RESUMEN
We hypothesized that after synovial injury, collagen V (Col V) expose occult antigens, and Col V autoantibodies develop, indicating the loss of immune tolerance against this molecule, thus leading to damage to mesenchymal-derived cells as well as the extracellular matrix in experimental arthritis. Thus, the present study investigated the effects of oral administration of Col V on the synovium after the development of inflammation in mBSA/CFA-induced arthritis. After fourteen days of intraarticular administration of mBSA, 10 male Lewis rats were orally administered Col V (500 µg/300 µL) diluted in 0.01 N acetic acid (IA-Col V group). The arthritic group (IA group, n = 10) received only intraarticular mBSA. An intra-articular saline injection (20 µL) was given to the control group (CT-Col V, n = 5). IA group presented damaged synovia, the expansion of the extracellular matrix by cellular infiltrate, which was characterized by T and B lymphocytes, and fibroblastic infiltration. In contrast, after Col V oral immunotherapy IA-Col V group showed a significant reduction in synovial inflammation and intense expression of IL-10+ and FoxP3+ cells, in addition to a reduction in Col V and an increase in Col I in the synovia compared to those in the IA group. Furthermore, an increase in IL-10 production was detected after IA-Col V group spleen cell stimulation with Col V in vitro. PET imaging did not differ between the groups. The evaluation of oral treatment with Col V, after mBSA/CFA-induced arthritis in rats, protects against inflammation and reduces synovial tissue damage, through modulation of the synovial matrix, showing an immunotherapeutic potential in inhibiting synovitis.
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Artritis Experimental , Colágeno Tipo V , Ratas Endogámicas Lew , Membrana Sinovial , Animales , Masculino , Administración Oral , Ratas , Artritis Experimental/inmunología , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/patología , Membrana Sinovial/inmunología , Membrana Sinovial/patología , Colágeno Tipo V/inmunología , Colágeno Tipo V/administración & dosificación , Adyuvante de Freund/administración & dosificación , Inmunoterapia/métodos , Interleucina-10 , Factores de Transcripción Forkhead/metabolismo , Albúmina Sérica BovinaRESUMEN
OBJECTIVES: Investigate whether and which synoviocytes would acquire trained immunity characteristics that could exacerbate joint inflammation following a secondary Staphylococcus aureus infection. METHODS: Lipopolysaccharide (LPS) and S. aureus were separately or double injected (21 days of interval) into the tibiofemoral joint cavity of male C57BL/6 mice. At different time points after these stimulations, mechanical nociception was analyzed followed by the analysis of signs of inflammation and damage in the affected joints. The trained immunity markers, including the glycolytic and mTOR pathway, were analyzed in whole tissue or isolated synoviocytes. A group of mice was treated with Rapamycin, an mTOR inhibitor before LPS or S. aureus stimulation. RESULTS: The double LPS - S. aureus hit promoted intense joint inflammation and damage compared to single joint stimulation, including markers in synoviocyte activation, production of proinflammatory cytokines, persistent nociception, and bone damage, despite not reducing the bacterial clearance. The double LPS - S. aureus hit joints increased the synovial macrophage population expressing CX3CR1 alongside triggering established epigenetic modifications associated with trained immunity events in these cells, such as the upregulation of the mTOR signaling pathway (p-mTOR and HIF1α) and the trimethylation of histone H3. Mice treated with Rapamycin presented reduced CX3CR1+ macrophage activation, joint inflammation, and bone damage. CONCLUSIONS: There is a trained immunity phenotype in CX3CR1+ synovial macrophages that contributes to the exacerbation of joint inflammation and damage during septic arthritis caused by S. aureus.
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Lipopolisacáridos , Macrófagos , Ratones Endogámicos C57BL , Infecciones Estafilocócicas , Staphylococcus aureus , Serina-Treonina Quinasas TOR , Animales , Masculino , Infecciones Estafilocócicas/inmunología , Macrófagos/inmunología , Serina-Treonina Quinasas TOR/inmunología , Staphylococcus aureus/inmunología , Receptor 1 de Quimiocinas CX3C/metabolismo , Membrana Sinovial/inmunología , Membrana Sinovial/patología , Artritis Infecciosa/inmunología , Artritis Infecciosa/microbiología , Ratones , Citocinas/inmunología , Citocinas/metabolismo , Sirolimus/farmacología , Inflamación/inmunología , Sinoviocitos/inmunología , Sinoviocitos/efectos de los fármacos , Nocicepción/efectos de los fármacos , Inmunidad EntrenadaRESUMEN
Pharmacological treatment of osteoarthritis is still inadequate due to the low efficacy of the drugs used. Dexmedetomidine via the intra-articular (i.a.) route might be an option for the treatment of osteoarthritis-associated pain. The present study assessed the analgesic and anti-inflammatory effects of dexmedetomidine administered via the i.a. route in different doses in an experimental model of rat knee osteoarthritis induced with monosodium iodoacetate. Rats were allocated to four groups with 24 animals in each group. The OA (osteoarthritis), DEX-1 (dexmedetomidine in dose of 1µg/kg) and DEX-3 (dexmedetomidine in dose of 3µg/kg) groups were subjected to induction of osteoarthritis through injection of monosodium iodoacetate (MIA) via the i.a. route on the right knee; the control group was not subjected to osteoarthritis induction. Clinical assessment was performed on day 0 (before osteoarthritis induction) and then on days 5, 10, 14, 21 and 28 after induction. Treatment was performed on day 7 via the i.a. route, consisting of dexmedetomidine in doses of 1 and 3 µg/kg, while group OA received 0.9% normal saline. The animals were euthanized on days 7, 14, 21 and 28. Samples of the synovial membrane were collected for histopathological analysis, and the popliteal lymph nodes were collected for measurement of cytokines (interleukin [IL] IL-6, tumor necrosis factor alpha [TNF-α]). Dexmedetomidine (1 and 3 µg/kg) significantly reduced the animals' weight distribution deficit during the chronic-degenerative stage of osteoarthritis and improved the pain threshold throughout the entire experiment. Histological analysis showed that dexmedetomidine did not cause any additional damage to the synovial membrane. The TNF-α levels decreased significantly in the DEX-3 group on day 28 compared with the OA group. Dexmedetomidine reduced pain, as evidenced by clinical parameters of osteoarthritis in rats, but did not have an anti-inflammatory effect on histological evaluation.
