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Can J Vet Res. 2014 Jan; 78(1): fifty–threescore.
Language: English | French
Relevance of synovial fluid chondroitin sulphate every bit a biomarker to monitor polo pony joints
Received 2012 Apr 18; Accepted 2012 Nov 1.
Abstract
Osteoarthritis (OA) of the metacarpophalangeal joint is the most common articular disease in polo ponies leading to early retirement. A biomarker that would discriminate between pathological and physiological changes secondary to practise could be helpful in OA prevention. The aim of this report was to investigate the effects of polo grooming on synovial fluid biomarkers of inflammation and cartilage turnover in polo ponies of dissimilar skill levels. Synovial fluid samples were collected from metacarpophalangeal joints of polo ponies before and during the polo flavour (320 d). Nucleated cells, soluble protein, prostaglandin Etwo (PGE2), glycosaminoglycans (GAG), and urea were measured. The main synovial fluid GAG are chondroitin sulphate (CS, ~25 μg/mL) and hyaluronic acid (HA, ~400 μg/mL). After a polo match, a transitory increase in poly peptide and PGE2, merely not CS and HA, occurred (expressed as urea ratio), returning to basal levels in 24 h. During the polo flavor, the number of synovial fluid nucleated cells was ever in the normal range. Increases in poly peptide and HA occurred during the initial twoscore to eighty d, returning to basal levels afterwards. In dissimilarity, in polo prospects the concentration of CS steadily increased during the season. Long-term follow-upward revealed that the synovial fluid CS was significantly higher in polo ponies that adult joint diseases within 24 months following our study. In decision, CS seems to be an early marker of articular cartilage damage.
Résumé
L'arthrose (OA) de l'articulation métacarpophalangienne est la maladie articulaire la plus fréquente chez les poneys de polo menant à la retraite anticipée. Un biomarqueur qui était discriminateur entre les changements pathologiques et physiologiques secondaires au exercice pourrait être utile cascade la prévention de 50'OA. 50'objectif de la présente étude était examiner les effets de l'activité de polo sur les biomarqueurs de l'inflammation et de le métabolisme de la cartilage dans le liquide synovial des poneys de polo de différents niveaux de qualification. Le SF était obtenu à partir de les articulations métacarpophalangiennes de poneys de polo, avant et pendant la saison de polo (320 jours). Les cellules nucléés, protéine soluble, prostaglandine Eastward2 (PGEii), glycosaminoglycanes (GAG) et l'urée ont été mesurés. Les principaux GAG de le liquide synovial sont le chondroïtine sulfate (CS, ~25 μg/mL) et fifty'acide hyaluronique (HA, ~400 μg/mL). Après united nations lucifer de polo, ocorru une augmentation transitoire de la protéine et de la PGE2, mais pas de CS et de HA (exprimé comme le raison d'urée), qui a retourné aux niveaux basal dans 24 h. Pendant la saison de polo, le numero de cellules nucléés dans le liquide synovial était toujours normaux. La protéine et le HA augmentaient pendant les premiers 40–80 jours, mais tous les deux sont retournés aux niveaux de base plus tard. En contraste, dans le group de jeunes poneys (G1), la concentration de CS a augmenté régulièrement pendant la saison. Accompagnant à long terme avait révéle que le CS de liquide synovial était significativement plus élevée chez les poneys de polo que, dans les 24 mois suivants, avaient developpé des maladies articulaires. En conclusion, le CS du liquide synovial semble être un marqueur précoce des destructions de la cartilage articulaire.
(Traduit par les auteurs)
Introduction
Articular cartilage is a specialized connective tissue that consists substantially of chondrocytes embedded in an arable extracellular matrix. It resists and redistributes impact loading of the articulation, while providing a resilient articular surface. These backdrop depend on the structural arrangement of the extracellular matrix of macromolecules. The extracellular matrix of cartilage is equanimous of a dense network of collagen fibers (i) that entrap a high concentration of proteoglycans (PG) and other noncollagenous proteins.
