Collagen is a structural protein found abundantly in the connective tissues of horses, including skin, tendons, ligaments, and cartilage. It contributes to the strength and elasticity of these tissues, playing a significant role in maintaining structural integrity. Collagen fibers are composed of amino acid chains, primarily glycine, proline, and hydroxyproline, which form a triple helix structure. In horses, collagen is involved in various physiological processes, including tissue repair and regeneration. Changes in collagen composition and structure can be associated with conditions such as joint disorders and tendon injuries. This page compiles peer-reviewed research studies and scholarly articles that explore the synthesis, function, and clinical implications of collagen in equine health.
MacLeod JN, Fubini SL, Gu DN, Tetreault JW, Todhunter RJ.To determine whether steady-state levels of type-II procollagen, aggrecan core protein, or fibronectin mRNA in articular chondrocytes are altered by synovitis or administration of methylprednisolone acetate (MPA). Methods: Articular cartilage specimens collected from 10 ponies, 2.5 to 3.5 years old and 200 to 300 kg. Methods: 4 experimental groups were compared, using the cartilage specimens: control, MPA-treated, lipopolysaccharide-induced synovitis, and lipopolysaccharide-induced synovitis with MPA treatment. RNA was isolated from articular cartilage and compared by northern blot analysis, u...
Fecteau KA, Haffner JC, Eiler H.The purpose of this study was to determine to what degree bacterial collagenase may digest human placentae compared to equine and bovine placentae. Placenta samples from human, equine and bovine were incubated with bacterial collagenase solution at various concentrations. The degree of hydrolysis and collagen breakdown was measured by the release of total proteins and hydroxyproline into the incubation media. Also, whole placentae were injected via umbilical cord arteries with collagenase solution (200 U/ml, 200 ml total volume in human and 1000 ml in equine) and hydrolysis determined chemical...
Todhunter RJ, Fubini SL, Vernier-Singer M, Wootton JA, Lust G, Freeman KP, MacLeod JN.To determine how acute synovitis, with and without intra-articular methylprednisolone acetate (MPA), affect synthesis of proteoglycan, total protein, and collagen in articular cartilage and total protein synthesis in synovial membrane. Methods: Synovitis was induced in 10 ponies by the injection of 0.5 ng lipopolysaccharide (LPS) into the left radiocarpal and midcarpal joints every 2 days for a total of four treatments. Synovitis was documented by clinical examination and synovial fluid analyses. Two days before euthanasia, MPA (0.1 mg/kg) was injected with the last dose of LPS into both the l...
Lynch TM, Caron JP, Arnoczky SP, Lloyd JW, Stick JA, Render JA.To evaluate the influence of exogenous hyaluronan (HA) on in vitro synthesis of HA and collagenase by equine synoviocytes from normal and inflamed joints. Methods: 9 adult horses. Methods: Synoviocytes for culture were taken from the middle carpal joint of 3 horses with normal joints (control) and 6 horses with osteochondral fractures (principal). Synoviocytes were propagated in monolayer cultures and were incubated with 3 commercial HA products at concentrations of 0, 200, 400, and 1,500 micrograms/ml. Newly synthesized HA was radiolabeled with [3H]glucosamine and quantified by cetylpyridiniu...
Johnson PJ, Tyagi SC, Katwa LC, Ganjam VK, Moore LA, Kreeger JM, Messer NT.Samples of connective tissue obtained from the hoof of six laminitic and eight non-laminitic adult horses were analysed zymographically to investigate whether connective tissue matrix metalloproteinases are activated or induced during laminitis. The activity or matrix metalloproteinases was substantially greater in the tissues from the laminitic horses than in the tissues from the non-laminitic horses. A comparison of the collagenolytic activity in the laminitic and control tissues showed that collagenolytic activities corresponding to the 92 kDa (P < 0.001), 72 kDa (P < 0.01) and 66 kDa (P < ...
Patterson-Kane JC, Wilson AM, Firth EC, Parry DA, Goodship AE.Injury to the core of the mid-metacarpal region of the superficial digital flexor tendon in Thoroughbred racehorses is a very frequent but poorly understood condition. It has been suggested that subclinical changes induced by galloping exercise weaken the collagen in this region of the tendon, predisposing it to rupture. The longitudinally arranged collagen fibrils in tendon follow a planar waveform, termed the crimp. Fibril bundles with a smaller crimp angle fail at a lower level of strain than those with a larger crimp angle. This study tested the hypothesis that a specific 18 month exercise...