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Cartílago Articular/inmunología , Dexmedetomidina/farmacología , Interleucina-6/inmunología , Osteoartritis/tratamiento farmacológico , Membrana Sinovial/inmunología , Factor de Necrosis Tumoral alfa/inmunología , Animales , Cartílago Articular/patología , Modelos Animales de Enfermedad , Inyecciones Intraarticulares , Masculino , Osteoartritis/inducido químicamente , Osteoartritis/inmunología , Osteoartritis/patología , Ratas , Ratas Wistar , Membrana Sinovial/patologíaRESUMEN
Inflammatory myofibroblastic tumor (IMT) is a lesion of intermediate biological potential with local recurrences and rare metastases found in multiple anatomical locations. We present a case of a pure intraarticular IMT of the knee, a location that has not been previously documented, with genetic confirmation of ALK-CARS fusion detected with next-generation sequencing. A 20-year-old healthy male was admitted to the orthopedic oncology department due to several months of pain and restriction in movement of his left knee. On magnetic resonance imaging, multiple intraarticular nodular lesions were seen. The patient underwent 2 synovectomies within the course of 1 year. The initial biopsy was interpreted as nodular fasciitis. The second biopsy revealed exuberant tissue displaying compact fascicles of spindle cells intermixed with myxoid areas in a background of inflammatory cells, highly suggestive for IMT. Due to the unusual intraarticular location, equivocal ALK immunostaining and the differential diagnosis with nodular fasciitis, we performed targeted next-generation sequencing using Archer FusionPlex Sarcoma panel, which can identify multiple fusions in a single assay. An ALK-CARS fusion was found, supporting the diagnosis of IMT. This report emphasizes the added value of broad molecular analysis in cases with unusual clinical presentation, equivocal immunohistochemistry, and a wide differential diagnosis.
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Articulación de la Rodilla/patología , Proteínas de Fusión Oncogénica/genética , Neoplasias de los Tejidos Blandos/diagnóstico , Membrana Sinovial/patología , Aminoacil-ARNt Sintetasas/genética , Quinasa de Linfoma Anaplásico/genética , Biopsia , Procedimientos Quirúrgicos de Citorreducción , Diagnóstico Diferencial , Fascitis/diagnóstico , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Articulación de la Rodilla/diagnóstico por imagen , Articulación de la Rodilla/inmunología , Articulación de la Rodilla/cirugía , Imagen por Resonancia Magnética , Masculino , Neoplasias de los Tejidos Blandos/genética , Neoplasias de los Tejidos Blandos/inmunología , Neoplasias de los Tejidos Blandos/cirugía , Sinovectomía , Membrana Sinovial/diagnóstico por imagen , Membrana Sinovial/inmunología , Adulto JovenRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Oenothera rosea (Onagraceae), commonly known as "hierba del golpe" in Mexico, is an herbaceous plant widely used in Mexican traditional medicine for the treatment of pain and inflammation. AIM OF THE STUDY: The aim of this study was to assess the effect of extracts and compounds isolated from O. rosea in kaolin-carrageenan induced arthritis. MATERIALS AND METHODS: Hydroalcoholic extract from aerial parts of O. rosea was obtained and chemically separated in order to obtain OrEA and isolated compounds using column chromatography, HPLC, UPLC and NMR analysis. O. rosea extract and derivatives were tested on the kaolin/carrageenan (K/C) induced arthritis model on ICR mice. Knee inflammation and paw withdrawal threshold were assessed following intraarticular administration of kaolin and carrageenan (4% and 2%, respectively) and subsequent oral administration of O. rosea. TNF-α, IL-1ß, IL-6 and IL-10 levels from synovial capsule were measured using ELISA kits. NF-κB activity was also measured using the RAWBlue™ cell line. Finally, spleen and lungs were dissected to investigate body index. RESULTS: Oral administration of the O. rosea ethyl acetate fraction (25, 50 and 100 mg/kg) and isolated compounds (2 mg/kg) reduced the edema induced by kaolin/carrageenan, similar to the effect of methotrexate (1 mg/kg). Hyperalgesia but not allodynia was observed during this experiment. O. rosea derivatives reduced this behavior. The quantification of cytokines showed a reduction in TNF-α, IL-1ß and IL-6, as well as an increase of IL-10. NF-κB production was also reduced by administering O. rosea derivatives. Chemical analysis of O. rosea derivatives showed that the major compounds present in the ethyl acetate fraction were phenolic compounds. Gallic acid, quercetin glucoside and quercetin rhamnoside were separated and identified by UPLC-UV-MS, and myricetin glycoside and tamarixetin glucoside using 1H and 13C NMR. CONCLUSIONS: O. rosea produces different phenolic compounds capable of reducing the inflammation and secondary mechanical hyperalgesia produced by K/C administration. They also reduced proinflammatory cytokines and increased anti-inflammatory cytokines. Finally, NF-κB modulation was reduced by the administration of O. rosea. Therefore, O. rosea could be considered of interest in inflammatory and painful diseases.
