Measurement of serum estrogen and estrogen metabolites in pre- and postmenopausal women with osteoarthritis using high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry

Although 17β-estradiol (E2) deficiency has been linked to the development of osteoarthritis (OA) in middle-aged women, there are few studies relating other estrogens and estrogen metabolites (EMs) to this condition. We developed a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) method to measure the levels of six EMs (i.e., estrone, E2, estriol, 2-hydroxyestrone, 2-hydroxyestradiol, and 16a-hydroxyestrone) in healthy pre- and postmenopausal women and women with OA. This method had a precision ranging from 1.1 to 3.1% and a detection limit ranging from 10 to 15 pg. Compared to healthy women, serum-free E2 was lower in the luteal and postmenopausal phases in women with OA, and total serum E2 was lower in postmenopausal women with OA. Moreover, compared to healthy women, total serum 2-hydroxyestradiol was higher in postmenopausal women with OA and total serum 2-hydroxyestrone was lower in both the luteal and follicular phases in women with OA. In conclusion, our HPLC-ESI-MS/MS method allowed the measurement of multiple biochemical targets in a single assay, and, given its increased cost-effectiveness, simplicity, and speed relative to previous methods, this method is suitable for clinical studies.

Plasma rico em plaquetas (PRP) aplicado na artroplastia total do joelho / Platelet-rich plasma (PRP) applied in total knee arthroplasty

Avaliar, através de um estudo clinico, randomizado, controlado e cego, a eficácia do plasma rico em plaquetas na cicatrização, dor e hemostasia após artroplastia total do joelho. Metodologia: Foram selecionados 40 pacientes que seriam submetidos a prótese total do joelho e randomizados. Em 20 destes pacientes foi aplicado o plasma rico em plaquetas antes do fechamento da cápsula articular. Foram realizadas dosagens de hemoglobina (mg/dL) e hematócrito (%) no pré-operatório, após 24 e 48 horas da cirurgia. Aplicado o questionário WOMAC, a escala verbal da dor e medidas as amplitudes de movimento do joelho até o 2o mês pós-operatório. A análise estatística comparou os resultados afim de comprovar haver diferença entre os grupos em cada um dos momentos da avaliação. Resultados: Medidas do valor da hemoglobina (mg/dL) e hematócrito (%) realizadas no pré-operatório, após 24 e 48 horas da cirurgia não mostraram diferenças significativas entre os grupos (p>0,05). O questionário WOMAC e a amplitude de movimento medidas no pré-operatorio e até os dois primeiros meses também não mostraram diferenças estatísticas entre os grupos (p>0,05). A avaliação da dor através da escala verbal, mostrou vantagem no grupo que utilizou o plasma rico em plaquetas após 24 e 48 horas, uma e três semanas e dois meses de pós-operatório (p<0,05). Conclusões: Da maneira que foi utilizado, o plasma rico em plaquetas não se mostrou efetivo para reduzir sangramento ou melhorar a função do joelho após a artroplastia em comparação aos controles. Houve vantagem na escala verbal de dor pós-operatória...

Purpose: To assess, through a clinical study, randomized, controlled ande singleblinded, the effectiveness of platelet-rich plasma in healing, pain and hemostasis after a total knee arthroplasty. Methods: 40 patients, that would be submitted to total knee arthroplasty, were selected and randomized. In 20 of these was applied platelet-rich plasma before the closure of the joint capsule. Hemoglobin and hematocrit levels were measured in the preoperative, after 24 and 48 hours of surgery. The WOMAC questionnaire and verbal pain scale were applied, measures of the range of motion of the knee were performed until the second postoperative month. Statistical analysis compared the results in order to prove if there is a difference between the groups at each time of evaluation times. Results: Hemoglobin and hematocrit levels performed preoperatively, after 24 and 48h of surgery showed no significant differences between groups (p>0,05). The WOMAC questionnaire and the measures of range of motion in the preoperative and even in the first two months also showed no statistic differences between groups (p>0,05). Pain assessement through verbal scale showed benefit in the group using platelet-rich plasma after 24 and 48 hours, one to three weeks and two months postoperative (p<0,05). Conclusions: The way it was used, the platelet-rich plasma was not effective for reducing bleeding or improve knee function after arthroplasty compared with controls. There was an advantage in the verbal scale of postoperative pain...

