Osteoarthritis: Trends and clinical implications in the standardization of a pathological induction protocol in rats using a single dose of monosodium iodoacetate.

Osteoarthritis: Trends and clinical implications in the standardization of a pathological induction protocol in rats using a single dose of monosodium iodoacetate. | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Osteoarthritis: Trends and clinical implications in the standardization of a pathological induction protocol in rats using a single dose of monosodium iodoacetate. Luis Angelo Santiago, Laércio Dias, Eduardo de Sousa This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8683676/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Osteoarthritis (OA) is a disease that causes progressive erosion of articular cartilage. Although radiographic imaging, histological and behavioral testing are widely used in preclinical research, few studies have systematically examined their interdependence. Induce OA using a single dose of MIA and to monitor the progression of joint damage over the course of 28 days, associating changes in clinical tests with radiographic and histological images. For that we evaluated the animals through clinical and radiological examinations in the 0, 3, 7, 14, 21, and 28 days and, after the end of each group period, the animals were euthanized, the joints being removed for histopathological analysis. Statistical analyzes employed for repeated measurements on outcome variables, ensuring methodological rigor with blinded assessors. We observe that a single dose of MIA in the rats had its peak of articular cartilage destruction on the 14th day of induction, as proven by clinical tests and radiographic and histological images. These findings suggest that a single-dose MIA induction model can be developed with high reproducibility and precision in an efficient experimental timeframe, which is valuable for future research assessing the therapeutic efficacy of potential agents for OA treatment. Statistical analyses employed analysis of variance with post hoc testing, ensuring methodological rigor with blinded evaluators. Health sciences/Pathogenesis Biological sciences/Physiology Osteoarthritis Induction Clinical Tests Radiographic Images Histological Images Figures Figure 1 Figure 2 Summary statement We analyzed and we follow the induction of OA with a single dose of MIA and monitored the progression of joint damage over 28 days, associating changes in clinical examinations with radiographic and histological images. INTRODUCTION Osteoarthritis (OA) is a chronic progressive disease that causes joint degeneration and loss of the articular cartilage surface, leading to an inflammatory process and localized pain, thus compromising joint integrity 1 . OA is a rheumatic disease with a high incidence, ranking among the most common causes of chronic pain worldwide. It has an enormous psychosocial impact, affecting not only the patients, but also their families and professional careers 1 . Experimental induction models play a key role in the development of new intervention strategies for OA, providing valuable insights into the mechanisms underlying the disease. One such model is monosodium iodoacetate ( MIA), first described by Kalbhen and Blum in 1977 and subsequently standardized as a model for joint arthrosis 2 . MIA is a reagent that modifies cysteine residues of proteins. When administered into the joints of animals, MIA causes a change in proteoglycan concentration in the articular cartilage, disrupting glycolysis and breaking down the metabolism of chondrocytes, thus resulting in joint degradation. Once administered intra-articularly into the knee and present in the synovial fluid, MIA is absorbed by chondrocytes together with specialized cells of the joint capsule, causing chronic degradation of the cartilage, apparently offering a more predictive model for the study of OA in humans. Therefore, the aim of this study was to induce OA using a single dose of MIA and to monitor the progression of joint damage over the course of 28 days, associating changes in clinical tests with radiographic and histological images. Although radiographic imaging, histological and behavioral testing are widely used in preclinical research, few studies have systematically examined their interdependence. Therefore, the aim of this study was to induce OA using a single dose of MIA and to monitor the progression of joint damage over the course of 28 days, associating changes in clinical tests with radiographic and histological images. RESULTS Clinical Trials - Joint edema Joint edema assessment was performed using circumferential measurements at multiple time points during the experimental protocol. On day 14, healthy the rats presented maintained the highest scores (mean 20.32 ± 3.62; p = 0.001) with statistically significant values ​​when compared with the pre-induction moment (mean 6.07 ± 0.14; p = 0.001), indicating a high degree of joint edema. (Fig. 1 A). Clinical Trials - Motor activity assessment (Rotarod test) Motor coordination was evaluated using the rotarod test at multiple time points during the experimental protocol. The assessment of the animals’ ambulation demonstrated impaired gait with statistically significant values when comparing day 0 (mean 5.0 ± 1.82; p = 0.001) with the other time points, day 7 (mean 1.0 ± 0.47; p = 0.001) and day 14 (mean 1.0 ± 0.62; p = 0.001), indicating gait impairment. (Fig. 1 B). Clinical course and pathophysiological changes in radiographic and histological images of MIA-induced OA Radiographic and histological examinations of rat knees showed differences in the images at the respective induction time points, indicating progression of joint damage in both images. Radiographic assessments using the Kellgren-Lawrence (KL) grading method for arthrosis revealed grade 0 immediately after induction (day 0), characterized by smooth and regular joint surfaces with normal opacity. The radiographs also showed a mineralized structure in the intra-articular space of the knee at the cranial and caudal aspects of the meniscus, with the patella overlapping its respective trochlear groove, and no signs of inflammatory changes or open fissure plates, compatible with the animal’s age (Fig. 2 : 1A ). On days 3 and 7 after induction, the joint exhibited a grade 3 score, with slight narrowing of the intra-articular space and slight sclerosis of the subchondral bone (Fig. 2 : 2B-2C ). On the 14th day of induction, joint damage reached its peak, with extensive joint destruction (grade 4 