Patterson-Kane JC, Firth EC, Parry DA, Wilson AM, Goodship AE.To determine the effect of a specific galloping exercise regimen on collagen fibril mass-average diameters (MAD) in the deep digital flexor tendon (DDFT) and suspensory ligament (SL) of young Thoroughbreds. Methods: 12 Thoroughbred fillies, 21 +/- 1 (mean +/- SD) months old. Methods: 6 horses underwent a specific 18-month treadmill training program involving galloping exercise. The remaining 6 horses served as controls, undertaking low-volume walking exercise over the same period. Sections were excised from the midpoint of the DDFT and SL, and small strips were dissected from central and perip...
Cochrane CA.Abstract Wound models attempt to simulate the natural healing processes in wounds. However, all models have significant limitations due to the complexity of the tissue repair process. Much can be learned from wound models in vitro by the use of cell culture techniques. The horse can provide a suitable naturally occurring model of chronic wound healing because it has many similarities to wound healing encountered in human medicine. The tissue architecture was investigated with regard to extracellular matrix and growth factor distribution during wound healing and growth factors were consisten...
Patterson-Kane JC, Parry DA, Goodship AE, Firth EC.One of the current concepts with regard to equine superficial digital flexor tendonitis is that cumulative subclinical microscopic damage weakens the structure, predisposing the tendon to partial or complete rupture. This microtrauma is likely to affect the waveform or crimp of the collagen fibrils, which are the units of tensile strength. Collagen fibril crimp morphology characteristics were determined in superficial digital flexor tendons of 18 horses presented for necropsy. Horses were separated into exercised (n = 9) and non-exercised groups (n = 9), based on recent function. Five of the e...
Henson FM, Davies ME, Jeffcott LB.This study describes (1) the histological appearance of dyschondroplasia, the primary lesion of osteochondrosis, in articular cartilage of the horse and (2) the localization of type VI collagen which is an important constituent of the extracellular matrix (ECM). Dyschondroplastic cartilage was identified on the basis of the presence of cartilage cores (i.e., cartilage extending into the subchondral bone) and confirmed with subsequent histological examination. Full-thickness cartilage samples from 57 horses were collected and paraffin embedded. Histological examination was used to examine the n...
Frisbie DD, Nixon AJ.To evaluate potential stimulatory or matrix-sparing effects of insulin-like growth factor 1 (IGF-1), alone or in combination with a corticosteroid, in an interleukin 1 (IL-1)-induced model of cartilage degradation. Methods: Cartilage from the weightbearing surfaces of trochlea and condyles of clinically normal 2-year-old male horses. Methods: Triamcinolone acetonide and IGF-1 effects were evaluated by assessing: matrix responses by sulfated glycosaminoglycan (GAG) assay and [35S]sulfated GAG synthesis; collagen content by hydroxyproline assay; and mitogenic response by [3H]thymidine incorporat...
Gillis C, Pool RR, Meagher DM, Stover SM, Reiser K, Willits N.To assess tendon morphology and non-reducible crosslink concentration, and associations of these findings with horse age and previously reported mechanical and ultrasonographic findings. Methods: Superficial digital flexor tendon samples were obtained from 23 horses aged 2 to 23 years. The tendons had undergone ultrasonography and were submitted to biomechanical testing in the physiologic range prior to sample acquisition. Methods: Samples were sectioned in a transverse plane; then dorsal, palmar, central, lateral, and medial regions were evaluated for fascicle cross-sectional area (CSA), sept...
Patterson-Kane JC, Wilson AM, Firth EC, Parry DA, Goodship AE.This study was undertaken to test the hypothesis that collagen fibrils, the submicroscopic units of strength in tendon, would hypertrophy in response to a specific defined training programme. Fibril diameters were measured in central and peripheral regions of superficial digital flexor tendon (SDFT) samples from five 18-month-old horses which underwent a subsequent 18 month training programme and 6 age- and sex-matched controls. Central region fibrils from the trained horses had a mass-average diameter (MAD) of 105.3 nm, which was significantly lower (P < 0.01) than that of 131.7 nm for the...
Sime A, McKellar Q, Nolan A.A serum-free cell culture method was developed for equine tracheal epithelial cells which allowed the growth and characterisation of the phenotypical properties of this cell type. Several variables influenced the efficacy of the attachment and growth of the isolated cells. Serum and a collagen matrix were essential components for efficient cell attachment. Once attachment had occurred, cell growth was enhanced by a serum-free medium containing bovine pituitary extract, retinoic acid, insulin, hydrocortisone, transferrin, epidermal growth factor, adrenaline and triiodothyronine. The mean time t...
Patterson-Kane JC, Firth EC, Goodship AE, Parry DA.To measure collagen fibril crimp angles and lengths as well as collagen fibril mass-average diameters in central and peripheral regions of the superficial digital flexor tendon of wild horses, to ascertain any age-related changes in either region in the absence of imposed galloping exercise. Methods: Measurements from a random cull of wild horses. Methods: Superficial digital flexor tendon samples were taken from 23 wild horses ranging in age from two to ten years. Methods: Horses were divided into 'young' (< 5 years, n = 10), 'middle-aged' (5 to < 10 years, n = 9) and 'ol...