Plasma rico em plaquetas (PRP) aplicado na artroplastia total do joelho / Platelet-rich plasma (PRP) applied in total knee arthroplasty

Avaliar, através de um estudo clinico, randomizado, controlado e cego, a eficácia do plasma rico em plaquetas na cicatrização, dor e hemostasia após artroplastia total do joelho. Metodologia: Foram selecionados 40 pacientes que seriam submetidos a prótese total do joelho e randomizados. Em 20 destes pacientes foi aplicado o plasma rico em plaquetas antes do fechamento da cápsula articular. Foram realizadas dosagens de hemoglobina (mg/dL) e hematócrito (%) no pré-operatório, após 24 e 48 horas da cirurgia. Aplicado o questionário WOMAC, a escala verbal da dor e medidas as amplitudes de movimento do joelho até o 2o mês pós-operatório. A análise estatística comparou os resultados afim de comprovar haver diferença entre os grupos em cada um dos momentos da avaliação. Resultados: Medidas do valor da hemoglobina (mg/dL) e hematócrito (%) realizadas no pré-operatório, após 24 e 48 horas da cirurgia não mostraram diferenças significativas entre os grupos (p>0,05). O questionário WOMAC e a amplitude de movimento medidas no pré-operatorio e até os dois primeiros meses também não mostraram diferenças estatísticas entre os grupos (p>0,05). A avaliação da dor através da escala verbal, mostrou vantagem no grupo que utilizou o plasma rico em plaquetas após 24 e 48 horas, uma e três semanas e dois meses de pós-operatório (p<0,05). Conclusões: Da maneira que foi utilizado, o plasma rico em plaquetas não se mostrou efetivo para reduzir sangramento ou melhorar a função do joelho após a artroplastia em comparação aos controles. Houve vantagem na escala verbal de dor pós-operatória...

Elevated Levels of T Helper 17 Cells Are Associated with Disease Activity in Patients with Rheumatoid Arthritis

BACKGROUND: Interleukin-17 (IL-17)-producing T helper (Th) 17 cells are considered as a new subset of cells critical to the development of rheumatoid arthritis (RA). We aimed to investigate the distribution of Th1 and Th17 cells and their association with disease activity, and determine the Th17-related cytokine levels in the peripheral blood of RA patients. METHODS: Peripheral blood mononuclear cells from 55 RA and 20 osteoarthritis (OA) patients were stimulated with mitogen, and the distributions of CD4+Interferon (INF)+IL-17- (Th1 cells) and CD4+INF-IL-17+ (Th17 cells) were examined by flow cytometry. Serum levels of IL-6, IL-17, IL-21, IL-23, and tumor necrosis factor (TNF)-alpha were measured by ELISA. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were recorded. The 28-joint disease activity score (DAS28) was also assessed. RESULTS: The median percentage of Th17 cells was higher in RA patients than in OA patients (P=0.04), and in active than in inactive RA (P=0.03), whereas that of Th1 cells was similar in both groups. Similarly, the levels of IL-17, IL-21, and IL-23 were detected in a significantly higher proportion of RA patients than OA patients and the frequencies of detectable IL-6, IL-17, and IL-21 were higher in active RA than in inactive RA group. The percentage of Th17 cells positively correlated with the DAS28, ESR, and CRP levels. CONCLUSIONS: These observations suggest that Th17 cells and Th17-related cytokines play an important role in RA pathogenesis and that the level of Th17 cells in peripheral blood is associated with disease activity in RA.

Effect of Combined Sex Hormone Replacement on Bone/Cartilage Turnover in a Murine Model of Osteoarthritis

BACKGROUND: Estrogens act on estrogen receptors distributed in articular cartilages, synovial membrane, and ligaments, which are thought to be related with degenerative changes. Meanwhile, progesterone is known to have a weak anabolic action on bone formation This study evaluates the effects of estrogen and progesterone hormone on bone/cartilage turnover in ovariectomized (OVX) rats. METHODS: Thirty-five 7-month-old female Sprague-Dawley rats were randomly divided into 5 groups and then ovariectomized bilaterally except the sham control group. The first and the second group acting as controls did not receive hormonal therapy, the third group received estrogen, the fourth group received progesterone, and the fifth group received combination of both hormones 10 weeks after surgery. Evaluations were done using the serum levels of cartilage oligomeric matrix protein (COMP) for cartilage turnover, collagen type I C-telopeptide (CTX-1) and osteocalcin (OC) for bone turnover at 11, 15, 19 weeks after OVX and histology using the Osteoarthritis Research Society International (OARSI) osteoarthritis (OA) cartilage histopathology assessment system. RESULTS: Significantly less cartilage degradation (decreased levels of COMP) was found in the combined hormone treated group in comparison with OVX group. Similarly, both hormonal treatment resulted in increased bone formation and decreased bone resorption i.e., a low overall bone turnover status (decrease in the serum OC and CTX-1 levels). CONCLUSIONS: Combined estrogen and progesterone therapy was found to be convincing in terms of reducing the severity of OA in this experimental model.