The major noncollagenous component of cartilage matrix is the PG aggrecan (2), composed of a ~200 kDa protein cadre containing 3 globular domains, about 100 chondroitin sulphate (CS) chains distributed along the CS1 and CS2 domains, and about 30 keratan sulphate chains attached to a repeat domain, located just N-final to the CS domains. Aggrecan forms large aggregates with hyaluronic acid (HA) and link poly peptide (iii), and also interacts with other macromolecules (four). Nigh 5% of the cartilage wet weight is aggrecan, which provides an extremely high stock-still accuse density that attracts and traps water, resulting in an expansion of the cartilage matrix. This is mainly acquired past the CS chains. Tightly packed collagen fibrils resist this expansion, giving the cartilage the capacity to resist compressive forces. Changes in PG and glycosaminoglycan (GAG) structure and concentration effect in changes of compressive stiffness and contribute to cartilage damage (5,6).
Articular cartilage impairment, whether traumatic or inflammatory in origin, can exist studied by means of the synovial fluid biomarkers in the cartilage matrix turnover and inflammatory mediators (7). Biomarkers reflecting aggrecan and collagen II turnover are capable of signaling changes in cartilage matrix homeostasis (8,9).
Prostaglandin Due eastii (PGE2) plays an intimate part in articular inflammatory and nociceptive pathways. Its release from both synovial cells and chondrocytes is stimulated by articulation inflammation, injury, vascular distension, and stress (10,11). Prostaglandin Due east2 is considered a sensitive predictor of articulation affliction (12), and its concentration of synovial fluid is high in most (if non all) joint diseases, including osteoarthritis (OA). The rise in PGE2 concentration is very rapid, peaking 2 to ix h afterwards the inflammatory stimulus (13,14). In horses, although PGE2 concentration was not correlated to the radiological signs of whatever joint disease (15), it is well correlated with lameness (16).
Polo is an equestrian team sport that demands quick bursts of speed, followed by a rapid turn and stop, in pursuit of the brawl through its many movements. The mounts used for polo are traditionally called "polo ponies," in reference to their agility. A polo match is divided into time periods named "chukka" or "chukkers," lasting most 7 min each, with the horse beingness on the movement throughout. So, polo ponies crave considerable training and ongoing workout due to the extreme demands placed on them.
A young equus caballus that has non nevertheless started any polo training is called a "polo prospect." A young equus caballus that is in training to go a polo pony is ofttimes referred to as a "green horse." Young polo prospects go through years of specialized preparation before they go what is called a "made pony," one that is set for apply in tournament polo.
The most mutual sources of lameness in polo ponies are tendonitis of the superficial digital flexor tendon and OA of the metacarpophalangeal joint. Furthermore, OA of the metacarpophalangeal joint is the most common articular problem leading to early retirement. The aim of this written report was to investigate possible changes in synovial fluid biomarkers within the polo season in polo prospects, green horses, and high handicap made ponies that could signal horses susceptible to OA.
Materials and methods
Materials
The post-obit materials were obtained and used throughout our study. Standard GAG chondroitin iv-sulphate (from whale cartilage), dermatan sulphate (from hog skin), and HA (from umbilical cord), and Streptomyces hyalurolyticus hyaluronate lyase (EC 4.two.2.1, HA lyase, or hyaluronidase) were obtained (Sigma-Aldrich, St. Louis, Missouri, The states). Heparan sulphate (from bovine pancreas) and chondroitin AC lyase (EC 4.2.2.v, from Flavobacterium heparinum) were prepared past methods previously described (17,xviii). Agarose (standard, low Mr) was obtained (Bio-Rad Laboratories, Hercules, California, USA).
Animals and sample collection
The present work was approved by the Upstanding Committees Universidade Federal de São Paulo — UNIFESP (CEP 0800/07) and Universidade de São Paulo — USP (1238/2007), and was carried out in accord with UNIFESP and USP guidelines, and also in accordance with EC Directive 86/609/EEC for beast experiments (http://ec.europa.european union/environment/chemicals/lab_animals/legislation_en.htm).