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Analgésicos/uso terapéutico , Antiinflamatorios/uso terapéutico , Artritis/tratamiento farmacológico , Hiperalgesia/tratamiento farmacológico , Oenothera , Fenoles/uso terapéutico , Extractos Vegetales/uso terapéutico , Analgésicos/química , Analgésicos/farmacología , Animales , Antiinflamatorios/química , Antiinflamatorios/farmacología , Artritis/inducido químicamente , Artritis/inmunología , Carragenina , Línea Celular , Citocinas/inmunología , Modelos Animales de Enfermedad , Femenino , Hiperalgesia/inmunología , Caolín , Ratones Endogámicos ICR , FN-kappa B/inmunología , Fenoles/análisis , Fenoles/farmacología , Fitoquímicos/análisis , Fitoquímicos/farmacología , Fitoquímicos/uso terapéutico , Componentes Aéreos de las Plantas , Extractos Vegetales/química , Extractos Vegetales/farmacología , Membrana Sinovial/efectos de los fármacos , Membrana Sinovial/inmunologíaRESUMEN
OBJECTIVE: To investigate the participation of canonical Wnt and NF-κB signaling pathways in an experimental model of chronic arthritis induced by methylated bovine serum albumin (mBSA) in rat temporomandibular joint (TMJ). MATERIALS AND METHODS: Wistar rats were sensitized by mBSA+Complete Freund Adjuvant (CFA)/Incomplete Freund Adjuvant (IFA) on the first 14 days (1 ×/week). Subsequently, they received 1, 2 or 3 mBSA or saline solution injections into the TMJ (1 ×/week). Hypernociceptive threshold was assessed during the whole experimental period. 24 h after the mBSA injections, the TMJs were removed for histopathological and immunohistochemical analyses for TNF-α, IL-1ß, NF-κB, RANKL, Wnt-10b, ß-catenin and DKK1. RESULTS: The nociceptive threshold was significantly reduced after mBSA injections. An inflammatory infiltrate and thickening of the synovial membrane were observed only after mBSA booster injections. Immunolabeling of TNF-α, IL-1ß and Wnt-10b was increased in the synovial membrane in arthritic groups. The immunoexpression of nuclear ß-catenin was significantly higher only in the group that received 2 booster TMJ injections. However, NF-κB, RANKL and DKK1 immunoexpression were increased only in animals with 3 mBSA intra-articular injections. CONCLUSION: Our results suggest that canonical Wnt and NF-κB signaling pathways participate in the hypernociception and inflammatory response in TMJ synovial membrane during the development of rheumatoid arthritis in rats.
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Artritis Experimental/inmunología , Artritis Reumatoide/inmunología , Hiperalgesia/inmunología , FN-kappa B/inmunología , Articulación Temporomandibular/inmunología , Vía de Señalización Wnt , Animales , Artritis Experimental/patología , Artritis Reumatoide/patología , Adyuvante de Freund , Hiperalgesia/patología , Péptidos y Proteínas de Señalización Intercelular/inmunología , Interleucina-1beta/inmunología , Lípidos , Masculino , Ligando RANK/inmunología , Ratas Wistar , Albúmina Sérica Bovina , Membrana Sinovial/inmunología , Membrana Sinovial/patología , Articulación Temporomandibular/patología , Factor de Necrosis Tumoral alfa/inmunología , Proteínas Wnt/inmunologíaRESUMEN
Because collagen type V (Col V) can be exposed in tissue injury, we hypothesized that oral administration of this collagen species modulates the inflammation and remodeling of experimental synovitis, avoiding joint destruction, and that the modulation may differ according to the temporal administration. Arthritis (IA, n = 20) was induced in Lewis rats by intraarticular (ia) injection of 500 µg of methylated bovine serum albumin (mBSA) emulsified in complete Freund's adjuvant (CFA) (10 µl) followed by an intraarticular booster of mBSA (50 µg) in saline (50 µl) administered at 7 and 14 days. The control group received saline (50 µl, ia). After the first intraarticular injection, ten IA animals were supplemented via gavage with Col V (500 µg/300 µl) daily for 30 days (IA/Suppl). The control group received saline (50 µL) and Col V supplement in the same way (Suppl). Col V oral administration in IA/Suppl led to 1) inhibited edema and severe inflammatory cell infiltration, 2) decreased collagen fiber content, 3) decreased collagen type I, 4) inhibited lymphocyte subpopulations and macrophages, 5) inhibited IL-1ß, IL-10, IL-17 and TNF-α production and 6) increased expression of caspase-9 in the synovial tissue. In conclusion, Col V supplementation decreased synovial inflammation and the fibrotic response, possibly by increased the apoptosis of inflammatory cells.
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Artritis Experimental/tratamiento farmacológico , Colágeno Tipo V/farmacología , Membrana Sinovial/efectos de los fármacos , Administración Oral , Animales , Artritis Experimental/inmunología , Artritis Experimental/patología , Caspasa 9/metabolismo , Citocinas/metabolismo , Edema/tratamiento farmacológico , Edema/inmunología , Edema/patología , Adyuvante de Freund , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Inflamación/patología , Masculino , Metaloproteinasa 2 de la Matriz/metabolismo , Ratas Endogámicas Lew , Albúmina Sérica Bovina , Membrana Sinovial/inmunología , Membrana Sinovial/patologíaRESUMEN
The induction of tolerance has been proposed as a therapeutic strategy for arthritis aiming to decrease progression of the pathology, probably by promoting suppressor mechanisms of the autoimmune response. This work aimed to confirm whether the treatment with vitamin D3 could synergize oral tolerance induced by hydrolyzed collagen peptides, in our experimental model of antigen induced arthritis in New Zealand rabbits. Clinical observation of the phenomenon indicates that simultaneous treatment with hydrolyzed collagen peptides and vitamin D3 was beneficial when compared with no treatment, for arthritic animals, and for arthritic animals that received treatment with only hydrolyzed collagen peptides or vitamin D3. Treatment with hydrolyzed collagen peptides caused diminished proinflammatory cytokine levels, an effect synergized significantly by the simultaneous treatment with vitamin D3. The anatomical-pathological studies of the animals that received both treatments simultaneously showed synovial tissues without lymphocytic and plasma cell infiltrates, and without vascular proliferation. Some of the synovial tissue of the animals of these groups showed a slight decrease in Galectin-3 expression. We propose that simultaneous oral treatment with vitamin D3 and hydrolyzed collagen peptides could increase the immunoregulatory effect on the process of previously triggered arthritis. We used articular cartilage hydrolysate and not collagen II because peptides best expose antigenic determinants that could induce oral tolerance. Oral tolerance may be considered in the design of novel alternative therapies for autoimmune disease and we have herein presented novel evidence that the simultaneous treatment with vitamin D3 may synergize this beneficial effect.
