FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Equine veterinary journal2022; 55(3); 534-550; doi: 10.1111/evj.13600

An in vitro model for discovery of osteoclast specific biomarkers towards identification of racehorses at risk for catastrophic fractures.

Abstract: Focal bone microcracks with osteoclast recruitment and bone lysis, may reduce fracture resistance in racehorses. As current imaging does not detect all horses at risk for fracture, the discovery of novel serum biomarkers of bone resorption or osteoclast activity could potentially address this unmet clinical need. The biology of equine osteoclasts on their natural substrate, equine bone, has never been studied in vitro and may permit identification of specific biomarkers of their activity. Objective: (1) Establish osteoclast cultures on equine bone, (2) Measure biomarkers (tartrate resistant acid phosphatase isoform 5b [TRACP-5b] and C-terminal telopeptide of type I collagen [CTX-I]) in vitro and (3) Study the effects of inflammation. Methods: In vitro experiments. Methods: Haematopoietic stem cells, from five equine sternal bone marrow aspirates, were differentiated into osteoclasts and cultured either alone or on equine bone slices, with or without a pro-inflammatory stimulus (IL-1β or LPS). CTX-I and TRACP-5b were immunoassayed in the media. Osteoclast numbers and bone resorption area were assessed. Results: TRACP-5b increased over time in osteoclast cultures without bone (p < 0.0001) and correlated with osteoclast number (r = 0.63, p < 0.001). CTX-I and TRACP-5b increased with time for cultures with bone (p = 0.002; p = 0.02 respectively), correlated with each other (r = 0.64, p < 0.002) and correlated with bone resorption (r = 0.85, p < 0.001; r = 0.82, p < 0.001 respectively). Inflammation had no measurable effects. Conclusions: Specimen numbers limited. Conclusions: Equine osteoclasts were successfully cultured on equine bone slices and their bone resorption quantified. TRACP-5b was shown to be a biomarker of equine osteoclast number and bone resorption for the first time; CTX-I was also confirmed to be a biomarker of equine bone resorption in vitro. This robust equine specific in vitro assay will help the study of osteoclast biology. Unassigned: Fokale Mikrofrakturen im Knochen mit Osteoklasten Rekrutierung und Knochen Lyse können die Frakturresistenz bei Rennpferden verringern. Da die derzeitige Bildgebung nicht alle Pferde mit Frakturrisiko erfasst, könnte die Entdeckung neuer Serum-Biomarker für die Knochenresorption oder Osteoklasten Aktivität diesen ungedeckten klinischen Bedarf möglicherweise decken. Die Biologie der Pferde Osteoklasten auf ihrem natürlichen Substrat, dem Pferdeknochen, wurde noch nicht in vitro untersucht und könnte die Identifizierung spezifischer Biomarker für ihre Aktivität ermöglichen. ZIELE: (1) Anlegen von Osteoklastenkulturen an Pferdeknochen, (2) Messung von Biomarkern (tartrate resistant acid phosphatase isoform 5b [TRACP-5b] und C-terminal telopeptide of type I collagen [CTX-I]) in vitro und (3) Untersuchung der Auswirkungen von Entzündungen. Methods: In vitro experimentelle Studie. Methods: Hämatopoetische Stammzellen aus fünf sternalen Knochenmark Aspiraten von Pferden wurden in Osteoklasten differenziert und entweder alleine oder auf Knochenscheiben von Pferden kultiviert, mit oder ohne proinflammatorischen Stimulus (IL 1β oder LPS). CTX-I und TRACP-5b wurden in den Medien immunoassayiert. Die Anzahl der Osteoklasten und die Fläche der Knochenresorption wurden bestimmt. Results: TRACP 5b nahm mit der Zeit ohne Knochen zu (p < 0.0001) und korrelierte mit der Osteoklasten Anzahl (r = 0.63, p < 0.001). CTX-I und TRACP-5b nahmen bei Kulturen mit Knochen mit der Zeit zu (p = 0.0018 bzw. p = 0.02), korrelierten miteinander (r = 0.64, p < 0.002) und korrelierten mit der Knochenresorption (r = 0.85, p < 0.001 bzw. r = 0.82, p < 0.001). Entzündungen hatten keine messbaren Auswirkungen. WICHTIGSTE EINSCHRÄNKUNGEN: Limitierte Probenanzahl. Unassigned: Osteoklasten von Pferden wurden erfolgreich auf Knochenscheiben von Pferden kultiviert und ihre Knochenresorption quantifiziert. TRACP-5b erwies sich zum ersten Mal als Biomarker für die Anzahl der Osteoklasten und die Knochenresorption bei Pferden; CTX-I wurde ebenfalls als Biomarker für die Knochenresorption bei Pferden in vitro bestätigt. Dieser robuste pferdespezifische in-vitro-Assay wird die Untersuchung der Osteoklasten Biologie unterstützen.
© 2022 EVJ Ltd.
Get Full Text
Publication Date: 2022-06-19 PubMed ID: 35616632DOI: 10.1111/evj.13600Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research focuses on developing in vitro models for studying osteoclast activity in racehorses. This study aims to discover novel serum biomarkers that might help identify horses at risk for serious fractures, a need not fully met by current imaging techniques.