A full of l healthy joints in 25 polo ponies, classified into 4 groups, were studied (Tabular array I). Only horses that were free from lameness and with normal metacarpophalangeal joints (past ultrasound and radiography) were included. These animals were submitted to clinical examinations of both forelimbs every 28 d, during the whole experimental period (320 d). These examinations included lameness grade (0 to 5), flexion test, and effusion grade of metacarpophalangeal joints. Furthermore, ultrasonographic and radiographic evaluations of both metacarpophalangeal joints were done on days 40, lxxx, 240, and 320 of the polo flavour. Clinical and radiographic examinations were repeated 24 mo afterward the end of the experimental period. Detailed results are presented elsewhere (nineteen)
Table I
Joints, gender, age, time of grooming, and groups of polo ponies
| Animate being | Joints | Gender | Age (years) | Training (years) | Group |
|---|---|---|---|---|---|
| ane | R and L | iii | 0 | ane | |
| 2 | R and L | Female person | four | 0 | 1 |
| three | R and L | Female | 4 | 0 | 1 |
| 4 | R and 50 | Female | 3 | 0 | ane |
| 5 | R and 50 | Female | four | 0 | 1 |
| 6 | R and L | Female | 3 | 0 | 1 |
| 7 | R and L | Male | four | 1–ii | 2 |
| eight | R and L | Female | 4 | 1–ii | 2 |
| 9 | R and L | Female | 4 | 1–2 | 2 |
| 10 | R and L | Female | 4 | i–2 | 2 |
| 11 | R and L | Female | iv | 1–2 | 2 |
| 12 | R and L | Male | 3 | 1–2 | 2 |
| 21 | R and L | Female | 11 | > 5 | 3 |
| 22 | R and L | Female person | 16 | > 5 | 3 |
| 23 | R and L | Female person | 14 | > 5 | 3 |
| 24 | R and L | Female | fourteen | > v | 3 |
| 26 | R and L | Female | 11 | > v | 3 |
| 27 | R and 50 | Female | 10 | > v | 3 |
| 28 | R and L | Male person | 12 | > v | 3 |
| 210 | R and L | Female person | ten | > 5 | 3 |
| I | R and Fifty | Female | 10 | > 5 | 24 h |
| Two | R and 50 | Female | 11 | > v | 24 h |
| Iii | R and L | Female person | x | > 5 | 24 h |
| 4 | R and L | Female | 12 | > 5 | 24 h |
| V | R and L | Female | nine | > 5 | 24 h |
To investigate the acute effect of exercises, synovial fluid samples (0.5 mL) were aspirated direct (neat) and without lavage from 10 metacarpophalangeal joints of 5 made polo ponies (numbers I to V, Tabular array I), nine to 12 y one-time, high handicap, all submitted to the same training scheme, 24 h earlier the polo friction match, and iii h, vi h, and 24 h after the end of the match.
To analyze the possible changes in synovial fluid during the polo season (320 d), horses were classified into 3 groups: group 1 — polo prospects, 6 polo ponies, 3 to 4 y old, who were offset the polo grooming with no previous participation in tournaments (12 joints); group 2 — dark-green horses, 6 polo ponies, three to 4 y quondam, who were nether specialized training for at least ane y (12 joints); and grouping 3 — high handicap made ponies, 8 polo ponies, 10 to 16 y onetime, who had been participating in tournaments for at to the lowest degree five y. Each group was submitted to a preparation scheme reflective of its corresponding skill level. This consisted of xxx min trot or amble and 1 h walk 5 d per wk with sporadic game preparation for grouping 1, and 40 min trot or canter and 1 h walk half dozen d per wk (except on game days), with either game grooming or polo match 2 to 3 times per wk, for groups 2 and iii.
