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.
van de Lest CH, Brama PA, van Weeren PR.This study aimed at the determination of the influence of exercise on the levels of a number of bone morphogenic enzymes in subchondral bone and at the comparison of these data with other (subchondral) bone-related parameters that have been investigated in the same experimental population.Forty-three foals were reared until weaning at 5 months of age under similar conditions, except for the type and amount of exercise. Fifteen foals remained at pasture (Pasture group and also control group), 14 foals were kept in box stalls (Box group) and 14 foals were kept in the same box stalls but were sub...
Schwartz AJ, Wilson DA, Keegan KG, Ganjam VK, Sun Y, Weber KT, Zhang J.To determine significant molecular and cellular factors responsible for differences in second-intention healing in thoracic and metacarpal wounds of horses. Methods: 6 adult mixed-breed horses. Methods: A full-thickness skin wound on the metacarpus and another such wound on the pectoral region were created, photographed, and measured, and tissue was harvested from these sites weekly for 4 weeks. Gene expression of type-I collagen, transforming growth factor (TGF)-beta1, matrix metalloproteinase (MMP)-1, and tissue inhibitor of metalloproteinase (TIMP)-1 were determined by quantitative in situ ...
Birch HL, Smith TJ, Poulton C, Peiffer D, Goodship AE.Partial rupture occurs most often to the superficial digital flexor tendon (SDFT) of the forelimb at the mid-metacarpal level. In this study, we tested the hypothesis that the mid-metacarpal region of the SDFT has the smallest cross sectional area (CSA) and a similar collagen content to other regions and, therefore, represents a weak point in the tendon. The SDFT was collected from the forelimbs of 9 horses. Each tendon was marked at 7 different levels from the origin of the accessory ligament to the phalangeal region. The CSA, water content and collagen content was measured at each level. The...
McKane SA, Slocombe RF.Necropsy studies of horses suffering exercise-induced pulmonary haemorrhage (EIPH) have identified mild inflammatory lesions with evidence of alveolar fibrosis and bronchiolitis. These lesions were thought to be the result of viral infections that predisposed the affected regions of the lung to EIPH. We have shown previously that during erythrophagocytosis in the alveolar space, there is a prolonged period of macrophage influx and activation. This present study used morphometric analysis to quantify the effects of macrophage activity during erythrophagocytosis, on the alveolar cell population ...
Jarvis GE, Atkinson BT, Snell DC, Watson SP.1. Various platelet membrane glycoproteins have been proposed as receptors for collagen, in some cases as receptors for specific collagen types. In this study we have compared the ability of a range of collagen types to activate platelets. 2. Bovine collagen types I-V, native equine tendon collagen fibrils and collagen-related peptide (CRP) all induced platelet aggregation and shape change. 3. Responses were abolished in FcRgamma chain-deficient platelets, which also lack GPVI, indicating a critical dependence on the GPVI/FcRgamma chain complex. 4. Responses to all collagens were unaffected in...
Magne D, Guicheux J, Weiss P, Pilet P, Daculsi G.Peritubular dentin (PTD) is a relatively dense mineralized tissue surrounding tooth dentin tubules, whose composition and mode of formation are still unclear. Fourier transform infrared microspectroscopic studies of the organic and mineral components of the highly developed horse PTD indicate that the peritubular matrix is less abundant than the intertubular matrix but is also mainly composed of collagen, which is more hydrated. These data suggest that most of the crystals are located outside the collagen fibrils and probably not associated with protein components. The crystals in PTD have nea...
Sun Y, Chen H, Kandel R, Hurtig M.Lesions in cartilage of equine weightbearing joints commonly result in lameness. Cell-based resurfacing techniques are currently being developed for human and veterinary applications. Biopsies of stifle joint cartilage (1 g) were harvested aseptically and chondrocytes were isolated by sequential enzyme digestion. The cells were grown in vitro on filter inserts. Analysis of cultures 8 weeks later showed that the cells had accumulated extracellular matrix and formed a continuous layer of cartilagenous tissue as determined histologically. The cells maintained their phenotype as they synthesised t...
Fortier LA, Mohammed HO, Lust G, Nixon AJ.Composites of chondrocytes and polymerised fibrin were supplemented with insulin-like growth factor-I (IGF-I) during the arthroscopic repair of full-thickness cartilage defects in a model of extensive loss of cartilage in horses. Repairs facilitated with IGF-I and chondrocyte-fibrin composites, or control defects treated with chondrocyte-fibrin composites alone, were compared before death by the clinical appearance and repeated analysis of synovial fluid, and at termination eight months after surgery by tissue morphology, collagen typing, and biochemical assays. The structure of cartilage was ...
