Effects of blood-derived products and sodium hyaluronate on equine synovial fluid cells and on synovial fluid from osteochondrotic joints of horses after arthroscopy and administration of treatment.

OBJECTIVE: To compare effects of platelet-rich plasma (PRP), interleukin-1 receptor antagonist protein (IRAP), autologous processed plasma (APP), and sodium hyaluronate treatments on synovial fluid cells in vitro and on synovial fluid obtained from osteochondrotic joints of horses. SAMPLE: Synovial fluid cells from 8 healthy equine tibiotarsal joints (in vitro experiment) and synovial fluid samples from 40 tibiotarsal joints of 25 horses with osteochondrosis dissecans (in vivo experiment). PROCEDURES: Effects of various treatments on concentrations of prostaglandin (PG) E2, interleukin (IL)-1ß, tumor necrosis factor-α, IL-10, and IL-1 receptor antagonist (IL-1ra) were analyzed in cell medium supernatant, and production of reactive oxygen species was analyzed by use of flow cytometry. In an in vivo experiment, synovial fluid samples were collected before and 48 hours after arthroscopy and treatment administration (8 joints/treatment) and evaluated to determine concentrations of hyaluronic acid, chondroitin sulfate, PGE2, tumor necrosis factor-α, IL-1, IL-10, and IL-1ra. RESULTS: All in vitro treatments reduced reactive oxygen species production, PRP increased PGE2 concentrations, and PRP, IRAP, and APP increased IL-1ra concentrations. Only IRAP and APP increased IL-1 concentrations. For the in vivo experiment, PRP increased and IRAP decreased PGE2 concentrations in synovial fluid after arthroscopy. All treatments increased IL-1ra concentrations, but only sodium hyaluronate resulted in a significant increase in concentration, compared with the concentration for untreated joints. Also, IRAP reduced hyaluronic acid breakdown in synovial fluid. CONCLUSIONS AND CLINICAL RELEVANCE: PRP should be used with caution in the period immediately after arthroscopy and treatment of osteochondrotic joints of horses. All treatments had antioxidant effects. Sodium hyaluronate, APP, and IRAP might help ameliorate joint inflammation.

Physiotherapy protocol during initial postoperative period of arthroscopy in horses / Protocolo fisioterápico durante o período pós-operatório inicial de cirurgia artroscópica em equinos

This study evaluated the effects of a physiotherapy protocol applied in joints with osteochondritis dissecans submitted to arthroscopy. Twelve horses totaling twenty joints were used and divided into two uniform groups, according to articular lesion grade. Treated Group (TG) received the physiotherapy protocol (cryotherapy, passive rage motion and controlled exercise) that initiate just after anesthetic recovery and extended for five days. Control Group (CG) remained resting in stall during the same period. Physical examination and synovial fluid analysis were used to evaluate the treatment. The synovial fluid examination consisted of physical analysis (color, aspect, and viscosity), mucin clot evaluation, Serum Amyloid A, Prostaglandin E2 and urea concentration. Synovial samples were collected by arthrocentesis at the beginning of the surgical procedure (D1), 48 hours (D3) and 96 hours (D5) after surgery. Before arthroscopy and daily during the postoperative period joints were evaluated by physical exam: superficial temperature (°C), range of motion (degrees) and circumference (centimeters). The joint physical examination showed no significant difference between groups and neither along the days for the same group. The parameters of synovial fluid showed difference over the moments in each group but didn't have difference between groups. Color and aspect had the same patterns across moments, in CG fluid had significant change when compared D1 with D3 (color and aspect: p<0.001) and D5 (color: p<0.001; aspect: p<0.05) becoming mostly bloody and cloudy in D3 and D5. However in TG the difference was significant just between D1 and D3 (color and aspect: p<0.05), showing an improvement of synovial fluid in D5 (color and aspect: p<0.05). Viscosity and mucin clot evaluation showed significant change in CG between D1 and D3 (viscosity: p<0.01; mucin clot: p<0.05) and between D1 and D5 (viscosity: p<0.01;mucin clot: p<0.01). In TG no significant difference of viscosity and mucin clot was observed over the moments, showing an early improvement of synovial fluid quality. The Serum Amyloid A concentration showed an extremely significant increase in CG (p<0.001) when compared D1 (1217.13±664.47µg/mL) and D3 (42423.80±52309.31µg/mL). The comparison between D1 and D5 in CG, and across moments in TG, had no statistical difference. The PGE2 eicosanoid remained statistically unchanged all over the time. Urea showed significant increase in D3 when compared to D1 (p<0.001) in CG, and had no variation in TG. The physiotherapy protocol minimized the inflammatory mediators and provided minor alterations in synovial fluid after arthroscopy.(AU)

