Clinical changes in sodium monoiodoacetate–induced stifle osteoarthritis model in dogs

In six dogs, experimental model of osteoarthritis (OA) was reproduced by intraarticular injection of sodium monoiodoacetate (MIA) in left stifle joints. Contralateral joints served as control. The clinical status and some goniometric parameters were monitored before MIA introduction and at post injection days 30, 60 and 105. The results showed convincingly that the used experimental chemical OA model reproduced successfully the disease in canine stifle joints. The studied clinical indices correlated with the severity of disease. Кey words: dogs, stifle joint, sodium monoiodoacetate, osteoarthritis Introduction this is the only means to confirm a working hypothesis. Orthopaedics uses frequently in vivo Osteoarthritis (ОА) is a degenerative articular experimental animal models (Bendele, 2001; cartilage disease encountered in all mammals, Murray, 2002). The reproduction of all disorders birds and men (Lipowitz, 1993). Ehrlich (2003) and symptoms of OA is however challenging, so and Haima (2005) describe it as a “wear-anda model possessing the most important signs of tear” process, while Bennett (1990) defines OA natural disease and that could be reproduced, is as the final stage of a prolonged accumulation of good enough for the purpose of research (Brandt, biochemical disorders, hence, as adaptation of 2002; Carlson, 2005). The interest towards the joint to abnormal stress. Gardner (1994); reproduction of OA of the knee is due to its Grainger & Cicuttini (2004) support the anatomical features and the increasing hypothesis for the multifactorial etiology of OA prevalence of natural degenerative events resulting in cartilage loss, bone remodeling, pain, (Bendele, 2001; Carlson, 2005). The numerous effusion and inactivity. existing experimental models of OA are The incidence of degenerative joint classified by Witter (1999) and Schaller et al. conditions in dogs is about 78%, with increasing (2005). share of primary OA with age (May, 1994). The metabolic inhibitor sodium monoioAccording to Anderson (1994) and Vasseur doacetate (MIA) destructs the joint cartilage by (1993), in small animals, the stifle joint is the blocking glyceraldehyde-3-phosphate dehydromost commonly affected (more than 20% of all genase in chondrocytes and inhibition of OA cases), a substantial part of alterations being glycolysis. The rapid depletion of ATP results in bilateral (Tirgari & Vaughan, 1975). cellular death. The number of cells is progressively The experimental reproduction of OA is of reduced, the synthesis of proteoglycans for the both scientific and applied importance. Sometimes, articular matrix is stopped (Kalbhen, 1987; Van Vet. World, 2012, Vol.5(3):138-144 RESEARCH www.veterinaryworld.org Veterinary World, Vol.5 No.3 March 2012 138 To cite this article : Goranov NV (2012) Clinical changes in sodium monoiodoacetate–induced stifle osteoarthritis model in dogs, Vet. World. 5(3):138-144, doi: 10.5455/vetworld.2012.138-144. Clinical changes in sodium monoiodoacetate–induced stifle osteoarthritis model in dogs der Kraan et al. 1989). MIA-induced degenerative higher doses and/or multiple injections would processes, typical for natural OA, appear within 6 result in more consistent OA changes in this to 8 weeks (Kalbhen & Blum, 1977; Janusz et al. animal species. 2001; Guzman et al. 2003). The time of occurrence The clinical evaluation of lameness, pain, of changes and the used MIA doses are very joint effusion and stifle joint motility is subjective different. Fourteen weeks after two intraarticular (Budsberg & Thomas, 2006). Therefore, several injections of 0.6 mg MIA in the stifle joints of authors (Cross et al., 1997; Grisneaux et al., hens, Kalbhen & Jansen (1990) detected 1999; Budsberg et al., 1999) introduced scoring radiological and gross anatomical changes. systems for cumulative assessment of these Gencosmanoglu et al. (2001) have achieved a parameters. The range of motion is an objective chondrotoxic effect in rats after 8 weekly although non-specific parameter, while injections of 1 mg MIA. Permanent histological goniometry is a non-invasive method for injuries were reported to occur after 2 to 8 weeks quantitative estimation of the range of motion by of application of 1 to 4 mg MIA in rats; they were measurement of specific angles formed by bones progressive and similar to OA