Measurement of Interleukin-33 (IL-33) and IL-33 Receptors (sST2 and ST2L) in Patients with Rheumatoid Arthritis

The interleukin-33 (IL-33)/ST2 pathway has emerged as an intercellular signaling system that participates in antigen-allergen response, autoimmunity and fibrosis. It has been suggested that IL-33/ST2 signaling has been involved in the pathogenesis of rheumatoid arthritis (RA), because IL-33 and its receptor have been specifically mapped to RA synovium. The aim of this study was to determine the levels of IL-33 and sST2 in sera and synovial fluids in patients with RA. The serum level of IL-33 was significantly higher in patients with RA (294.9 +/- 464.0 pg/mL) than in healthy controls (96.0 +/- 236.9 pg/mL, P = 0.002). The synovial fluid level of IL-33 was significantly higher in RA patients than in osteoarthritis patients. The level of serum sST2 was higher in RA patients than in healthy controls (P = 0.042). A significant relationship was found between the levels of IL-33 and IL-1beta (r = 0.311, P = 0.005), and IL-33 and IL-6 (r = 0.264, P = 0.017) in 81 RA patients. The levels of IL-33, sST2 and C-reactive protein decreased after conventional disease-modifying antirheumatic drugs treatment in 10 patients with treatment-naive RA. Conclusively, IL-33 is involved in the pathogenesis of RA and may reflect the degree of inflammation in patients with RA.

Determination of oxidant stress in plasma of rheumatoid arthritis and primary osteoarthritis patients.

Determination of oxidant stress in plasma of rheumatoid arthritis (RA) and primary osteoarthritis (POA) patients is important in understanding the pathogenesis of these diseases. In this study, we examined the relationship between oxidant stress and inflammation by measuring protein carbonyl content, thiol levels and plasma protein fractions as the oxidation markers and erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) tests as inflammation markers. Protein carbonyls content was higher in RA and POA patients, as compared to controls (p<0.0001), while the plasma thiol levels in both groups of patients were significantly lower than controls (p<0.0001). Increased levels of proteins under 40 kDa molecular mass were detected in the RA and POA patients compared to that of controls (p<0.0001) both in HPLC and SDS-PAGE analysis. Total protein concentration in plasma of RA patients was higher than the controls (p<0.001), while in POA patients was lower than that of controls (p<0.001). ESR and CRP levels were higher in both the patient groups than the normal group (p<0.001). These results suggested that alterations in the oxidant stress markers could be the cause of inflammation in these diseases. Thus, while working for RA/POA treatment strategies, consideration of the relationship between oxidant stress and inflammation would be worth evaluating.

[Comparison of serum ferritin level between patients with systemic lupus erythematosus, rheumatoid arthritis and osteoarthritis]

Systemic Lupus Erytematosus [SLE] is a chronic and relapsing disorder affecting several organs. This cross-sectional study was conducted on 60 patients with SLE [divided into two groups with low and high disease activity], 20 patients with osteoarthritis [OA], and 20 patients with rheumatoid arthritis [RA]. After having received informed consent, the blood and urine samples were collected from all patients to be tested for iron, TIBC, and serum ferritin levels and also to calculate the lupus disease activity. Patients with iron deficiency anemia were excluded. Data were analyzes by independent t-test, ANOVA, logistic regression and correlation tests. There was no significant difference between three study groups regarding the sex however, the age and duration of disease showed significant differences among 3 groups [p<0.001]. High serum ferritin level was seen in 61.7% of SLE, 15% of RA, and 5% of OA patients [p<0.001]. Increase in ESR levels demonstrated significant differences among study groups [p<0/001]. In patients with SLE, no significant correlation between serum ferritin and the decreased serum complement level and increased anti dsDNA titer was observed. Both serum ferritin and CRP levels increased in patients with serositis [p=0.019] and neurological complication [p=0.04]. Serum ferritin level was significantly higher in patients with SLE compared to those with OA and RA. There was no significant difference between serum ferritin level and SLE activity

Evaluation of serum chondroitin sulfate and hyaluronan: biomarkers for osteoarthritis in canine hip dysplasia