Brama PA, TeKoppele JM, Bank RA, Barneveld A, van Weeren PR.Subchondral bone provides structural support to the overlying articular cartilage, and plays an important role in osteochondral diseases. There is growing insight that the mechanical features of bone are related to the biochemistry of the collagen network and the mineral content. In the present study, part of the normal developmental process and the influence of physical activity on biochemical composition of subchondral bone was studied. Water content, calcium content and characteristics of the collagen network (collagen, hydroxylysine, lysylpyridinoline (LP) and hydroxylysylpyridinoline (HP)...
Dahm AM, de Bruin A, Linat A, von Tscharner C, Wyder M, Suter MM.We describe the establishment and characterisation of equine keratinocyte cultures with maintenance of a high proliferative capacity up to the second passage. Improved attachment and growth were obtained by seeding primary cells on equine feeder layers. Subcultured keratinocytes showed optimal growth when seeded on collagen type I. The proliferation rate of cells on this substrate exceeded that seen for cells seeded on equine feeder layers. By immunohistochemistry, epithelial origin and state of differentiation of the equine keratinocytes were determined. They expressed keratin and desmoplakin...
Fortier LA, Nixon AJ, Lust G.To assess the effects of supraphysiologic concentrations of insulin-like growth factor-1 (IGF-1) on morphologic and phenotypic responses of chondrocytes. Methods: Articular cartilage obtained from 2 young horses. Methods: Chondrocytes were suspended in fibrin cultures and supplemented with 25, 12.5, or 0 mg of IGF-1/ml of fibrin. Chondrocyte morphology and phenotypic expression were assessed histologically, using H&E and Alcian blue stains, immunoreaction to collagen type I and II, and in situ hybridization. Proteoglycan content, synthesis, and monomer size were analyzed. The DNA content w...
Price JS, Jackson BF, Gray JA, Harris PA, Wright IM, Pfeiffer DU, Robins SP, Eastell R, Ricketts SW.This study describes longitudinal changes in serum levels of biochemical markers of bone cell activity in a group of 24 thoroughbred foals from birth to 18 months of age. The markers of bone formation included the type I collagen carboxy-terminal propeptide (PICP), the bone-specific isoenzyme of alkaline phosphatase (BAP), and osteocalcin (OC). Levels of the cross-linked telopeptide of type I collagen (ICTP), a marker of bone resorption, and the N-terminal propeptide of type III collagen (PNIIIP), a marker of soft tissue turnover, were also measured. Levels of all markers fell significantly be...
Murray RC, Birch HL, Lakhani K, Goodship AE.It was hypothesized that cartilage macro-molecular characteristics are influenced by exercise intensity and by location within a joint. Objective: To determine the macromolecular characteristics of carpal articular cartilage at common and uncommon sites of pathology in horses undergoing high or low intensity exercise, and to compare this composition between exercise groups. Methods: Twelve horses (19.3+/-0.9 years) were assigned to exercise groups. Each group underwent 19 weeks high-intensity treadmill training (N=6) or low-intensity exercise (N=6). Dorsal and palmar test sites were identified...
Dahlgren LA, Nixon AJ, Brower-Toland BD.To investigate effects of beta-aminopropionitrile and a combination of insulin-like growth factor (IGF)-I and beta-aminopropionitrile on metabolism of equine tendon fibroblasts. Methods: Flexor tendon explants from 3 horses. Methods: Explants received 1 of 4 treatments (control, IGF-I, beta-aminopropionitrile, and IGF-I/beta-aminopropionitrile) for 10 days, and message expression for collagen types I and III was assessed by use of in situ hybridization. Histologic findings, new protein production, and quantitative determinations of glycosaminoglycan, DNA, and de novo collagen synthesis were ma...
Cherdchutham W, Becker CK, Spek ER, Voorhout WF, van Weeren PR.To determine the effects of exercise on collagen fibril diameter distribution in the superficial digital flexor tendon (SDFT) of foals. Methods: 43 Dutch Warmblood foals. Methods: From 1 week until 5 months of age, group-1 foals (n = 14) were housed in stalls and not exercised, group-2 foals (14) were housed in stalls but were exercised, and group-3 foals (15) were maintained at pasture. Biopsy specimens were collected from the SDFT at 2 months, and 8 foals in each group were euthanatized at 5 months. Remaining foals were housed together in a loose stall and paddock until euthanatized at 11 mo...
