Exploring Ankle Sprain Dynamics: A New Experimental Model in WISTAR Rats
Main Article Content
Abstract
Background: Ankle sprain is one of the most common orthopedic injuries that results in pain, swelling and ankle instability, all of which greatly affect functional activities and gait. The present study aims to develop an experimental model of grade 2 ankle sprain in WISTAR rats in a cost-economical surgical setting, while ensuring good animal welfare and humane surgical interventions according to established guidelines and after obtaining ethical approval. Methods: Twelve four-week-old female WISTAR rats were divided into two groups namely, control (n=6) and surgery (n=6). Rats in the control group were used as reference, while rats in the surgery group underwent a minor surgical procedure to induce ankle sprain under a magnifying glass. The swelling was assessed using the figure-of-eight measurement and MRI. Functional activities were evaluated using open field test and spontaneous rears, and gait changes were analyzed and compared using step length asymmetry, stride length asymmetry, paw overlap asymmetry, hindfoot base width and forefoot base width during gait of the reference and ankle sprain rats. Results: Gait analysis revealed that ankle sprain rats demonstrated poorer locomotion, reduced spontaneous rears, reduction in contralateral step length, ipsilateral stride length, greater ipsilateral paw asymmetry and greater forelimb and hindlimb base width. Conclusion: We have successfully developed an ankle sprain model in WISTAR rats using our surgical protocol based on our outcome measures on inflammation, reduced functional activities and impairment of gait.
Downloads
Article Details
References
Gan QF, Foo CN, Leong PP, Cheong SK (2021) Incorporating regenerative medicine into rehabilitation programmes: a potential treatment for ankle sprain. https://doi.org/1012968/ijtr20190119. https://doi.org/10.12968/IJTR.2019.0119
Dalmau-Pastor M, El-Daou H, Stephen JM, Vega J, Malagelada F, Calder J (2023) Clinical Relevance and Function of Anterior Talofibular Ligament Superior and Inferior Fascicles: A Robotic Study. American Journal of Sports Medicine 51:2169–2175
Fu GQ, Wah YC, Sura S, Jagadeesan S, Chinnavan E, Judson JPE (2018) Influence of rearfoot alignment on static and dynamic postural stability. https://doi.org/1012968/ijtr20182512628 25:628–635
Xue X, Yu L, Zheng S, et al (2024) Insufficient recovery of proprioception in chronic ankle instability after surgical restabilization: A systematic review. Asia Pac J Sports Med Arthrosc Rehabil Technol 35:48–58
Kovaleski JE (2024) How Much Should I Be Concerned About Persistent Laxity Associated With the Lateqral Ankle Ligaments? Quick Questions in Ankle Sprains 165–169
Wisthoff BA, Docherty CL, Glutting J, Gustavsen G, Royer TD, Swanik C, Kaminski TW (2021) Identifying Range-of-Motion Deficits and Talocrural Joint Laxity After an Acute Lateral Ankle Sprain. J Athl Train 56:408–417
Fu Gan Q, Pooi Leong P, Keng Cheong S, Nien Foo C, Pooi Pooi L (2020) Regenerative Medicine as a Potential and Future Intervention for Ankle Sprain. Malaysian Journal of Medicine and Health Sciences 16:2636–9346
Gan QF, Leong PP, Cheong SK, Foo CN (2024) Incorporating Stem Cells Into Physical Rehabilitation. Reference Module in Biomedical Sciences. https://doi.org/10.1016/B978-0-443-15717-2.00075-5
Gan QF, Choy KW, Foo CN, Leong PP, Cheong SK (2021) Incorporating insulin growth Factor-1 into regenerative and personalised medicine for musculoskeletal disorders: A systematic review. J Tissue Eng Regen Med 15:419–441