Research Objectives

  • The main objectives of this research were to establish osteoclast cultures on equine bone, to measure biomarkers in vitro, and to study the effects of inflammation.

Research Methodology

  • The research implemented various in vitro experiments. Haematopoietic stem cells sourced from equine sternal bone marrow aspirates were differentiated into osteoclasts.
  • These osteoclasts were then cultured either alone or on equine bone slices, with or without a pro-inflammatory stimulus (IL-1β or LPS).
  • The presence and levels of two biomarkers, tartrate resistant acid phosphatase isoform 5b (TRACP-5b) and C-terminal telopeptide of type I collagen (CTX-I), were quantified using an immunoassay conducted on the media.
  • Osteoclast numbers and bone resorption area were also assessed.

Research Results

  • Results showed that TRACP-5b increased over time in osteoclast cultures without bone and correlated with osteoclast number.
  • Both CTX-I and TRACP-5b increased over time for cultures with bone, with a correlation between the two as well as with bone resorption.
  • The researchers did not observe any measurable effects of inflammation.

Research Conclusions

  • The conclusions made were that equine osteoclasts could be successfully cultured on equine bone slices, and their bone resorption could be quantified.
  • TRACP-5b was identified as a biomarker of equine osteoclast number and bone resorption for the first time; CTX-I was also validated as a biomarker of equine bone resorption in vitro.
  • However, this study was limited by its sample size.
  • Overall, the creation of this robust equine-specific in vitro assay will aid future research into osteoclast biology.

Cite This Article

APA
Malek G, Richard H, Beauchamp G, Laverty S. (2022). An in vitro model for discovery of osteoclast specific biomarkers towards identification of racehorses at risk for catastrophic fractures. Equine Vet J, 55(3), 534-550. https://doi.org/10.1111/evj.13600

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 55
Issue: 3
Pages: 534-550

Researcher Affiliations

Malek, Gwladys
  • Comparative Orthopaedic Research Laboratory, Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Montreal, St-Hyacinthe, Q, Canada.
Richard, Hélène
  • Comparative Orthopaedic Research Laboratory, Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Montreal, St-Hyacinthe, Q, Canada.
Beauchamp, Guy
  • Comparative Orthopaedic Research Laboratory, Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Montreal, St-Hyacinthe, Q, Canada.
Laverty, Sheila
  • Comparative Orthopaedic Research Laboratory, Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Montreal, St-Hyacinthe, Q, Canada.

MeSH Terms

  • Horses
  • Animals
  • Osteoclasts
  • Tartrate-Resistant Acid Phosphatase / pharmacology
  • Acid Phosphatase / pharmacology
  • Isoenzymes / pharmacology
  • Biomarkers
  • Bone Resorption / veterinary
  • Fractures, Bone / veterinary
  • Inflammation / veterinary
  • Horse Diseases / diagnosis

Grant Funding

  • CF00142067 / Natural Sciences and Engineering Research Council of Canada
  • DF1_130184 / Quebec Cell, Tissue and Gene Therapy Network -Thu00e9Cell (a thematic network supported by the Fonds de recherche du Quu00e9bec-Santu00e9)

