartmann D.-J. Schmutz A. Denoix J.-M. et al. An Experimentally Induced Osteoarthritis Model in Horses Performed on Both Metacarpophalangeal and Metatarsophalangeal Joints: Technical, Clinical, Imaging, Biochemical, Macroscopic and Microscopic Characterization PLoS ONE 2020 15 e0235251 10.1371/journal.pone.0235251