Wilmink JM, Nederbragt H, van Weeren PR, Stolk PW, Barneveld A.The contribution of wound contraction to wound closure determines the speed of second intention wound healing and it has been shown that significant differences exist with regard to both contraction and inflammatory response between horses and ponies and between various areas of the body. In this study, the contraction capacity of fibroblasts from limbs and buttocks of 4 Dutch Warmblood horses and 4 Shetland ponies was studied in vitro, in order to determine whether differences in wound contraction are due to differences in the inherent contraction capacity of the fibroblasts or to differences...
Fubini SL, Todhunter RJ, Burton-Wurster N, Vernier-Singer M, MacLeod JN.Experimental evidence suggests that recommended dosages of some corticosteroids used clinically as antiinflammatory agents for treating arthropathies damage articular cartilage, but low dosages may be chondroprotective. The purpose of this study was to evaluate how different concentrations of methylprednisolone affect chondrocyte function and viability. Articular cartilage and chondrocytes were obtained from young adult horses, 1.5-3.5 years of age. Corticosteroid-induced changes in collagen expression were studied at the transcriptional level by Northern blot analyses and at the translational...
Fortier LA, Balkman CE, Sandell LJ, Ratcliffe A, Nixon AJ.This study evaluated the constitutive insulin-like growth factor-I (IGF-I) gene expression pattern in spontaneously healing cartilage defects over the course of 16 weeks, and correlated the tissue morphology and matrix gene expression with IGF-I mRNA levels. Full-thickness 15 mm cartilage defects were debrided in the femoral trochlea of both femoropatellar joints of 8 horses and the healing defects examined 2, 4, 8, or 16 weeks after surgery. Samples were harvested for histologic assessment of tissue healing using H&E staining, toluidine blue histochemical reaction for proteoglycan deposition,...
Worster AA, Brower-Toland BD, Fortier LA, Bent SJ, Williams J, Nixon AJ.This study evaluated chondrogenesis of mesenchymal progenitor stem cells (MSCs) cultured initially under pre-confluent monolayer conditions exposed to transforming growth factor-beta1 (TGF-beta1), and subsequently in three-dimensional cultures containing insulin-like growth factor I (IGF-I). Bone marrow aspirates and chondrocytes were obtained from horses and cultured in monolayer with 0 or 5 ng of TGF-beta 1 per ml of medium for 6 days. TGF-beta 1 treated and untreated cultures were distributed to three-dimensional fibrin disks containing 0 or 100 ng of IGF-I per ml of medium to establish fou...
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...
Brounts SH, Rashmir-Raven AM, Black SS.This case report describes a distinctive deep cutaneous lesion in a 1-year-old Quarter Horse filly with hyperelastosis cutis. The horse had a typical clinical presentation of hyperelastic skin associated with a 6-month history of cutaneous wounds that developed following minor cutaneous trauma. Punch biopsies of skin from the affected horse were thinner than similar biopsies from an age- and breed-matched control. Significant microscopic lesions were not seen in cutaneous punch biopsies stained with haematoxylin and eosin and Masson's trichrome stains, but the ultrastructure of the dermis from...
Skiöldebrand E, Lorenzo P, Zunino L, Rucklidge GJ, Sandgren B, Carlsten J, Ekman S.The aim of the present investigation was to study the metabolic activity of the third carpal bone and the release of COMP, aggrecan and collagen type II molecules in the synovial fluid as a result of injury. Cartilage oligomeric matrix protein (COMP), aggrecan and collagen type II or fragments of these molecules released to the synovial fluid and serum (COMP) were quantified in samples from 73 left equine middle carpal joints from 2 breeds with different activity profiles (52 Standardbred trotters [STB] and 21 Swedish Warmblood riding horses [SWH]) and different articular cartilage lesions. Sy...
Raulo SM, Sorsa TA, Kiili MT, Maisi PS.To determine collagenase activity and evaluate matrix metalloproteinase (MMP)-8 and MMP-13 in horses with chronic obstructive pulmonary disease (COPD). Methods: 12 horses with COPD and 12 healthy control horses. Methods: Collagenase activity was determined by use of an assay for degradation of type-I collagen. Western immunoblot analysis was used to identify interstitial collagenases MMP-8 and MMP-13 in tracheal epithelial lining fluid (TELF). Immunocytochemistry and in situ hybridization were used to determine cellular expression of these 2 collagenases in cells in bronchoalveolar lavage flui...
Billinghurst RC, Buxton EM, Edwards MG, McGraw MS, McIlwraith CW.To develop an antibody that specifically recognizes collagenase-cleaved type-II collagen in equine articular cartilage. Methods: Cartilage specimens from horses euthanatized for problems unrelated to the musculoskeletal system. Methods: A peptide was synthesized representing the carboxy- (C-) terminus (neoepitope) of the equine type-II collagen fragment created by mammalian collagenases. This peptide was used to produce a polyclonal antibody, characterized by western analysis for reactivity to native and collagenase-cleaved equine collagens. The antibody was evaluated as an antineoepitope anti...