References

This article includes 128 references
  1. The Jockey Club. The Jockey Club Releases Data from the Equine Injury Database for 2020 - Supplemental Tables of Equine Injury Database Statistics for Thoroughbreds. Available from: http://jockeyclub.com/pdfs/eid_12_year_tables.pdf. 2021. Accessed 3 June 2021.
  2. Johnson BJ, Stover SM, Daft BM, Kinde H, Read DH, Barr BC. Causes of death in racehorses over a 2 year period. Equine Vet J 1994;26:327-30.
  3. Parkin TD, Clegg PD, French NP, Proudman CJ, Riggs CM, Singer ER. Analysis of horse race videos to identify intra-race risk factors for fatal distal limb fracture. Prev Vet Med 2006;74:44-55.
  4. Hitchens PL, Hill AE, Stover SM. Jockey falls, injuries, and fatalities associated with thoroughbred and quarter horse racing in California, 2007-2011. Orthop J Sports Med 2013;1:1-7.
  5. Boyde A, Firth EC. Musculoskeletal responses of 2-year-old thoroughbred horses to early training. 8. Quantitative back-scattered electron scanning electron microscopy and confocal fluorescence microscopy of the epiphysis of the third metacarpal bone. N Z Vet J 2005;53:123-32.
  6. Cornelissen BP, van Weeren PR, Ederveen AG, Barneveld A. Influence of exercise on bone mineral density of immature cortical and trabecular bone of the equine metacarpus and proximal sesamoid bone. Equine Vet J 1999;31(Suppl. 31):79-85.
  7. Firth EC. The response of bone, articular cartilage and tendon to exercise in the horse. J Anat 2006;208:513-26.
  8. Firth EC, Rogers CW, Jopson N. Effects of racetrack exercise on third metacarpal and carpal bone of New Zealand thoroughbred horses. J Musculoskelet Neuronal Interact 2000;1:145-7.
  9. Holmes JM, Mirams M, Mackie EJ, Whitton RC. Thoroughbred horses in race training have lower levels of subchondral bone remodelling in highly loaded regions of the distal metacarpus compared to horses resting from training. Vet J 2014;202:443-7.
  10. Noble P, Singer ER, Jeffery NS. Does subchondral bone of the equine proximal phalanx adapt to race training?. J Anat 2016;229:104-13.
  11. Norrdin RW, Stover SM. Subchondral bone failure in overload arthrosis: a scanning electron microscopic study in horses. J Musculoskelet Neuronal Interact 2006;6:251-7.
  12. Bertuglia A, Lacourt M, Girard C, Beauchamp G, Richard H, Laverty S. Osteoclasts are recruited to the subchondral bone in naturally occurring post-traumatic equine carpal osteoarthritis and may contribute to cartilage degradation. Osteoarthr Cartil 2016;24:555-66.
  13. Lacourt M, Gao C, Li A, Beachamp G, Henderson JE, Laverty S. Relationship between cartilage and subchondral bone lesions in repetitive impact trauma-induced equine osteoarthritis. Osteoarthr Cartil 2012;20:572-83.
  14. Whitton RC, Ayodele BA, Hitchens PL, Mackie EJ. Subchondral bone microdamage accumulation in distal metacarpus of Thoroughbred racehorses. Equine Vet J 2018;50:766-73.
  15. Norrdin RW, Kawcak CE, Capwell BA, McIlwraith CW. Subchondral bone failure in an equine model of overload arthrosis. Bone 1998;22:133-9.
  16. Colopy SA, Benz-Dean J, Barrett JG, Sample SJ, Lu Y, Danova NA. Response of the osteocyte syncytium adjacent to and distant from linear microcracks during adaptation to cyclic fatigue loading. Bone 2004;35:881-91.
  17. Muir P, Peterson AL, Sample SJ, Scollay MC, Markel MD, Kalscheur VL. Exercise-induced metacarpophalangeal joint adaptation in the thoroughbred racehorse. J Anat 2008;213:706-17.
  18. Gittens RA, Olivares-Navarrete R, Schwartz Z, Boyan BD. Implant osseointegration and the role of microroughness and nanostructures: lessons for spine implants. Acta Biomater 2014;10:3363-71.
  19. Riggs CM, Whitehouse GH, Boyde A. Pathology of the distal condyles of the third metacarpal and third metatarsal bones of the horse. Equine Vet J 1999;31:140-8.
  20. Martig S, Chen W, Lee PV, Whitton RC. Bone fatigue and its implications for injuries in racehorses. Equine Vet J 2014;46:408-15.
  21. Muir P, McCarthy J, Radtke CL, Markel MD, Santschi EM, Scollay MC. Role of endochondral ossification of articular cartilage and functional adaptation of the subchondral plate in the development of fatigue microcracking of joints. Bone 2006;38:342-9.