lesions in humans (Lipowitz, 1993; Jaegger et al., 2002). They (Saied et al. 1997; Guinamp et al. 1997; Bove et could be also used to assess the efficacy of al. 2003; Guzman et al. 2003). The rat OA model therapy (Crook et al., 2007). These measurements (2 mg MIA) was found useful for monitoring of are routinely performed in men, but literature chronic OA pain over 10 weeks (Combe et al. data about dogs are very limited (Millis & 2004). In mice, Boileau et al. (2004) have used a Lavine, 1997; Jaegger et al. (2002). single dose of 0.1 mg MIA. The histological The purpose of the present study was to changes were detected 7-14 days later. attempt to reproduce experimentally stifle In rabbits, the described events occurred osteoarthritis in dogs by means of intraarticular th administration of sodium monoiodoacetate and until the 12 week (Regling et al. 1989), with to evaluate the success or failure of the model by correlative changes in alkaline phosphatase and means of gait and pain analyses. lactate dehydrogenase activities (Horn et al. 1988). Gustafson et al. (1992) have evaluated the Materials and methods changes in equine carpal joints with regard to the Experimental animals: Six clinically healthy applied MIA dose as mild (0.09 mg/kg), mongrel dogs from both sexes were used (body moderate (0.12 mg/kg), and severe (0.16 mg/kg) weight 15±2 kg). They were housed indoor in after a 12-week period. Moreover, it turned out individual boxes and had free access to drinking that high MIA doses (60-100 mg/ml in 2 ml) were water and dry canine food for maintenance. The able to provoke a chemical arthrodesis of equine experiment was approved by the Committee on tarsal joint after 13-51 months (Penraat et al. Animal Experimentation at the Trakia University, 2000; Bohanon, 1995). Therefore, the MIA dose Stara Zagora, Bulgaria and was performed in and the number of intraarticular applications strict compliance with animal welfare regulations were essential for the reproduction of the (Directive 86/609/EEC). generative process. Stobie et al. (1994) were the only researchers Reproduction of osteoarthritis: In the left stifle having applied twice MIA in dogs at doses of joint of each dog, ten intraarticular injections of 0.375 mg/kg and 0.500 mg/kg MIA at a 2-week sodium monoiodoacetate (MIA) (MERCK # interval, but the result was an insignificant S05800 228) were performed once weekly at lameness without permanently affected doses of 0.12, 0.14, 0.16, 0.26, 0.36, 0.96, 1.28, th locomotion and biochemical profile until the 12 3.00, 5.00 and 10.00 mg/kg in 1 ml 0.9% saline post injection week. The authors did not support solution. The right joint served as control. The their findings with histological evidence, but clinical parameters were monitored on days 0, 30, nevertheless they suggested that most probably, 60 and 105. www.veterinaryworld.org Veterinary World, Vol.5 No.3 March 2012 139 Clinical changes in sodium monoiodoacetate–induced stifle osteoarthritis model in dogs Table-3. Scoring system for evaluation of hindlimb use Clinical evaluation and goniometric analysis: in dogs with ОА (Budsberg et al., 1999). Three clinical scoring systems – according to Grisneaux et al. (1999), Cross et al. (1997) and Budsberg et al. (1999), originally used for drug therapy efficacy evaluation, were parallelly used for gait, behaviour and pain analysis (Tables 1, 2, 3). Table-1. Criteria for pain and behaviour evaluation of dogs with osteoarthritis (Grisneaux et al., 1999). * response of the affected hindlimb; ** replacement of the contralateral limb in < 10 s *** replacement of the contralateral limb in < 5 s The thigh circumference (TC), the stifle joint circumference (SC) and the range of motion of the stifle joint (ROM) between full flexion and extension were measured in all dogs (Robins, 1990; Millis & Levine, 1997) with goniometer and a band. The animals were in lateral recumbency, with the studied limb on the top. The Table-2. Kinetic gait analysis system for clinical thigh circumference was determined in the evaluation of lameness, pain and joint effusion of knee middle of the thigh, the stifle joint circumference osteoarthritis in dogs (Cross et al., 1997) o – in 90 flexion, ROM was measured between the Parameter Score Clinical sign longitudinal axes of the femur through trochanter Standing 1 Normal weight-bearing lameness 2 Partial weight-bearing major and the tibia through maleolus lateralis. All 3 Intermittent toe touching results were compared to the contralateral joint. 