Semevolos SA, Nixon AJ, Brower-Toland BD.To determine molecular changes in the expression of insulin-like growth factor-I (IGF-I) and transforming growth factor-beta1 (TGF-beta1) in horses with osteochondrosis, and to characterize expression of matrix aggrecan and collagen types I, II, and X in articular cartilage of affected joints. Methods: Articular cartilage from affected stifle or shoulder joints of 11 horses with naturally acquired osteochondrosis and corresponding joints of 11 clinically normal horses. Methods: Harvested specimens were snap frozen in liquid nitrogen, and total RNA was isolated. Specimens were fixed in 4% paraf...
Brama PA, Bank RA, Tekoppele JM, Van Weeren PR.Subchondral bone provides structural support to the overlying articular cartilage and plays an important role in osteochondral diseases. There is growing insight that the mechanical features of bone are related to the biochemistry of the collagen network. In this study the effect of exercise on water, calcium and the collagen network (total collagen, lysyl-hydroxylation, hydroxylysylpyridinoline, and lysylpyridinoline crosslinks) of subchondral bone at two differently loaded sites (site 1: intermittently loaded; site 2: constantly loaded) is investigated in foals. Exercise influenced calcium c...
Brama PA, Barneveld A, Karssenberg D, Van Kampen GP, van Weeren PR.A recently developed indenter system that aims at determination of local structural properties of the cartilage surface was evaluated for suitability in the horse. To this aim, maximum indenter force was measured of the articular surface and related to biochemical characteristics of the cartilage at different sites of the distal metacarpal bone (MC). Significant topographical variation exists in structural properties of the articular surface of the MC. The dorsal margin showed a significantly lower maximum indenter force than more centrally located areas, indicating an increased compliance und...
Thompson CC, Clegg PD, Carter SD.Cartilage destruction in osteoarthritis (OA) is associated with increased levels of several matrix metalloproteinases (MMPs), including the gelatinases MMP-2 and MMP-9. While increases in some MMPs may be destructive, up-regulation of others may result from increases in normal tissue turnover. The production of MMP-2 and MMP-9 by the anabolic transforming growth factor beta-1 (TGF-beta1) in normal equine chondrocytes was investigated. Methods: Equine chondrocytes from clinically normal femoropatellar joints were maintained in alginate beads. After serum deprivation, cells were exposed to TGF-b...
Brooks DE, Komaromy AM, Garcia-Fernandez MC, Cutler TJ, Samuelson DA, Kallberg ME.Purpose To use immunohistochemical techniques to identify and localize the structural macromolecules of the extracellular matrix (ECM) of the normal adult equine lamina cribrosa in order to make comparisons to the extracellular matrix of the lamina cribrosa of horses with glaucoma. METHODS: Normal eyes of five adult horses between 5 and 10 years of age were fixed in 10% neutral buffered formalin and embedded in paraffin. Polyclonal rabbit-derived antibodies against human elastin, laminin, fibrillin-1, and collagen types I, III and IV, and polyclonal goat-derived antibodies against collagen typ...
Egerbacher M, Edinger J, Tschulenk W.To study chondrotoxic effects of enrofloxacin (ENR) and ciprofloxacin hydrochloride (CFX) on canine and equine articular chondrocytes in culture and to compare the effects with that of cultivation in Mg2+-free medium. Methods: Chondrocytes from articular cartilage of 4- and 6 -month old dogs and 2- to 4- year-old horses. Methods: Chondrocytes were cultivated with 10, 40, 80, and 160 microg of CFX/ml, 10, 50, 100, and 150 microg of ENR/ml, or in Mg2+-free medium. A live-to-dead test was performed to test cytotoxic effects. Morphologic changes were evaluated by electron microscopy. An attachment...
Pritchard A, Nielsen BD, Robison C, Manfredi JM.As osteoarthritis is a major cause of lameness in horses in the United States, improving collagen health prior to onset and increasing collagen turnover within affected joints could improve health- and welfare-related outcomes. Through its positive effects on bone mineral content and density and its role in increasing collagen synthesis, silicon (Si) may slow the development and progression of osteoarthritis, thereby reducing lameness. This study evaluated the hypothesis that Si supplementation would increase cartilage turnover through increased collagen degradation and formation markers, as w...