  22. Stover SM. The epidemiology of thoroughbred racehorse injuries. Clin Tech Equine Pract 2003;2:312-22.
  23. Stover SM. Nomenclature, classification, and documentation of catastrophic fractures and associated preexisting injuries in racehorses. J Vet Diagn Invest 2017;29:396-404.
  24. Powell SE. Low-field standing magnetic resonance imaging findings of the metacarpo/metatarsophalangeal joint of racing Thoroughbreds with lameness localised to the region: a retrospective study of 131 horses. Equine Vet J 2012;44:169-77.
  25. Ramzan PH, Palmer L, Powell SE. Unicortical condylar fracture of the thoroughbred fetlock: 45 cases (2006-2013). Equine Vet J 2015;47:680-3.
  26. Morgan JW, Santschi EM, Zekas LJ, Scollay-Ward MC, Markel MD, Radtke CL. Comparison of radiography and computed tomography to evaluate metacarpo/metatarsophalangeal joint pathology of paired limbs of thoroughbred racehorses with severe condylar fracture. Vet Surg 2006;35:611-7.
  27. Trope GD, Ghasem-Zadeh A, Anderson GA, Mackie EJ, Whitton RC. Can high-resolution peripheral quantitative computed tomography imaging of subchondral and cortical bone predict condylar fracture in thoroughbred racehorses?. Equine Vet J 2015;47:428-32.
  28. Simon V, Dyson SJ. Radiographic and scintigraphic evaluation of the third carpal bone of control horses and horses with carpal lameness. Vet Radiol Ultrasound 2012;53:465-73.
  29. Spriet M, Espinosa-Mur P, Cissell DD, Phillips KL, Arino-Estrada G, Beylin D. (18) F-sodium fluoride positron emission tomography of the racing thoroughbred fetlock: validation and comparison with other imaging modalities in nine horses. Equine Vet J 2019;51:375-83.
  30. Page AE, Adam E, Arthur R, Barker V, Franklin F, Friedman R. Expression of select mRNA in Thoroughbreds with catastrophic racing injuries. Equine Vet J 2022;54:63-73.
  31. Turlo AJ, Cywinska A, Frisbie DD. Revisiting predictive biomarkers of musculoskeletal injury in thoroughbred racehorses: longitudinal study in polish population. BMC Vet Res 2019;15:66.
  32. Parfitt AM. Osteonal and hemi-osteonal remodeling: the spatial and temporal framework for signal traffic in adult human bone. J Cell Biochem 1994;55:273-86.
  33. Stefaniuk-Szmukier M, Ropka-Molik K, Piórkowska K, Bugno-Poniewierska M. The expression profile of genes involved in osteoclastogenesis detected in whole blood of Arabian horses during 3 years of competing at race track. Res Vet Sci 2019;123:59-64.
  34. Stefaniuk-Szmukier M, Ropka-Molik K, Piórkowska K, Żukowski K, Bugno-Poniewierska M. Transcriptomic hallmarks of bone remodelling revealed by RNA-Seq profiling in blood of Arabian horses during racing training regime. Gene 2018;676:256-62.
  35. Troen BR. The regulation of cathepsin K gene expression. Ann N Y Acad Sci 2006;1068:165-72.
  36. Ono T, Nakashima T. Recent advances in osteoclast biology. Histochem Cell Biol 2018;149:325-41.
  37. Garnero P, Borel O, Byrjalsen I, Ferreras M, Drake FH, McQueney MS. The collagenolytic activity of cathepsin K is unique among mammalian proteinases. J Biol Chem 1998;273:32347-52.
  38. Garnero P, Ferreras M, Karsdal MA, Nicamhlaoibh R, Risteli J, Borel O. The type I collagen fragments ICTP and CTX reveal distinct enzymatic pathways of bone collagen degradation. J Bone Miner Res 2003;18:859-67.
  39. Breuil V, Cosman F, Stein L, Horbert W, Nieves J, Shen V. Human osteoclast formation and activity in vitro: effects of alendronate. J Bone Miner Res 1998;13:1721-9.
  40. Foged NT, Delaissé JM, Hou P, Lou H, Sato T, Winding B. Quantification of the collagenolytic activity of isolated osteoclasts by enzyme-linked immunosorbent assay. J Bone Miner Res 1996;11:226-37.
  41. Rosenquist C, Fledelius C, Christgau S, Pedersen BJ, Bonde M, Qvist P. Serum CrossLaps one step ELISA. First application of monoclonal antibodies for measurement in serum of bone-related degradation products from C-terminal telopeptides of type I collagen. Clin Chem 1998;44:2281-9.
  42. Alatalo SL, Halleen JM, Hentunen TA, Mönkkönen J, Väänänen HK. Rapid screening method for osteoclast differentiation in vitro that measures tartrate-resistant acid phosphatase 5b activity secreted into the culture medium. Clin Chem 2000;46:1751-4.