Clarkson JM, Martin JE, McKeegan DEF (2022) A review of methods used to kill laboratory rodents: issues and opportunities. Lab Anim 56:419–436
Bloem JL, Reijnierse M, Huizinga TWJ, Van Der Helm-Van Mil AHM (2018) Viewpoint: MR signal intensity: staying on the bright side in MR image interpretation. RMD Open 4:728
Bakewell R, Gerety EL, Grainger AJ (2020) Basic Imaging Principles of Tendons and Ligaments. Med Radiol 99–119
Kang Y, Dianat S, Bencardino JT (2023) Anterior Cruciate Ligament Injuries: MR Imaging Diagnosis with Surgical Implications. ,;10(3):1-12 10:1–12
Võikar V, Stanford SC (2023) The Open Field Test. Neuromethods 190:9–29
Ferreira JS, Leite Junior JB, de Mello Bastos JM, Samuels RI, Carey RJ, Carrera MP (2022) A new method to study learning and memory using spontaneous locomotor activity in an open-field arena. J Neurosci Methods 366:109429
Gnanasegaran N, Govindasamy V, Simon C, Gan QF, Vincent-Chong VK, Mani V, Krishnan Selvarajan K, Subramaniam V, Musa S, Abu Kasim NH (2017) Effect of dental pulp stem cells in MPTP-induced old-aged mice model. Eur J Clin Invest 47:403–414
Simon C, Gan QF, Kathivaloo P, et al (2019) Deciduous DPSCs Ameliorate MPTP-Mediated Neurotoxicity, Sensorimotor Coordination and Olfactory Function in Parkinsonian Mice. International Journal of Molecular Sciences 2019, Vol 20, Page 568 20:568
Hubbard-Turner T, Wikstrom EA, Guderian S, Turner MJ (2013) Acute ankle sprain in a mouse model. Med Sci Sports Exerc 45:1623–1628
Garrick JM, Costa LG, Cole TB, Marsillach J (2021) Evaluating Gait and Locomotion in Rodents with the CatWalk. Curr Protoc 1:e220
Akula SK, McCullough KB, Weichselbaum C, Dougherty JD, Maloney SE (2020) The trajectory of gait development in mice. Brain Behav 10:e01636
Warmink K, Rios JL, van Valkengoed DR, Korthagen NM, Weinans H (2021) Sprague dawley rats develop more joint damage compared to wistar-han rats in a high fat-high sucrose groove model of osteoarthritis. Osteoarthritis Cartilage 29:S138
van Hest A, van Haaren F, van de Poll NE (1987) Behavioral differences between male and female Wistar rats on DRL schedules: effect of stimuli promoting collateral activities. Physiol Behav 39:255–261
Sudakov SK, Alekseeva E V., Nazarova GA, Bashkatova VG (2021) Age-Related Individual Behavioural Characteristics of Adult Wistar Rats. Animals (Basel). https://doi.org/10.3390/ANI11082282
Sengupta P (2013) The Laboratory Rat: Relating Its Age With Human’s. Int J Prev Med 4:624
Tatro-Adams D, Forrester McGann S (1995) Reliability of the Figure-of-Eight Method of-Ankle ~easurement. Journal of Orthopaedic & Sports Physical Therapy® Downloaded from www.jospt.org at on 22:
Watson CP, Boland RA, Refshauge KM (2008) Measurement Reliability of Swelling in the Acute Ankle Sprain. The Foot & Ankle Journal. https://doi.org/10.3827/faoj.2008.0112.0004
Haridasan S, Acharya D (2022) Role of 3 Tesla MRI in Evaluation of Ankle Joint and Foot Pathologies. European Journal of Molecular and Clinical Medicine 9:5195–5216
Pogarell T, Heiss R, Janka R, Nagel AM, Uder M, Roemer FW (2024) Modern low-field MRI. Skeletal Radiol 53:1751–1760
Khodarahmi I, Fritz J (2021) The Value of 3 Tesla Field Strength for Musculoskeletal Magnetic Resonance Imaging. Invest Radiol 56:749–763
Franettovich Smith MM, Elliott JM, Al-Najjar A, Weber KA, Hoggarth MA, Vicenzino B, Hodges PW, Collins NJ (2021) New insights into intrinsic foot muscle morphology and composition using ultra‐high‐field (7-Tesla) magnetic resonance imaging. BMC Musculoskelet Disord 22:1–14
