Production of Cytotoxic Antibodies After Intra-Articular Injection of Allogeneic Synovial Membrane Mesenchymal Stem Cells With and Without LPS Administration.

Allogeneic mesenchymal stem cells (MSC) are widely used in clinical routine due to the shorter expansion time and reliability of its quality. However, some recipients can produce alloantibodies that recognize MSCs and activate the immune system, resulting in cell death. Although antibody production was already described after MSC injection, no previous studies described the immune response after intra-articular MSC injection in acute synovitis. This study aimed to evaluate the influence of inflammation on immune response after single and repeated intra-articular injections of synovial membrane MSC (SMMSC). Horses were divided in three groups: control group (AUTO) received autologous synovial membrane MSCs; whereas group two (ALLO) received allogeneic SMMSCs and group three (ALLO LPS) was submitted to acute experimental synovitis 8 h before SMMSCs injection. The procedure was repeated for all groups for 28 days. Physical and lameness evaluations and synovial fluid analysis were performed. Sera from all animals were obtained before and every 7 days after each injection up to 4 weeks, to perform microcytotoxicity assays incubating donor SMMSCs with recipients' sera. The first injection caused a mild and transient synovitis in all groups, becoming more evident and longer in ALLO and ALLO LPS groups after the second injection. Microcytotoxicity assays revealed significant antibody production as soon as 7 days after SMMSC injection in ALLO and ALLO LPS groups, and cytotoxicity scores of both groups showed no differences at any time point, being equally different from AUTO group. Although inflammation is capable of inducing MHC expression in MSCs, which enhances immune recognition, cytotoxicity scores were equally high in ALLO and ALLO LPS groups, making it difficult to determine the potentiation effect of inflammation on antibody production. Our findings suggest that inflammation does not display a pivotal role in immune recognition on first allogeneic MSC injection. In a translational way, since specific antibodies were produced against MSCs, patients that need more than one MSC injection may benefit from a first allogeneic injection followed by subsequent autologous injections.

Hyaluronic acid has chondroprotective and joint-preserving effects on LPS-induced synovitis in horses.

The intra-articular use of hyaluronic acid (HA) for the treatment of synovitis and osteoarthritis is still controversial. As a consequence, corticosteroids remain the most frequently employed therapeutic agents, despite their potential systemic and local deleterious effects. This study examined the anti-inflammatory, antioxidant, and chondroprotective activities of low and high molecular weight hyaluronic acid (LMW-HA and HMW-HA) on lipopolysaccharide (LPS)-induced synovitis in horses compared to triamcinolone acetonide (TA). LPS was injected in the metacarpophalangeal joints, which were treated intra-articularly with either TA (as control) or LMW-HA or HMW-HA. Joint clinical evaluation and synovial fluid (SF) analysis were performed at 0, 8, 24, and 48 h. The white blood cell counts (WBC), prostaglandin E2 (PGE2), interleukin (IL)-1, IL-6, IL-10, tumor necrosis factor-α, chondroitin sulfate (CS) and HA concentrations, oxidative burst, and HA molecular weights were measured. TA reduced the lameness, swelling, and PGE2 release but increased the SF CS concentrations enormously at 24h and 48h, and decreased the SF HA modal molecular weight. These results indicate the breakdown of articular cartilage aggrecan and SF HA. In contrast, LMW-HA and HMW-HA were less effective in reducing the inflammation symptoms, but preserved the joints because only a modest increase in CS occurred at 24 h, decreasing at 48 h, and the SF HA was maintained. The HA-treatment also had anti-inflammatory actions, and LMW-HA was the most effective in reducing the release of cytokine. In summary, the HA treatment inhibited efficiently the digestion of cartilage proteoglycans and SF HA breakdown.

Pro-inflammatory mediators and neutrophils are increased in synovial fluid from heifers with acute ruminal acidosis.