Murphy DJ, Nixon AJ.To examine the site-specific and dose-dependent effects of insulin-like growth factor I (IGF-I) on normal equine tendon in vitro. Methods: Superficial digital flexor tendon explants derived from a euthanatized 3-year-old horse. Methods: Explants in culture were treated with 0, 100, 250, or 500 ng of IGF-I/ml for 14 days with an end-stage radiolabel of 20 microCi of [3H]proline/ml or 5 microCi of [3H]thymidine/ml. The tendon tissues were then analyzed biochemically for hydroxyproline content by reverse-phase high-performance liquid chromatography, DNA content by fluorometry, and glycosaminoglyc...
Fortier LA, Nixon AJ, Mohammed HO, Lust G.To determine the effects of transforming growth factor-beta 1 (TGF-beta 1) on the synthesis of DNA, collagen, and proteoglycans (PG) by equine chondrocytes. Methods: Articular cartilage obtained from multiple joints of a 4-month-old foal. Methods: Chondrocytes were isolated by collagenase digestion, cultured in monolayer, trypsinized, and implanted at a cellular density of 10 x 10(6) chondrocytes/ml in a three-dimensional fibrin matrix. Chondrocytes in culture were supplemented with TGF-beta 1 at concentrations of 0, 1, 5, or 10 ng/ml in serum-free medium or medium containing fetal bovine seru...
Patterson-Kane JC, Parry DA, Birch HL, Goodship AE, Firth EC.The superficial digital flexor tendon is the most commonly injured tendon in the racing Thoroughbred. Despite the clinical significance of this structure, only limited data exist regarding normal age-related morphology of the tensile units, the collagen fibrils. The age at which these collagen fibrils become mature in composition and structure may be of importance. Consequently, the association of age and collagen fibril crosslink composition, diameter distribution and crimp morphology in the superficial and deep digital flexor tendons of Thoroughbreds up to and including three years of age ha...
Koivunen AL, Maisi P, Konttinen YT, Prikk K, Sandholm M.The collagenolytic activity and its sensitivity to doxycycline inhibition in tracheal aspirates (TA) of horses with chronic obstructive pulmonary disease (COPD) was analyzed with SDS-PA gel electrophoresis (SDS-PAGE), using Type 1 collagen as the substrate. Both autoactive and total collagenase activities were significantly higher in TAs of horses with symptomatic COPD than in TAs of healthy horses. Doxycycline inhibition studies suggest that most of the TA collagenase is of the neutrophil type (MMP-8), but some is derived from other cells such as fibroblasts and monocyte/macrophages (MMP-1) a...
Martin RB, Lau ST, Mathews PV, Gibson VA, Stover SM.We studied birefringence as an indicator of collagen fiber orientation in the diaphysis of the equine third metacarpal bone. We had previously shown that tissue from the lateral cortex of this bone is stronger monotonically, but less fatigue resistant, than tissue from the medial and dorsal regions. To learn whether collagen fiber orientation might play a role in this regional specialization, we tested three hypotheses using the same specimens: (1) collagen fiber orientation is regionally dependent; (2) remodeling changes collagen fiber orientation; (3) longitudinal collagen fibers correlate p...
Skedros JG, Mason MW, Nelson MC, Bloebaum RD.Functionally induced strains provide epigenetic signaling for bone modeling and remodeling activities. Strain gauge documentation of the equine third metacarpal reveals a neutral axis passing through the craniolateral cortex, resulting in a narrow band of cortex loaded predominantly in tension, with the remainder of the cortex experiencing a wide range of compression strain magnitudes that are maximal in the caudomedial cortex. This predictable strain pattern provides a model for examining the hypothesis that strain mode, magnitude, and strain energy density are potential correlates of compact...
Todhunter RJ, Fubini SL, Wootton JA, Lust G.The effect of different doses of methylprednisolone acetate (MPA) on proteoglycan and collagen metabolism of articular cartilage from normal equine joints was tested in vitro. Methods: Cultured explants were treated with 0, 0.0004, 0.004, 0.04, 0.4 and 4.0 mg/ml (approximately 10(-6)-10(-2) M) MPA for 72 h. Proteoglycan synthesis was measured by incorporation of sodium [35S]sulfate into proteoglycans and proteoglycan degradation was measured by release of total and radiolabeled proteoglycan into the culture media. The size of the proteoglycans was assessed with size exclusion chromatography un...