score). Notable findings included prominent marginal osteophytes, narrowing of the intra-articular spaces, subchondral bone sclerosis, cartilage erosion, partial loss of the normal appearance of the structures, increased synovial volume, and medial patellar dislocation, also observed on days 21 and 28 (Fig. 2 : 2D-2E-2F ). In the histological analysis, using the OARSI score, days 0 and 3 of induction exhibited structural integrity of the joint (score of 0), showing a regular surface, preserved mark lines, chondrocytes cells viable with presence of a nucleus (Fig. 2 : 1A-1B ). From day 7 onwards, joint damage showed a high peak, classified as grade 4 according to the OARSI score, in addition to erosion and excavation with loss of the superficial layer of the matrix and intermediate zone, loss of the Mark line, and further progression in the subsequent two days after induction. This score was maintained on days 14, 21, and 28 (Fig. 2 : 1C-1D-1E-1F ). DISCUSSION Guzman 7 stated OA induction was first described by Kalbhen and Blum in 1977. The findings of this experimental OA study, we were able to accurately observe all the stages of progression of OA induced by a single dose of MIA, both in clinical tests and in radiographic and histological images. Tou our knowledge, previous studies have implemented induction protocols that primarily focused on either clinical tests or radiographic and histological images, but rarely in combination. Our study is the first to investigate the progression and sequential evolution of OA induced by a single dose of MIA in the tibiofemoral joint of rats and to correlate clinical tests with radiographic and histological images. The clinical assessment of joint edema revealed a peak on the 14th day of induction. A peak was also observed for functional gait impairment between the 7th and 14th days of induction. The radiographic and histological images corroborated the findings of the clinical tests, in which destruction of the tibiofemoral joint peaked between the 7th and 14th days of induction. We were then able to reproduce the progression of OA from the moment of induction with a single dose of MIA, from day 0, including the peak of the highest incidence of the lesion, to day 28. Many researchers have described either the clinical changes or the radiographic and histological evidence of damage to the tibiofemoral articular cartilage after MIA injection. 8–13 . Therefore, while previous studies that induced OA using MIA injections have observed joint damage from a clinical/behavioral perspective or in radiographic and histological images, they have not provided chronological, sequential, and precise findings such as those described by Pitcher, Sousa-Valente, and Malcangio 14 . These authors investigated behavioral changes associated with hypersensitivity during the various stages of OA induction using three MIA concentrations (0.5 mg, 0.75 mg, and 1.0 mg) and over 28 days after induction. The authors concluded that a dose of 1.0 mg was ideal for producing similar joint damage in humans and that the peak in behavioral changes, as assessed by the clinical sensitivity test, occurred on the 10th day of induction. Their finding is consistent with what we founda, peak between the 7th and 14th days of induction for the clinical tests. Takahashi 15 conducted a study that analyzed only histological images and the progression of joint damage using two concentrations of MIA (0.2 mg and 1.0 mg) to induce OA in rat knee joints This is also in line with our findings. The authors concluded that 1 mg of MIA caused larger joint damage than the dose of 0.2 mg, histological images at 4 weeks of induction compatible with OA, in addition to an increase in the progression of OA at 12 weeks of induction, with no specification of when the peak occurred. Therefore, we observed an interrelationship between clinical tests, radiological images and histological images from the 14th day of induction. The findings demonstrated that a single dose of MIA in the rat had its peak of articular cartilage destruction on the 14th day of induction, proven through by clinical tests (joint edema and impaired walking)and radiographic and histological images (presence of large marginal osteophyte formation, narrowing of the intra-articular spaces, subchondral bone sclerosis, cartilage erosion, and erosion and excavation with loss of the superficial layer of the matrix and of the intermediate zone, and loss of Mark lines). These findings suggest that a single-dose MIA induction model can be developed with high reproducibility and precision in an efficient experimental timeframe, which is valuable for future research assessing the therapeutic efficacy of potential agents for OA treatment. MATERIALS AND METHODS Animals and ethical aspects Eighteen 8-week-old female Wistarrats ( Rattus norvegicus ) weighing 250-300 g, obtained from the UFMA Central Animal Facility, were used. The rats were fed standard chow and received water ad libitum throughout the experimental period.They were kept under controlled conditions of temperature (23 ± 1º C) and humidity (40-60%) and under a 12-hour light-dark cycle. At the end of each experimental group, euthanasia was performed by deep anesthesia via the intraperitoneal route at a dosage of 0.3 ml of Xylazine (80mL/kg) + 0.3 ml of Ketamine (10mg/kg). Ethical statement The research adhered to the ARRIVE standards and complied with the U.K. Animals (Scientific Procedures) Act of 1986 and the relevant EU Directive 2010/63/EU for animal experimentation. All experimental evaluation protocols were approved and authorized by the Animal Use Ethics Committee (CEUA) of the Federal University of Maranhão (UFMA), process n o . 