  43. Alatalo SL, Ivaska KK, Waguespack SG, Econs MJ, Väänänen HK, Halleen JM. Osteoclast-derived serum tartrate-resistant acid phosphatase 5b in Albers-Schonberg disease (type II autosomal dominant osteopetrosis). Clin Chem 2004;50:883-90.
  44. Chu P, Chao TY, Lin YF, Janckila AJ, Yam LT. Correlation between histomorphometric parameters of bone resorption and serum type 5b tartrate-resistant acid phosphatase in uremic patients on maintenance hemodialysis. Am J Kidney Dis 2003;41:1052-9.
  45. Halleen JM, Tiitinen SL, Ylipahkala H, Fagerlund KM, Väänänen HK. Tartrate-resistant acid phosphatase 5b (TRACP 5b) as a marker of bone resorption. Clin Lab 2006;52:499-509.
  46. Janckila AJ, Yam LT. Biology and clinical significance of tartrate-resistant acid phosphatases: new perspectives on an old enzyme. Calcif Tissue Int 2009;85:465-83.
  47. Rissanen JP, Suominen MI, Peng Z, Halleen JM. Secreted tartrate-resistant acid phosphatase 5b is a marker of osteoclast number in human osteoclast cultures and the rat ovariectomy model. Calcif Tissue Int 2008;82:108-15.
  48. Rissanen JP, Ylipahkala H, Fagerlund KM, Long C, Väänänen HK, Halleen JM. Improved methods for testing antiresorptive compounds in human osteoclast cultures. J Bone Miner Metab 2009;27:105-9.
  49. Halleen JM, Alatalo SL, Janckila AJ, Woitge HW, Seibel MJ, Väänänen HK. Serum tartrate-resistant acid phosphatase 5b is a specific and sensitive marker of bone resorption. Clin Chem 2001;47:597-600.
  50. Halleen JM, Ylipahkala H, Alatalo SL, Janckila AJ, Heikkinen JE, Suominen H. Serum tartrate-resistant acid phosphatase 5b, but not 5a, correlates with other markers of bone turnover and bone mineral density. Calcif Tissue Int 2002;71:20-5.
  51. Hannon RA, Clowes JA, Eagleton AC, Al Hadari A, Eastell R, Blumsohn A. Clinical performance of immunoreactive tartrate-resistant acid phosphatase isoform 5b as a marker of bone resorption. Bone 2004;34:187-94.
  52. Billinghurst RC, Brama PA, van Weeren PR, Knowlton MS, McIlwraith CW. Significant exercise-related changes in the serum levels of two biomarkers of collagen metabolism in young horses. Osteoarthr Cartil 2003;11:760-9.
  53. Frisbie DD, McIlwraith CW, Arthur RM, Blea J, Baker VA, Billinghurst RC. Serum biomarker levels for musculoskeletal disease in two- and three-year-old racing thoroughbred horses: a prospective study of 130 horses. Equine Vet J 2010;42:643-51.
  54. Jackson BF, Dyson PK, Lonnell C, Verheyen KL, Pfeiffer DU, Price JS. Bone biomarkers and risk of fracture in two- and three-year-old Thoroughbreds. Equine Vet J 2009;41:410-3.
  55. Kellerhouse PL, Brown C, Newhall K, Judd K, Thompson D. Assessment of bone resorption marker assays in thoroughbred horses. 2000.
  56. Carstanjen B, Hoyle NR, Gabriel A, Hars O, Sandersen C, Amory H. Evaluation of plasma carboxy-terminal cross-linking telopeptide of type I collagen concentration in horses. Am J Vet Res 2004;65:104-9.
  57. Delguste C, Amory H, Guyonnet J, Thibaud D, Garnero P, Detilleux J. Comparative pharmacokinetics of two intravenous administration regimens of tiludronate in healthy adult horses and effects on the bone resorption marker CTX-1. J Vet Pharmacol Ther 2008;31:108-16.
  58. Filipović N, Stojević Z, Plevnik N, Mašek T, Prvanović N, Tuček Z. The influence of age on bone metabolism in mares during late pregnancy and lactation. Res Vet Sci 2014;97:194-8.
  59. Hussein H, Ishihara A, Menendez M, Bertone A. Pharmacokinetics and bone resorption evaluation of a novel Cathepsin K inhibitor (VEL-0230) in healthy adult horses. J Vet Pharmacol Ther 2014;37:556-64.
  60. van de Lest CH, Brama PA, van Weeren PR. The influence of exercise on bone morphogenic enzyme activity of immature equine subchondral bone. Biorheology 2003;40:377-82.
  61. Kawaguchi T, Nakano T, Sasagawa K, Ohashi T, Miura T, Komoda T. Tartrate-resistant acid phosphatase 5a and 5b contain distinct sugar moieties. Clin Biochem 2008;41:1245-9.
  62. Gray AW, Davies ME, Jeffcott LB. In vitro generation of equine osteoclasts from bone marrow cells using a novel culture system. Res Vet Sci 1998;65:155-60.
  63. Gray AW, Davies ME, Jeffcott LB. Equine osteoclast-like cells generated in vitro demonstrate similar characteristics to directly isolated mature osteoclasts. Res Vet Sci 2000;68:161-7.