Henson FMD, Davies ME, Schofield PN, Jeffcott LB.The synthesis and expression of collagen types II, VI and X were investigated in growth cartilage selected from a group of 31 horses and ponies in the age range 157 days of gestation to 12 years. Collagen isolation, immunolocalisation and in situ hybridisation techniques were used in order to provide information on the pattern of synthesis of these 3 collagens during endochondral ossification in normal horses. Type II collagen immunoreactivity and mRNA expression was found in each of the 3 zones of growth cartilage chondrocytes in all samples studied, whereas the localisation of both collagen ...
Donald PJ, Deckard-Janatpour K, Sharkey N, Lagunas-Solar M.Various centers report irradiated cartilage graft absorption rates that differ quite widely. We postulated that a major factor governing this phenomenon might be irradiation dose. Irradiation produces collagen cross-binding and increased resistance to absorption of such material when implanted. Since cross-binding produces stiffening of collagen, cartilage grafts were exposed to increasing doses of irradiation and their elastic modulus was measured. The postulate was that increasing radiation doses will produce grafts of increasing stiffness. Sternal cartilage, harvested from horses, was cut i...
Farndale RW, Napthine CS, Evans RJ, Hayes LJ, Heath MF.Equine platelet aggregation was stimulated by collagen fibres or platelet-activating factor. The action of both ligands was blocked by forskolin or prostaglandin E(1) agents which are known to activate adenylate cyclase. Equine platelet membranes were found to contain adenylate cyclase activity which was inhibited in dose-dependent fashion by both collagen and platelet-activating factor. Platelet-activating factor-induced inhibition was antagonised by WEB2086.
Miles CA, Fursey GA, Birch HL, Young RD.The velocity, attenuation and apparent backscattering coefficient of 6-11-MHz ultrasound were measured in three orthogonal directions in equine deep digital flexor (DDF) and superficial digital flexor (SDF) tendons at 0 degree C. Ultrasonic measurements were examined for correlation with tendon water, collagen, DNA and glycosaminoglycans contents, determined by chemical analyses and with structure observed by scanning electron microscopy. The SDF tendon contained more water, more DNA (i.e., more cells), less collagen and less glycosaminoglycans and exhibited lower velocities and attenuations t...
Davankar SP, Deane NJ, Davies AS, Firth EC, Hodge H, Parry DA.Since horses bred for the racing industry are subject to rigorous training procedures there is a real need to understand how the stresses experienced by their tendons and ligaments in vivo relate to the major load-bearing elements-the collagen fibrils. Consequently, an age-related study has been made of the collagen fibril diameter distributions of nine ligaments in and around the equine carpus. This is the first stage of a larger study aimed at understanding the ultrastructural changes that occur as a result of exercise. Most of the ligaments showed a bimodal diameter distribution at maturity...
Keg PR, Schamhardt HC, van Weeren PR, Barneveld A.Controversy exists with respect to the innervation of the suspensory ligament (SL) in the fore limb of the horse. It is uncertain whether this structure is exclusively innervated by branches of the ulnar nerve or also to some extent by median nerve branches. Ground Reaction Forces (GRF) were determined in horses before and after the induction of a tendonitis in the lateral branch of the SL by the injection of collagenase, and before and after a high palmar and an ulnar block respectively. The high palmar block succeeded in bringing all GRF variables back to their original values which the ulna...
Yvorchuk-St Jean K, Gaughan E, St Jean G, Frank R.To evaluate the effect of a porous bovine-derived collagen membrane (PBCM) on the rates of wound healing, cellular events, presence of granulation tissue, and appearance at termination of the study in surgically created full-thickness cutaneous wounds of the distal portion of the extremities of horses. Methods: Treated wounds (n = 12) received a PBCM dressing and control wounds were covered with a nonadherent dressing. Forelimbs and hind limbs were cross paired; the right forelimb and left hind limb always received the same dressing application, as did the left forelimb and right hind limb. Li...
van den Berg MJ, Rijkenhuizen AB, Németh F, Gruys E.Chronic changes of several structures in around the fetlock tunnel can be a cause of the so-called fetlock tunnel syndrome (FTS) in the horse. Forty-nine annular ligaments (AL) from dead horses without a known history or clinical evidence of lameness and/or digital tendon sheath problems in these legs and 30 AL biopsies from horses suffering from FTS were studied macroscopically and microscopically. Macroscopically, the normal AL had a shiny white appearance, whereas the affected AL were often thicker and less white. Microscopically, the normal AL were about +/- 1 mm thick and were composed of...
Visser MB, Pollitt CC.The basement membrane (BM) is a thin layer of extracellular matrix that regulates cell functions as well as providing support to tissues of the body. Primary components of the BM of epithelial tissues are laminin-332 (Ln-332) and collagen type IV. Equine laminitis is a disease characterized by destruction and dislocation of the hoof lamellar BM. Immunohistochemistry was used to characterize the distribution of Ln-332 and collagen type IV in the organs of normal horses and these proteins were found to be widespread. Analysis of a panel of tissue samples from horses with experimentally-induced l...