23115.031386/2019-28. The experiments were conducted in accordance with the Brazilian College of Animal Experimentation (COBEA) guidelines for the use of animals in research. OA induction by MIA The rats were previously sedated with 10% ketamine hydrochloride ® (90 mg/kg) and then anesthetized using 2% xylazine hydrochloride ® (5 mg/kg) intraperitoneally. Once the anesthesia was confirmed, the rat knee was flexed at a 90° angle, a topical solution of 10% povidone-iodine was applied for local asepsis, and OA was induced by administering 2 mg/25 μL of MIA (Sigma-Aldrich, St. Louis, MO, USA) into the right tibiofemoral joint, through the patellar ligament 3 . Experimental design The experimental groups were assessed at 0, 3, 7, 14, 21, and 28 days after induction, with three animals (n=3) per group. On day 0, OA was induced in all groups by a single intra-articular injection of MIA at 2 mg/kg. The animals were then monitored for 28 days. After induction, each group underwent clinical assessments, consisting of joint circumference measurements and walk tests, followed by radiographic imaging in the groups 0, 3, 7, 14, 21 and 28 days. After reaching their respective time points (0, 3, 7, 14, 21, or 28 days). After the end of each group period, the animals were euthanized for histological analysis. The tibiofemoral joints were excised and fixed in 10% formaldehyde + phosphate buffered saline (PBS) for slide preparation and subsequent analysis. Clinical tests Joint edema Joint edema was measured using a stainless steel universal digital caliper (MTX ® ) for measurements up to 150 mm, quantifying joint thickness resulting from the inflammatory response in MIA-induced OA. Following the experimental schedule, the animals were evaluated at 0, 3, 14, 21, and 28 days after induction. Knee joint thickness was measured three times, with the animals under anesthesia during preparation for euthanasia. Motor activity assessment - forced walking ( Rotarod test ) The animals were placed on a Rotarod (IITC Life Science, CA, USA), a rotor-like device, at a speed of 4 to 40 rpm for 300s. Two preliminary training tests were carried out followed by two recorded evaluations. After induction, three evaluations were performed and the average latency was recorded. The animals rested for 15 min between assessments. The data were expressed as walk values and were coded by a single examiner. Observational analysis was conducted using a 5-point numerical scale, where 1 indicated normal limb use; 2, slight limping; 3, severe limping; 4, intermittent disuse of the affected limb; and 5, complete disuse of the limb 4 . Radiographic imaging Radiographic images were acquired in the anteroposterior and laterolateral planes, using a portable digital X-ray machine equipped with an image capture sensor (Diox ® ). The images were analyzed by a professional veterinarian with expertise in animal imaging. After evaluation, each knee was graded according to the Kellgren-Lawrence (KL) system for OA (0 - normal, 1 - doubtful, 2 - minimal, 3 - moderate, and 4 - severe). The average score was then calculated for comparison 5 . Histological analysis The animals were euthanized as scheduled, i.e., at 0, 3, 7, 14, 21, and 28 days, after completion of the clinical assessments and radiographic imaging. For histological analysis, the tibiofemoral joint was excised and then fixed in 10% formaldehyde + PBS. The specimens were decalcified by immersion in 20% ethylenediaminetetraacetic acid (EDTA) for 20 days. The specimens were then embedded in paraffin blocks, cut into 5 µm sections, and the organic cartilage matrices were stained with hematoxylin-eosin (H&E). The analysis was carried out blindly by a veterinary histopathologist. The histopathological assessment of cartilage followed the protocols adapted from the Osteoarthritis Research Society International (OARSI) guidelines 6 . The parameters correspond to the stages of OA, focusing on the extent of the articular cartilage surface, area, or volume involved in local OA. The degree of OA was determined according to the depth of the pathology within the cartilage, with points assigned on a scale ranging from 0 (intact surface) to 6 (loss of full-thickness cartilage and bone deformation). Statistical analysis Statistical analysis was performed using GraphPad Prism 9.0 software (San Diego, CA, USA). The Shapiro-Wilk normality test was performed, indicating a p > 0.05 for parametric tests. All experiments were performed independently at least three times, and data were presented as mean ± standard deviation (SD). Statistical analyses were performed using GraphPad Prism 9.0 software (San Diego, CA, USA). One-way ANOVA for repeated measurements was applied for moments 3, 7, 14, 21 and 28 induction days. Abbreviations Hematoxylin-Eosin (H&E); Kellgren-Lawrence (KL); Monosodium Iodoacetate , MIA; Osteoarthritis, OA; Osteoarthritis Research Society International, OARSI. Declarations Conflicts of interest The authors declare that there are no conflicts of interest in the collection, analysis or interpretation of the data; in the writing of the manuscript, or in the decision to publish the results. Data declaration Data is available upon request. Author Contributions All authors were involved with the design of the study, interpretation of the data, critical review of the manuscript and approval of the final version for submission. LÂMS, LSD and EM de S : Conceptualization, Methodology, Software. LÂMS, LSD and EM de S : Data curation, Writing - Preparation of the original draft. LÂM S and L S D Visualization. LÂMS , ( [email protected] ), LSD ( [email protected] ) and EM de S ( [email protected] ) take full responsibility for the integrity of the work from the beginning to the finished manuscript. Acknowledgements This research was funded by the Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão (FAPEMA) under grant numbers FAPEMA-INFRA-02012/21. The funding bodies had no role in the design of the study; in the collection, analysis, or interpretation of data; in the writing of the manuscript; or in the decision to submit the manuscript for publication. DATA AVAILABILITY The data is in Excel documents and stored in a Google Drive affiliated with the Federal University of Maranhão and can be made available to anyone upon request for access. References Carvalho, F.M. Costa, M.C. & Silva, T.C.D. Doenças reumáticas no Brasil: revisão de estudos epidemiológicos. Efdeportes.com, Revista Digital, 184(18), 1-1, 2018. Kalbhem, D. A.; Blum, U. [Hypothesis and experimental confirmation of a new pharmacological model of osteoarthrosis. Arzneimittel-Forschung, v. 27, n. 3, p. 527–31, jan. 1977. Silva, A.; Andersen, M.L; Tufik, S. Padrão de sono em um modelo experimental de osteoartrite. Dor. 2008,140, 446-455. Tonussi, C.R.; Ferreira, S.H. Rat knee-joint carrageenin incapacitation test: an objective screen for central and peripheral analgesics. Pain. 1992, 48, 421–427. Keyes, et al . The radiographic classification of medial gonarthrosis. The radiographic classification of medial gonarthrosis. Correlation with operation methods in 200 knees . Acta Orthop Scand. 1992, 63-5,497-501. Pritzker, K.P.H.; Gay, S.; Jimenez, S.A.; Ostergaard, K.; Pelletier, J.P.; Revell, P.A.; Salter, D.; Pathxxx, F.R.C. and Van den Berg, W.B. Osteoarthritis cartilage histopathology: grading and staging. Osteoarthritis and Cartilage. 