Este estudo avaliou os efeitos de um protocolo fisioterápico, aplicado em articulações com osteocondrite dissecante, submetidas à artroscopia. Foram utilizados 12 cavalos, totalizando 20 articulações, divididas em dois grupos homogêneos de acordo com a graduação da lesão articular. O grupo tratado (GT) recebeu o protocolo fisioterápico (crioterapia, movimentação passiva e exercício controlado) que se iniciou imediatamente após a recuperação anestésica e se estendeu por cinco dias. O grupo controle (GC) permaneceu em repouso na baia, pelo mesmo período. Exame físico da articulação e análise do líquido sinovial foram utilizados para avaliar o tratamento. O exame do líquido sinovial consistiu em análise física (cor, aspecto e viscosidade), avaliação do coágulo de mucina e concentrações de amiloide sérica A, prostaglandina E2 e ureia. Amostras de líquido sinovial foram colhidas por artrocentese no início do procedimento cirúrgico (D1) e após 48 (D3) e 96 horas (D5) do procedimento cirúrgico. Antes da artroscopia e diariamente no período pós-operatório, as articulações foram avaliadas por exame físico: temperatura superficial (°C), ângulo de flexão (graus), circunferência (centímetros). A avaliação física das articulações não apresentou diferença significativa entre os grupos nem ao longo dos dias em cada grupo. Nas análises do líquido sinovial, observou-se uma variação diferente entre os momentos em cada grupo porém sem diferença significativa entre os grupos. A cor e o aspecto tiveram resultados semelhantes ao longo do tempo, no GC houve uma alteração significativa quando comparados D1 e D3 (cor e aspecto: p<0,001) e D1 e D5 (cor: p<0,001; aspecto: p<0,05) tornando-se sanguinolento e turvo na maioria das amostras em D3 e D5. Já no GT, houve diferença significativa apenas entre D1 e D3 (cor e aspecto: p<0,05), demonstrando melhora no líquido sinovial em D5 (cor e aspecto: p<0,05). A viscosidade e o coágulo de mucina apresentou alteração significativa no GC entre D1 e D3 (viscosidade: p<0,01; coágulo de mucina: p<0,05) e entre D1 e D5 (viscosidade e coágulo de mucina: P<0,01). No grupo tratado não foram observadas alterações significativas em viscosidade e coágulo de mucina, ao longo dos momentos, demonstrando uma melhora precoce na qualidade do líquido sinovial. A amiloide sérica A apresentou um aumento extremamente significante no GC (p<0,001) quando comparados D1 (1217,13±664,47µg/dL) e D3 (42423,80±52309,31µg/dL). Quando comparados D1 e D5 no GC e ao longo do tempo no GT não foram observadas diferenças significativas. A concentração de PGE2 permaneceu sem alterações. As mensurações de ureia apresentaram aumento significativo em D3 quando comparado a D1 (p<0,001) no GC e não apresentou variação no GT. O protocolo fisioterápico minimizou os mediadores inflamatórios e proporcionou menor alteração do líquido sinovial após artroscopia.(AU)

Short- and long-term effects of platelet-rich plasma upon healthy equine joints: Clinical and laboratory aspects.

This study aimed to verify whether transient inflammatory reactions incited by the administration of intra-articular platelet-rich plasma (PRP) affected joint components through short- and long-term in vivo evaluation of inflammatory biomarkers and extracellular matrix degradation products in synovial fluid. The effects of PRP were analyzed in a short phase protocol (SPP) and in a prolonged phase protocol (PPP), using saline-injected joints as controls. In the SPP, higher white blood cell counts and prostaglandin E2 and total protein concentrations were observed in the synovial fluid of PRP-treated joints (P < 0.05). There were no differences between the interleukin-1ß, interleukin-1 receptor antagonist protein, tumor necrosis factor-α, chondroitin sulfate, or hyaluronic acid concentrations between PRP and saline injected joints. In the PPP, there were no differences in evaluated parameters between groups. PRP injection elicits a mild and self-limiting inflammatory response shortly after administration, without long-term deleterious effects on joint homeostasis.