Synovial fluid samples (2 to five mL) were aspirated directly (peachy) from metacarpophalangeal joints (left and right) at the finish of tourn-out (a period of rest or vacations for polo ponies prior to a polo season, basal levels), and on days 40, 80, 240, and 320 of the polo season, ever 24 h after the stop of a polo match.
Synovial fluid cell count, soluble protein, PGEii, and urea concentrations
For nucleated jail cell count, 1 mL of each synovial fluid sample was centrifuged at 100 × g for 10 min. The supernatant was removed, and the pellet resuspended in 1 mL of phosphate-buffered saline (PBS) solution. The cells were counted in a Neubauer counting chamber, and the differential cell count was determined in smears stained using May Grunwald-Giemsa.
The supernatant was divided in aliquots that were used to measure out soluble proteins, PGEtwo, urea, and GAG. Soluble proteins were measured in 4 μL aliquots by biuret reaction in an automated biochemical analyzer (Labmax 240; Tokyo Boeki Machinery Ltd., Tokyo, Nippon). The PGEii was quantified by an enzyme immunoassay kit (True cat #514010; Cayman Chemic, Ann Arbor, Michigan, USA) (xx). Urea was determined in synovial fluid samples (v μL) by completely automatic enzymatic reactions (urease-glutamate dehydrogenase; Randox RX, Crumlin, County Antrim, Britain). Urea concentration was used to right the protein, PGEii and GAG concentrations for fluid volume (21), because urea is a robust indicator of correction for dilution of synovial fluid volume due to joint effusion (22).
Glycosaminoglycans: HA and CS
A simple and reliable method was used for the identification and quantification of synovial fluid GAG. This procedure was based on: i) migration of GAG in agarose gel electrophoresis in a special buffer organization (PDA), that allows the complete segregation of CS and HA; ii) differential staining of sulphated and not-sulphated GAG by using toluidine blue in different pH levels; and three) susceptibility to specific GAG lyases. This procedure also avoids interference from other macromolecular and ionic components.
Synovial fluid supernatant samples (100 μL) were submitted to proteolysis (4 mg/mL maxatase, detergent alkaline family of serine endopeptidases isolated from Bacillus subtilis, EC 3.4.21.62, in 0.05 M Tris-HCl, pH 8.0, 200 μL). After overnight incubation at fifty°C, maxatase was estrus inactivated (15 min, 100°C), and debris was removed past centrifugation (3000 × k, x min, room temperature). The supernatant was freeze-dried and resuspended in l μL of water.
Aliquots (5 μL) were submitted to 0.5% agarose gel electrophoresis in 0.05 Chiliad 1,3-diaminopropane-acetate buffer, pH ix (PDA), as previously described (23). This is the optimum condition to separate HA from sulphated GAG. Afterward fixation with 0.1% cetyltrimethylam-monium bromide, sulphated GAG were stained by toluidine blue in an acrid solution (0.1% toluidine blue in 1% acetic acid and 50% ethanol, fifteen min, room temperature). The excess dye was removed with 1% acetic acid in 50% ethanol solution. The gel slab was stale at room temperature, and and then non-sulphated GAG (especially HA) were stained in the same slabs by toluidine blue at pH five.0 (0.1% toluidine blue in 0.025M sodium acetate buffer, pH 5.0). The excess dye was removed using the same buffer. The synovial fluid GAGs were quantified by densitometry of the gel slabs using a densitometer (CS-9000 Shimadzu; Shimadzu, Kyoto, Nippon).
These compounds were further characterized by enzymatic degradation using specific GAG lyases: F. heparinum chondroitin Air-conditioning lyase (24) and Streptomyces hyalurolyticus HA lyase (25), as described (9).
Statistical analysis
Data were evaluated for normality using the Kolmogorov-Smirnov exam, which revealed that the sample distributions were parametric. Afterwards, the unpaired t-examination was used to compare each indicate to the basal levels (B) of each group. Differences between groups were analyzed by analysis of variance (ANOVA). Software (GraphPad Instat 3; GraphPad Software, San Diego, California, U.s.) was used to exercise the statistical analysis. Results are presented as mean ± standard deviation (SD). Values of P < 0.05 were considered meaning.