2006, 14, 1. Guzman RE, Evans MG, Bove S, Morenko B, Kilgore K. Mono-iodoacetate-induced histologic changes in subchondral bone and articular cartilage of rat femorotibial joints: an animal model of osteoarthritis. Toxicol Pathol. 2003; 31: 619–624. https://doi.org/10.1080/01926230390241800 PMID: 14585729. Bove SE, Calcaterra SL, Brooker RM, Huber CM, Guzman RE, Juneau PL, et al. Weight bearing as a measure of disease progression and efficacy of anti-inflammatory compounds in a model of monoso- dium iodoacetate-induced osteoarthritis. Osteoarthritis Cartilage. 2003; 11: 821–830. PMID: 14609535. Janusz MJ, Hookfin EB, Heitmeyer SA, Woessner JF, Freemont AJ, Hoyland JA, et al. Moderation of iodoacetate-induced experimental osteoarthritis in rats by matrix metalloproteinase inhibitors. Osteoarthritis Cartilage. 2001; 9: 751–760. https://doi.org/10.1053/joca.2001.0472 PMID: 11795995. Dunham J, Hoedt-Schmidt S, Kalbhen DA. Structural and metabolic changes in articular cartilage induced by iodoacetate. Int J Exper Pathol. 1992; 73: 455–464. Moon SJ, Woo YJ, Jeong JH, Park MK, Oh HJ, Park JS, et al. Rebamipide attenuates pain severity and cartilage degeneration in a rat model of osteoarthritis by downregulating oxidative damage and cata- bolic activity in chondrocytes. Osteoarthritis Cartilage. 2012; 20: 1426–1438. https://doi.org/10.1016/j. joca.2012.08.002 PMID: 22890185. Chaplan, S. R. et al . Quantitative assessment of tactile allodynia in the rat paw. Journal of neuroscience methods. 1994, 53-1, 55–63. Servin, E.T.N.; Czeczko, N.G.; Malafaia, O.; Torres, O. J. M.; Lima, F.C.V.M.; Silva, G. E. B.; Cartágenes, M. S. S.; Garcia, J. B. S. Effect of Arrabidaea Chica Verlot Hydroalcoholic Extract on Monosodium Iodoacetate-Induced Osteoarthritis of Rat Knees. Brazilian Journal of health Review. 2020, 3-3, 7038-7057. Pitcher T.; Sousa-Valente J. e Malcangio M. The Monoiodoacetate Model of Osteoarthritis Pain in the Mouse. JOVE, Journal of Visualized Experiments, 2016. http://www.jove.com/video/53746 Takahashi, I; Matsuzaki, T; Kuroki, H Hoso, M. Induction of osteoarthritis by injecting monosodium iodoacetate into the patellofemoral joint of an experimental rat model. PLOS ONE, 2018. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8683676","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":582154052,"identity":"3d46d3f2-46c1-4889-b48c-0eb41c13433e","order_by":0,"name":"Luis Angelo Santiago","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA2UlEQVRIie3OMQrCMBSA4VcCdYl2jSh4hbp0UnuVhEA9gKMOgUKctLPgOdwlUA8RB0VwcugkTmJQ7NhkFMw/hDfk4z0An+8H60AgPlMLghOMzEAsJKwJAhRD5kS+IQiJGyE8r2BxnBQIlXNMdwPorU4WwiSB8so3eZhpTPVQ9A+xlZhX8VjhRLcfOhAksx3GzGFPxVNDZmZL6kIECaSaxAgnyBBmJ/gsCVsrSlTIu1uqueyXzSRqTS9VdVdptMz31Y3qcdGTzeQdBWCi3usA3qWuH30+n+8PewETkz9bpMdibAAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-3943-8670","institution":"Universidade Federal do Maranhão","correspondingAuthor":true,"prefix":"","firstName":"Luis","middleName":"Angelo","lastName":"Santiago","suffix":""},{"id":582154053,"identity":"035300cb-87c5-48ca-89ad-86763005e1b4","order_by":1,"name":"Laércio Dias","email":"","orcid":"","institution":"Federal University of Maranhão","correspondingAuthor":false,"prefix":"","firstName":"Laércio","middleName":"","lastName":"Dias","suffix":""},{"id":582154054,"identity":"5f8031dc-b33c-4bc0-83c0-a795745c2747","order_by":2,"name":"Eduardo de Sousa","email":"","orcid":"","institution":"Universidade UniCEUMA","correspondingAuthor":false,"prefix":"","firstName":"Eduardo","middleName":"","lastName":"de Sousa","suffix":""}],"badges":[],"createdAt":"2026-01-24 04:25:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8683676/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8683676/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":103439742,"identity":"6093882e-5386-440f-a425-06b1b03ef2e1","added_by":"auto","created_at":"2026-02-25 17:05:53","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":506554,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eA and 1B \u003c/strong\u003e- \u003cstrong\u003eClinical tests for joint edema and gait in rats subjected to the monosodium iodoacetate-induced experimental osteoarthritis protocol. Analysis was conducted at experimental time points 0 (pre-induction), 3, 7, 14, 21, and 28 days post-induction. Data are presented as mean values, standard deviations and statistic (p \u0026lt; 0.001)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"CLINICALTESTS.001.png","url":"https://assets-eu.researchsquare.com/files/rs-8683676/v1/335420135012aabd1e69d7ca.png"},{"id":103439743,"identity":"f24bde57-3b93-4286-a368-b1b52b927a0b","added_by":"auto","created_at":"2026-02-25 17:05:53","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1081667,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eRadiographic and histopathological changes in the articular cartilage of the right knee of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eRattus norvegicus \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eup to 28 days after MIA-induced OA.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Radiographicandhistopathologicaltiff.png","url":"https://assets-eu.researchsquare.com/files/rs-8683676/v1/43e7952bcc6a375312250f63.png"},{"id":103439762,"identity":"01b69428-0e38-4cd2-ab0e-7fe2987e4bec","added_by":"auto","created_at":"2026-02-25 17:05:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2750922,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8683676/v1/17fdd511-a2b1-4fe5-940e-e4eab38011f6.pdf"},{"id":103439744,"identity":"a9819e0e-68dd-4b36-8d97-c47e7261accf","added_by":"auto","created_at":"2026-02-25 17:05:53","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":2098232,"visible":true,"origin":"","legend":"Highlights","description":"","filename":"Highlights3.docx","url":"https://assets-eu.researchsquare.com/files/rs-8683676/v1/8b6f5f9105153c8de3767df1.docx"}],"financialInterests":"There is no conflict of interest","formattedTitle":"Osteoarthritis: Trends and clinical implications in the standardization of a pathological induction protocol in rats using a single dose of monosodium iodoacetate.","fulltext":[{"header":"Summary statement","content":"\u003cp\u003eWe analyzed and we follow the induction of OA with a single dose of MIA and monitored the progression of joint damage over 28 days, associating changes in clinical examinations with radiographic and histological images.