BMC research notesNovember 8, 2012
Volume 5 626 doi: 10.1186/1756-0500-5-626
Boudko SP, Ishikawa Y, Lerch TF, Nix J, Chapman MS, Bächinger HP.Hyperelastosis cutis is an inherited autosomal recessive connective tissue disorder. Affected horses are characterized by hyperextensible skin, scarring, and severe lesions along the back. The disorder is caused by a mutation in cyclophilin B. Results: The crystal structures of both wild-type and mutated (Gly6->Arg) horse cyclophilin B are presented. The mutation neither affects the overall fold of the enzyme nor impairs the catalytic site structure. Instead, it locally rearranges the flexible N-terminal end of the polypeptide chain and also makes it more rigid. Conclusions: Interactions of th...
Todhunter RJ, Fubini SL, Wootton JA, Lust G.The effect of different doses of methylprednisolone acetate (MPA) on proteoglycan and collagen metabolism of articular cartilage from normal equine joints was tested in vitro. Methods: Cultured explants were treated with 0, 0.0004, 0.004, 0.04, 0.4 and 4.0 mg/ml (approximately 10(-6)-10(-2) M) MPA for 72 h. Proteoglycan synthesis was measured by incorporation of sodium [35S]sulfate into proteoglycans and proteoglycan degradation was measured by release of total and radiolabeled proteoglycan into the culture media. The size of the proteoglycans was assessed with size exclusion chromatography un...
Kim TI, Chung CP, Heo MS, Park YJ, Rhee SH.This study was performed to evaluate the periodontal wound healing effect of particulate equine bone mineral on canine alveolar bone defects. Methods: Twelve adult male beagle dogs were used as study subjects. The mandibular second and fourth premolars were extracted prior to the experimental surgery, and the extraction sites were allowed to heal for 8 weeks. After periodontal probing, two-walled defects were created at the mesial and distal sides of the mandibular third premolars bilaterally, and the defects were filled with equine particulate bone with collagen membrane or bovine particulate...
Stack A, Derksen FJ, Sordillo LM, Williams KJ, Stick JA, Brandenberger C, Steibel JP, Robinson NE.To determine the effects of 2 weeks of intense exercise on expression of markers of pulmonary venous remodeling in the caudodorsal and cranioventral regions of the lungs of horses. Methods: 6 horses. Methods: Tissue samples of the caudodorsal and cranioventral regions of lungs were obtained before and after conditioning and 2 weeks of intense exercise. Pulmonary veins were isolated, and a quantitative real-time PCR assay was used to determine mRNA expression of matrix metalloproteinase-2 and -9, tissue inhibitor of metalloproteinase-1 and -2, collagen type I, tenascin-C, endothelin-1, platelet...
Brinkman EL, Weed BC, Patnaik SS, Brazile BL, Centini RM, Wills RW, Olivier B, Sledge DG, Cooley J, Liao J, Rashmir-Raven AM.OBJECTIVE To compare biomechanical and histologic features of heart valves and echocardiographic findings between Quarter Horses with and without heritable equine regional dermal asthenia (HERDA). DESIGN Prospective case-control study. ANIMALS 41 Quarter Horses. PROCEDURES Ultimate tensile strength (UTS) of aortic and mitral valve leaflets was assessed by biomechanical testing in 5 horses with HERDA and 5 horses without HERDA (controls). Histologic evaluation of aortic and mitral valves was performed for 6 HERDA-affected and 3 control horses. Echocardiography was performed in 14 HERDA-affected...
Barr AR, Wotton SF, Dow SM, Waterman AE, Goodship AE, Duance VC.The effects of osteochondral defect location and post-operative walking exercise on structural repair and recovery of joint function were examined in the midcarpal joints of ponies. Functional recovery was monitored by measuring ground reaction forces using a force plate. Structural repair was evaluated histologically and by measuring the total collagen and uronic acid content and relative proportions of Type I and II collagen in the repair tissue. Central defects tended to cause a more marked functional disturbance but were repaired with fibrocartilage rather than fibrous tissue in 3 out of 6...
Micklethwaite L, Wood AK, Sehgal CM, Polansky M, Dowling BA, Dart AJ, Rose RJ, Hodgson DR.To determine whether quantitative analysis of sonographic brightness could be used to detect healing of an induced injury of the superficial digital flexor tendon in horses and whether rate of healing was influenced by equine recombinant growth hormone. Methods: 8 clinically normal Standardbreds. Methods: A localized injury was created in the left and right superficial digital flexor tendons of each horse by injection of 2,000 units of collagenase. After injury, 4 horses received equine recombinant growth hormone, a possible promoter of tendon healing. Sonographic images (7.5 MHz) of the flexo...