  64. Gray AW, Davies ME, Jeffcott LB. Localisation and activity of cathepsins K and B in equine osteoclasts. Res Vet Sci 2002;72:95-103.
  65. Gray AW, Davies ME, Jeffcott LB. Generation and activity of equine osteoclasts in vitro: effects of the bisphosphonate pamidronate (APD). Res Vet Sci 2002;72:105-13.
  66. Hussein H, Boyaka P, Dulin J, Russell D, Smanik L, Azab M. Cathepsin K localizes to equine bone in vivo and inhibits bone marrow stem and progenitor cells differentiation in vitro. J Stem Cells Regen Med 2017;13:45-53.
  67. Fortin-Trahan R, Lemirre T, Santschi EM, Janes JG, Richard H, Fogarty U. Osteoclast density is not increased in bone adjacent to radiolucencies (cysts) in juvenile equine medial femoral condyles. Equine Vet J 2021;00:1-10.
    doi: 10.1111/evj.13530google scholar: lookup
  68. Gilday R, Richard H, Beauchamp G, Fogarty U, Laverty S. Abundant osteoclasts in the subchondral bone of the juvenile thoroughbred metacarpus suggest an important role in joint maturation. Equine Vet J 2020;52:733-42.
  69. Stover SM, Pool RR, Martin RB, Morgan JP. Histological features of the dorsal cortex of the third metacarpal bone mid-diaphysis during postnatal growth in thoroughbred horses. J Anat 1992;181(Pt 3):455-69.
  70. Robling AG, Castillo AB, Turner CH. Biomechanical and molecular regulation of bone remodeling. Annu Rev Biomed Eng 2006;8:455-98.
  71. Murray RC, Vedi S, Birch HL, Lakhani KH, Goodship AE. Subchondral bone thickness, hardness and remodelling are influenced by short-term exercise in a site-specific manner. J Orthop Res 2001;19:1035-42.
  72. Whitton RC, Trope GD, Ghasem-Zadeh A, Anderson GA, Parkin TDH, Mackie EJ. Third metacarpal condylar fatigue fractures in equine athletes occur within previously modelled subchondral bone. Bone 2010;47:826-31.
  73. Whitton RC, Mirams M, Mackie EJ, Anderson GA, Seeman E. Exercise-induced inhibition of remodelling is focally offset with fatigue fracture in racehorses. Osteoporos Int 2013;24:2043-8.
  74. Désévaux C, Laverty S, Doizé B. Sternal bone biopsy in standing horses. Vet Surg 2000;29:303-8.
  75. Boraschi-Diaz I, Komarova SV. The protocol for the isolation and cryopreservation of osteoclast precursors from mouse bone marrow and spleen. Cytotechnology 2016;68:105-14.
  76. Rueden CT, Schindelin J, Hiner MC, DeZonia BE, Walter AE, Arena ET. ImageJ2: ImageJ for the next generation of scientific image data. BMC Bioinform 2017;18:1-26.
  77. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T. Fiji: an open-source platform for biological-image analysis. Nature 2012;9:676-82.
  78. Vesprey A, Yang W. Pit assay to measure the bone Resorptive activity of bone marrow-derived osteoclasts. Bio Protoc 2016;6:1-10.
  79. Durand M, Komarova SV, Bhargava A, Trebec-Reynolds DP, Li K, Fiorino C. Monocytes from patients with osteoarthritis display increased osteoclastogenesis and bone resorption: the in vitro osteoclast differentiation in arthritis study. Arthritis Rheum 2013;65:148-58.
  80. James IE, Lark MW, Zembryki D, Lee-Rykaczewski E, Hwang SM, Tomaszek TA. Development and characterization of a human in vitro resorption assay: demonstration of utility using novel antiresorptive agents. J Bone Miner Res 1999;14:1562-9.
  81. Krisher T, Bar-Shavit Z. Regulation of osteoclastogenesis by integrated signals from toll-like receptors. J Cell Biochem 2014;115:2146-54.
  82. Kudo O, Fujikawa Y, Itonaga I, Sabokbar A, Torisu T, Athanasou NA. Proinflammatory cytokine (TNFalpha/IL-1alpha) induction of human osteoclast formation. J Pathol 2002;198:220-7.
  83. Hussein H, Boyaka P, Dulin J, Bertone A. Cathepsin K inhibition renders equine bone marrow nucleated cells hypo-responsive to LPS and unmethylated CpG stimulation in vitro. Comp Immunol Microbiol Infect Dis 2016;45:40-7.
  84. Noé B, Poole AR, Mort JS, Richard H, Beauchamp G, Laverty S. C2K77 ELISA detects cleavage of type II collagen by cathepsin K in equine articular cartilage. Osteoarthr Cartil 2017;25:2119-26.
  85. Vézina Audette R, Lavoie-Lamoureux A, Lavoie JP, Laverty S. Inflammatory stimuli differentially modulate the transcription of paracrine signaling molecules of equine bone marrow multipotent mesenchymal stromal cells. Osteoarthr Cartil 2013;21:1116-24.