BACKGROUND: Acute ruminal acidosis (ARA) is a metabolic disease of cattle characterized by an aseptic synovitis. ARA is the result of an increased intake of highly fermentable carbohydrates that frequently occurs in dairy cattle subjected to high production requirements. In human joint diseases such as rheumatoid arthritis and gout, several pro-inflammatory molecules are increased in the synovial fluid, including cytokines, prostaglandin E2 (PGE2), metalloproteinases, and neutrophil extracellular traps (NETs). The aim of this study was to identify the presence of proinflammatory mediators and neutrophils in the synovial fluid of heifers with ARA, induced by an oligofructose overload. Five heifers were challenged with an oligofructose overload (13 g/kg BW) dissolved in water. As a control, a similar vehicle volume was used in four heifers. Synovial fluid samples were collected from the tarso-crural joint and PGE2, IL-6, IL-1ß, ATP, lactate dehydrogenase (LDH), albumin, glucose, matrix metalloproteinase-9 (MMP-9), cellular free DNA, NETs, and serpin B1 were analyzed at 0, 9, and 24 h post treatment. RESULTS: At 9 h post oligofructose overload, an increase of IL-1ß, IL-6, PGE2, serpin B1 and LDH was detected in the joints when compared to the control group. At 24 h, the synovial fluid was yellowish, viscous, turbid, and contained abundant neutrophils. An increase of DNA-backbone-like traps, histone 3 (H3cit), aggregated neutrophil extracellular traps (aggNETs), and serpin B1 were observed 24 h post treatment. Furthermore, albumins, LDH, ATP, MMP-9, IL-6, and IL-1ß were increased after 24 h. CONCLUSIONS: The overall results indicate that IL-1ß, IL-6 and PGE2, were the earliest proinflammatory parameters that increased in the synovial fluid of animals with ARA. Furthermore, the most sever inflammatory response in the joint was observed after 24 h and could be associated with a massive presence of neutrophils and release of aggNETs.

P2X3 and P2X2/3 Receptors Play a Crucial Role in Articular Hyperalgesia Development Through Inflammatory Mechanisms in the Knee Joint Experimental Synovitis.

Osteoarthritis (OA) is a degenerative and progressive disease characterized by cartilage breakdown and by synovial membrane inflammation, which results in disability, joint swelling, and pain. The purinergic P2X3 and P2X2/3 receptors contribute to development of inflammatory hyperalgesia, participate in arthritis processes in the knee joint, and are expressed in chondrocytes and nociceptive afferent fibers innervating the knee joint. In this study, we hypothesized that P2X3 and P2X2/3 receptors activation by endogenous ATP (adenosine 5'-triphosphate) induces articular hyperalgesia in the knee joint of male and female rats through an indirect sensitization of primary afferent nociceptors dependent on the previous release of pro-inflammatory cytokines and/or on neutrophil migration. We found that the blockade of articular P2X3 and P2X2/3 receptors significantly attenuated carrageenan-induced hyperalgesia in the knee joint of male and estrus female rats in a similar manner. The carrageenan-induced knee joint inflammation increased the expression of P2X3 receptors in chondrocytes of articular cartilage. Further, the blockade of articular P2X3 and P2X2/3 receptors significantly reduced the increased concentration of TNF-α, IL-6, and CINC-1 and the neutrophil migration induced by carrageenan. These findings indicate that P2X3 and P2X2/3 receptors activation by endogenous ATP is essential to hyperalgesia development in the knee joint through an indirect sensitization of primary afferent nociceptors dependent on the previous release of pro-inflammatory cytokines and/or on neutrophil migration.

Intra-Articular Blockade of P2X7 Receptor Reduces the Articular Hyperalgesia and Inflammation in the Knee Joint Synovitis Especially in Female Rats.

Synovitis is a key factor in joint disease pathophysiology, which affects a greater proportion of women than men. P2X7 receptor activation contributes to arthritis, but whether it plays a role in articular inflammatory pain in a sex-dependent manner is unknown. We investigated whether the P2X7 receptor blockade in the knee joint of male and female rats reduces the articular hyperalgesia and inflammation induced by a carrageenan knee joint synovitis model. Articular hyperalgesia was quantified using the rat knee joint incapacitation test and the knee joint inflammation, characterized by the concentration of cytokines tumor necrosis factor-α, interleukin-1ß, interleukin-6, and cytokine-induced neutrophil chemoattractant-1, and by neutrophil migration, was quantified using enzyme-linked immunosorbent assay and by myeloperoxidase enzyme activity measurement, respectively. P2X7 receptor blockade by the articular coadministration of selective P2X7 receptor antagonist A740003 with carrageenan significantly reduced articular hyperalgesia, pro-inflammatory cytokine concentrations, and myeloperoxidase activity induced by carrageenan injection into the knee joint of male and estrus female rats. However, a lower dose of P2X7 receptor antagonist was sufficient to significantly induce the antihyperalgesic and anti-inflammatory effects in estrus female but not in male rats. These results suggest that P2X7 receptor activation by endogenous adenosine 5'-triphosphate is essential to articular hyperalgesia and inflammation development in the knee joint of male and female rats. However, female rats are more responsive than male rats to the antihyperalgesic and anti-inflammatory effects induced by P2X7 receptor blockade. PERSPECTIVE: P2X7 receptors could be promising therapeutic targets in the treatment of knee joint disease symptoms, especially in women, who are more affected than men by these conditions.