Effets à court et à long terme de le plasma riche en plaquettes sur les articulations sains chez les équines : aspects cliniques et laboratoiren. Cette étude a pour but de vérifier si les réactions inflammatoires passagères induites par l'administration intra articulaire de Plasma Enrichi en Plaquettes (PRP) affectent les composants articulaires. Les bio-marqueurs de l'inflammation et les produits de dégradation de la matrice extracellulaire ont été évalués in vivo, dans le liquide synovial, à court et long terme. Les effets du PRP ont été analysés lors d'un protocole court terme et lors d'un protocole long terme et les articulations contrôles ont été injectées avec du liquide physiologique. Le protocole court terme a révélé des comptages de globules blancs et de prostaglandine E2, ainsi que des concentrations en protéines totales plus élevés dans le liquide synovial des articulations traitées au PRP (P < 0,05). Cependant, aucune différence de concentration en interleukine-1 bêta, en protéine antagoniste des récepteurs à l'interleukine-1, en facteur de nécrose tumorale alpha, en chondroïtine sulfate et en acide hyaluronique n'a été notée entre les articulations injectées au PRP et les articulations contrôles. Le protocole long terme n'a démontré aucune différence des paramètres évalués entre les deux groupes. L'injection de PRP provoque une réponse inflammatoire légère et auto-limitante rapidement après l'administration, sans effet délétère sur l'homéostasie de l'articulation à long terme.(Traduit par les auteurs).

Platelet-rich plasma in orthopedic therapy: a comparative systematic review of clinical and experimental data in equine and human musculoskeletal lesions.

BACKGROUND: This systematic review aimed to present and critically appraise the available information on the efficacy of platelet rich plasma (PRP) in equine and human orthopedic therapeutics and to verify the influence of study design and methodology on the assumption of PRP's efficacy. We searched Medline, PubMed, Embase, Bireme and Google Scholar without restrictions until July 2013. Randomized trials, human cohort clinical studies or case series with a control group on the use of PRP in tendons, ligaments or articular lesions were included. Equine clinical studies on the same topics were included independently of their design. Experimental studies relevant to the clarification of PRP's effects and mechanisms of action in tissues of interest, conducted in any animal species, were selected. RESULTS: This review included 123 studies. PRP's beneficial effects were observed in 46.7% of the clinical studies, while the absence of positive effects was observed in 43.3%. Among experimental studies, 73% yielded positive results, and 7.9% yielded negative results. The most frequent flaws in the clinical trials' designs were the lack of a true placebo group, poor product characterization, insufficient blinding, small sampling, short follow-up periods, and adoption of poor outcome measures. The methods employed for PRP preparation and administration and the selected outcome measures varied greatly. Poor study design was a common feature of equine clinical trials. From studies in which PRP had beneficial effects, 67.8% had an overall high risk of bias. From the studies in which PRP failed to exhibit beneficial effects, 67.8% had an overall low risk of bias. CONCLUSIONS: Most experimental studies revealed positive effects of PRP. Although the majority of equine clinical studies yielded positive results, the human clinical trials' results failed to corroborate these findings. In both species, beneficial results were more frequently observed in studies with a high risk of bias. The use of PRP in musculoskeletal lesions, although safe and promising, has still not shown strong evidence in clinical scenarios.

Autologous processed plasma: cytokine profile and effects upon injection into healthy equine joints.

This experimental controlled study was performed to evaluate the composition of autologous processed plasma (APP), and the effects of APP intra-articular injection into healthy equine metacarpophalangeal joints. The effects on joints were analysed with a short-phase protocol and a prolonged-phase protocol using saline-injected joints as controls. For the short protocol, horses received one intra-articular APP injection. Synovial fluid samples were collected prior to the injection and 3, 6, 24, 48, and 16 h after treatment. For the prolonged protocol, the joints received three weekly injections of APP, and samples were collected at 0, 7, 14, 21, and 28 days before APP administration. IL1-ra level was found to be increased in APP compared to plasma. Upon intra-articular administration of APP, transient (up to 24 h) increases in white blood cell (WBC) counts along with elevated protein and prostaglandin E2 (PGE2) concentrations were observed in the treated joints. Over the 28-day observation period, APP did not elicit changes relative to baseline levels, but WBC counts, PGE2 and chondroitin sulphate concentrations were lower than those found in the control. In conclusion, APP intra-articular injection induced a mild and transitory inflammatory response but no inflammation reaction was observed over a longer period of treatment and observation.

Relevance of synovial fluid chondroitin sulphate as a biomarker to monitor polo pony joints.