Wise LM, Stuart GS, Sriutaisuk K, Adams BR, Riley CB, Theoret CL.Fibroproliferative disorders occur in both humans and horses following skin injury. In horses, wound healing on the limb is often complicated by the formation of fibroproliferative exuberant granulation tissue, characterized by persistent expression of pro-fibrotic transforming growth factor-beta1 (TGF-β1) and deficient expression of anti-inflammatory interleukin-10 (IL-10). IL-10 has been shown to directly modulate fibrotic gene expression in human fibroblasts, so we hypothesized that equine IL-10 (eIL-10) may exert similar anti-fibrotic effects on equine dermal fibroblasts. Cell-lines were ...
Johnson SA, Biscoe EW, Eilertson KE, Lutter JD, Schneider RK, Roberts GD, Cary JA, Frisbie DD.Elastography is an emerging imaging modality for characterizing tendon injury in horses, but its ability to differentiate tissue deformability relative to treatment group and biochemical properties using a prospective, experimental study design remain unknown. Objectives of the current study were to (a) to investigate differences in glycosaminoglycan, DNA, and soluble collagen levels in mesenchymal stem cell (MSC) treated limbs compared to untreated control limbs utilizing a collagenase model of tendinopathy; (b) compare elastographic features between treatment groups; and (c) determine tissue...
Kobrina Y, Isaksson H, Sinisaari M, Rieppo L, Brama PA, van Weeren R, Helminen HJ, Jurvelin JS, Saarakkala S.The collagen phase in bone is known to undergo major changes during growth and maturation. The objective of this study is to clarify whether Fourier transform infrared (FTIR) microspectroscopy, coupled with cluster analysis, can detect quantitative and qualitative changes in the collagen matrix of subchondral bone in horses during maturation and growth. Equine subchondral bone samples (n = 29) from the proximal joint surface of the first phalanx are prepared from two sites subjected to different loading conditions. Three age groups are studied: newborn (0 days old), immature (5 to 11 months ol...
Chalmers HJ, Caswell J, Perkins J, Goodwin D, Viel L, Ducharme NG, Piercy RJ.A unilateral neurectomy model was used to study the relationship between histologic and ultrasonographic tissue characteristics during muscle atrophy over time. Methods: This investigation was an in vivo experimental study in an equine model (n = 28). Mean pixel intensity of ultrasonographic images was measured, a muscle appearance grade was assigned weekly, and muscles were harvested from 4 to 32 weeks. Minimum fiber diameter, fiber density per unit area, percent collagen, percent fat, and fiber type profile were measured from muscle cryosections and correlated with the ultrasonographic param...
Osborne AC, Carter SD, May SA, Bennett D.An investigation was made into the possible contribution of autoimmune mechanisms to equine arthropathies. Serum and synovial fluid (SF) immune complexes and anti-collagen Type II antibodies were measured, by ELISA, in groups of horses with naturally occurring osteoarthritis (OA), osteochondritis dissecans (OCD), bone fracture, traumatised joints, synovitis, infected joints and non-diseased (control) joints. Significantly raised anti-collagen Type II antibodies were found in osteoarthritic (P < 0.02) and traumatised joint synovial fluids (P < 0.01) compared with the control, where ten of...
Brunso L, Segura D, Monreal L, Escolar G, White JG, Diaz-Ricart M.Studies in animal models are useful to understand the basic mechanisms involved in hemostasis and the functional differences among species. Ultrastructural observations led us to predict differences in the activation and secretion mechanisms between equine and human platelets. The potential mechanisms involved have been comparatively explored in the present study. Equine and human platelets were activated with thrombin (0.5 U/ml) and collagen (20 µg/ml), for 90 seconds, and samples processed to evaluate: i) ultrastructural changes, by electron microscopy, ii) actin polymerization and cy...
Skiöldebrand E, Ekman S, Heinegård D, Hultenby K.The present study was designed to delineate the presence of COMP at the ultrastructural level comparing concentrations between two areas of articular cartilage from the equine third carpal bone, subjected to different loading, from trained and untrained horses. We also analyzed the fibril thickness of collagen type II in the same compartments and zones. Samples were collected from high load-bearing areas of the dorsal radial facet (intermittent high load) and an area of the palmar condyle (low constant load) in five non-trained and three trained young racehorses. The data show that COMP is muc...