4 No weight-bearing Trotting 1 Normal weight-bearing lameness 2 Marked lameness with partial weight-bearing Statistical analysis: The results were statistically 3 Marked lameness with intermittent toe touching processed by the non-parametric Friedman and 4 No weight-bearing Pain response 1 Absence of pain and response Mann-Whitney tests using statistical software 2 Slight pain, allowing manipulations of the (Statmost for Windows, Datamost Corp., 1994limb within the normal range of motility, manifested by turning the head and pulling 1995). Differences were accepted as statistically the limb away 3 Moderate pain, not allowing manipulations of significant at p<0.05. Relationships between the limb within the normal range of motility, parameters were estimated by the Pearson manifested as described for score 2 4 Significant pain, not allowing manipulations correlation analysis test. of the limb Joint effusion 1 Normal – palpatory compression upon the Results patellar ligament 2 Weak – slight increase, the patellar ligament The average Grisneaux's score increased is palpated 3 Moderate – marked increase, slightly statistically significantly at p<0.01 from 61 perceptible ligament points in the beginning of the experiment to 14+2 4 Significant – the patellar ligament is not palpated (day 30) and 12+2 (day 60) (Table 4). The Parameter Score Clinical sign Lameness 1 Stands and walks normally 2 Stands normally, slightly lame at walk 3 Stands normally, severely lame at walk 4 Abnormal stance, slightly lame at walk 5 Abnormal stance, severely lame at walk Weightbearing 1 Normal at both rest and walk 2 Normal at rest, favours affected limb at walk 3 Partial at both rest and walk 4 Partial at rest, no weightbearing at walk Parameter Score Clinical sign 5 No weightbearing at rest and walk Response to 1 Accepts displaced weight Behaviour 1 Apathetic or indifferent contralateral 2 Mild resistance to displaced weight 2 Friendly limb lift* 3 Moderate resistance to displaced weight** 3 Nervous, submissive behaviour 4 Strong resistance to displaced weight*** 4 Very nervous, tries to move away 5 Refusal to lift the contralateral limb 5 Aggressive Response to 1 No response Compliance 0 No objection affected limb 2 Mild response (turning head toward the with restraint 1 Recognizes manipulations, no complaint extension affected limb) 2 Objects but does not try to bite 3 Moderate response (withdrawal of affected limb) 3 Tries to bites and struggles 4 Severe response (vocalization, aggression) Heart rate 0 0 to 10% greater than normal 5 Disallows manipulation or palpation of 1 11 to 30% greater than normal affected limb 2 31 to 50% greater than normal 3 >50% greater than normal Respiratory 0 Normal rate 1 Mild abdominal assistance 2 Marked abdominal assistance Vocalization 0 No crying 1 Crying but responds to calm voice 2 Crying but does not respond to calm voice Agitation 0 Asleep or calm 1 Mild agitation 2 Moderate agitation 3 Severe agitation Response to 0 No response manipulation 1 Minimal response 2 Turns head toward site, slight vocalization 3 Turns head to bite, howls www.veterinaryworld.org Veterinary World, Vol.5 No.3 March 2012 140 Clinical changes in sodium monoiodoacetate–induced stifle osteoarthritis model in dogs respective score according to Cross et al. yielded Budsberg's scores; r=–0.63, p<0.001 with Cross' 4+0 points (day 0), 13+1 (day 30), 12+1 (day 60) score. The same relationships were observed for and 9+1 (day 105) (p<0.01 vs baseline). Average thigh and joint circumferences (Table 5). Budsberg score before the first MIA adminisDiscussion tration was 4+0 with considerable increase by The obtained results showed that OA in dogs day 30 to 17+1, followed by reduction to 13+1 could be successfully reproduced by intraarticular and 11+1 by days 60 and 10+5, respectively injection of the glycolysis inhibitor sodium (p<0.01 vs day 0). A strong positive correlation monoiodoacetate in higher doses and multiple was observed between scores obtained by the applications unlike Stobie et al. (1994), which did three systems (Table 5). not manage to induce OA with 0.5 mg/kg and Goniometric analysis provided evidence for 0.375 mg/kg MIA in this animal species. thigh muscles atrophy of the left hindlimb (Table According to Beyreuther et al. (2007), a single 4) as thigh circumference