Kuwano A, Ueno T, Katayama Y, Nishiyama T, Arai K.Between the laminar epidermis and the laminar dermis of laminar region (LR) in equine foot, it can be observed the basement membrane zone (BMZ), which is composed of a basement membrane and its accompaniments like the hemidesmosome and anchoring fibril. Alteration in the BMZ in equine laminitis is possibly related with not only development but also recovery outcome and recurrence of this disease. However, there is little known about the structure of the BMZ during the recovery phase of this disease. To assess the condition of the BMZ of LR affected by chronic laminitis, the tissue was examined...
Bone reportsJanuary 5, 2023
Volume 18 101653 doi: 10.1016/j.bonr.2023.101653
Grissom SK, Semevolos SA, Duesterdieck-Zellmer K.The objective of this study is to better understand the pathogenesis of early equine osteochondrosis (OC) by identifying differences in gene and protein expression of extracellular matrix components and regulators in normal and diseased cartilage and bone, focusing on the osteochondral junction and cells surrounding the cartilage canals. We expected to find an upregulation of matrix metalloproteinases and a decrease in extracellular matrix constituent expression along the osteochondral junction and cells surrounding the cartilage canals in OC samples. Paraffin-embedded osteochondral samples (6...
Grigolo B, Desando G, Cavallo C, Zini N, Ghisu S, Facchini A.Association of biomaterials with autologous cells can provide a new generation of implantable devices for cartilage repair. An ideal scaffold should possess a preformed three-dimensional shape, fix the cells to the damaged area and prevent their migration into the articular cavity. Furthermore, the constructs should have sufficient mechanical strength to facilitate handling in a clinical setting and stimulate the uniform spreading of cells and a phenotype re-differentiation process. The aim of this study was to verify the ability of an equine collagen membrane to support the growth of human ch...
Olivry T, Borrillo AK, Xu L, Dunston SM, Slovis NM, Affolter VK, Demanuelle TC, Chan LS.Bullous pemphigoid (BP) is an autoimmune subepithelial blistering dermatosis of humans, dogs, cats and pigs. It is characterized by skin-fixed and circulating IgG autoantibodies that target one or both BP antigens. An immunological homologue of BP in humans was diagnosed in two horses with cutaneous and mucosal ulcerations as well as microscopic subepithelial vesiculation. Immunological investigations revealed similar findings for both the horses. Direct immunofluorescence demonstrated the presence of IgG deposited linearly at the dermoepidermal junction in mucosal and skin biopsy specimens. I...
Hellings IR, Dolvik NI, Ekman S, Olstad K.Some epiphyseal growth cartilage canals are surrounded by a ring of hypereosinophilic matrix consisting of collagen type I. Absence of the collagen type I ring may predispose canal vessels to failure and osteochondrosis, which can lead to fragments in joints (osteochondrosis dissecans). It is not known whether the ring develops in response to programming or biomechanical force. The distribution that may reveal the function of the ring has only been described in the distal femur of a limited number of foals. It is also not known which cells are responsible for producing the collagen ring. The a...
Fortier LA, Schnabel LV, Mohammed HO, Mayr KG.To determine the effects of matrix metalloproteinase (MMP)-13, compared with interleukin (IL)-1alpha, on cartilage matrix molecule gene expression in a coculture system of equine cartilage explants and synoviocytes. Methods: Articular cartilage and synovium specimens harvested from femoropatellar joints of 4 horses, aged 3 to 5 years. Methods: Synoviocytes were isolated and cocultured with cartilage explants. Cultures were treated with human recombinant MMP-13 (1, 25, or 100 ng/mL) or IL-1alpha (0.01, 0.1, 1.0, or 10 ng/mL) for 96 hours, with medium exchange at 48 hours. Cartilage extracts and...
Spiers S, May SA, Harrison LJ, Bennett D, Edwards GB.Significant amounts of collagenase and caseinase activity were detected in infected synovial fluid samples. Partial characterisation of the enzymes by gel filtration suggested that synovial fluid from cases of infectious arthritis may contain enzymes from both the synovial cells and neutrophils. This finding was also supported by analysis of sequential synovial fluid samples from 4 infected joints. In 3 joints the concentration of caseinase and in 1 joint collagenase paralleled the decline in total nucleated cell count. However, in 3 joints the concentration of collagenase remained high after ...