Physiological and analgesic effects of continuous-rate infusion of morphine, butorphanol, tramadol or methadone in horses with lipopolysaccharide (LPS)-induced carpal synovitis.

BACKGROUND: Continuous-rate infusion (CRI) of drugs results in more stable plasma drug concentrations than administration of intermittent boluses, thus providing greater stability of physiological parameters. The aim of this study was to evaluate the physiologic and analgesic effects of the administration of morphine, butorphanol, tramadol or methadone by CRI in horses with induced synovitis of the radiocarpal joint. RESULTS: Increased values of cardiorespiratory parameters and body temperature were observed in all groups after initiation of opioid administration, and these increases were sustained throughout the CRI period. Morphine, butorphanol and methadone each caused a reduction in gut sounds, and this effect was greatest in animals that received morphine. Administration of morphine or methadone reduced the degree of lameness after the end of intravenous infusion. Administration of tramadol did not alter the degree of lameness in the animals. CONCLUSIONS: CRI of morphine or methadone, but not butorphanol or tramadol, provided analgesia in horses with carpal synovitis. All of these opioids increased cardiovascular and respiratory parameters and reduced gut sounds during CRI.

Beneficial effects of Anadenanthera colubrina (Vell.) Brenan extract on the inflammatory and nociceptive responses in rodent models.

ETHNOPHARMACOLOGICAL RELEVANCE: Anadenanthera colubrina (Vell.) Brenan, popularly known as "angico", is a plant that has been widely used in folk medicine due to its anti-inflammatory property. To evaluate the pharmacological activities of this plant, studies were performed on its antinociceptive and anti-inflammatory properties. MATERIALS AND METHODS: The AE of Anadenanthera colubrina, made from the bark, was used in rodents via oral route (p.o.), at 100, 200, and 400mg/kg in classical models of nociception (acetic acid-induced writhing and hot-plate test) and inflammation evoked by carrageenan (e.g., paw edema, peritonitis, and synovitis). RESULTS: The acetic acid-induced abdominal writhes in mice were significantly reduced (P<0.001) by oral treatment with the extract (100, 200, and 400mg/kg), but the extract did not significantly increase the latency in the nociceptive hot-plate test. Anadenanthera colubrina aqueous extract reduced significantly the edema and, besides, diminished the mieloperoxidase activity (200 and 400mg/kg, P<0.01). The carrageenan-induced peritonitis was significantly reduced (P<0.05) by the aqueous extract at 100, 200, and 400mg/kg. The aqueous extract (200mg/kg) reduces the synovial leukocyte infiltration on carrageenan-induced synovitis in rats (P<0.01), but failed to significantly affect joint swelling and impaired mobility. CONCLUSIONS: We show for the first time that the anti-inflammatory and peripheral antinociceptive activities of Anadenanthera colubrina are consistent, at least in part, with the use of this plant in popular medicine practices.

Epidural analgesia with morphine or buprenorphine in ponies with lipopolysaccharide (LPS)-induced carpal synovitis.

This study evaluated the analgesia effects of the epidural administration of 0.1 mg/kg bodyweight (BW) of morphine or 5 µg/kg BW of buprenorphine in ponies with radiocarpal joint synovitis. Six ponies were submitted to 3 epidural treatments: the control group (C) received 0.15 mL/kg BW of a 0.9% sodium chloride (NaCl) solution; group M was administered 0.1 mg/kg BW of morphine; and group B was administered 5 µg/kg BW of buprenorphine, both diluted in 0.9% NaCl to a total volume of 0.15 mL/kg BW administered epidurally at 10 s/mL. The synovitis model was induced by injecting 0.5 ng of lipopolysaccharide (LPS) in the left or right radiocarpal joint. An epidural catheter was later introduced in the lumbosacral space and advanced up to the thoracolumbar level. The treatment started 6 h after synovitis induction. Lameness, maximum angle of carpal flexion, heart rate, systolic arterial pressure, respiratory rate, temperature, and intestinal motility were evaluated before LPS injection (baseline), 6 h after LPS injection (time 0), and 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, and 24 h after treatments. Although the model of synovitis produced clear clinical signs of inflammation, the lameness scores in group C were different from the baseline for only up to 12 h. Both morphine and buprenorphine showed a reduction in the degree of lameness starting at 0.5 and 6 h, respectively. Reduced intestinal motility was observed at 0.5 h in group M and at 0.5 to 1 h in group B. Epidural morphine was a more effective analgesic that lasted for more than 12 h and without side effects. It was concluded that morphine would be a valuable analgesic option to alleviate joint pain in the thoracic limbs in ponies.