\u003c/p\u003e"},{"header":"INTRODUCTION","content":"\u003cp\u003eOsteoarthritis (OA) is a chronic progressive disease that causes joint degeneration and loss of the articular cartilage surface, leading to an inflammatory process and localized pain, thus compromising joint integrity\u003csup\u003e\u003cb\u003e1\u003c/b\u003e\u003c/sup\u003e. OA is a rheumatic disease with a high incidence, ranking among the most common causes of chronic pain worldwide. It has an enormous psychosocial impact, affecting not only the patients, but also their families and professional careers\u003csup\u003e\u003cb\u003e1\u003c/b\u003e\u003c/sup\u003e. Experimental induction models play a key role in the development of new intervention strategies for OA, providing valuable insights into the mechanisms underlying the disease. One such model is monosodium iodoacetate \u003cb\u003e(\u003c/b\u003eMIA), first described by Kalbhen and Blum in 1977 and subsequently standardized as a model for joint arthrosis\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e. MIA is a reagent that modifies cysteine residues of proteins. When administered into the joints of animals, MIA causes a change in proteoglycan concentration in the articular cartilage, disrupting glycolysis and breaking down the metabolism of chondrocytes, thus resulting in joint degradation. Once administered intra-articularly into the knee and present in the synovial fluid, MIA is absorbed by chondrocytes together with specialized cells of the joint capsule, causing chronic degradation of the cartilage, apparently offering a more predictive model for the study of OA in humans.\u003c/p\u003e \u003cp\u003eTherefore, the aim of this study was to induce OA using a single dose of MIA and to monitor the progression of joint damage over the course of 28 days, associating changes in clinical tests with radiographic and histological images.\u003c/p\u003e \u003cp\u003eAlthough radiographic imaging, histological and behavioral testing are widely used in preclinical research, few studies have systematically examined their interdependence. Therefore, the aim of this study was to induce OA using a single dose of MIA and to monitor the progression of joint damage over the course of 28 days, associating changes in clinical tests with radiographic and histological images.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cp\u003e \u003cb\u003eClinical Trials -\u003c/b\u003e Joint edema\u003c/p\u003e \u003cp\u003eJoint edema assessment was performed using circumferential measurements at multiple time points during the experimental protocol. On day 14, healthy the rats presented maintained the highest scores (mean 20.32\u0026thinsp;\u0026plusmn;\u0026thinsp;3.62; p\u0026thinsp;=\u0026thinsp;0.001) with statistically significant values ​​when compared with the pre-induction moment (mean 6.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14; p\u0026thinsp;=\u0026thinsp;0.001), indicating a high degree of joint edema. (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eClinical Trials -\u003c/b\u003e Motor activity assessment (Rotarod test)\u003c/p\u003e \u003cp\u003eMotor coordination was evaluated using the rotarod test at multiple time points during the experimental protocol. The assessment of the animals\u0026rsquo; ambulation demonstrated impaired gait with statistically significant values when comparing day 0 (mean 5.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.82; p\u0026thinsp;=\u0026thinsp;0.001) with the other time points, day 7 (mean 1.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47; p\u0026thinsp;=\u0026thinsp;0.001) and day 14 (mean 1.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.62; p\u0026thinsp;=\u0026thinsp;0.001), indicating gait impairment. (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eClinical course and pathophysiological changes in radiographic and histological images of MIA-induced OA\u003c/h2\u003e \u003cp\u003eRadiographic and histological examinations of rat knees showed differences in the images at the respective induction time points, indicating progression of joint damage in both images. Radiographic assessments using the Kellgren-Lawrence (KL) grading method for arthrosis revealed grade 0 immediately after induction (day 0), characterized by smooth and regular joint surfaces with normal opacity. The radiographs also showed a mineralized structure in the intra-articular space of the knee at the cranial and caudal aspects of the meniscus, with the patella overlapping its respective trochlear groove, and no signs of inflammatory changes or open fissure plates, compatible with the animal\u0026rsquo;s age (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e: \u003cb\u003e1A\u003c/b\u003e). On days 3 and 7 after induction, the joint exhibited a grade 3 score, with slight narrowing of the intra-articular space and slight sclerosis of the subchondral bone (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e: \u003cb\u003e2B-2C\u003c/b\u003e). On the 14th day of induction, joint damage reached its peak, with extensive joint destruction (grade 4 score). Notable findings included prominent marginal osteophytes, narrowing of the intra-articular spaces, subchondral bone sclerosis, cartilage erosion, partial loss of the normal appearance of the structures, increased synovial volume, and medial patellar dislocation, also observed on days 21 and 28 (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e: \u003cb\u003e2D-2E-2F\u003c/b\u003e). In the histological analysis, using the OARSI score, days 0 and 3 of induction exhibited structural integrity of the joint (score of 0), showing a regular surface, preserved mark lines, chondrocytes cells viable with presence of a nucleus (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e: \u003cb\u003e1A-1B\u003c/b\u003e). From day 7 onwards, joint damage showed a high peak, classified as grade 4 according to the OARSI score, in addition to erosion and excavation with loss of the superficial layer of the matrix and intermediate zone, loss of the Mark line, and further progression in the subsequent two days after induction. This score was maintained on days 14, 21, and 28 (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e: \u003cb\u003e1C-1D-1E-1F\u003c/b\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eGuzman\u003csup\u003e\u003cb\u003e7\u003c/b\u003e\u003c/sup\u003e stated OA induction was first described by Kalbhen and Blum in 1977. The findings of this experimental OA study, we were able to accurately observe all the stages of progression of OA induced by a single dose of MIA, both in clinical tests and in radiographic and histological images.