Pluim M, Heier A, Plomp S, Boshuizen B, Gröne A, van Weeren R, Vanderperren K, Martens A, Dewulf J, Chantziaras I, Koene M, Luciani A, Oosterlinck M....High-power laser therapy gained popularity recently as a regenerative treatment for tendinitis and desmitis in the horse. However, studies evaluating the effects of laser therapy on tissue repair at the histological level in large mammals are lacking. Objective: To evaluate the effects of high-power laser therapy on suspensory desmitis healing, using a model of suspensory ligament branch injury. Methods: In vivo experiments. Methods: Standardised lesions were surgically induced in all four lateral suspensory branches of 12 healthy Warmblood horses. Laser therapy (class 4, 15W) was applied dail...
Maldonado-Cabrera B, Sánchez-Machado DI, López-Cervantes J, Osuna-Chávez RF, Ibarra-Zazueta C, Robles-Zepeda RE.Consultation was requested for a 7-year-old Gypsy Vanner male horse with a 2-year history of foreskin injury. Upon revision, an ulcer, 153 cm2 in size, with yellowish granules was observed; a RESVECH 2.0 evaluation revealed a score of 32/35 points. Medical history confirmed multiple failed deworming, anti-inflammatory, and antibiotic treatments with different topical therapies and recurrence in summer. Laboratory results confirmed elevated total proteins (8.8 g/dL) and globulins (5.5 g/dL), negative bacterial and fungal cultures, as well as negative coproparasitoscopic findings, and finally, i...
Skiöldebrand E, Heinegård D, Olofsson B, Rucklidge G, Ronéus N, Ekman S.This study evaluates how strenuous training, age and lameness influence the release of cartilage oligomeric matrix protein (sf-COMP), aggrecan and collagen type II into synovial fluid in 28 (19.5-40 months) Standardbred trotters (STB), during a long-term training programme (24 months). All the horses were trained by the same trainer and were healthy on entering the training programme. Synovial fluid (sf) from the left middle carpal joint in each subject was sampled every third month. Enzyme-linked immunosorbent assay was used to determine the concentrations of sf-COMP, sf-aggrecan and sf-colla...
Nixon AJ, Stashak TS, Smith FW, Norrdin RW.Carbon fibre-polylactic acid composites and monofilament non-absorbable suture material were compared for the repair of surgically transected superficial digital flexor tendons in 10 horses. All surgical wounds healed by first intention. The repaired tendons were enlarged, the carbon implanted tendons being larger than those sutured. The horses were killed at six, eight, 12,20 or 24 weeks. Greater fibrous thickening occurred in tendons repaired with carbon fibre, especially at 12 weeks postoperatively. Carbon fibre incited a greater histological response with macrophages, lymphocytes, plasma c...
Andrews RK, Booth WJ, Bendall LJ, Berndt MC.von Willebrand Factor (vWF) is a multifunctional glycoprotein in plasma and vascular subendothelial matrix which plays a major role in cellular adhesion. vWFdependent adhesion of platelets to the subendothelium at high shear rates involves a specific platelet membrane receptor, the glycoprotein (GP) Ib-IX complex. We have previously purified a 39/34-kiloDalton (kDa) dispase fragment of vWF (Leu-480/Val-481 to Gly-718) and demonstrated that this fragment contains the binding site for the GP Ib-IX complex [Andrews R K, et al. Biochemistry 1989; 28: 8326-83361. vWF also mediates agglutination of ...
Sese M, Ueda H, Watanabe T, Yamamoto E, Hosaka Y, Tangkawattana P, Takehana K.The superficial digital flexor tendon (SDFT) is one of the longest tendons in the horse. In racehorses, disturbance of the locomotor functions of the SDFT occurs most frequently in the central area of the mid-metacarpal region. While many studies have investigated the equine SDFT, there are no reports to date of the morphological characteristics of collagen fibrils in the central and peripheral areas of each of the three regions that comprise the entire tendon: the myotendinous junction (MTJ), the mid-metacarpal region (mM) and the osteotendinous junction (OTJ). Mass average diameter (MAD), wh...
Barr AR, Duance VC, Wotton SF, Waterman AE, Holt PE.Cyanogen bromide was used to solubilise and specifically fragment purified equine Type I and II collagen and equine articular surface repair tissue. The resultant peptides were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and quantified by densitometric scanning. Measurement of the relative amounts of the peptides alpha 2(I) CB3, 5 and alpha 1(II)CB10 provided an accurate method of establishing the ratio of Type I to Type II collagen in mixtures of purified equine collagens. The method was sensitive to 6% Type II collagen when the band areas were corrected for peptid...