Hip dysplasia (HD) is one of the most important bone and joint diseases in dogs. Making the radiographic diagnosis is sometime possible when the disease has markedly progressed. Chondroitin sulfate (CS) and hyaluronan (HA) are the most important cartilage biomolecules that are elevated in the serum taken from dogs with osteoarthritis. The serum CS and HA can be detected by an ELISA technique, with using monoclonal antibodies against CS epitope 3B3 and WF6 and the HA chain as the primary antibodies. The aim of this study was to compare the levels of serum CS (both epitopes) and HA in non-HD and HD dogs. All 123 dogs were categorized into 2 groups. The non-HD group was composed of 98 healthy dogs, while the HD group was comprised of 25 HD dogs. Blood samples were collected for analyzing the serum CS and HA levels with using the ELISA technique. The results showed that the average serum level of the CS epitope WF6 in the HD group (2,594 +/- 3,036.10 ng/ml) was significantly higher than that in the non-HD group (465 +/- 208.97 ng/ml) (p < 0.01) while the epitope 3B3 in the HD group (105 +/- 100.05 ng/ml) was significantly lower than that in the non-HD group (136 +/- 142.03 ng/ml) (p < 0.05). The amount of serum HA in the HD group (134.74 +/- 59.71 ng/ml) was lower than that in the non HD group (245.45 +/- 97.84 ng/ml) (p < 0.05). The results indicate that the serum CS and HA levels might be used as biomarkers for osteoarthritis in HD dogs.

Serum and synovial fluid levels of MMP-3 and TIMP-1 in rheumatoid arthritis and osteoarthritis

Matrix metalloproteinases [MMPs] are cytokine-modulated enzymes that play an important role in the pathogenesis of RA by inducing bone resorption and cartilage destruction. Tissue inhibitors of metalloproteinases [TIMPs] are naturally occurring MMPs inhibitors. In rheumatoid arthritis [RA], there is a disturbed balance between MMPs and TIMPs favoring proteolysis. This study was performed to evaluate the significance of measuring the serum and synovial fluid [SF] levels of MMP-3 and TIMPs in RA and osteoarthritis [OA] patients in an attempt to provide more insight in their role in the pathogenesis of those two diseases. Serum levels of MMP-3 and TIMP-1 were measured from 30 RA, 20 OA patients and 20 healthy controls using double-antibody ELISA. Also, their levels were measured in the SF of 14 RA and 9 OA patients. RA disease activity was assessed using the Multivariate Analysis of Mallya and Mace and joint erosions were assessed using Larsen score. Serum and SF levels of MMP-3 and TIMP-1 were significantly higher in RA than OA patients and in OA patients than controls. Their levels were significantly higher in the SF than in the serum. Serum and SF TIMP-1: MMP-3 ratio was significantly lower in RA as compared to OA patients and normal controls and this ratio was significantly lower in the SF than in the serum of RA patients. Serum levels of MMP-3 and TIMP-1 correlated strongly with clinical and laboratory parameters of rheumatoid disease activity, and the serum levels of MMP-3 showed a significant positive correlation with the number of joint erosions but TIMP-1 levels did not show this positive correlation. Serum and SF MMP-3 and TIMP-1 levels were significantly higher in RA than OA patients and normal controls. They appear to play a critical role in joint inflammation and destruction, especially MMP-3, which may serve as an additional marker for assessment of RA disease activity and severity

Serum and synovial fluid levels of vascular endothelial growth factor in patients with inflammatory and degenerative arthritis

This work conducted to evaluate the clinical utility of measuring vascular endothelial growth factor [VEGF] by enzyme-linked immunosorbent assay in serum and synovial fluid [SF] samples from patients with early rheumatoid arthritis [RA], other recent-onset inflammatory arthritides, established RA and osteoarthritis [OA], as well as non-arthritic healthy controls. The study, also, investigated whether serum and SF VEGF had any relation with the standard clinical, laboratory and radiological markers of RA disease. The results showed that serum VEGF levels were elevated in patients with RA, inflammatory arthritis and OA in comparison with non-arthritic controls. Patients with recent-onset RA had the highest VEGF values both in serum and SF. In early and established RA groups, VEGF levels were higher in SF than in the serum. In the early inflammatory arthritides and the OA groups, the opposite occurred. Serum and SF VEGF were higher in active than inactive RA disease, with a positive correlation between their levels and each of ESR and disease activity grade. Moreover, values of VEGF in SF, but not in serum correlated well with the development of radiological damage in the established RA group

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

Osteoartritis de la articulación temporomandibular. Parte III: manifestaciones histopatológicas, clínicas, serológicas y radiográficas, tratamiento y pronóstico / Osteoarthritis affecting the temporomandibular joint. Part III

