Summary

Application of Acupotomy in a Knee Osteoarthritis Model in Rabbit

Published: October 20, 2023
doi:

Summary

In this protocol, a knee osteoarthritis model was prepared using the modified Videman method, and the operation procedures and precautions of acupotomy are detailed. The effectiveness of acupotomy has been demonstrated by testing the mechanical properties of quadriceps femoris and tendon and the mechanical and morphological properties of cartilage.

Abstract

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department, which seriously reduces the quality of life of people with KOA. Among several pathogenic factors, the biomechanical imbalance of the knee joint is one of the main causes of KOA. Acupotomology believes that restoring the mechanical balance of the knee joint is the key to treating KOA. Clinical studies have shown that acupotomy can effectively reduce pain and improve knee mobility by reducing adhesion, contracture of soft tissues, and stress concentration points in muscles and tendons around the knee joint.

In this protocol, we used the modified Videman method to establish a KOA model by immobilizing the left hindlimb in a straight position. We have outlined the method of operation and the precautions related to acupotomy in detail and evaluated the efficacy of acupotomy in conjunction with the theory of “Modulating Muscles and Tendons to Treat Bone Disorders” through the detection of the mechanical properties of quadriceps femoris and tendon, as well as cartilage mechanics and morphology. The results show that acupotomy has a protective effect on cartilage by adjusting the mechanical properties of the soft tissues around the knee joint, improving the cartilage stress environment, and delaying cartilage degeneration.

Introduction

Knee osteoarthritis (KOA) is the most frequent form of osteoarthritis, often recognized as a whole-joint disease characterized by articular cartilage degeneration, which manifests clinically as pain, swelling, and limited movement of the affected joints1. According to recent epidemiological statistics, KOA is reported to have affected 654.1 million individuals globally who were 40 years of age or older by 2020. The prevalence and incidence of KOA rise with age, are the highest in middle-aged and older adults, and affect more women than men2. The prevalence of KOA is likely to increase due to the aging population and obesity epidemic worldwide, posing a growing threat to global public health. Age, sex, obesity, trauma, and other complicated risk factors associated with KOA all directly impact knee instability, making a biomechanical imbalance in knee joints one of the primary causes of KOA3.

Under normal physiological conditions, the knee joint is in a state of mechanical balance, ensuring that the mechanical loads in the joint are evenly distributed on the cartilage. Any mechanical imbalance in the knee joint can lead to abnormal stress in cartilage, resulting in cartilage degeneration and the onset of KOA4. The muscle-tendon system is the main dynamic system that maintains the mechanical balance of the knee joint. The coordinated movement of the extensor and flexor muscle-tendon system can evenly distribute the load generated by the movement on the cartilage surface, avoiding the metabolic imbalance of local cartilage stresses beyond its physiological load that results in cartilage loss5. Decreased muscle strength is the main cause of intramuscular movement disorder and cartilage damage, which may occur before symptomatic KOA.

KOA can also induce arthrogenous muscle inhibition (AMI), manifesting as muscle weakness and decreased muscle strength around the knee6. Among these muscles, the quadriceps femoris group functions as the only knee extensor, an important structure in maintaining knee joint stability. Studies have shown that a decrease in quadriceps cross-sectional area and muscle strength is significantly and positively correlated with KOA progression7. The decline in quadriceps strength affects the gait pattern, knee stability, movement patterns, and many other functions. Moreover, the decline in muscle strength impairs tendon function, manifested as a decrease in tendon stiffness, elastic modulus, and other biomechanical properties8. In long-term strain repair, changes such as adhesion and contracture may occur in the muscles and tendons of the knee joint, damaging their mechanical properties, causing joint instability, and ultimately forming a vicious cycle of pathological changes of KOA. It is, therefore, crucial for KOA treatment to improve the mechanical properties of the muscle-tendon system and restore the joint mechanical balance.