\u003c/p\u003e \u003cp\u003eTou our knowledge, previous studies have implemented induction protocols that primarily focused on either clinical tests or radiographic and histological images, but rarely in combination. Our study is the first to investigate the progression and sequential evolution of OA induced by a single dose of MIA in the tibiofemoral joint of rats and to correlate clinical tests with radiographic and histological images. The clinical assessment of joint edema revealed a peak on the 14th day of induction. A peak was also observed for functional gait impairment between the 7th and 14th days of induction. The radiographic and histological images corroborated the findings of the clinical tests, in which destruction of the tibiofemoral joint peaked between the 7th and 14th days of induction. We were then able to reproduce the progression of OA from the moment of induction with a single dose of MIA, from day 0, including the peak of the highest incidence of the lesion, to day 28.\u003c/p\u003e \u003cp\u003eMany researchers have described either the clinical changes or the radiographic and histological evidence of damage to the tibiofemoral articular cartilage after MIA injection.\u003csup\u003e\u003cb\u003e8\u0026ndash;13\u003c/b\u003e\u003c/sup\u003e. Therefore, while previous studies that induced OA using MIA injections have observed joint damage from a clinical/behavioral perspective or in radiographic and histological images, they have not provided chronological, sequential, and precise findings such as those described by Pitcher, Sousa-Valente, and Malcangio\u003csup\u003e\u003cb\u003e14\u003c/b\u003e\u003c/sup\u003e. These authors investigated behavioral changes associated with hypersensitivity during the various stages of OA induction using three MIA concentrations (0.5 mg, 0.75 mg, and 1.0 mg) and over 28 days after induction. The authors concluded that a dose of 1.0 mg was ideal for producing similar joint damage in humans and that the peak in behavioral changes, as assessed by the clinical sensitivity test, occurred on the 10th day of induction. Their finding is consistent with what we founda, peak between the 7th and 14th days of induction for the clinical tests.\u003c/p\u003e \u003cp\u003eTakahashi\u003csup\u003e\u003cb\u003e15\u003c/b\u003e\u003c/sup\u003e conducted a study that analyzed only histological images and the progression of joint damage using two concentrations of MIA (0.2 mg and 1.0 mg) to induce OA in rat knee joints This is also in line with our findings. The authors concluded that 1 mg of MIA caused larger joint damage than the dose of 0.2 mg, histological images at 4 weeks of induction compatible with OA, in addition to an increase in the progression of OA at 12 weeks of induction, with no specification of when the peak occurred.\u003c/p\u003e \u003cp\u003eTherefore, we observed an interrelationship between clinical tests, radiological images and histological images from the 14th day of induction.\u003c/p\u003e \u003cp\u003eThe findings demonstrated that a single dose of MIA in the rat had its peak of articular cartilage destruction on the 14th day of induction, proven through by clinical tests (joint edema and impaired walking)and radiographic and histological images (presence of large marginal osteophyte formation, narrowing of the intra-articular spaces, subchondral bone sclerosis, cartilage erosion, and erosion and excavation with loss of the superficial layer of the matrix and of the intermediate zone, and loss of Mark lines). These findings suggest that a single-dose MIA induction model can be developed with high reproducibility and precision in an efficient experimental timeframe, which is valuable for future research assessing the therapeutic efficacy of potential agents for OA treatment.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003e\u003cstrong\u003eAnimals and ethical aspects\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEighteen 8-week-old female Wistarrats (\u003cem\u003eRattus norvegicus\u003c/em\u003e) weighing 250-300 g, obtained from the UFMA Central Animal Facility, were used. The rats were fed standard chow and received water \u003cem\u003ead libitum\u0026nbsp;\u003c/em\u003ethroughout the experimental period.They were kept under controlled conditions of temperature (23 ± 1º C) and humidity (40-60%) and under a 12-hour light-dark cycle. At the end of each experimental group, euthanasia was performed by deep anesthesia via the intraperitoneal route at a dosage of 0.3 ml of Xylazine (80mL/kg) + 0.3 ml of Ketamine (10mg/kg).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe research adhered to the ARRIVE standards and complied with the U.K. Animals (Scientific Procedures) Act of 1986 and the relevant EU Directive 2010/63/EU for animal experimentation. All experimental evaluation protocols were approved and authorized by the Animal Use Ethics Committee (CEUA) of the Federal University of Maranhão (UFMA), process n\u003csup\u003eo\u003c/sup\u003e. 23115.031386/2019-28. The experiments were conducted in accordance with the Brazilian College of Animal Experimentation (COBEA) guidelines for the use of animals in research.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eOA induction by MIA\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe rats were previously sedated with 10% ketamine hydrochloride\u003csup\u003e®\u003c/sup\u003e (90 mg/kg) and then anesthetized using 2% xylazine hydrochloride\u003csup\u003e®\u003c/sup\u003e (5 mg/kg) intraperitoneally. Once the anesthesia was confirmed, the rat knee was flexed at a 90° angle, a topical solution of 10% povidone-iodine was applied for local asepsis, and OA was induced by administering 2 mg/25 μL of MIA (Sigma-Aldrich, St. Louis, MO, USA) into the right tibiofemoral joint, through the patellar ligament\u003cstrong\u003e\u003csup\u003e3\u003c/sup\u003e\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental design\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe experimental groups were assessed at 0, 3, 7, 14, 21, and 28 days after induction, with three animals (n=3) per group. On day 0, OA was induced in all groups by a single intra-articular injection of MIA at 2 mg/kg. The animals were then monitored for 28 days. After induction, each group underwent clinical assessments, consisting of joint circumference measurements and walk tests, followed by radiographic imaging in the groups 0, 3, 7, 14, 21 and 28 days. After reaching their respective time points (0, 3, 7, 14, 21, or 28 days). After the end of each group period, the animals were euthanized for histological analysis. The tibiofemoral joints were excised and fixed in 10% formaldehyde + phosphate buffered saline (PBS) for slide preparation and subsequent analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical tests \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eJoint edema\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;Joint edema was measured using a stainless steel universal digital caliper (MTX\u003csup\u003e®\u003c/sup\u003e) for measurements up to 150 mm, quantifying joint thickness resulting from the inflammatory response in MIA-induced OA. Following the experimental schedule, the animals were evaluated at 0, 3, 14, 21, and 28 days after induction. Knee joint thickness was measured three times, with the animals under anesthesia during preparation for euthanasia.