A influência da crioterapia na dor e edema induzidos por sinovite experimental / The cryotherapy influence on pain and edema induced by experimental synovitis

O objetivo deste estudo foi analisar a influência da crioterapia na dor e edema advindos de sinovite induzida em ratos. Foram utilizados 12 ratos, distribuídos em dois grupos: Controle (GC) – submetido à indução de sinovite no joelho direito, e não tratado; e Tratamento (GT) – submetido à sinovite no joelho direito, e tratado com crioterapia. Para induzir a lesão, foi injetado no espaço tíbio-femoral formalina 5%. Para avaliação da dor foi utilizado o teste de incapacidade funcional, que avaliou a dor durante a marcha do animal (tempo de elevação da pata – TEP); e para quantificar o edema foi utilizado um paquímetro metálico, na região da interlinha do joelho. As avaliações ocorreram antes da injeção de formalina (AV1), 1 (AV2) e 2 horas (AV3) após. Após 10 minutos da lesão, o membro posterior direito foi submerso em água com gelo, à 5ºC por 20 minutos. A avaliação do TEP mostrou aumento de 194,03% (AV2) e 169,26% (AV3) para GC; e 134,25% (AV2) e 103,13% (AV3) para GT, com relação à AV1. Na comparação entre os grupos, em AV3, houve diminuição significativa para GT. A avaliação do edema mostrou aumento do diâmetro, para GC de 39,15% (AV2) e 42,39% (AV3); e 27,91% (AV2) e 14,50% (AV3) para GT, tendo como referência AV1; sendo que apenas GT apresentou diminuição significativa entre AV2 e AV3. Conclui-se que os efeitos em curto prazo, da crioterapia, foram significativos para reduzir a dor e edema, em ratos submetidos à indução de sinovite.

The aim of this study was to examine the influence of cryotherapy on pain and swelling in an induced synovitis in rats. A total of 12 rats were allocated into two groups: the control (CG) – underwent the synovitis induction in his right knee, and not treated, and treatment (TG) – synovitis in his right knee, and treated with cryotherapy. To induce injury, was injected into the tibio-femoral joint space 5% formalin. For the pain assessment was used the functional incapacitation test, which assessed pain during gait of the animal (paw elevation time – PET) and to quantify the swelling was used a metal caliper in the region of the interline knee. Assessments occurred prior to injection of formalin (EV1), 1 (EV2) and 2 hours (EV3) after. After 10 minutes of the injury, the right hind limb was immersed in ice water, 5ºC for 20 minutes. The evaluation of PET revealed increased 194.03% (EV2) and 169.26% (EV3) for CG, and 134.25% (EV2) and 103.13% (EV3) to TG, with respect to EV1. Comparing the groups, EV3, a significant decrease in GT. The edema assessment showed increased diameter of 39.15% for CG (EV2) and 43.39% (EV3) and 27.91% (EV2) and 14.5% (EV3) to TG, with reference EV1; and only TG significantly decreased between EV2 and EV3. We conclude that the effects in the short term, cryotherapy, were significant to reduce pain and edema in rats with induced synovitis.

Differing effects of exogenous and endogenous hydrogen sulphide in carrageenan-induced knee joint synovitis in the rat.