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Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/inmunología , Colecalciferol/administración & dosificación , Péptidos/administración & dosificación , Administración Oral , Animales , Colágeno/química , Citocinas/inmunología , Femenino , Humanos , Conejos , Membrana Sinovial/inmunologíaRESUMEN
INTRODUCTION: Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by neutrophil articular infiltration, joint pain and the progressive destruction of cartilage and bone. IL-22 is a key effector molecule that plays a critical role in autoimmune diseases. However, the function of IL-22 in the pathogenesis of RA remains controversial. In this study, we investigated the role of IL-22 in the early phase of antigen-induced arthritis (AIA) in mice. METHODS: AIA was induced in C57BL/6, IL-22(-/-), ASC(-/-) and IL-1R1(-/-) immunized mice challenged intra-articularly with methylated bovine serum albumin (mBSA). Expression of IL-22 in synovial membranes was determined by RT-PCR. Articular hypernociception was evaluated using an electronic von Frey. Neutrophil recruitment and histopathological analyses were assessed in inflamed knee joint. Joint levels of inflammatory mediators and mBSA-specific IgG concentration in the serum were measured by ELISA. RESULTS: The IL-22 mRNA expression and protein levels in synovial tissue were increased during the onset of AIA. In addition, pharmacological inhibition (anti-IL-22 antibody) and genetic deficiency (IL-22(-/-) mice) reduced articular pain and neutrophil migration in arthritic mice. Consistent with these findings, recombinant IL-22 joint administration promoted articular inflammation per se in WT mice, restoring joint nociception and neutrophil infiltration in IL-22(-/-) mice. Moreover, IL-22-deficient mice showed reduced synovitis (inflammatory cell influx) and lower joint IL-1ß levels, whereas the production of IL-17, MCP-1/CCL2, and KC/CXCL1 and the humoral immune response were similar, compared with WT mice. Corroborating these results, the exogenous administration of IL-22 into the joints induced IL-1ß production in WT mice and reestablished IL-1ß production in IL-22(-/-) mice challenged with mBSA. Additionally, IL-1R1(-/-) mice showed attenuated inflammatory features induced by mBSA or IL-22 challenge. Articular nociception and neutrophil migration induced by IL-22 were also reduced in ASC(-/-) mice. CONCLUSIONS: These results suggest that IL-22 plays a pro-inflammatory/pathogenic role in the onset of AIA through an ASC-dependent stimulation of IL-1ß production.
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Artritis Experimental/inmunología , Interleucina-1beta/inmunología , Interleucinas/inmunología , Articulación de la Rodilla/inmunología , Animales , Antígenos/inmunología , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/inmunología , Proteínas Reguladoras de la Apoptosis/metabolismo , Artralgia/genética , Artralgia/inmunología , Artralgia/metabolismo , Artritis Experimental/genética , Artritis Experimental/metabolismo , Proteínas Adaptadoras de Señalización CARD , Movimiento Celular/inmunología , Ensayo de Inmunoadsorción Enzimática , Expresión Génica/inmunología , Interleucina-1beta/metabolismo , Interleucinas/biosíntesis , Interleucinas/genética , Articulación de la Rodilla/metabolismo , Ratones Endogámicos C57BL , Ratones Noqueados , Neutrófilos/citología , Neutrófilos/inmunología , Neutrófilos/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Membrana Sinovial/inmunología , Membrana Sinovial/metabolismo , Sinovitis/genética , Sinovitis/inmunología , Sinovitis/metabolismo , Zimosan/inmunología , Interleucina-22RESUMEN
Brazilin is an active compound of Caesalpinia sappan L. (Leguminosae), which possesses pro-apoptotic and anti-inflammation potentials depending on the specific cell type. However, it is largely unknown whether autophagy is implicated in the mechanism underlying its chemotherapeutic and anti-inflammatory effects in rheumatoid arthritis (RA). Here, we show that treatment of RA fibroblast-like synoviocytes (FLS) with brazilin results in enhanced level of autophagic flux, evidenced by accumulation of autophagosome and increased level of lipidated LC3 (LC3-II), which is mainly mediated by enhanced production of reactive oxygen species (ROS). Interestingly, long-term exposure of brazilin was able to restore cell survival against the cytotoxity, exclusively in RA FLS, but not in normal fibroblast. Importantly, such a restoration from brazilin-induced cytotoxity in RA FLS was completely abrogated after co-treatment with autophagy inhibitors including NH4Cl or chloroquine. Furthermore, we found that the pretreatment of RA FLS with brazilin reduced LPS- or TNF-induced NF-κB activation and the secretion of inflammatory cytokines in parallel with the enhanced autophagic flux. Such anti-NF-κB potentials of brazilin were drastically masked in RA FLS when autophagy was suppressed. These results suggest that brazilin is capable of activating autophagy exclusively in RA FLS, and such inducible autophagy promotes cell survival and limits inflammatory response.