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMotor activity assessment - forced walking (\u003cem\u003eRotarod test\u003c/em\u003e)\u003c/p\u003e\n\u003cp\u003eThe animals were placed on a Rotarod (IITC Life Science, CA, USA), a rotor-like device, at a speed of 4 to 40 rpm for 300s. Two preliminary training tests were carried out followed by two recorded evaluations. After induction, three evaluations were performed and the average latency was recorded. The animals rested for 15 min between assessments. The data were expressed as walk values and were coded by a single examiner. Observational analysis was conducted using a 5-point numerical scale, where 1 indicated normal limb use; 2, slight limping; 3, severe limping; 4, intermittent disuse of the affected limb; and 5, complete disuse of the limb\u003cstrong\u003e\u003csup\u003e\u0026nbsp;4\u003c/sup\u003e\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRadiographic imaging\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eRadiographic images were acquired in the anteroposterior and laterolateral planes, using a portable digital X-ray machine equipped with an image capture sensor (Diox\u003csup\u003e®\u003c/sup\u003e\u003cstrong\u003e).\u0026nbsp;\u003c/strong\u003eThe images were analyzed by a professional veterinarian with expertise in animal imaging. After evaluation, each knee was graded according to the Kellgren-Lawrence (KL) system for OA (0 - normal, 1 - doubtful, 2 - minimal, 3 - moderate, and 4 - severe). The average score was then calculated for comparison \u003cstrong\u003e\u003csup\u003e5\u003c/sup\u003e\u003c/strong\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHistological analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe animals were euthanized as scheduled, i.e., at 0, 3, 7, 14, 21, and 28 days, after completion of the clinical assessments and radiographic imaging. For histological analysis, the tibiofemoral joint was excised and then fixed in 10% formaldehyde + PBS. The specimens were decalcified by immersion in 20% ethylenediaminetetraacetic acid (EDTA) for 20 days. The specimens were then embedded in paraffin blocks, cut into 5 µm sections, and the organic cartilage matrices were stained with hematoxylin-eosin (H\u0026amp;E). The analysis was carried out blindly by a veterinary histopathologist. The histopathological assessment of cartilage followed the protocols adapted from the Osteoarthritis Research Society International (OARSI) guidelines\u003cstrong\u003e\u003csup\u003e6\u003c/sup\u003e\u003c/strong\u003e. The parameters correspond to the stages of OA, focusing on the extent of the articular cartilage surface, area, or volume involved in local OA. The degree of OA was determined according to the depth of the pathology within the cartilage, with points assigned on a scale ranging from 0 (intact surface) to 6 (loss of full-thickness cartilage and bone deformation).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStatistical analysis was performed using GraphPad Prism 9.0 software (San Diego, CA, USA). The Shapiro-Wilk normality test was performed, indicating a p \u0026gt; 0.05 for parametric tests. All experiments were performed independently at least three times, and data were presented as mean ± standard deviation (SD). Statistical analyses were performed using GraphPad Prism 9.0 software (San Diego, CA, USA). One-way ANOVA for repeated measurements was applied for moments 3, 7, 14, 21 and 28 induction days.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eHematoxylin-Eosin (H\u0026amp;E); Kellgren-Lawrence (KL); Monosodium Iodoacetate\u003cstrong\u003e,\u0026nbsp;\u003c/strong\u003eMIA; Osteoarthritis, OA; Osteoarthritis Research Society International, OARSI.\u0026nbsp;\u003c/p\u003e"},{"header":"Declarations","content":" \u003ch2\u003eConflicts of interest\u003c/h2\u003e \u003cp\u003eThe authors declare that there are no conflicts of interest in the collection, analysis or interpretation of the data; in the writing of the manuscript, or in the decision to publish the results.\u003c/p\u003e \u003ch2\u003eData declaration\u003c/h2\u003e \u003cp\u003eData is available upon request.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eAuthor Contributions\u003c/h2\u003e \u003cp\u003eAll authors were involved with the design of the study, interpretation of the data, critical review of the manuscript and approval of the final version for submission. \u003cb\u003eL\u0026Acirc;MS, LSD and EM de S\u003c/b\u003e: Conceptualization, Methodology, Software. \u003cb\u003eL\u0026Acirc;MS, LSD and EM de S\u003c/b\u003e: Data curation, Writing - Preparation of the original draft. \u003cb\u003eL\u0026Acirc;M S and L S D\u003c/b\u003e Visualization. \u003cb\u003eL\u0026Acirc;MS\u003c/b\u003e, ([email protected]), \u003cb\u003eLSD\u003c/b\u003e ([email protected]) and \u003cb\u003eEM de S\u003c/b\u003e ([email protected]) take full responsibility for the integrity of the work from the beginning to the finished manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e \u003cp\u003eThis research was funded by the Funda\u0026ccedil;\u0026atilde;o de Amparo \u0026agrave; Pesquisa e ao Desenvolvimento Cient\u0026iacute;fico e Tecnol\u0026oacute;gico do Maranh\u0026atilde;o (FAPEMA) under grant numbers FAPEMA-INFRA-02012/21. The funding bodies had no role in the design of the study; in the collection, analysis, or interpretation of data; in the writing of the manuscript; or in the decision to submit the manuscript for publication.\u003c/p\u003e\u003ch2\u003eDATA AVAILABILITY\u003c/h2\u003e \u003cp\u003eThe data is in Excel documents and stored in a Google Drive affiliated with the Federal University of Maranh\u0026atilde;o and can be made available to anyone upon request for access.