BACKGROUND AND PURPOSE: Recent findings suggest that the noxious gas H(2)S is produced endogenously, and that physiological concentrations of H(2)S are able to modulate pain and inflammation in rodents. This study was undertaken to evaluate the ability of endogenous and exogenous H(2)S to modulate carrageenan-induced synovitis in the rat knee. EXPERIMENTAL APPROACH: Synovitis was induced in Wistar rats by intra-articular injection of carrageenan into the knee joint. Sixty minutes prior to carrageenan injection, the rats were pretreated with indomethacin, an inhibitor of H(2)S formation (DL-propargylglycine) or an H(2)S donor [Lawesson's reagent (LR)]. KEY RESULTS: Injection of carrageenan evoked knee inflammation, pain as characterized by impaired gait, secondary tactile allodynia of the ipsilateral hindpaw, joint swelling, histological changes, inflammatory cell infiltration, increased synovial myeloperoxidase, protein nitrotyrosine residues, inducible NOS (iNOS) activity and NO production. Pretreatment with LR or indomethacin significantly attenuated the pain responses, and all the inflammatory and biochemical changes, except for the increased iNOS activity, NO production and 3-NT. Propargylglycine pretreatment potentiated synovial iNOS activity (and NO production), and enhanced macrophage infiltration, but had no effect on other inflammatory parameters. CONCLUSIONS AND IMPLICATIONS: Whereas exogenous H(2)S delivered to the knee joint can produce a significant anti-inflammatory and anti-nociceptive effect, locally produced H(2)S exerts little immunomodulatory effect. These data further support the development and use of H(2)S donors as potential alternatives (or complementary therapies) to the available anti-inflammatory compounds used for treatment of joint inflammation or relief of its symptoms.

Aplicação do laser 808nm na dor articular crônica de ratos Wistar / Aplication of 808nm laser in chronic joint pain in Wistar rats

Durante o processo inflamatório, alguns tipos de células predominam e estão envolvidos com a produção de dor. O laser de baixa potência mostra-se útil no controle de mediadores inflamatórios, mas a ampla divergência nos parâmetros dosimétricos, conduz a variações nos resultados positivos. O objetivo deste estudo foi analisar o uso do laser 808nm, na dor articular crônica e edema de ratos Wistar submetidos a sinovite. Foram utilizados 10 ratos Wistar, divididos em dois grupos: GC - indução de sinovite em joelho direito e tratamento placebo; GT - indução de sinovite e irradiado com laser 10J/cm². Para a produção de sinovite foram injetados 100µl de formalina a 5 por cento, no espaço articular tibiofemoral direito. A avaliação da dor ocorreu pelo tempo de elevação da pata (TEP). No 10º dia após a indução de sinovite, iniciou-se o tratamento com laser sobre a face articular medial do joelho, procedimento mantido no decorrer de 10 dias. Os resultados mostraram que o laser apresentou efeitos analgésicos, visto que, para o grupo tratado, houve redução significativa nos valores de TEP, indicando redução da dor, quando comparados os valores pós-tratamento com os pré-tratamento, o que não ocorreu para o grupo que recebeu tratamento placebo; para o edema, o laser também mostrou redução significativa do mesmo. Concluiu-se que o laser 808nm, com 10J/cm², é eficaz na redução da dor articular crônica e edema, em animais submetidos a sinovite experimental.

During the inflammatory process some kinds of cells are predominant and are involved with pain production. Low power laser seems to be useful in the control of inflammatory mediators, but wide divergence in the dosemetric parameters leads to variations in its positive results. The aim of this study was to assess the use of the 808nm laser in the chronic joint pain and edema of Wistar rats submitted to synovitis. Sample was composed of 10 Wistar rats divided in two groups: CG - synovitis induction on right knee and placebo treatment; TG - synovitis induction and irradiated with 10J/cm² laser. 100µl of 5 percent phormaline were injected in the right tibiofemoral joint space. Pain assessment occurred by the time of paw elevation (TPE). On the 10th day after synovitis induction, laser treatment began on the knee medial joint surface and this procedure was kept for 10 days. The outcomes showed that laser presented analgesic effects, since significant reduction of the TPE values was observed for the treated group, hence indicating pain reduction when compared with the pre-treatment values; the same situation did not occur to the group which received the placebo treatment. Concerning edema, laser has also shown significant reduction. It can be concluded that laser 808nm with 10J/cm² is efficient in reduction of chronic joint pain and edema in animals submitted to experimental synovitis.

Effects of intraarticular ropivacaine and morphine on lipopolysaccharide-induced synovitis in horses.