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Antiinflamatorios/farmacología , Artritis Reumatoide/tratamiento farmacológico , Autofagia/efectos de los fármacos , Benzopiranos/farmacología , Fibroblastos/efectos de los fármacos , Fibroblastos/patología , Membrana Sinovial/citología , Animales , Antiinflamatorios/química , Artritis Reumatoide/inmunología , Artritis Reumatoide/patología , Benzopiranos/química , Caesalpinia/química , Células Cultivadas , Fibroblastos/citología , Fibroblastos/inmunología , Humanos , Ratones , FN-kappa B/inmunología , Células 3T3 NIH , Especies Reactivas de Oxígeno/inmunología , Membrana Sinovial/efectos de los fármacos , Membrana Sinovial/inmunología , Membrana Sinovial/patologíaRESUMEN
BACKGROUND: Platelet-rich plasma (PRP) preparations are a common treatment in osteoarthritis (OA) and inflammatory synovitis. However, there is ambiguity regarding the ideal concentration of leukocytes and platelets in these preparations necessary to induce an adequate anti-inflammatory and anabolic response in joint tissues, such as the synovial membrane. This research aimed to study, in normal synovial membrane explants (SME) challenged with lipopolysaccharide (LPS), the temporal effects (at 48 and 96h) of leukocyte- and platelet-rich gel (L-PRG) and pure platelet-rich gel (P-PRG) supernatants on the production and degradation of platelet associated growth factors (GF) (platelet derived GF isoform BB (PDGF-BB) and transforming growth factor beta-1 (TGF-ß1)), pro-inflammatory (tumour necrosis factor alpha (TNF-α)) and anti-inflammatory cytokines (interleukin 4 (IL-4) and IL-1 receptor antagonists (IL-1ra)) and hyaluronan (HA). METHODS: Synovial membrane explants (SMEs) from 6 horses were challenged with LPS and cultured for 96h with L-PRG and P-PRG supernatants at concentrations of 25 and 50 %, respectively. The SME culture medium was changed every 48h and used for determination by ELISA of PDGF-BB, TGF-ß1, TNF-α, IL-4, IL-1ra and HA. These molecules were also determined in synovial fluid from the horses. RESULTS: Both the 25 and 50 % PRG supernatants produced a molecular profile in the culture media unlike that of the SME challenged with LPS only. They presented GF, cytokine and HA concentrations very near to the concentrations of these molecules in normal synovial fluid when compared with the SME control groups (either with LPS or without LPS). However, in comparison with the rest of the SME treated groups, the 25 % L-PRG produced the most IL-1ra, and the 50 % P-PRG induced the sustained production of IL-4 and HA. CONCLUSIONS: These in vitro findings suggest that anabolic and anti-inflammatory joint responses depend on the leukocyte and platelet concentration of the PRP preparation and on the volume of this substance injected. Moreover, it is possible, that leukoreduced PRP preparations are more effective for the medical treatment of patients with OA and inflammatory synovitis.
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Plaquetas/metabolismo , Citocinas/metabolismo , Ácido Hialurónico/metabolismo , Mediadores de Inflamación/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Lipopolisacáridos/farmacología , Plasma Rico en Plaquetas/metabolismo , Membrana Sinovial/efectos de los fármacos , Animales , Plaquetas/inmunología , Fraccionamiento Celular , Medios de Cultivo/metabolismo , Geles , Caballos , Leucocitos/efectos de los fármacos , Leucocitos/inmunología , Leucocitos/metabolismo , Plasma Rico en Plaquetas/inmunología , Líquido Sinovial/inmunología , Líquido Sinovial/metabolismo , Membrana Sinovial/inmunología , Membrana Sinovial/metabolismo , Factores de Tiempo , Técnicas de Cultivo de TejidosRESUMEN
Seaweed lectins have been widely investigated as anti-nociceptive and anti-inflammatory agents. This study analyzed the anti-nociceptive and anti-inflammatory responses of a lectin from the green seaweed Caulerpa cupressoides (CcL) on zymosan-induced arthritis of the rat temporomandibular joint (TMJ). Rats received i.v. CcL 30 min prior to injection of zymosan (2mg/art.) or 0.9% saline into the left TMJ. Mechanical hyper-nociception was measured by the electronic von Frey method at baseline and 4h after zymosan injection. Animals were euthanized 6h after zymosan injection and the synovial fluid was collected for leukocyte counting and myeloperoxidase activity assessment. Other animals were treated with ZnPP-IX (3mg/kg; s.c.), a specific heme oxygenase-1 pathway inhibitor, and naloxone (10 µg/art.), a nonselective opioid receptor antagonist. TMJ tissues were excised to perform histopathological and immunohistochemistry analyses. CcL (0.1, 1 or 10mg/kg) significantly reduced zymosan-induced hyper-nociception (81, 83 and 89.5%, respectively) and inhibited the leukocyte influx (77.3, 80.7 and 98.5%, respectively) compared with the zymosan-only group, as confirmed by myeloperoxidase activity; however, treatment with naloxone or ZnPP-IX did not revert the effects of CcL (10mg/kg), suggesting that the naloxone-sensitive opioid and heme oxygenase-1 pathways are not involved. CcL also reduced the leukocyte influx and the expression of IL-1ß and TNF-α in the TMJ, based on histopathological and immunohistochemistry analyses, respectively. Therefore, CcL reduces TMJ hyper-nociception and inflammation with a mechanism that is partially dependent on TNF-α and IL-1ß inhibition. CcL reveals a potentially valuable alternative tool for future studies of TMJ disorders.
Asunto(s)
Artritis Experimental/tratamiento farmacológico , Caulerpa/inmunología , Inflamación/tratamiento farmacológico , Leucocitos/efectos de los fármacos , Lectinas de Plantas/administración & dosificación , Membrana Sinovial/efectos de los fármacos , Articulación Temporomandibular/efectos de los fármacos , Administración Intravenosa , Animales , Artritis Experimental/inducido químicamente , Movimiento Celular/efectos de los fármacos , Inflamación/inducido químicamente , Interleucina-1beta/metabolismo , Leucocitos/inmunología , Masculino , Nocicepción/efectos de los fármacos , Peroxidasa/metabolismo , Lectinas de Plantas/efectos adversos , Ratas , Ratas Wistar , Membrana Sinovial/inmunología , Articulación Temporomandibular/patología , Factor de Necrosis Tumoral alfa/metabolismo , Zimosan/administración & dosificaciónRESUMEN
A Leishmaniose Visceral Canina (LVC) é uma zoonose sistêmica grave, causada pelo protozoário intracelular obrigatório Leishmania infantum. Os cães são considerados os principais reservatórios da doença e podem desenvolver desde a infecção assintomática, a uma variedade de sinais clínicos, tais como linfadenopatia periférica, perda de peso, letargia, lesões ulcerativas de pele e febre. Alterações oculares, ortopédicas e neurológicas também podem ser observadas associadas à LVC, tais como ceratoconjuntivite, uveíte, convulsões, alterações comportamentais, artralgia e claudicação. Tal fato inclui a doença no diagnóstico diferencial das afecções oftálmicas, osteoarticulares e neurológicas, principalmente em áreas consideradas endêmicas para a infecção. O objetivo deste trabalho foi realizar o diagnóstico da leishmaniose visceral canina utilizando amostras de conjuntiva, humor aquoso, líquido sinovial, membrana sinovial e líquido cefalorraquidiano (LCR). O teste ELISA foi aplicado em amostras de LCR, líquido sinovial e humor aquoso; e avaliações histológica e imunohistoquímica foram realizadas na conjuntiva e na membrana sinovial. As amostras foram coletadas de 20 cães sorologicamente positivos para leishmaniose visceral. Através do Ensaio Imunoenzimático foi possível detectar anticorpos IgG anti-Leishmania em uma das amostras de LCR e de humor aquoso, e em seis amostras de líquido sinovial. A imunohistoquímica apresentou marcação positiva de formas amastigotas de L. infantum em cinco membranas sinoviais e cinco conjuntivas estudadas. Diante dos resultados, conclui-se que formas amastigotas de Leishmania sp. são encontradas parasitando a membrana sinovial e a conjuntiva de cães sorologicamente positivos para a doença, bem como anticorpos IgG específicos no humor aquoso, no LCR e no líquido sinovial desses animais.