Results
Astute effect of polo lucifer upon synovial fluid components
Protein, PGE2, urea, and GAG were measured in synovial fluid samples obtained from 10 metacarpophalangeal joints from 5 polo ponies, high handicap, 24 h earlier (B = basal levels), and iii h, 6 h, and 24 h after the finish of a polo match. The urea concentration did not vary significantly (Figure i). This concentration was used to correct protein, PGEtwo, and GAG for fluid book. A transitory increase in protein and PGE2 was observed iii h and 6 h after the end of the polo lucifer, returning to basal levels at 24 h (Figure i).
Urea, soluble poly peptide, and prostaglandin Eii (PGE2) concentrations in synovial fluid samples of loftier handicap polo ponies, collected 24 h earlier a polo match (B = basal levels) and 3 h, 6 h, and 24 h after the end of the match. Protein and PGE2 are presented as urea ratios, to right for fluid volume (mean ± SD). *Statistically pregnant in comparison to basal levels, P < 0.05.
A representative agarose gel electrophoresis of synovial fluid GAG from ponies I through V is shown in Figure 2A, and also from 3 other polo ponies (numbers 1, 7, and 21). Ane abrupt band, migrating as CS, and a broad band, migrating as HA, are clearly visible in all samples. These compounds were further characterized by incubation with specific lyases, and quantified by densitometry of the agarose gel slabs. The results, expressed both as concentration (mg/mL) and as GAG/urea ratios, are shown in Effigy 2B. The GAG concentration did not significantly vary within the 24 h period after a polo lucifer.
Synovial fluid glycosaminoglycans (GAG) from loftier handicap polo ponies collected 24 h earlier a polo friction match (Basal), and 3 h, 6 h, and 24 h afterward the cease of the friction match. Sulphated and nonsulphated GAG were analyzed by agarose gel electrophoresis. A — Shows a representative agarose gel electrophoresis of basal synovial fluid GAG. B — Shows quantitative results, expressed both as concentration (mg/mL) and as GAG/urea ratios (hateful ± SD). I, Two, III, IV, Five, ane, 7, and 27 refers to the polo pony numbers. CS — chondroitin sulphate; DS — dermatan sulphate; HS — heparan sulphate; HA — hyaluronic acid; Or — origin; S — mixture of standard glycosaminoglycans.
Changes in synovial fluid components during the polo season
In order to investigate possible changes during the polo season, synovial fluid samples were collected before the showtime of polo season (0 = basal), and on days twoscore, 80, 240, and 320, e'er 24 h later on a polo match, from polo prospects (group 1), greenish horses (group ii), and high handicap made ponies (group iii). The nucleated cell count was maintained in the normal range for all groups (Effigy 3). Mononuclear cells were always the most abundant cells. These data did not show any inflammatory reaction.
Nucleated cell counts of synovial fluid samples collected during the polo season from polo ponies of different skill levels. Synovial fluid samples were collected before the beginning of polo season (B, basal levels) and on days 40, eighty, 240, and 320 of flavor from all iii groups. A — Shows total cell count (mean ± SD). B — Relative numbers of different jail cell type are shown.
This finding is in understanding with the poly peptide and PGE2 concentrations (Figure 4). An increase in protein concentration was observed in the first twoscore to 80 d for all groups, while PGE2 did not significantly vary, over again suggesting no inflammation.
Synovial fluid urea, soluble poly peptide, and prostaglandin Etwo (PGE2) concentrations in polo ponies of different skill levels during the polo season. *Statistically significant in comparison to basal levels, P < 0.05.
For HA, an early increase occurred (compare basal to 40 d), especially for groups ane and 2, suggesting adaptation to the exercises (Figure 5).