  86. Kylmäoja E, Nakamura M, Turunen S, Patlaka C, Andersson G, Lehenkari P. Peripheral blood monocytes show increased osteoclast differentiation potential compared to bone marrow monocytes. Heliyon 2018;4:e00780.
  87. Shiratori T, Kyumoto-Nakamura Y, Kukita A, Uehara N, Zhang J, Koda K. IL-1β induces pathologically activated osteoclasts bearing extremely high levels of resorbing activity: a possible pathological subpopulation of osteoclasts, accompanied by suppressed expression of Kindlin-3 and Talin-1. J Immunol 2018;200:218-28.
  88. Strålberg F, Kassem A, Kasprzykowski F, Abrahamson M, Grubb A, Lindholm C. Inhibition of lipopolysaccharide-induced osteoclast formation and bone resorption in vitro and in vivo by cysteine proteinase inhibitors. J Leukoc Biol 2017;101:1233-43.
  89. Zou W, Bar-Shavit Z. Dual modulation of osteoclast differentiation by lipopolysaccharide. J Bone Miner Res 2002;17:1211-8.
  90. Cohen J. A power primer. Psychol Bull 1992;112:155-9.
  91. Schober P, Boer C, Schwarte LA. Correlation coefficients: appropriate use and interpretation. Anesth Analg 2018;126:1763-8.
  92. Collar E, Huber M, Iwaniec U, Duesterdieck-Zellmer K, Stover S. Third Carpal bone fracture is associated with focal subchondral bone porosity in racehorses. 2016.
  93. Merrild DM, Pirapaharan DC, Andreasen CM, Kjaersgaard-Andersen P, Møller AMJ, Ding M. Pit- and trench-forming osteoclasts: a distinction that matters. Bone Res 2015;3:15032.
  94. Susa M, Luong-Nguyen NH, Cappellen D, Zamurovic N, Gamse R. Human primary osteoclasts: in vitro generation and applications as pharmacological and clinical assay. J Transl Med 2004;2:6.
  95. Kingsmill VJ, Gray C, Boyde A. Osteoclastic resorption of equine cranial and postcranial bone in vitro. J Bone Miner Metab 2000;18:148-52.
  96. Perrotti V, Nicholls BM, Piattelli A. Human osteoclast formation and activity on an equine spongy bone substitute. Clin Oral Implants Res 2009;20:17-23.
  97. Hadjidakis DJ, Androulakis II. Bone remodeling. Ann N Y Acad Sci 2006;1092:385-96.
  98. Mira-Pascual L, Patlaka C, Desai S, Paulie S, Näreoja T, Lång P. A novel Sandwich ELISA for tartrate-resistant acid phosphatase 5a and 5b protein reveals that both isoforms are secreted by differentiating osteoclasts and correlate to the type I collagen degradation marker CTX-I in vivo and in vitro. Calcif Tissue Int 2020;106:194-207.
  99. Søe K, Delaissé JM. Glucocorticoids maintain human osteoclasts in the active mode of their resorption cycle. J Bone Miner Res 2010;25:2184-92.
  100. Bonde M, Qvist P, Fledelius C, Riis BJ, Christiansen C. Immunoassay for quantifying type I collagen degradation products in urine evaluated. Clin Chem 1994;40:2022-5.
  101. Garnero P, Gineyts E, Riou JP, Delmas PD. Assessment of bone resorption with a new marker of collagen degradation in patients with metabolic bone disease. J Clin Endocrinol Metab 1994;79:780-5.
  102. Alatalo SL, Peng Z, Janckila AJ, Kaija H, Vihko P, Vaananen HK. A novel immunoassay for the determination of tartrate-resistant acid phosphatase 5b from rat serum. J Bone Miner Res 2003;18:134-9.
  103. Gerdhem P, Ivaska KK, Alatalo SL, Halleen JM, Hellman J, Isaksson A. Biochemical markers of bone metabolism and prediction of fracture in elderly women. J Bone Miner Res 2004;19:386-93.
  104. Smith LL. Tissue trauma: the underlying cause of overtraining syndrome?. J Strength Cond Res 2004;18:185-93.
  105. Huldani PI, Massi MN, Idris I, Bukhari A, Widodo ADW, Achmad H. Research reviews on effect of exercise on DAMP's, HMGB1, proinflammatory cytokines and leukocytes. Sys Rev Pharm 2020;11:306-12.
  106. Walsh NP, Gleeson M, Shephard RJ, Gleeson M, Woods JA, Bishop NC. Position statement. Part one: Immune function and exercise. Exerc Immunol Rev 2011;17:6-63.
  107. Capomaccio S, Cappelli K, Spinsanti G, Mencarelli M, Muscettola M, Felicetti M. Athletic humans and horses: comparative analysis of interleukin-6 (IL-6) and IL-6 receptor (IL-6R) expression in peripheral blood mononuclear cells in trained and untrained subjects at rest. BMC Physiol 2011;11:3.