Amaral A, Fernandes C, Szóstek-Mioduchowska A, Lukasik K, Rebordão MR, Pinto-Bravo P, Skarzynski DJ, Ferreira-Dias G.Cathepsin G (CAT) is a protease released by neutrophils when forming neutrophil extracellular traps that was already associated with inducing type I collagen (COL1) in equine endometrium in vitro. Endometrosis is a fibrotic condition mainly characterized by COL1 deposition in the equine endometrium. The objective was to evaluate if noscapine (an alkaloid for cough treatment with anti-neoplastic and anti-fibrotic properties) would reduce transcription (evaluated by qPCR) and COL1 protein relative abundance (evaluated by western blot) induced by CAT in equine endometrial explants from follicula...
Watkins JP, Auer JA, Morgan SJ, Gay S.The effect of pulsing electromagnetic field (PEMF) therapy on the healing of surgically created defects in equine superficial digital flexor tendons was evaluated. Defects were created in both front superficial digital flexor tendons of 20 horses. The defect in 1 limb was exposed to a PEMF for 2 hours daily. The other limb served as a control. Histologic and immunofluorescent evaluations were done in horse killed at postsurgical weeks 2, 4, 8, 12, and 24. Therapy with the PEMF significantly (P less than 0.05) delayed the maturation of the tissue formed within the defect at postsurgical weeks 8...
Faramarzi B, Lantz L, Lee D, Khamas W.The digital cushion (DC) plays a role in absorbing and dampening forces applied to the foot and therefore supports internal structures such as navicular bone; yet, its architecture is not well-known. The goal of this study was to characterize the microanatomical structure of the DC in horses with clinically sound hooves. Both forefeet from the cadavers of 12 adult Quarter horses were cut and sectioned and samples of the following 4 regions of the DC were obtained: axial proximal (AxProx), axial distal (AxDis), abaxial lateral (AbxLat), and abaxial medial (AbxMed). The samples were processed an...
Tulleners E, Mann P, Raker CW.Epiglottic augmentation with injectable bovine collagen or an autogenous or allogenous auricular cartilage graft was performed in 12 horses with endoscopically and radiographically normal epiglottises. The grafting procedures were easy to perform and did not cause apparent discomfort. Cartilage graft extrusion or resorption may have occurred, but was not seen by endoscopy and lateral laryngeal radiography. Only collagen implants remained evident endoscopically, as smooth round submucosal bulges ventral to the epiglottic cartilage. Two horses with collagen implants, and all horses with cartilag...
Souza SS, Aguiar FLN, Alves BG, Alves KA, Brandão FAS, Brito DCC, Raposo RDS, Gastal MO, Rodrigues APR, Figueiredo JR, Teixeira DÍA, Gastal EL.Ovarian tissue transplantation methods using cooled and cryopreserved samples have been attractive options for fertility preservation in animal models and humans. The aim of this study was to evaluate the impact of previous exposure to cooling, cryopreservation, and VEGF on the overall efficiency of equine ovarian tissue after heterotopic xenotransplantation in mice. The end points evaluated were follicular morphology and development, follicular and stromal cell densities, angiogenesis (i.e. the density of new and mature blood vessels), collagen types I and III fiber densities, and total fibro...
Tizzoni R, Tizzoni M.The present case illustrates how a tooth, which had a highly questionable prognosis, was preserved by carrying out a periodontal regeneration surgery. Background: Treatment of periodontitis involves a careful consideration of all the factors that may allow the achievement of a favorable outcome; among those, the skillful use of guided tissue regeneration (GTR) membranes is of paramount importance. Methods: A 39-year-old patient presented with a mobile central upper incisor due to severe periodontitis and was treated according to GTR principles using a collagen membrane. A collagen-preserving b...
Depuydt E, Chiers K, Van Hecke L, Saunders J, Martens A, Pille F, Spaas JH.Injuries to equine tendons and ligaments are career-compromising, causing reduced performance and premature retirement. Promising treatment alternatives have been investigated in the field of mesenchymal stem cells (MSCs). In this study, the tissue adherence and protein expression of tenogenic primed mesenchymal stem cells (tpMSCs) after administration to ex vivo tendon and ligament explants is investigated. First, collagen type I (COL I) and smooth muscle actin (SMA) expression was assessed in cytospins prepared from native MSCs and tpMSCs. Second, equine superficial digital flexor tendon and...
Cissell JM, Milton SC, Dahlgren LA.To evaluate the effects of prostaglandin E₂ (PGE₂) treatment on the metabolism of equine tendon fibroblasts in vitro to aid in investigating the response of tendon fibroblasts to injury and novel therapeutics. Methods: Superficial digital flexor tendon fibroblasts isolated via collagenase digestion from six young adult horses were grown in monolayer in four concentrations of PGE₂ (0, 10, 50, 100 ng/ml) for 48 hours. Cells and medium were harvested for gene expression (collagen types I and III, cartilage oligomeric matrix protein [COMP], decorin, and matrix metalloproteinase-1, -3, and -1...