Glycosaminoglycans (GAG) in synovial fluid samples nerveless during the polo season from polo ponies of different skill levels. A — Representative agarose gel electrophoresis of samples obtained on days xl, 80, 240 and 320 and besides before the polo season (B, basal) from polo ponies #10 (correct metacarpophalangeal articulation) and #22 (right and left metacarpophalangeal joints) are shown. Samples (100 μL) of the synovial fluid samples described in Figure 4 were submitted to proteolysis, and aliquots (5 μL) were analyzed for sulphated and non-sulphated GAGs by agarose gel electrophoresis, as described in Methods. S — mixture of standard glycosaminoglycans; DS — dermatan sulphate; HS — heparan sulphate; Or — Origin. Individual data for B — Private data for chondroitin sulphate (CS) and (C) hyaluronic acid (HA) concentrations (expressed both equally μg/mL and CS/urea and HA/urea ratios) in synovial fluid of ponies during the polo season of Polo Projects (G1), Green Horses (G2) and loftier handicap Made Ponies (G3) during the polo flavor. Horizontal lines point mean ± standard divergence (SD). *Statistically significant in comparison to basal (B) levels, P < 0.05.
Apropos CS (Effigy 5), similar results were obtained for groups 2 and iii, simply non for group 1. In the group 1 animals, who were beginning training, the CS concentration steadily increased up to the 240th twenty-four hours, and remained high until the end of the polo season (in comparison to basal levels). This upshot was mainly due to 3 animals (numbers 1, 2, and 4), in which the CS concentration became very high (200 to 250 μg/mL, 0.5 to 1 CS/urea ratios), in contrast to other animals of the same group (xv to xxx μg/mL, ~0.one CS/urea ratios).
Biomarkers and joint diseases
Follow-up data of the polo ponies, obtained by interviewing the referring veterinary, revealed that iii horses from group 1 (numbers ane, 2, and 4), 2 from group 2 (numbers 7 and eight), and 1 from grouping 3 (number 27, run into Tabular array I), had developed OA in the metacarpophalangeal joints within 24 mo after the end of our study. These animals presented increased joint volume and/or history of articulation lameness, associated with characteristic radiographic imaging (26). The synovial fluid CS levels of these animals on the 320th day of polo season were higher than in synovial fluid of polo ponies that remained good for you and in full preparation (Figure 6). In contrast, the urea, HA, protein, and PGE2 levels did not vary significantly amidst animals that remained healthy and animals that after developed OA. It is of import to emphasize that, during our study, no clinical, ultrasonographic, or radiographic signs of OA were observed.
Synovial fluid chondroitin sulphate (CS), urea, hyaluronic acid (HA), protein, and prostaglandin Eastward2 (PGEtwo) in polo ponies that developed osteoarthritis (OA) in the metacarpophalangeal joints inside 24 mo later on our study ended, in comparison to those that remained salubrious. Individual values of synovial fluid urea (day 320, mg/mL), CS, HA, poly peptide and PGE2 (urea ratios to correct for fluid volume) in polo ponies that developed OA inside 24 months subsequently the cease of our study (OA), in comparison to polo ponies that remained healthy at the end of this period (NL). Each indicate is the mean of 3 measurements. Horizontal lines indicate hateful ± SD of all points. *Statistically significant if P < 0.05. NL — synovial fluid from normal, healthy joints; JD — synovial fluid from joints that developed osteoarthritis.
Discussion
Polo was the outset equestrian sport in recorded history. Information technology originated in Persia (Iran) in the 6th century BC (27), equally a training game for cavalry units. Despite the antiquity of polo playing, nowadays, the sport has spread around the world. Polo ponies are subjected to complex physical challenges and at that place are few studies on the physiological furnishings of polo specialized grooming upon them (28), especially on their joints. In this report, biomarkers of inflammatory response and cartilage turnover were analyzed in synovial fluid samples from the metacarpophalangeal joints of polo ponies of different skill levels. The short-term (24 h) effect of polo preparation was also evaluated in high handicap made ponies.