Canine Visceral Leishmaniasis (CVL) is a severe systemic zoonosos caused by the obligate intracellular protozoan Leishmania infantum. The dogs are the major reservoir of the disease and may develop since asymptomatic infection to a variety of clinical signs, such as peripheral lymphadenopathy, weight loss, lethargy, ulcerative lesions of the skin and fever. Ocular, orthopedic and neurological disorders can also be seen associated with CVL, such as keratitis, seizures, behavioral changes, joint pain and lameness. This fact includes the disease in the differential diagnosis of ophthalmic disorders, osteoarticular and nervous system, especially in endemic areas to infection. The aim of this study was the diagnosis of canine visceral leishmaniasis using samples of conjunctiva, aqueous humor, synovial fluid, synovial membrane and cerebrospinal fluid (CSF). The ELISA test was applied to samples of CSF, synovial fluid and aqueous humor, and histological and immunohistochemical evaluations were performed in the conjunctiva and in the synovial membrane of these animals. The samples were collected from 20 dogs serologically positive for visceral leishmaniasis. Through the immunoenzymatic assay could detect anti-Leishmania IgG antibodies in a sample of CSF and aqueous humor, and six samples of synovial fluid. Immunohistochemistry showed positive staining of amastigotes of L. infantum five synovial membranes and connective five studied. From the results, we conclude that amastigotes of Leishmania sp. are found parasitizing the synovial membrane and the conjunctiva of dogs serologically positive for the disease, as well as specific IgG antibodies in aqueous humor, CSF and synovial fluid of these animals.
Asunto(s)
Animales , Perros , Humor Acuoso/inmunología , Leishmaniasis Visceral/diagnóstico , Líquido Cefalorraquídeo/inmunología , Líquido Sinovial/inmunología , Membrana Sinovial/inmunología , Conjuntiva/inmunología , Diagnóstico Diferencial , Ensayo de Inmunoadsorción Enzimática/veterinariaRESUMEN
OBJECTIVE: Deposition of monosodium urate monohydrate (MSU) crystals in the joints promotes an intense inflammatory response and joint dysfunction. This study evaluated the role of the NLRP3 inflammasome and 5-lipoxygenase (5-LOX)-derived leukotriene B(4) (LTB(4) ) in driving tissue inflammation and hypernociception in a murine model of gout. METHODS: Gout was induced by injecting MSU crystals into the joints of mice. Wild-type mice and mice deficient in NLRP3, ASC, caspase 1, interleukin-1ß (IL-1ß), IL-1 receptor type I (IL-1RI), IL-18R, myeloid differentiation factor 88 (MyD88), or 5-LOX were used. Evaluations were performed to assess neutrophil influx, LTB(4) activity, cytokine (IL-1ß, CXCL1) production (by enzyme-linked immunosorbent assay), synovial microvasculature cell adhesion (by intravital microscopy), and hypernociception. Cleaved caspase 1 and production of reactive oxygen species (ROS) were analyzed in macrophages by Western blotting and fluorometric assay, respectively. RESULTS: Injection of MSU crystals into the knee joints of mice induced neutrophil influx and neutrophil-dependent hypernociception. MSU crystal-induced neutrophil influx was CXCR2-dependent and relied on the induction of CXCL1 in an NLRP3/ASC/caspase 1/IL-1ß/MyD88-dependent manner. LTB(4) was produced rapidly after injection of MSU crystals, and this was necessary for caspase 1-dependent IL-1ß production and consequent release of CXCR2-acting chemokines in vivo. In vitro, macrophages produced LTB(4) after MSU crystal injection, and LTB(4) was relevant in the MSU crystal-induced maturation of IL-1ß. Mechanistically, LTB(4) drove MSU crystal-induced production of ROS and ROS-dependent activation of the NLRP3 inflammasome. CONCLUSION: These results reveal the role of the NLRP3 inflammasome in mediating MSU crystal-induced inflammation and dysfunction of the joints, and highlight a previously unrecognized role of LTB(4) in driving NLRP3 inflammasome activation in response to MSU crystals, both in vitro and in vivo.