Neto da Silva AC, Costa AL, Teixeira A, Alpoim-Moreira J, Fernandes C, Fradinho MJ, Rebordão MR, Silva E, Ferreira da Silva J, Bliebernicht M....In older mares, increasing collagen fibers (fibrosis) in the endometrium and oviduct predisposes to sub-fertility and infertility. In this study, (i) gene transcription of collagen (qPCR: COL1A1, COL1A2, COL3A1, COL5A1); (ii) total collagen protein (hydroxyproline); (iii) collagen distribution (Picrosirius red staining; polarized light microscopy); and (iv) microvascular density (Periodic acid-Schiff staining), were evaluated in mares' placenta, and related to mares age, and placenta and neonate weights. Samples were collected from the gravid horn, non-gravid horn, and body of the placenta fro...
Smith MA, Dyson SJ, Murray RC.Collagen fibers oriented at 55 degrees to the static magnetic field (B0) are characterized by an artifactual increase in signal intensity due to the magic angle effect. We hypothesized that there would be increased signal intensity in the collateral ligaments of the distal interphalangeal joint and oblique sesamoidean ligaments when these ligaments were at angles approaching 55 degrees to a horizontal B0 during standing magnetic resonance (MR) imaging. MR imaging was performed on four cadaver forelimbs in a 0.27 T standing system. Transverse and dorsal images were obtained using various sequen...
Leonida A, Todeschini G, Lomartire G, Cinci L, Pieri L.To histologically assess the effectiveness of a socket-preservation technique using enzyme-treated equine bone granules as a bone-graft material in combination with an equine collagen matrix as a scaffold for soft-tissue regeneration. Background: Enzyme-treated equine bone granules and equine collagen matrix recently have been developed to help overcome alveolar bone deficiencies that develop in the wake of edentulism. Methods: The patient had one mandibular molar extracted and the socket grafted with equine bone granules. The graft was covered with the equine collagen matrix, placed in a doub...
Pien N, Van de Maele Y, Parmentier L, Meeremans M, Mignon A, De Schauwer C, Peeters I, De Wilde L, Martens A, Mantovani D, Van Vlierberghe S....Hand tendon injuries represent a major clinical problem and might dramatically diminish a patient's life quality. In this study, a targeted solution for flexor tendon repair was developed by combining a mechanical and biological approach. To this end, a novel acrylate-endcapped urethane-based polymer (AUP) was synthesized and its physico-chemical properties were characterized. Next, tubular repair constructs were developed using electrospinning of the AUP material with incorporated naproxen and hyaluronic acid (i.e. anti-inflammatory and anti-adhesion compounds, respectively), and with a tubul...
Haffner JC, Fecteau KA, Held JP, Eiler H.Under laboratory conditions and in clinical experiments, bacterial collagenase has proven to be effective in hydrolyzing placenta and detaching cotyledon from caruncle in the bovine species. Laboratory studies in which placental samples were incubated with collagenase have also demonstrated that collagenase is 3.7 times more effective in hydrolyzing equine placenta than bovine placenta. This led to the hypothesis that collagenase may be a potential treatment for mares with retained placenta. However, that collagenase may hydrolyze the uterine wall and perforate the uterus was a concern. It was...
Schmidbauer SM, Venner M, von Samson-Himmelstjerna G, Drommer W, Gruber AD.Interstitial lung disease with chronic fibrosis is a frequent cause of reduced performance in horses. The aim of this study was to establish a model of acute alveolar damage and interstitial lung disease in horses that could be used to monitor the histopathological lesions and changes in expression levels of genes relevant to pulmonary fibrosis. Six adult horses were given a single intravenous injection (6 mg per kg body weight) of perilla mint ketone (PMK). Transthoracic lung biopsy samples (1 x 0.2 x 0.2 cm) were collected before and after (days 1, 4, 8, 11, 15, 18, 22, 25 and 29) the admini...
Rashmir-Raven A, Lavagnino M, Sedlak A, Gardner K, Arnoczky S.Hereditary equine regional dermal asthenia (HERDA) is a genetic disorder of collagen resulting in fragile, hyper-extensible skin and ulcerative lesions. The predominance of skin lesions have been shown to occur on the dorsum of HERDA-affected horses. While this has been postulated to be due to increased exposure to sunlight of these areas, the precise pathological mechanism which causes this to occur is unclear. Objective: We hypothesized that an increase in collagenase activity, that has been associated with the exposure of dermal fibroblasts to sunlight, will significantly degrade the materi...