Duesterdieck-Zellmer KF, Moneta L, Ott JF, Larson MK, Gorman EM, Hunter B, Löhr CV, Payton ME, Morré JT, Maier CS.To determine effects of intraarticularly administered tiludronate on articular cartilage in vivo, eight healthy horses were injected once with tiludronate (low dose tiludronate [LDT] 0.017 mg, n = 4; high dose tiludronate [HDT] 50 mg, n = 4) into one middle carpal joint and with saline into the contralateral joint. Arthrocentesis of both middle carpal joints was performed pre-treatment, and 10 min, 24 h, 48 h, 7 and 14 days after treatment. Synovial nucleated cell counts and total solids, tiludronate, sulfated glycosaminoglycan (sGAG), chondroitin sulfate 846 epitope (CS-846, a measure of aggr...
Vachon AM, Keeley FW, McIlwraith CW, Chapman P.Articular cartilage specimens from the distal articular surface of 32 radiocarpal bones from 24 2- to 5-year-old horses were analyzed. The total collagen content was determined on the basis of the 4-hydroxyproline content, using a colorimetric method. A method for estimating the proportions of types-I and -II collagen by measuring spectrophotometric densities of specific cyanogen bromide peptide bands from mixtures of types-I and -II collagen on sodium dodecyl sulfate-polyacrylamide gels was used. The cyanogen bromide peptides representative of each collagen types-I and -II were identified. Th...
Todhunter RJ, Fubini SL, Vernier-Singer M, Wootton JA, Lust G, Freeman KP, MacLeod JN.To determine how acute synovitis, with and without intra-articular methylprednisolone acetate (MPA), affect synthesis of proteoglycan, total protein, and collagen in articular cartilage and total protein synthesis in synovial membrane. Methods: Synovitis was induced in 10 ponies by the injection of 0.5 ng lipopolysaccharide (LPS) into the left radiocarpal and midcarpal joints every 2 days for a total of four treatments. Synovitis was documented by clinical examination and synovial fluid analyses. Two days before euthanasia, MPA (0.1 mg/kg) was injected with the last dose of LPS into both the l...
Endoh C, Matsuda K, Okamoto M, Tsunoda N, Taniyama H.Aortic rupture is a well recognized cause of sudden death in thoroughbred horses. Some microscopic lesions, such as those caused by cystic medial necrosis and medionecrosis, can lead to aortic rupture. However, these microscopic lesions are also observed in normal horses. On the other hand, a previous study of aortic rupture suggested that underlying elastin and collagen deposition disorders might be associated with aortic rupture. Therefore, the purpose of this study was to compare the structural components of the tunica media of the aortic arch, which is composed of elastin, collagen, smooth...
Carstanjen B, Hoyle NR, Gabriel A, Hars O, Sandersen C, Amory H, Remy B.To evaluate a human assay for quantification of carboxy-terminal cross-linking telopeptide of type I collagen (CTX-I), assess the influence of age on plasma CTX-I concentration, investigate the relationship between plasma CTX-I and serum osteocalcin concentrations, and determine whether concentrations of plasma CTX-I or serum osteocalcin fluctuate in circadian manner in horses. HORSES: 75 clinically normal horses. Methods: Cross-reactivity between equine serum CTX-I and CTX-I antibodies in an automated electrochemiluminescent sandwich antibody assay (ECLIA) was evaluated via a specificity test...
van den Berg MJ, Rijkenhuizen AB, Németh F, Gruys E.Chronic changes of several structures in around the fetlock tunnel can be a cause of the so-called fetlock tunnel syndrome (FTS) in the horse. Forty-nine annular ligaments (AL) from dead horses without a known history or clinical evidence of lameness and/or digital tendon sheath problems in these legs and 30 AL biopsies from horses suffering from FTS were studied macroscopically and microscopically. Macroscopically, the normal AL had a shiny white appearance, whereas the affected AL were often thicker and less white. Microscopically, the normal AL were about +/- 1 mm thick and were composed of...
Patan-Zugaj B, Gauff FC, Egerbacher M, Licka TF.OBJECTIVE To investigate the effect of lipopolysaccharide (LPS) on type VII collagen- cleaving matrix metalloproteinases (MMPs) in the lamellar tissue of extracorporeally perfused equine limbs. SAMPLE 10 right forelimbs and 3 left forelimbs collected from 10 adult horses after slaughter at a licensed abattoir. PROCEDURES Extracorporeal perfusion of the isolated equine limbs was performed for 10 hours under physiologic conditions (control-perfused limbs; n = 5) and with the addition of 80 ng of LPS/L of perfusate (LPS-perfused limbs; 5). Lamellar tissue specimens were then collected from the do...