OBJECTIVE: To compare the intraarticular (IA) analgesic effects of ropivacaine and morphine in horses with experimentally induced synovitis. STUDY DESIGN: Randomized, blinded cross-over design. ANIMALS: Twelve healthy mixed breed horses between 8-15 years old. METHODS: Synovitis was induced in the left radio-carpal joint with an injection of lipopolysaccharide (Escherichia coli 055:B5). Six hours later, the horses were treated with an IA injection of 40 mg of ropivacaine (ROPI), 40 mg of morphine (MOR), 20 mg of ropivacaine added to 20 mg of morphine in saline (RM) or 4 mL of saline (SAL), as control. Analgesia was measured subjectively using a numerical rating scale, a simple descriptive scale, pain upon maximal flexion of the carpus and by the range of motion exhibited by the affected joint. Data are reported as mean +/- SD and were analyzed using anova. Blood and synovial data were analyzed by split plots in time with units (treatments: SAL, ROPI, MOR and RM) and subunits (times: T0-24), in a completely randomized design with six replicates. Mean comparisons were made by Tukey's test; differences were considered significant at p < 0.05. RESULTS: Ropivacaine had a clinical analgesic effect with a relative short duration ( approximately 2.5 to 3.5 hours). Morphine had a slower onset of action than ROPI, but a stronger analgesic effect of longer duration. The RM showed an earlier onset of action than MOR and had a strong analgesic effect for the 24-hour post-injection period. All treatments caused a significant decrease in total nucleated cells compared with the control, 24 hours after administration. CONCLUSIONS AND CLINICAL RELEVANCE: Morphine alone or in combination with ropivacaine produced a strong analgesic effect of prolonged duration, which may offer pain relief for acute synovitis for at least 24 hours.

Análise morfológica e bioquímica da sinóvia de coelhos imunizados com colágeno do tipo V / Morphological and biochemical analysis of the synovia of rabbits immunized with type V collagen

Descrevemos modelo original de sinovite experimental em coelhos imunizados com colágeno V com escasso processo inflamatório, intenso remodelamento matricial e vasculite. Analise morfológica e bioquímica foi realizada em coelhas Nova Zelândia (N=20) imunizadas com colágeno do tipo V, comparadas com controles. Foi observado o aumento dos colágenos I, III e V, oclusão do lúmen vascular e escasso processo inflamatório. A análise bioquímica confirmou a fibrose com aumento da síntese de colágeno. Nós postulamos que as alterações sinoviais descritas neste modelo foram conseqüência das particularidades do colágeno V, que promove manifestações imunológicas e clínicas semelhantes à esclerodermia.

We described an original model of experimental synovitis in rabbits immunized with collagen V with scant cellular infiltration, intense matrix remodeling and vasculitis. Morphological and biochemical analysis were realized in New Zealand female rabbits (N=20) immunization with type V collagen, compared with control rabbits. It was observed increase of collagen I, III and V, vascular lumen occlusion and scant inflammatory process. Biochemical analysis confirmed the fibrosis with increased synthesis of collagen. We postulate that synovial changes described in this model are consequence of collagen V particularities, which promotes immunologic and clinical manifestations similar to scleroderma.

Reactive nitrogen species scavenging, rather than nitric oxide inhibition, protects from articular cartilage damage in rat zymosan-induced arthritis.

1. The contribution of nitric oxide (NO) and peroxynitrite (PN) to inflammation in a zymosan-induced (1 mg, intra-articular, i.art.) rat model of arthritis was assessed by histopathology and by measuring the glycosaminoglycan (GAG) content of the articular cartilage. 2. Progression of the chronic synovitis in zymosan-induced arthritis (ZYA) was associated with increased nitrite and nitrotyrosine (3-NT) levels in the joint exudates that paralleled a progressive loss of the GAG content. An increase in 3-NT was also observed after i.art. PN. 3. The nonselective nitric oxide synthase (NOS) inhibitor l-N(G)-nitroarginine methyl ester (25-75 mg x kg(-1)day(-1)) or the selective inducible NOS inhibitor aminoguanidine (50-100 mg x kg(-1)day(-1)) given 1 h before (prophylactic) or 3 days after (therapeutic) injection of the zymosan ameliorated the synovitis, but worsened the GAG loss, as measured at the end of the experiment (day 7). 4. The PN scavenger uric acid (100-250 mg x kg(-1) i.p. four times daily) given prophylactically until the end of the experiment (day 14), in a dose compatible with its PN scavenging activity, significantly decreased both the synovitis and the GAG loss. 5. In conclusion, PN formation is associated with cartilage damage in addition to proinflammatory activity in ZYA. NOS inhibitors and a PN scavenger were able to reduce the cellular infiltration, while displaying opposite effects on cartilage homeostasis either by enhancing or ameliorating the damage, respectively.