Osteoarthritis (OA) of the metacarpophalangeal joint is the most common articular disease in polo ponies leading to early retirement. A biomarker that would discriminate between pathological and physiological changes secondary to exercise could be helpful in OA prevention. The aim of this study was to investigate the effects of polo training on synovial fluid biomarkers of inflammation and cartilage turnover in polo ponies of different skill levels. Synovial fluid samples were collected from metacarpophalangeal joints of polo ponies before and during the polo season (320 d). Nucleated cells, soluble protein, prostaglandin E2 (PGE2), glycosaminoglycans (GAG), and urea were measured. The main synovial fluid GAG are chondroitin sulphate (CS, ~25 µg/mL) and hyaluronic acid (HA, ~400 µg/mL). After a polo match, a transitory increase in protein and PGE2, but not CS and HA, occurred (expressed as urea ratio), returning to basal levels in 24 h. During the polo season, the number of synovial fluid nucleated cells was always in the normal range. Increases in protein and HA occurred during the initial 40 to 80 d, returning to basal levels afterwards. In contrast, in polo prospects the concentration of CS steadily increased during the season. Long-term follow-up revealed that the synovial fluid CS was significantly higher in polo ponies that developed joint diseases within 24 months following our study. In conclusion, CS seems to be an early marker of articular cartilage damage.

L'arthrose (OA) de l'articulation métacarpophalangienne est la maladie articulaire la plus fréquente chez les poneys de polo menant à la retraite anticipée. Un biomarqueur qui était discriminateur entre les changements pathologiques et physiologiques secondaires au exercice pourrait être utile pour la prévention de l'OA. L'objectif de la présente étude était examiner les effets de l'activité de polo sur les biomarqueurs de l'inflammation et de le métabolisme de la cartilage dans le liquide synovial des poneys de polo de différents niveaux de qualification. Le SF était obtenu à partir de les articulations métacarpophalangiennes de poneys de polo, avant et pendant la saison de polo (320 jours). Les cellules nucléés, protéine soluble, prostaglandine E2 (PGE2), glycosaminoglycanes (GAG) et l'urée ont été mesurés. Les principaux GAG de le liquide synovial sont le chondroïtine sulfate (CS, ~25 µg/mL) et l'acide hyaluronique (HA, ~400 µg/mL). Après un match de polo, ocorru une augmentation transitoire de la protéine et de la PGE2, mais pas de CS et de HA (exprimé comme le raison d'urée), qui a retourné aux niveaux basal dans 24 h. Pendant la saison de polo, le numero de cellules nucléés dans le liquide synovial était toujours normaux. La protéine et le HA augmentaient pendant les premiers 40­80 jours, mais tous les deux sont retournés aux niveaux de base plus tard. En contraste, dans le group de jeunes poneys (G1), la concentration de CS a augmenté régulièrement pendant la saison. Accompagnant à long terme avait révéle que le CS de liquide synovial était significativement plus élevée chez les poneys de polo que, dans les 24 mois suivants, avaient developpé des maladies articulaires. En conclusion, le CS du liquide synovial semble être un marqueur précoce des destructions de la cartilage articulaire.(Traduit par les auteurs).

Urinary glycosaminoglycans in horse osteoarthritis. Effects of chondroitin sulfate and glucosamine.

Our objectives were to characterize the urinary excretion of glycosaminoglycans (GAGs) in horse osteoarthritis, and to investigate the effects of chondroitin sulfate (CS) and glucosamine (GlcN) upon the disease. Urinary GAGs were measured in 47 athletic horses, 20 healthy and 27 with osteoarthritis. The effects of CS and GlcN were investigated in mild osteoarthritis. In comparison to normal, urinary GAGs were increased in osteoarthritis, including mild osteoarthritis affecting only one joint. Treatment with CS+GlcN led to a long lasting increase in the urinary CS and keratan sulfate (KS), and significant improvement in flexion test of tarsocrural and metacarpophalangeal joints was observed. In conclusion, urinary CS and KS seems to reflect the turnover rates of cartilage matrix proteoglycans, and the measurement of these compounds could provide objective means of evaluating and monitoring joint diseases.

Bilateral sinus cysts in a filly treated by endoscopic sinus surgery.

Bilateral cysts in the frontal and caudal maxillary sinuses in a filly treated by endoscopic sinus surgery (sinoscopy) and triangulation technique are described. Sinoscopy has the advantage of being a minimally invasive technique and permits a complete inspection of the frontal and maxillary sinuses, which is not possible through flap sinusotomy.