Wintz LR, Lavagnino M, Gardner KL, Sedlak AM, Arnoczky SP.To describe the effect of systemically administered oxytetracycline on the viscoelastic properties of rat tail tendon fascicles (TTfs) to provide a mechanistic rationale for pharmacological treatment of flexural limb deformities in foals. Methods: TTfs from ten 1-month-old and ten 6-month-old male Sprague-Dawley rats. Methods: 5 rats in each age group were administered oxytetracycline (50 mg/kg, IP, q 24 h) for 4 days. The remaining 5 rats in each age group served as untreated controls. Five days after initiation of oxytetracycline treatment, TTfs were collected and their viscoelastic properti...
Amaral A, Cebola N, Szóstek-Mioduchowska A, Rebordão MR, Kordowitzki P, Skarzynski D, Ferreira-Dias G.Myeloperoxidase is an enzyme released by neutrophils when neutrophil extracellular traps (NETs) are formed. Besides myeloperoxidase activity against pathogens, it was also linked to many diseases, including inflammatory and fibrotic ones. Endometrosis is a fibrotic disease of the mare endometrium, with a large impact on their fertility, where myeloperoxidase was shown to induce fibrosis. Noscapine is an alkaloid with a low toxicity, that has been studied as an anti-cancer drug, and more recently as an anti-fibrotic molecule. This work aims to evaluate noscapine inhibition of collagen type 1 (C...
Okumura M, Tagami M, Fujinaga T.Keratan sulphate (KS) concentration and anticollagen type II antibody levels were measured in the serum and synovial fluid (SF) of clinically normal horses and horses with osteoarthritis (OA). Serum KS in OA was significantly higher than that in normal horses, while no significant difference was found in KS levels of SF between normal and OA. Assays of antibody to collagen type II showed no significant increase in sera and SF of OA. It was suggested that levels of serum KS would be of value in the pathological detection of OA in the joint, although there was no evidence that the measurable aut...
Hauspie S, Forsyth R, Vanderperren K, Declercq J, Martens A, Saunders JH.The objective of this study is to describe the normal histological appearance of the dorsoproximal aspect of the sagittal ridge of the third metacarpal/metatarsal bone in young Warmblood horses, and to compare it to the different radiographic variations (irregular, indentation, lucency, notch) described at this level. A total of 25 metacarpo/metatarsophalangeal joints of 12 Warmblood horses were used. Five samples of each radiographically described group were selected for histological processing. Each category was compared with the normal control group. Each group showed a bone cortex, covered...
Wells HC, Sizeland KH, Kirby N, Haverkamp RG.Thin, strong scaffold materials are needed for surgical applications. New materials are required, particularly those readily available, such as from non-human sources. Bovine amniotic membrane (antepartum) and equine amniotic membrane (postpartum) were characterized with tear and tensile tests. The structural arrangement of the collagen fibrils was determined by small-angle X-ray scattering, scanning electron microscopy, and ultrasonic imaging. Bovine amnion had a thickness-normalized tear strength of 12.6 (3.8) N/mm, while equine amnion was 14.8 (5.3) N/mm. SAXS analysis of the collagen fibri...
Kamr AM, Dembek KA, Gilsenan W, Bozorgmanesh R, Hassan HY, Rosol TJ, Toribio RE.Hypocalcemia is a common finding in critically ill equine patients. Parathyroid hormone (PTH) helps to maintain calcium homeostasis in hypocalcemic patients by promoting renal calcium reabsorption and bone resorption. Increased serum PTH concentrations have been reported in critically ill people and animals, including horses and foals. It is unknown whether increased secretion of PTH is associated with markers of bone turnover in hospitalized foals. The goals of this study were to measure markers of bone resorption (C-terminal telopeptide of type I collagen [CTX-I]) and bone formation (osteoca...
Khatibzadeh SM, Menarim BC, Nichols AEC, Werre SR, Dahlgren LA.Extracellular matrix (ECM) is responsible for tendon strength and elasticity. Healed tendon ECM lacks structural integrity, leading to reinjury. Porcine urinary bladder matrix (UBM) provides a scaffold and source of bioactive proteins to improve tissue healing, but has received limited attention for treating tendon injuries. The objective of this study was to evaluate the ability of UBM to induce matrix organization and tenogenesis using a novel in vitro model. We hypothesized that addition of UBM to tendon ECM hydrogels would improve matrix organization and cell differentiation. Hydrogels se...
Gurusinghe S, Hilbert B, Trope G, Wang L, Bandara N, Strappe P.Immortalization of chondrocytes enables long term in vitro culture; however, the chondrogenic capacity of transformed cells varies, thus highlighting the need to develop a proliferative and tuneable chondrocyte cell line where hypertrophic differentiation can be controlled. In this study the SV40 large T antigen and human telomerase reverse transcriptase were employed to immortalize pooled equine chondrocytes through lentiviral vector mediated transduction either singly or on combination. Transformed chondrocytes proliferated stably over multiple passages, but resulted in significantly lower e...