decreased significantly low MIA dose results only in transient synovitis, from 32+1 cm in the beginning to 23+1 cm by the th th resolving by the 14 day. Our experiments 105 day (p<0.01). The values between left and evidenced that higher and repeated doses (5-10 right limb were also statistically significantly mg/kg) succeeded to reproduced all symptoms of different (p<0.05 by days 30 and 60 and p<0.01 osteoarthritis: acute inflammation at onset, by day 105). The stifle joint circumference did progressive degeneration, transition to chronic not show significant differences with time. The atrophic phase, similarly to conclusions of range of motion (ROM) of left joints decreased Guinamp et al. (1997), that only a sufficient considerably from 115+2 in the beginning to amount of MIA resulted in rapid decline in the 95+7, 91+2, and 83+4 by days 30, 60 and 105, locomotor function of rat stifles and that low respectively (p<0.01). ROM of the left joint doses provoked a transient effect. Bove et al. exhibited a negative correlation with clinical (2003) demonstrated a relationship between the scores: r=–0.61, p<0.001 with Grisneaux's and www.veterinaryworld.org Veterinary World, Vol.5 No.3 March 2012 141 Parameter Days after the first MIA injection 0 30 60 105 Grisneaux' score 6±1 14±2** 12±2** 11 ± 3 Cross' score 4±0 13±1** 12±1** 9±1** Budsberg' score 4±0 17±1** 13±1** 11 ±1** Thigh circumference, (cm) left 32±1 27±1* 27±1** 23±1** right 32±1 32±1# 30±1# 30±1## Stifle joint circumference, (cm) left 25±1 27±1 26±1 25±1 right 25±1 23±1 23±1 23±1 Range of motion, (o) left 115±2 95±7** 91±2** 83±4** right 115±2 111±5 113±3## 110 ±4## Таble-4. Clinical scores and goniometric parameters in dogs with experimental monoiodoacetate (MIA) model of stifle joint osteoarthritis (mean + SEM; n=6). # ## *p<0.05;**p<0.01 vs baseline (day 0); p<0.05; p<0.01 between left (OA) and right (control) joints GSc CSc BSc TCir SCir ROM GSc ------r = 0.88 p<0.001 r = 0.83 p<0.001 r = –0.27 r = –0.61 p<0.001 r = 0.30 p<0.001 Csc r = 0.88 p<0.001 ------r = 0.92 p<0.001 r = –0.37 p<0.001 r = –0.63 p<0.001 r = 0.25 Bsc r = 0.83 p<0.001 r = 0.92 *** ------r = –0.32 p<0.01 r = –0.61 p<0.001 r = 0.33 p<0.001 Tcir r = –0.27 r = –0.37 *** r = –0.32 ** ------r = 0.36 p<0.001 r = 0.46 p<0.001 Scir r = –0.61 p<0.001 r = –0.63 *** r = –0.61 *** r = 0.36 p<0.001 ------r = –0.33 p<0.001 ROM r = 0.30 p<0.01 r = 0.26 r = 0.33 *** r = 0.46 p<0.001 r = –0.33 p<0.001 ------Таble-5. Correlation coefficients between clinical and goniometric parameters in dogs with experimental monoiodoacetate model of osteoarthritis GSc – clinical score according to Grisneaux et al. (1999); CSc clinical score according to Cross et al. (1997); BSc clinical score according to Budsberg et al. (1999); TCir – thigh circumference; SCir – stifle joint circumference; ROM – range of motion. Clinical changes in sodium monoiodoacetate–induced stifle osteoarthritis model in dogs locomotor impairment in rats, MIA concentrations goniometric data and the correlation between them.and the cumulative effect occurring with time.AcknowledgementsAnimal behaviour trials showed that jointMany thanks to the team of the Smallpain and allodynamic were commonly observedAnimal Clinic at the Faculty of Veterinaryafter experimentally induced OA and therefore,Medicine, Stara Zagora for their assistance inthey were recommended for OA pain assessmentperforming the experiments.(Bove, 2006). They are even more appropriate for chemical OA model, as the one used in the present Conflict of intereststudy, because the joint instability obtained afterAuthors declare that they have no conflict ofthe mechanical models could be ruled out as ainterest.cause for occurring kinematic changes.Since the structural integrity of cartilage Referencesrelies on the normal functioning of chondrocytes,1. Anderson J. (2011). The Stifle. Surgical anatomy.intra-articular injection with MIA producesApproaches to the stifle. Specific conditionscartilage degeneration and subchondral bone affecting the stifle joint. In: Houlton J. anddisorders corresponding to histopathology of Collinson R. Editors. Manual of Small AnimalOA. As this degenerative model progresses, the Arthrology, BSAVA, ch. 16:267-280.subchondral bone becomes exposed joint 2. Bendele A. (2001) Animal models of osteo-impairment and mechanical hypersensitivity arthritis. 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