While a transitory increment in synovial fluid volume during exercise was reported by Persson (29) in horses, Kingston et al (30) showed no changes in humans. Also Frisbie et al (31) showed no meaning increment in synovial fluid volume following treadmill practise in horses, but Hardy et al (32) reported increased synovial fluid volume in inflamed joints, and urea levels can be used to gauge the changes in fluid book due to joint effusion (21,22). In this study, we measured synovial fluid urea to right for fluid volume. Although no meaning fluctuations in the urea concentration were observed, the concentrations of poly peptide, PGE2, HA, and CS were presented every bit urea ratios.
Important inflammatory reactions do not seem to have occurred, since the total cell count and the profiles of nucleated cells inside synovial fluid, as well equally the protein and PGE2 concentrations, were maintained inside the normal range throughout the polo flavor for all groups. According to Hardy et al (14), PGE2 is an important inflammatory mediator in joints. During the first 12 h after an injury, PGEtwo, half-dozen-keto-PGF1, tromboxane B2, and leukotriene B4 are present, while at 24 h only PGEtwo remains (33). In the nowadays study, protein and PGE2 levels were increased a few hours after the cease of the polo match, but returned to basal levels within 24 h, again suggesting no inflammation.
With respect to HA, it is believed that virtually of the synovial fluid HA is synthesized past the fibroblast-like synoviocytes (or B-synoviocytes). Due to its high molecular weight, HA forms viscous solutions, and is largely responsible for the viscosity of the synovial fluid (34). The HA concentration increased during the first xl to 80 d. This increase could be due to either increased synthesis or decreased degradation (or both). The HA is synthesized by HA synthases (HAS1, HAS2, and HAS3) (35). It was shown that the synthesis of HA is up-regulated in fibroblast-like synoviocytes exposed in vitro to CS, due to increased expression of HAS1 and HAS2, but not HAS3 (36).
The CS concentration steadily increased in polo projects (grouping i), with very loftier values recorded in 3 horses (in comparison to basal levels). Interviewing the referring veterinarian revealed that these animals, and also 2 ponies from group ii and 1 from group 3, had developed OA in the metacarpophalangeal joints within 24 mo of the end of our study, confirmed by clinical and radiographic examinations. Information technology is important to note that these animals did not present whatever signs of disease during our study.
Increased synovial fluid GAG has been previously reported in chronic and astute joint diseases (37), while contradictory results were obtained by different authors regarding do (31,38). Still, in these studies GAG were measured by using either alcian bluish or 1,9-dimethylmethylene bluish (DMMB) dye bounden assays, which are non specific and does not allow for the identification of CS in a mixture of GAG (39). We have previously shown that DMMB assay is inappropriately used to quantify the synovial fluid GAG because HA interferes with CS quantification (nine). We have also shown that synovial fluid CS indicates abnormal articulation metabolism in osteochondritis dissecans, since information technology is increased both in asymptomatic and in symptomatic joints (9). Therefore, CS seems to be a good biomarker to evaluate the cartilage turnover, although the analysis of aggrecan biomarkers, such equally alanine-arginine-glycine-serine (ARGS)-aggrecan (31), could besides give clues apropos the origin of synovial fluid CS.
Our information have shown that synovial fluid CS seems to be a useful biomarker to evaluate cartilage turnover and homeostasis. In dissimilarity, protein and PGEtwo are useful to evaluate brusque-term changes, since their concentration increased a few hours after the game and returned to basal levels within 24 h.
Acknowledgments
The authors thank Dr. Roberto P. de Pádua Foz Filho, the referring veterinarian who allowed the collection of synovial fluid samples from polo ponies. The authors are also thankful to Dr. João Roberto M. Martins and Ms C. Aline Mendes for helpful discussions and technical assistance in many experiments. This enquiry was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), São Paulo, SP, Brazil; Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brasília, DF, Brazil; and Fundação Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brasília, SP, Brazil.
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Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3878009/