Asunto(s)
Proteínas Portadoras/metabolismo , Gota/metabolismo , Hiperalgesia/metabolismo , Inflamasomas/metabolismo , Leucotrieno B4/metabolismo , Infiltración Neutrófila/fisiología , Neutrófilos/metabolismo , Animales , Caspasa 1/metabolismo , Citocinas/metabolismo , Gota/inducido químicamente , Gota/inmunología , Hiperalgesia/inmunología , Inflamasomas/inmunología , Inflamación/inmunología , Inflamación/metabolismo , Interleucina-1beta/metabolismo , Leucotrieno B4/inmunología , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones , Ratones Noqueados , Proteína con Dominio Pirina 3 de la Familia NLR , Neutrófilos/inmunología , Especies Reactivas de Oxígeno/metabolismo , Membrana Sinovial/inmunología , Membrana Sinovial/metabolismo , Ácido Úrico/farmacologíaRESUMEN
Arthritis is one of the most common complications of human brucellosis, but its pathogenic mechanisms have not been elucidated. Fibroblast-like synoviocytes (FLS) are known to be central mediators of joint damage in inflammatory arthritides through the production of matrix metalloproteinases (MMPs) that degrade collagen and of cytokines and chemokines that mediate the recruitment and activation of leukocytes. In this study we show that Brucella abortus infects and replicates in human FLS (SW982 cell line) in vitro and that infection results in the production of MMP-2 and proinflammatory mediators (interleukin-6 [IL-6], IL-8, monocyte chemotactic protein 1 [MCP-1], and granulocyte-macrophage colony-stimulating factor [GM-CSF]). Culture supernatants from Brucella-infected FLS induced the migration of monocytes and neutrophils in vitro and also induced these cells to secrete MMP-9 in a GM-CSF- and IL-6-dependent fashion, respectively. Reciprocally, culture supernatants from Brucella-infected monocytes and neutrophils induced FLS to produce MMP-2 in a tumor necrosis factor alpha (TNF-α)-dependent fashion. The secretion of proinflammatory mediators and MMP-2 by FLS did not depend on bacterial viability, since it was also induced by heat-killed B. abortus (HKBA) and by a model Brucella lipoprotein (L-Omp19). These responses were mediated by the recognition of B. abortus antigens through Toll-like receptor 2. The intra-articular injection of HKBA or L-Omp19 into the knee joint of mice resulted in the local induction of the proinflammatory mediators MMP-2 and MMP-9 and in the generation of a mixed inflammatory infiltrate. These results suggest that FLS, and phagocytes recruited by them to the infection focus, may be involved in joint damage during brucellar arthritis through the production of MMPs and proinflammatory mediators.
Asunto(s)
Artritis Infecciosa/inmunología , Brucella abortus/inmunología , Brucelosis/inmunología , Articulaciones/microbiología , Articulaciones/patología , Metaloproteinasas de la Matriz/biosíntesis , Membrana Sinovial/inmunología , Animales , Antígenos Bacterianos/inmunología , Artritis Infecciosa/enzimología , Artritis Infecciosa/microbiología , Artritis Infecciosa/patología , Proteínas de la Membrana Bacteriana Externa/inmunología , Brucella abortus/crecimiento & desarrollo , Brucella abortus/patogenicidad , Brucelosis/enzimología , Brucelosis/microbiología , Brucelosis/patología , Línea Celular , Movimiento Celular/efectos de los fármacos , Quimiocinas/biosíntesis , Medios de Cultivo Condicionados , Citocinas/biosíntesis , Citocinas/metabolismo , Inducción Enzimática , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Articulación de la Rodilla/microbiología , Lipoproteínas/inmunología , Activación de Linfocitos , Metaloproteinasas de la Matriz/metabolismo , Ratones , Ratones Endogámicos BALB C , Monocitos/fisiología , Neutrófilos/fisiología , Membrana Sinovial/citología , Membrana Sinovial/microbiología , Receptor Toll-Like 2/metabolismoRESUMEN
OBJECTIVE: Neutrophil accumulation contributes to the pathogenesis of rheumatoid arthritis. This study was undertaken to examine the ability of H2O2 to influence neutrophilic inflammation in a model of antigen-induced arthritis (AIA) in mice. METHODS: AIA was induced by administration of antigen into the knee joints of previously immunized mice. Neutrophil accumulation was measured by counting neutrophils in the synovial cavity and assaying myeloperoxidase activity in the tissue surrounding the mouse knee joint. Apoptosis was determined by morphologic and molecular techniques. The role of H2O2 was studied using mice that do not produce reactive oxygen species (gp91phox-/- mice) and drugs that enhance the generation or enhance the degradation of H2O2. RESULTS: Antigen challenge of immunized mice induced neutrophil accumulation that peaked at 12-24 hours after challenge. H2O2 production peaked at 24 hours, after which time, the inflammation resolved. Neutrophil recruitment was similar in wild-type and gp91phox-/- mice, but there was delayed resolution in gp91phox-/- mice or after administration of catalase. In contrast, administration of H2O2 or superoxide dismutase (SOD) resolved neutrophilic inflammation. The resolution of inflammation induced by SOD or H2O2 was accompanied by an increase in the number of apoptotic neutrophils. Apoptosis was associated with an increase in Bax and caspase 3 cleavage and was secondary to phosphatidylinositol 3-kinase (PI3K)/Akt activation. CONCLUSION: Our findings indicate that levels of H2O2 increase during neutrophil influx and are necessary for the natural resolution of neutrophilic inflammation. Mechanistically, enhanced levels of H2O2 (endogenous or exogenous) inhibit p-Akt/NF-κB and induce apoptosis of migrated neutrophils. Modulation of H2O2 production may represent a novel strategy for controlling neutrophilic inflammation in the joints.
Asunto(s)
Artritis Experimental/inmunología , Peróxido de Hidrógeno/metabolismo , Neutrófilos/inmunología , Membrana Sinovial/metabolismo , Animales , Artritis Experimental/metabolismo , Artritis Experimental/patología , Artritis Reumatoide/inmunología , Artritis Reumatoide/metabolismo , Artritis Reumatoide/patología , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/patología , Articulaciones/inmunología , Articulaciones/metabolismo , Articulaciones/patología , Masculino , Ratones , Neutrófilos/metabolismo , Neutrófilos/patología , Membrana Sinovial/inmunología , Membrana Sinovial/patologíaRESUMEN
Mast cells are involved in immune disorders so that many of the proinflammatory and tissue destructive mediators produced by these cells have been implicated in the pathogenesis of rheumatoid arthritis. This scenario prompted us to investigate the correlation between mast cell degranulation and neutrophil influx within the digits and knees joints of arthritic mice assessing what could be the functional role(s) of joint mast cells in the response to collagen immunization. DBA/1J mice were submitted to collagen-induced arthritis and disease was assessed on day 21, 32 and 42 post-immunization. Pharmacological treatment with the glucocorticoid prednisolone, commonly used in the clinic, and nedocromil, a mast cell stabilizer, was performed from day 21 to 30. Arthritis develop after immunization, gradually increased up to day 42. Neutrophil infiltration peaked on day 32 and 21, in the digits and knees, respectively, showing an unequal pattern of recruitment between these tissues. This difference emerged for mast cells: they peaked in the digits on day 21, but a higher degree of degranulation could be measured in the knee joints. Uneven modulation of arthritis occurred after treatment of mice with prednisolone or nedocromil. Neutrophils migration to the tissue was reduced after both therapies, but only prednisolone augmented mast cell migration to the joints. Nedocromil exerted inhibitory properties both on mast cell proliferation and migration, more effectively on the digit joints. Thus, collagen induced an inflammatory process characterized by tissue mast cells activation and degranulation, suggesting a potential driving force in propagating inflammatory circuits yielding recruitment of neutrophils. However, the different degree of affected joint involvement suggests a time-related implication of digits and knees during collagen-induced arthritis development. These results provide evidence for local alterations whereby mast cells contribute to the initiation of inflammatory arthritis and may be targeted in intervention strategies.
Asunto(s)
Artritis Experimental/inmunología , Mastocitos/inmunología , Infiltración Neutrófila/inmunología , Neutrófilos/inmunología , Animales , Antiinflamatorios/farmacología , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/patología , Miembro Anterior/efectos de los fármacos , Miembro Anterior/patología , Glucocorticoides/farmacología , Miembro Posterior/efectos de los fármacos , Miembro Posterior/patología , Articulación de la Rodilla/efectos de los fármacos , Articulación de la Rodilla/inmunología , Articulación de la Rodilla/patología , Recuento de Leucocitos , Mastocitos/efectos de los fármacos , Mastocitos/patología , Ratones , Ratones Endogámicos DBA , Nedocromil/farmacología , Infiltración Neutrófila/efectos de los fármacos , Neutrófilos/efectos de los fármacos , Neutrófilos/patología , Prednisolona/farmacología , Membrana Sinovial/efectos de los fármacos , Membrana Sinovial/inmunología , Membrana Sinovial/patología , Factores de TiempoRESUMEN
Autoimmune diseases result from an interplay between susceptibility genes and environmental factors. These interacting etiopathogenetic components converge in a critical step preceding disease, the loss of tolerance to self. In this review, we examine the evidences linking tobacco smoking with the initiation and perpetuation of inflammation affecting both the synovial membrane and the endothelial lining in patients with rheumatoid arthritis. This disease is a compelling argument for the decisive role of environment in the triggering of a human autoimmune disease in genetically prone individuals.
Asunto(s)
Artritis Reumatoide/etiología , Aterosclerosis/etiología , Citrulina/metabolismo , Endotelio Vascular/metabolismo , Procesamiento Proteico-Postraduccional , Fumar/efectos adversos , Membrana Sinovial/metabolismo , Animales , Artritis Reumatoide/genética , Artritis Reumatoide/inmunología , Artritis Reumatoide/metabolismo , Aterosclerosis/genética , Aterosclerosis/inmunología , Aterosclerosis/metabolismo , Autoinmunidad , Endotelio Vascular/inmunología , Predisposición Genética a la Enfermedad , Humanos , Medición de Riesgo , Factores de Riesgo , Autotolerancia , Membrana Sinovial/inmunologíaRESUMEN
OBJECTIVES: Interleukin 33 (IL-33) is a new member of the IL-1 family of cytokines which signals via its receptor, ST2 (IL-33R), and has an important role in Th2 and mast cell responses. This study shows that IL-33 orchestrates neutrophil migration in arthritis. METHODS AND RESULTS: Methylated bovine serum albumin (mBSA) challenge in the knee joint of mBSA-immunised mice induced local neutrophil migration accompanied by increased IL-33R and IL-33 mRNA expression. Cell migration was inhibited by systemic and local treatments with soluble (s)IL-33R, an IL-33 decoy receptor, and was not evident in IL-33R-deficient mice. IL-33 injection also induced IL-33R-dependent neutrophil migration. Antigen- and IL-33-induced neutrophil migration in the joint was dependent on CXCL1, CCL3, tumour necrosis factor alpha (TNFalpha) and IL-1beta synthesis. Synovial tissue, macrophages and activated neutrophils expressed IL-33R. IL-33 induces neutrophil migration by activating macrophages to produce chemokines and cytokines and by directly acting on neutrophils. Importantly, neutrophils from patients with rheumatoid arthritis successfully treated with anti-TNFalpha antibody (infliximab) expressed significantly lower levels of IL-33R than patients treated with methotrexate alone. Only neutrophils from patients treated with methotrexate alone or from normal donors stimulated with TNFalpha responded to IL-33 in chemotaxis. CONCLUSIONS: These results suggest that suppression of IL-33R expression in neutrophils, preventing IL-33-induced neutrophil migration, may be an important mechanism of anti-TNFalpha therapy of inflammation.
Asunto(s)
Antirreumáticos/farmacología , Artritis Reumatoide/inmunología , Interleucinas/inmunología , Infiltración Neutrófila/inmunología , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Animales , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/inmunología , Artritis Reumatoide/tratamiento farmacológico , Factores Quimiotácticos/inmunología , Quimiotaxis de Leucocito/inmunología , Citocinas/inmunología , Regulación de la Expresión Génica/inmunología , Humanos , Proteína 1 Similar al Receptor de Interleucina-1 , Interleucina-33 , Interleucinas/biosíntesis , Interleucinas/genética , Activación de Macrófagos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , ARN Mensajero/genética , Receptores de Interleucina , Membrana Sinovial/inmunologíaRESUMEN
The Wnt signaling pathways play a key role in cell renewal, and there are two such pathways. In patients with rheumatoid arthritis (RA), the synovial membrane expresses genes such as Wnt and Fz at higher levels than those observed in patients without RA. The Wnt proteins are glycoproteins that bind to receptors of the Fz family on the cell surface. The Wnt/Fz complex controls tissue formation during embryogenesis, as well as throughout the process of limb development and joint formation. Recent studies have suggested that this signaling pathway plays a role in the pathophysiology of RA. Greater knowledge of the role of the Wnt signaling pathway in RA could improve understanding of the differences in RA clinical presentation and prognosis. Further studies should also focus on Wnt family members as molecular targets in the treatment of RA.