En la osteoartritis de la articulación temporomandibular el aspecto macroscópico del cartílago sufre algunas modificaciones, histológicamente hay una pérdida de proteoglicanos, una desintegración de la red de fibras colágeno y una degeneración grasa. La zona del hueso subyacente sufre modificaciones, ocurren microfracturas y un aumento en la densidad ósea. Se pueden formar osteofitos. La formación de nuevo hueso puede ser liso, esclerótico o eburnado. Se discuten otros cambios estructurales. Los hallazgos clínicos pueden incluir dolor a la palpación, limitación, crepitación de los movimientos con desviación hacia el lado afectado y evidencia radiográfica de cambios estructurales. El mejor manejo de la mayoría de los pacientes con osteoartritis de la articulación temporomandibular es a través de un tratamiento conservador y no invasivo

Serum and synovial fluid transforming growth factor beta, interleukin-1 beta and hyaluronan in osteoarthritis and rheumatoid arthritis

Serum and synovial fluid levels of transforming growth factor beta [TGF-beta], interleukin-1 beta [IL-1 beta] and hyaluronan in osteoarthritis and rheumatoid arthritis were estimated. Twenty- three and twenty-two serum and synovial fluid [SF] samples were obtained from patients with osteoarthritis [OA] and rheumatoid arthritis [RA] and thirteen serum samples from age and sex matched controls. Serum TGF-beta, IL-1 beta and HA levels were significantly elevated in OA and RA patients compared with control group. The significant increase of these previous serum markers was observed in RA patients compared with OA patients, whereas no significant differences were obtained in SF TGF-beta levels between the two groups. On the other hand, SFHA levels were reduced significantly in OA and RA patients with high index radiological progression, whereas serum HA and SF IL-1 beta levels were elevated significantly in RA patients with high index radiological progression. In RA patients, SF IL-1 beta levels were also correlated significantly with the clinical variables. Concerning the biochemical markers interrelationship, SFHA levels were negatively correlated with SF IL-1 beta level and RA, respectively. Also, SFHA levels were correlated with SF TGF-beta-levels in OA and RA, respectively. Moreover, negative correlation was observed between serum HA and SFHA in RA patients

Serum levels of insulin-like growth factor-1 in patients with osteoarthritis of the knee

Insulin related growth factor-1 [IGF-1] is an important growth promoting peptide which has anabolic effects on cartilage metabolism. The fact that serum levels tend to decrease in elderly subjects while osteoarthritis [OA] increases in prevalence suggests that suboptimal levels of IGF-1 may play a part in the development of OA. This study therefore considers the hypothesis that IGF-1 levels are low in subjects with OA of the knee. Serum levels of 48 patients with symptomatic knee OA and radiographic changes of different grade were compared with normal controls. No significant difference was found in serum levels between cases and controls [mean [SD] 20,67 [7.73] for cases and 23.91 [12.0] mmol for controls]. No significant correlations were shown between IGF-1 levels, age and total radiographic score. The study provides no evidence that serum levels of IGF-1 are low in OA

Lack of expression of c-myc oncogene in peripheral blood mononuclear cells of patients with active rheumatoid arthritis.

We looked for the expression of c-myc oncogene, one of the genes that enhance apoptosis, in 11 patients with active rheumatoid arthritis (RA) along with five patients with osteoarthritis (OA) knee as disease controls and six healthy volunteers. A dot-blot assay using a probe specific for c-myc oncogene was performed on total RNA obtained from peripheral blood mononuclear cells. There was no expression in patients with active RA and healthy volunteers. One patient with OA expressed c-myc. Lack of expression of c-myc suggests that in active RA circulating lymphocytes are not in the replicative phase despite ongoing disease activity.

Statistics for clinicians. 5. Interval data (I).

Interval data may be discrete or continuous. They are usually summarized by the average (arithmetic mean). Sometimes, for example when the possible values in a series change by a constant multiple, we need to use the geometric mean. To obtain the overall or mean percentage of a series of percentage values, we need to calculate their weighted mean. The variability of observations in a sample is measured by the standard deviation, and the variability of sample means is measured by the standard error of mean. Confidence interval is a range which contains the population mean with a known probability. It is obtained by deducting from the sample mean, and adding to it, "t" times the SEM, the value of "t" depending on the desired confidence level (1-P) and the sample size (N). The significance of difference between the mean of two sets of unpaired interval data (MA-MB) is tested by Student's t-test. If the data are paired, the significance of the mean difference (MD) is tested by paired t-test. Ordinal data, ie, grades and ranks, may be analyzed by means of the t-test which is more sensitive and allows more refined analyses if needed.