Enantiospecific pharmacokinetics of ketoprofen in plasma and synovial fluid of horses with acute synovitis.

Pharmacokinetic parameters were established for enantiomers of the nonsteroidal anti-inflammatory drug (NSAID) ketoprofen (KTP) administered as the racemic mixture at a dose of 2.2 mg/kg and as separate enantiomers, each at a dose of 1.1 mg/kg to a group of six horses (five mares and one gelding). A four-period cross-over study in a LPS-induced model of acute synovitis was used. After administration of the racemic mixture S(+)KTP was the predominant enantiomer in plasma as well as in synovial fluid. Unidirectional inversion of R(-) to S(+)KTP was demonstrated but the inversion was less marked than previously reported. It is suggested that this reduction could be because of the influence of the inflammatory reaction on hepatic metabolism. The disposition of KTP enantiomers after administration of the racemic mixture was similar to those observed after administration of S(+) and R(-)KTP. The S(+) and R(-)KTP concentrations in synovial fluid were low and short lasting. After administration of R(-)KTP significant concentrations of the optical antipode were detected in synovial fluid.

Zymosan modulates CD44 isoform expression in a murine model of inflammation resembling rheumatoid arthritis synovitis.

OBJECTIVE: To study local inflammation induced by zymosan in the murine air pouch, considered a model of synovial-like tissue inflammation, we investigated the time-course synthesis of CD44 and tumor necrosis factor-alpha (TNF-alpha) mRNA and established a relationship with leukocyte migration into the air pouch and CD44 expression on the leukocyte membrane. METHODS: Leukocytes from the air pouch exudate were collected and counted at 1, 4, 12, 24, 48, and 72 h after zymosan or saline injection. CD44 and TNF-alpha mRNA were studied by RT-PCR. CD44 variable exon analysis was assessed by Southern blot and CD44 membrane expression by flow cytometry. RESULTS: Leukocyte accumulation after zymosan injection was significantly higher than in saline injected controls. CD44 standard and variable isoforms including at least variable exons v6 and v9 were highly expressed in leukocytes from the zymosan air pouch exudate. In contrast, only the CD44 mRNA standard isoform was present in leukocytes from saline air pouch. Maximal TNF-alpha mRNA level was observed at 48 h after zymosan injection, whereas CD44 mRNA was constantly expressed throughout the whole term of the experiment, although variations in leukocyte count and relative formula were observed. CONCLUSION: Expression of CD44 variable isoform in leukocytes was specifically induced by zymosan, since none was detected in saline controls. TNF-alpha mRNA expression and leukocyte count at every time point served as markers for local inflammation. The presence of variable isoforms, including at least exons v6 and v9, consistently expressed throughout the assay suggests that they could play a role in this arthritis-like inflammation induced under zymosan stimulus.

Modelo experimental de sinovite transitória do quadril, produzida por reagente de fenol: estudo em coelhos jovens / Experimental model of transient hip synovitis, produced by phenol reagent: study in young rabbits

Foi produzido um método experimental de sinovite transitória do quadril em coelhos jovens, através de injeçao intra-articular de Reagente de Fenol. Foram utilizados sessenta e sete coelhos, dos quais vinte e sete serviram às observaçoes do grupo piloto. Dos quarenta coelhos estudados na fase definitiva, dez serviram de controle através de injeçao intra-articular de soluçao fisiológica. Atributos clínicos, radiológicos e anátomo-pathológicos foram analisados nos tempos de 1 hora, 6 horas, 24 horas, 48 horas e 7 dias, após a injeçao intra-articular. Os resultados obtivos revelaram que, tanto clínica como anátomo-patologicamente, houve a produçao de sinovite aguda que se iniciava no grupo de seis horas, intensificando-se até 48 horas e com regressao da inflamaçao ao 7§ dia, sem resultar em necrose intra-articular, nem em cronificaçao da sinovite. Radiologicamente foram observados: aumento dos espaço articular e ausência de lesoes ósseas nos quadris tratados. Foram discutidos o método e as técnicas empregadas, com especiais referências à movimentaçao passiva relacionada com a intensidade da sinovite, ao resultado radiológico do método e ao quadro anátomo-patológico obtido. Conclui-se tratar de método válido na produçao de modelo experimental de sinovite transitória do quadril.