  108. Capomaccio S, Vitulo N, Verini-Supplizi A, Barcaccia G, Albiero A, D'Angelo M. RNA sequencing of the exercise transcriptome in equine athletes. PLoS One 2013;8:e83504.
  109. Cappelli K, Felicetti M, Capomaccio S, Nocelli C, Silvestrelli M, Verini-Supplizi A. Effect of training status on immune defence related gene expression in thoroughbred: are genes ready for the sprint?. Vet J 2013;195:373-6.
  110. Cywinska A, Turło A, Witkowski L, Szarska E, Winnicka AJMW. Changes in blood cytokine concentrations in horses after long-distance endurance rides. Med Weter 2014;70:568-71.
  111. Donovan DC, Jackson CA, Colahan PT, Norton N, Hurley DJ. Exercise-induced alterations in pro-inflammatory cytokines and prostaglandin F2alpha in horses. Vet Immunol Immunopathol 2007;118:263-9.
  112. Horohov DW, Sinatra ST, Chopra RK, Jankowitz S, Betancourt A, Bloomer RJ. The effect of exercise and nutritional supplementation on proinflammatory cytokine expression in young racehorses during training. J Equine Vet Sci 2012;32:805-15.
  113. Liburt NR, Adams AA, Betancourt A, Horohov DW, McKeever KH. Exercise-induced increases in inflammatory cytokines in muscle and blood of horses. Equine Vet J 2010;42(Suppl 38):280-8.
  114. Sloan AJ, Taylor SY, Smith EL, Roberts JL, Chen L, Wei XQ. A novel ex vivo culture model for inflammatory bone destruction. J Dent Res 2013;92:728-34.
  115. Crisan M, Dzierzak E. The many faces of hematopoietic stem cell heterogeneity. Development 2016;143:4571-81.
  116. Carter-Arnold JL, Neilsen NL, Amelse LL, Odoi A, Dhar MS. In vitro analysis of equine, bone marrow-derived mesenchymal stem cells demonstrates differences within age- and gender-matched horses. Equine Vet J 2014;46:589-95.
  117. Schnabel LV, Pezzanite LM, Antczak DF, Felippe MJ, Fortier LA. Equine bone marrow-derived mesenchymal stromal cells are heterogeneous in MHC class II expression and capable of inciting an immune response in vitro. Stem Cell Res Ther 2014;5:1-13.
  118. Mora S, Prinster C, Proverbio MC, Bellini A, de Poli SC, Weber G. Urinary markers of bone turnover in healthy children and adolescents: age-related changes and effect of puberty. Calcif Tissue Int 1998;63:369-74.
  119. Rauchenzauner M, Schmid A, Heinz-Erian P, Kapelari K, Falkensammer G, Griesmacher A. 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;92:443-9.
  120. Walsh JS, Paggiosi MA, Eastell R. Cortical consolidation of the radius and tibia in young men and women. J Clin Endocrinol Metab 2012;97:3342-8.
  121. Estberg L, Stover SM, Gardner IA, Johnson BJ, Case JT, Ardans A. Fatal musculoskeletal injuries incurred during racing and training in Thoroughbreds. J Am Vet Med Assoc 1996;208:92-6.
  122. Estberg L, Stover SM, Gardner IA, Johnson BJ, Jack RA, Case JT. Relationship between race start characteristics and risk of catastrophic injury in Thoroughbreds: 78 cases (1992). J Am Vet Med Assoc 1998;212:544-9.
  123. Hatipoglu MG, Inal S, Kabay S, Cayci MK, Deger A, Kuru HI. The influence of different nonsteroidal anti-inflammatory drugs on alveolar bone in rats: an experimental study. Acta Stomatol Croat 2015;49:325-30.
  124. Shi J, Wang Z, Guo X, Shen J, Sun H, Bai J. Aspirin inhibits osteoclast formation and wear-debris-induced bone destruction by suppressing mitogen-activated protein kinases. J Cell Physiol 2020;235:2599-608.
  125. Son HS, Lee J, Lee HI, Kim N, Jo Y-J, Lee G-R. Benzydamine inhibits osteoclast differentiation and bone resorption via down-regulation of interleukin-1 β expression. Acta Pharm Sin B 2020;10:462-74.
  126. Zhang S, Huo S, Li H, Tang H, Nie B, Qu X. Flufenamic acid inhibits osteoclast formation and bone resorption and act against estrogen-dependent bone loss in mice. Int Immunopharmacol 2020;78:106014.
  127. Mashiba T, Turner CH, Hirano T, Forwood MR, Johnston CC, Burr DB. Effects of suppressed bone turnover by bisphosphonates on microdamage accumulation and biomechanical properties in clinically relevant skeletal sites in beagles. Bone 2001;28:524-31.
  128. Sloan AV, Martin JR, Li S, Li J. Parathyroid hormone and bisphosphonate have opposite effects on stress fracture repair. Bone 2010;47:235-40.

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,824 horse owners like you who receive our email updates on horse health and nutrition.