Among the causes of KOA, biomechanical imbalance is the main inducing factor for knee pain, dysfunction, inflammatory lesions, and cartilage degeneration9. Therefore, the key to treating KOA is to restore the biomechanical balance of the knee joint. Acupotomology believes that the etiology and pathogenesis of KOA are “mechanical imbalance.” When the mechanical characteristics of the soft tissues around the knee change abnormally, the knee joint loses its mechanical balance, and the abnormal mechanical stress environment of the joint accelerates degeneration, causing inflammatory stimulation to further aggravate the soft tissue adhesions, contractures, and further decline in joint stability. This vicious cycle eventually develops into KOA. By loosening soft tissue adhesions and contractures, as well as reducing stress concentration in the muscles and tendons, acupotomy in conjunction with the theory of “Modulating Muscles and Tendons to Treat Bone Disorders” improves the soft tissue mechanics and “modulates muscles and tendons,” which balances the mechanical stress of the joint, effectively alleviating cartilage degeneration and “treating bone disorders”10. In terms of animal model selection, based on the purpose of this study, we prepared the KOA model by the modified Videman method of left hindlimb extension immobilization.

This paper details the establishment of the KOA model using the modified Videman method of left hind limb extension immobilization and the method of operation and precautions of acupotomy. We demonstrate the effectiveness of acupotomy by testing the mechanical properties of quadriceps femoris and tendon and detecting changes in articular cartilage stress and morphology.

Protocol

All animal experiments were reviewed and approved by the Animal Ethics Committee of Beijing University of Chinese Medicine (No. BUCM-4-2022010101-1097). In this protocol, 24 6-week-old male New Zealand rabbits were housed under a specific condition, namely, 20-25 °C, 50-60% humidity, and a 12 h light/12 h dark circadian cycle, with free access to a regular chow diet. The rabbits were anesthetized and sacrificed by combining deep anesthesia and air embolization. Pain is one of the typical pathological features of KOA…

Representative Results

Experimental results of mechanical properties of quadriceps femoris and tendon To evaluate the effect of acupotomology on the mechanical properties of quadriceps femoris in rabbits with KOA, we used real-time shear wave elastic ultrasound imaging and a muscle tension transducer, respectively. Compared with the control group, Young's modulus of the quadriceps femoris in the KOA group was decreased (P < 0.05). Compared with the KOA group, Young's modulus of the acupotomy group wa…

Discussion

An appropriate animal model is one of the key factors to achieve experimental objectives and clarify a specific scientific question. This study was based on the theories of "Zongjin controlling bones and lubricating joints" and "mechanical imbalance" in acupotomology, aiming to explain the scientific connotation behind the treatment of KOA by "modulating muscles and tendons to treat bone disorders" in acupotomy therapy. In other words, acupotomy improves the abnormal mechanical environment of the …

Disclosures

The authors have nothing to disclose.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (No.82074523,82104996).

Materials

Acupotomy Beijing Zhuoyue Huayou Medical Devices Co., Ltd. 0.4 x 40 mm
Connect Cast Orthopedic Casting Tape Suzhou Connect Medical Technology Co.,Ltd. KCP06 15.0 cm x 360 cm
Double-sided Foam Tape Deli Group Co.,Ltd. NO.30416 36 mm x 5 yard x 2.5 mm
Environmental Dewaxing Solution Wuhan Servicebio Technology Co.,Ltd. G1128
Ethanol absolute Beijing Hengkangda Medicine Co., Ltd.
Fast Green solution Wuhan Servicebio Technology Co.,Ltd. G1031
Fast grenn FCF Sigma,America 2353-45-9
Fatigue testing machine BOSE, America Bose Electro Force 3300
Four-channel physiological recorder Chengdu Instrumeny Frctory RM-6420
FPD-305E Fuji, Japan
FPD-306E Fuji, Japan
Hematoxylin solution Wuhan Servicebio Technology Co.,Ltd. G1005
Medical iodophor disinfectant Shan Dong Lircon Medical Technology Co., Ltd.
Medical Tape Shandong Rongjian Sanitary Products Co., Ltd. 200402 1.5 x 500 cm
Muscle tension transducer  Chengdu Instrumeny Frctory JH-2204005, 50 g
Prescale Fuji, Japan
Real-time SWE ultrasound diagnostic instrument SuperSonic Imagine SA,France SuperSonic Imagine AixPlorer
Rhamsan gum Wuhan Servicebio Technology Co.,Ltd. WG10004160
Safranine O Sigma,America 477-73-6
Safranine O solution Wuhan Servicebio Technology Co.,Ltd. G1015
Statistical Package for the Social Sciences (SPSS) IBM, America

References

  1. Roseti, L., Desando, G., Cavallo, C., