Evaluation of glucosamine and snail mucin on the progression of experimental knee osteoarthritis in dogs / Evaluación de la glucosamina y mucina de caracol en la progresión de la osteoartritis experimental de rodilla en perros

This study evaluated the effect of oral glucosamine and intramuscular injection (IM) of snail mucin on the progression of experimental osteoarthritis (OA) in dogs. Twenty adult mongrels with mean body weight (12.4±1.8 kg) were used. Experimental OA was induced surgically using the groove model. The dogs were randomly divided into three groups following radiographic evidence of OA. Group one (control) comprised of ten dogs treated with normal saline twice weekly for four weeks following OA. Group two comprised of five dogs treated with 10mg/kg of oral glucosamine daily for four weeks. Group three comprised of five dogs treated with 5mg/kg intramuscular injection of 5% solution of snail mucin twice weekly for four weeks. Blood was obtained from the cephalic vein before surgical arthrotomy, after surgical arthrotomy, immediately after radiographic confirmation of OA (Week 0) and at two weeks interval up to 4 weeks of treatment. Efficacy of the drugs was assessed by changes in plasma IL-6 and MMP-3, while safety was determined using the changes in packed cell volume (PCV), total white blood cell counts (WBC) and observable adverse reactions associated with the administration of the drugs. In this study, the PCV and WBC did not differ significantly (P> 0.05) from the control group. Plasma IL-6 and MMP-3 were significantly (P< 0.05) lower both in glucosamine-treated and snail mucin-treated dogs up to week 4 of treatment when compared with the control group. However, there were no significant (P > 0.05) differences in IL-6 and MMP-3 between the two treatment groups. In addition, painful swelling at the site of injection was observed in dogs treated with snail mucin, while no adverse reaction was observed in dogs treated with oral glucosamine. It was therefore concluded that both oral glucosamine and IM injection of snail mucin comparably modified the progression of OA. However, owing to the adverse reaction noted with IM injection of snail mucin, further study is required to determine the most appropriate route of administration.

Se evaluaron los efectos de la glucosamina oral y la inyección intramuscular (IM) de mucina de caracol en la progresión de la osteoartritis (OA) experimental en perros. Fueron utilizados 20 perros mestizos adultos con un peso medio de 12,4±1,8 kg. La OA experimental se indujo quirúrgicamente mediante el modelo de ranura. Los animales se dividieron aleatoriamente en tres grupos después de la evidencia radiográfica de OA. El grupo 1 (control, 10 perros) fue tratado con una solución salina normal dos veces por semana durante cuatro semanas. El grupo 2 (5 perros) fue tratado con 10 mg/kg de glucosamina oral al día por cuatro semanas, y el grupo 3 (5 perros) fue tratado con 5 mg/kg IM de una solución de mucina de caracol al 5% dos veces por semana durante cuatro semanas. Se obtuvieron muestras de sangre desde la vena cefálica previo a la artrotomía quirúrgica, después de la artrotomía e inmediatamente después de la confirmación radiográfica de OA (semana 0), y en el intervalo de dos semanas hasta cuatro semanas de tratamiento. La eficacia de los fármacos se evaluó por los cambios plasmáticos de IL-6 y MMP-3, mientras que la seguridad, se determinó por los cambios en el volumen del hematocrito (VH), el recuento total de glóbulos blancos (RGB), y la observación de reacciones adversas asociadas a la administración de fármacos. El VH y RGB no difirieron significativamente (P>0,05) en el grupo control. Los niveles de IL-6 y MMP-3 plasmática fueron significativamente más bajas (P<0,05) en los perros tratados con glucosamina y mucina de caracol hasta 4 semanas, en comparación con el grupo control. Sin embargo, no hubo diferencias significativas (P>0,05) en la IL-6 y MMP-3 entre los dos grupos de tratamiento. Además, se observó un edema doloroso en el sitio de inyección de los perros tratados con mucina de caracol. En los perros tratados con glucosamina oral no se observó reacción adversa. Se concluye que tanto la glucosamina oral y la inyección IM de mucina de caracol modifican comparablemente la progresión de OA. Sin embargo, debido a la reacción adversa observada con la inyección IM de mucina caracol, se necesitan estudios adicionales para determinar la vía de administración más adecuada.

Over-expression of extracellular superoxide dismutase in mouse synovial tissue attenuates the inflammatory arthritis

Oxidative stress such as reactive oxygen species (ROS) within the inflamed joint have been indicated as being involved as inflammatory mediators in the induction of arthritis. Correlations between extracellular-superoxide dismutase (EC-SOD) and inflammatory arthritis have been shown in several animal models of RA. However, there is a question whether the over-expression of EC-SOD on arthritic joint also could suppress the progression of disease or not. In the present study, the effect on the synovial tissue of experimental arthritis was investigated using EC-SOD over-expressing transgenic mice. The over-expression of EC-SOD in joint tissue was confirmed by RT-PCR and immunohistochemistry. The degree of the inflammation in EC-SOD transgenic mice was suppressed in the collagen-induced arthritis model. In a cytokine assay, the production of pro-inflammatory cytokines such as, IL-1beta, TNFalpha, and matrix metalloproteinases (MMPs) was decreased in fibroblast-like synoviocyte (FLS) but not in peripheral blood. Histological examination also showed repressed cartilage destruction and bone in EC-SOD transgenic mice. In conclusion, these data suggest that the over-expression of EC-SOD in FLS contributes to the activation of FLS and protection from joint destruction by depressing the production of the pro-inflammatory cytokines and MMPs. These results provide EC-SOD transgenic mice with a useful animal model for inflammatory arthritis research.

Doxiciclina em pacientes com linfangioleiomiomatose: segurança e eficácia no bloqueio de metaloproteinases / Doxycycline use in patients with lymphangioleiomyomatosis: safety and efficacy in metalloproteinase blockade

OBJETIVO: A linfangioleiomiomatose (LAM) é caracterizada pela presença de cistos pulmonares, cuja formação está associada à hiperreatividade de metaloproteinases de matriz (MMP), principalmente MMP-2 e MMP-9. Objetivamos comparar os níveis dessas MMPs entre pacientes com LAM e controles saudáveis, assim como avaliar, nas pacientes com LAM, a segurança e a eficácia do tratamento com doxiciclina, um potente inibidor de MMPs. MÉTODOS: Estudo clínico prospectivo no qual as pacientes com LAM receberam doxiciclina (100 mg/dia) por seis meses, coletando-se amostras de urina e sangue para a dosagem de MMP-2 e MMP-9 antes e ao final do período. Foram ainda obtidas amostras de 10 mulheres saudáveis. RESULTADOS: De 41 pacientes com LAM que iniciaram o tratamento, 34 concluíram o protocolo. Os níveis de MMP-9 sérica e urinária foram significativamente inferiores no grupo controle (p < 0,0001). Comparando-se os valores antes e após o tratamento, a mediana do nível sérico da MMP-9 reduziu de 919 ng/mL para 871 ng/mL (p = 0,05), enquanto a mediana da dosagem urinária de MMP-9 diminui de 11.558 pg/mL para 7.315 pg/mL (p = 0,10). A mediana da MMP-2 sérica apresentou um decréscimo significativo após o tratamento (p = 0,04). Não foram detectados níveis de MMP-2 urinária. Epigastralgia, náuseas e diarreia foram os efeitos adversos mais prevalentes, e geralmente autolimitados. Apenas 1 paciente interrompeu o tratamento devido a efeitos colaterais. CONCLUSÕES: Pela primeira vez, conseguiu-se evidenciar em pacientes com LAM a redução dos níveis séricos e urinários de MMPs após o uso de doxiciclina, que se mostrou uma medicação segura, com efeitos colaterais leves e toleráveis.

OBJECTIVE: Lymphangioleiomyomatosis (LAM) is characterized by lung cysts, whose development is associated with matrix metalloproteinase (MMP) hyperactivity, principally that of MMP-2 and MMP-9. Our objective was to compare LAM patients and controls in terms of the levels of these MMPs, as well as to determine the safety and efficacy of treatment with doxycycline, a potent MMP inhibitor. METHODS: Prospective clinical study involving female LAM patients who received doxycycline (100 mg/day) for six months. Urine and blood samples were collected for the quantification of MMP-2 and MMP-9 before and after the treatment period. Samples from 10 healthy women were also collected. RESULTS:Of the 41 LAM patients who started the treatment, 34 completed the protocol. Serum and urinary MMP-9 levels were significantly lower in the controls than in the LAM patients (p < 0.0001). Comparing pre- and post-treatment values, we found that the median level of MMP-9 in serum decreased from 919 ng/mL to 871 ng/mL (p = 0.05), whereas that of MMP-9 in urine decreased from 11,558 pg/mL to 7,315 pg/mL (p = 0.10). After treatment, the median level of MMP-2 in serum was significantly lower (p = 0.04) and urinary MMP-2 levels were undetectable. Nausea, diarrhea, and epigastric pain were the most prevalent adverse affects and were often self-limiting. There was only one case in which the patient discontinued the treatment because of side effects. CONCLUSIONS: We have demonstrated, for the first time, a decrease in serum and urine levels of MMPs in LAM patients treated with doxycycline, which proved to be a safe medication, with mild and well-tolerated side effects.

Role of pro-inflammatory cytokines, and matrix metalloprotinases in the pathogenesis of heart failure

Extracellular matrix remodeling is thought to play an important role in the progression of heart failure [HF]. Matrix metalloproteinases [MMPs] and tissue inhibitors of metalloproteinases [TIMPs] are matrix-degrading enzymes that have been demonstrated to influence left ventricular properties and serve as targets of potential anti-remodeling agents. It has been reported that MMPs concentration and activity are upregulated in the failing human heart. However, there are few reports describing the role of elevated level of circulating MMPs in severe congestive heart failure [CHF] patients. This study examined whether circulating MMPs are also related to the pathogenesis of CHF. The study involved 50 patients with severe CHF and 20 apparently healthy subjects, with matched age and sex were selected as a control group. Two Dimensional echocardiography, Doppler and colour flow mapping were done for the patients. Left ventricular dimensions [LVD] and cardiac size were measured. LV mass [LVM] was calculated from Interventricular septum [IVS], Left ventricular end diastolic dimensions [LVEDd], Left ventricular wall thickness [LVWT] and Left ventricular end systolic dimension [LVESd]. The serum levels of MMP-2, MMP-9 and TIMP-l as well as IL-18, TNF-alpha as pro-inflammatory cytokines were measured in patients with CHF and control subjects. The serum levels of MMP-2, MMP-9, TIMP-1, IL-18 and TNF-alpha were significantly higher in the CHF patients than control group. Moreover, MMP-2, MMP-9 and TIMP-1 serum levels were positively correlated with the levels of IL-8, TNF-alpha, cholesterol, triglyceride and CRP. Furthermore, MMP-2, MMP-9 andTIMP-1 were positively coffelated with LVM, LVED[d] and LVWT. We conclude that, the increasing serum levels of MMP-2, MMP-9 and TIMP-l were associated with increased LV diastolic dimensions and increased wall thickness in patients with CHF. These observations indicate that MMPs and TIMP- I serum levels may be markers for cardiac extracellular matrix degradation, a process involved in LV remodelling. These findings may open a new avenue for therapy that ameliorating heart failure especially high risk patients

Clinical significance of levels of matrix metalloproteinases MMP-3 and MMP-1 in sera and synovial fluids in patients with rheumatoid arthritis and osteoarthritis

To investigate whether serum levels of matrix metalloproteinases [MMP-3, stromelysin] and [MMP-1, collagenase] are specifically elevated in joint disease as rheumatoid arthritis [RA] compared to osteoarthritis [OA] and to assess how these markers reflect the clinical activity of RA compared to circulating cytokine as tumor necrosis factor-alpha [TNF-alpha] as well as established variables as [C-reactive protein [CRP] and erythrocyte sedimentation rate [ESR]]. This study included 22 patients with RA, 10 patients with OA and 10 healthy control subjects matched for age and sex. Patients with superimposed infection were excluded. Serum levels of MMP-3, MMP-1, TNF-alpha and CRP were assayed. Synovial fluid [SF] levels of MMP-3 and MMP-1 were also assayed. Serum levels of TNF-alpha and CRP in RA patients were significantly higher than normal subjects. Serum MMP-1 was significantly elevated in patients with RA and OA, compared to healthy controls but there were no significant differences between patients with RA and those with OA. Serum MMP-3 levels did not differ between OA patients and normal sera. However, RA patients displayed significantly elevated levels of this enzyme, compared to OA and control sera. Levels of MMP-3 and MMP-1 in the SF of RA patients were significantly higher than in OA fluids. CRP, ESR, TNF-alpha and MMP-3 correlated significantly with the swollen joint count, The strongest positive correlations existed between rheumatoid activity as assessed by the levels of CRP and circulating levels of MMP-3. Similar correlations between TNF-alpfa concentration and CRP, MMP-1 and MMP-3 were observed in RA patients. Serum levels of MMP-3 correlated significantly with serum concentrations of MMP-l in RA patients [r = 0.487, p < 0.05]. There was close correlation between serum and SF concentrations of MMP-3 in RA patients [r = 0.619, p < 0.01]. In the same patients there was highly significant correlation between SF concentrations of MMP-3 and MMP-l [r= 0.732, p <0.001]. Our data suggested that elevated MMP-3 levels reflected disease activity of RA better than cytokine levels. MMP-3 may be seen as a constitutive marker of the pathological process underlying joint tissue degradation in RA

Serum levels of matrix metalloproteinases in type II diabetes mellitus

Hyperglycemia is associated with excessive non-enzymatic glycosylation of the glomerular matrix thus contributing to the pathogenesis of diabetic nephropathy. Matrix metallopoteinases [mmp[s]] are responsible for matrix degradation. This study was conducted to unvestigate the potential diagnostic value of serum levels of two of the known mmp[s]], namely mmp-1 and mmp-2 in type ii diabetes and their relationship to vascular complications. This study was concluted on 30 female type ii niddm diabetes mellitus [dm] patients recruited from the diabetic clinic, kasr al aini hospital, cairo university. A control group including 10 apparently healthy age matched females with no family history of diabetes was also studied. All patients were subjected to full history taking and full clinical examination. Laboratory investigations included fasting blood sugar, liver function tests, kidney functions, lipid profiles, glycosylated hemoglobin, according to which, patients were divided into 2 groups, fairly and poorly controlled and microalbuminuria, according to which, patents were divided into with and without diabetic nephropathy. Mmp-1 and mmp-2 were measured using an elisa technique. The ages of the patients under study ranged from 40-55, with a mean of 48.83 +/- 4.36. The duration of diabetes ranged between 1-20 years with a mean of 110-170 with a mean of 129.33 +/- 18.56 and their diastolic blood pressure ranged from 60-110 with a mean of 79.33 +/- 8.68. The mean serum mmp-1 level was significantly higher 16.63 +/- 9.13 and 5.48 +/- 2.37 respectively [p= < 0.01]. Mmp-1 was significantly higher in patients with nephropathy compared to controls, however it was significantly decreased than its mean value in patients without nephropathy 14.3 +/- 8.55 and 21.3 +/- 8.83 respectively [p= < 0.05]. The mean serum mmp-1 level was significantly higher in the poorly controlled and fairly controlled diabetics [16.71 ng/ml +/- 9.23 and 16.52 ng/ml +/- 9.37] as compared to the control group [5.48 ng/l +/- 2.37] [p= < 0.01]. The mean serum mmp-1 was significantly higher in the hypertensive group compared with control group 13.49 +/- 8.52 and 5.48 +/- 2.37 respectively [p= < 0.01]. On the other mean serum mmp-2 level was significantly reduced in hypertensive cases compared to the non-hypertensive ones 197.4 +/- 56.76 and 24.741 +/- 57.53 respectively [p= < 0.05]. The mean mmp-2 levels also showed a positive correlation with the diastolic blood pressure [r=0.756, p< 0.01]. Mmp-1 serum levels may be implicated in the complications of DM

Clinical Utility of Serum Vascular Endothelial Growth factor and Matrix Metalloprpteinase - 9 in breast Cancer patients

This study included 36 females [aged 48.9 +/- 9.5 years] with breast cancer; 28 had localized disease with or without locoregional lymphatic spread [localized subgroup] and 8 with distant visceral and/or bone metastasis [distant metastasis subgroup]. They were staged according to the American Joint Committee on Cancer and graded according to the histological examination of the studied biopsies. The results were compared with those of 15 age-matched females suffering from benign mastopathies and 19 healthy age-matched females. Serum aspartate aminotransferase [AST], alanine aminotransferase [ALT], alkaline phosphatase [ALP], total calcium and phosphorus were assayed on Synchron Cx-7 autoanalyzer. Moreover, CA15-3, VEGF and MMP-9 were measured using an enzyme linked immunosorbent assay. In conclusion, the results suggest the implication of MMP-9 and VEGF in enhancement of tumor invasion and progression, with the superior prognostic significance of MMP-9. Hence, their addition to the routine battery of investigations, could improve the laboratory work up of breast cancer patients and have its implication on the design of new treatment modalities employing anti-MMP-9 therapeutic agents and anti-VEGF which block the receptor signaling pathway