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eCarvalho, F.M. Costa, M.C. \u0026amp; Silva, T.C.D. Doen\u0026ccedil;as reum\u0026aacute;ticas no Brasil: revis\u0026atilde;o de estudos epidemiol\u0026oacute;gicos. Efdeportes.com, Revista Digital, 184(18), 1-1, 2018.\u003c/li\u003e\n\u003cli\u003eKalbhem, D. A.; Blum, U. [Hypothesis and experimental confirmation of a new pharmacological model of osteoarthrosis. Arzneimittel-Forschung, v. 27, n. 3, p. 527\u0026ndash;31, jan. 1977.\u003c/li\u003e\n\u003cli\u003eSilva, A.; Andersen, M.L; Tufik, S. Padr\u0026atilde;o de sono em um modelo experimental de osteoartrite. Dor. 2008,140, 446-455.\u003c/li\u003e\n\u003cli\u003eTonussi, C.R.; Ferreira, S.H. Rat knee-joint carrageenin incapacitation test: an objective screen for central and peripheral analgesics. Pain. 1992, 48, 421\u0026ndash;427.\u003c/li\u003e\n\u003cli\u003eKeyes, \u003cem\u003eet al\u003c/em\u003e. The radiographic classification of medial gonarthrosis. The radiographic classification of medial gonarthrosis. \u003cem\u003eCorrelation with operation methods in 200 knees\u003c/em\u003e. Acta Orthop Scand. 1992, 63-5,497-501.\u003c/li\u003e\n\u003cli\u003ePritzker, K.P.H.; Gay, S.; Jimenez, S.A.; Ostergaard, K.; Pelletier, J.P.; Revell, P.A.; Salter, D.; Pathxxx, F.R.C. and Van den Berg, W.B. Osteoarthritis cartilage histopathology: grading and staging. Osteoarthritis and Cartilage. 2006, 14, 1.\u003c/li\u003e\n\u003cli\u003eGuzman RE, Evans MG, Bove S, Morenko B, Kilgore K. Mono-iodoacetate-induced histologic changes in subchondral bone and articular cartilage of rat femorotibial joints: an animal model of osteoarthritis. Toxicol Pathol. 2003; 31: 619\u0026ndash;624. https://doi.org/10.1080/01926230390241800 PMID: 14585729.\u003c/li\u003e\n\u003cli\u003eBove SE, Calcaterra SL, Brooker RM, Huber CM, Guzman RE, Juneau PL, et al. Weight bearing as a measure of disease progression and efficacy of anti-inflammatory compounds in a model of monoso- dium iodoacetate-induced osteoarthritis. Osteoarthritis Cartilage. 2003; 11: 821\u0026ndash;830. PMID: 14609535.\u003c/li\u003e\n\u003cli\u003eJanusz MJ, Hookfin EB, Heitmeyer SA, Woessner JF, Freemont AJ, Hoyland JA, et al. Moderation of iodoacetate-induced experimental osteoarthritis in rats by matrix metalloproteinase inhibitors. Osteoarthritis Cartilage. 2001; 9: 751\u0026ndash;760. https://doi.org/10.1053/joca.2001.0472 PMID: 11795995.\u003c/li\u003e\n\u003cli\u003eDunham J, Hoedt-Schmidt S, Kalbhen DA. Structural and metabolic changes in articular cartilage induced by iodoacetate. Int J Exper Pathol. 1992; 73: 455\u0026ndash;464.\u003c/li\u003e\n\u003cli\u003eMoon SJ, Woo YJ, Jeong JH, Park MK, Oh HJ, Park JS, et al. Rebamipide attenuates pain severity and cartilage degeneration in a rat model of osteoarthritis by downregulating oxidative damage and cata- bolic activity in chondrocytes. Osteoarthritis Cartilage. 2012; 20: 1426\u0026ndash;1438. https://doi.org/10.1016/j. joca.2012.08.002 PMID: 22890185.\u003c/li\u003e\n\u003cli\u003eChaplan, S. R. \u003cem\u003eet al\u003c/em\u003e. Quantitative assessment of tactile allodynia in the rat paw. Journal of neuroscience methods. 1994, 53-1, 55\u0026ndash;63.\u003c/li\u003e\n\u003cli\u003eServin, E.T.N.; Czeczko, N.G.; Malafaia, O.; Torres, O. J. M.; Lima, F.C.V.M.; Silva, G. E. B.; Cart\u0026aacute;genes, M. S. S.; Garcia, J. B. S. Effect of Arrabidaea Chica Verlot Hydroalcoholic Extract on Monosodium Iodoacetate-Induced Osteoarthritis of Rat Knees. Brazilian Journal of health Review. 2020, 3-3, 7038-7057.\u003c/li\u003e\n\u003cli\u003ePitcher T.; Sousa-Valente J. e Malcangio M. The Monoiodoacetate Model of Osteoarthritis Pain in the Mouse. JOVE, Journal of Visualized Experiments, 2016. http://www.jove.com/video/53746\u003c/li\u003e\n\u003cli\u003eTakahashi, I; Matsuzaki, T; Kuroki, H Hoso, M. Induction of osteoarthritis by injecting monosodium iodoacetate into the patellofemoral joint of an experimental rat model. PLOS ONE, 2018.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Osteoarthritis, Induction, Clinical Tests, Radiographic Images, Histological Images","lastPublishedDoi":"10.21203/rs.3.rs-8683676/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8683676/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eOsteoarthritis (OA) is a disease that causes progressive erosion of articular cartilage. Although radiographic imaging, histological and behavioral testing are widely used in preclinical research, few studies have systematically examined their interdependence. Induce OA using a single dose of MIA and to monitor the progression of joint damage over the course of 28 days, associating changes in clinical tests with radiographic and histological images. For that we evaluated the animals through clinical and radiological examinations in the 0, 3, 7, 14, 21, and 28 days and, after the end of each group period, the animals were euthanized, the joints being removed for histopathological analysis. Statistical analyzes employed for repeated measurements on outcome variables, ensuring methodological rigor with blinded assessors. We observe that a single dose of MIA in the rats had its peak of articular cartilage destruction on the 14th day of induction, as proven by clinical tests and radiographic and histological images. These findings suggest that a single-dose MIA induction model can be developed with high reproducibility and precision in an efficient experimental timeframe, which is valuable for future research assessing the therapeutic efficacy of potential agents for OA treatment. Statistical analyses employed analysis of variance with post hoc testing, ensuring methodological rigor with blinded evaluators.\u003c/p\u003e","manuscriptTitle":"Osteoarthritis: Trends and clinical implications in the standardization of a pathological induction protocol in rats using a single dose of monosodium iodoacetate.","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-25 17:05:43","doi":"10.21203/rs.3.rs-8683676/v1","editorialEvents":[{"type":"communityComments","content":1}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"aec2b15d-8d5c-4875-8673-1626074758f0","owner":[],"postedDate":"February 25th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":61934989,"name":"Health sciences/Pathogenesis"},{"id":61934990,"name":"Biological sciences/Physiology"}],"tags":[],"updatedAt":"2026-03-10T00:21:36+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-25 17:05:43","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8683676","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8683676","identity":"rs-8683676","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}