Ching SH, Chiu YC, Liao YC, Yang SH, Tsai YJ (2022) A new mouse model of ankle instability induced by multiple mechanical sprains with controlled inversion angle and speed. Front Bioeng Biotechnol 10:927987
Gan QF, Foo CN, Leong PP, Cheong SK (2020) Regenerative Medicine as a Potential and Future Intervention for Ankle Sprain. Malaysian Journal of Medicine and Health Sciences 16:290–299
Du Y, Wang S, Yang F, Xu H, Cheng Y, Yu J (2024) Effects of chronic ankle instability after grade I ankle sprain on the post-traumatic osteoarthritis. Arthritis Res Ther 26:168
Gan QF, Foo CN, Choy KW, Yu CW, Lwin TS, Dalayi N, Lim YT (2020) Frequency and types of recreational physical activity determines static and dynamic stability. Gazzetta Medica Italiana Archivio per le Scienze Mediche. https://doi.org/10.23736/s0393-3660.19.04168-8
Gould TD, Dao DT, Kovacsics CE (2009) The Open Field Test. Neuromethods 42:1–20
Georgopoulos M, Pavlopoulos A, Zerva N, Kokkonakis A, Mourouzis I, Plesnila N, Pantos C, Lourbopoulos A, Georgopoulos M (2025) The Mouse Stroke Unit Protocol with Standardized Neurological Scoring for Translational Mouse Stroke Studies. J Vis Exp e66847
Kamal OM-F, Ojeda-Hernández DD, Selma-Calvo B, et al (2025) Technical Assessment of Motor and Behavioral Tests in Rodent Models of Multiple Sclerosis. Journal of Integrative Neuroscience 2025, 24(2), 26429 24:26429
Mann A, Chesselet MF (2015) Techniques for Motor Assessment in Rodents. Movement Disorders: Genetics and Models: Second Edition 139–157
Shi X, Bai H, Wang J, et al (2021) Behavioral Assessment of Sensory, Motor, Emotion, and Cognition in Rodent Models of Intracerebral Hemorrhage. Front Neurol 12:667511
Li J, Chen Z, Cheng Y, et al (2022) Ligamentous injury-induced ankle instability causing posttraumatic osteoarthritis in a mouse model. BMC Musculoskelet Disord 23:1–10
Refshauge K (1999) Effects of reduced ankle dorsiflexion following lateral ligament sprain on temporal and spatial gait parameters. Gait & Posture
Lakes EH, Allen KD (2016) Gait analysis methods for rodent models of arthritic disorders: reviews and recommendations. Osteoarthritis Cartilage 24:1837–1849
Lin Y, Rankin JW, Luı́s L, Lamas P, Moazen M, Hutchinson JR (2024) Hindlimb kinematics, kinetics and muscle dynamics during sit-to-stand and sit-to-walk transitions in emus (Dromaius novaehollandiae). Journal of Experimental Biology. https://doi.org/10.1242/JEB.247519
Quan Fu G, Chye Wah Y, Sura S, Jagadeesan S, Chinnavan E, Judson JPE (2018) Influence of rearfoot alignment on static and dynamic postural stability. Int J Ther Rehabil 25:628–635
Hong CC, Lee JC, Tsuchida A, Katakura M, Jones M, Mitchell AW, Dalmau-Pastor M, Calder J (2023) Individual fascicles of the ankle lateral ligaments and the lateral fibulotalocalcaneal ligament complex can be identified on 3D volumetric MRI. Knee Surgery, Sports Traumatology, Arthroscopy 31:2192–2198
Berwouts D, Lootens T, De Vil J, Mertens J (2022) Ligament and Tendon Injuries. Clinical Atlas of Bone SPECT/CT 1–16
Quan Fu G, Pooi Pooi L, Soon Keng C, Chai Nien F (2024) Incorporating stem cells into physical rehabilitation. Comprehensive Hematology and Stem Cell Research
Kamemoto K, Yamada M, Matsuda T, Ogata H, Ishikawa A, Kanno M, Miyashita M, Sakamaki-Sunaga M (2022) Effects of menstrual cycle on appetite-regulating hormones and energy intake in response to cycling exercise in physically active women. J Appl Physiol 132:224–235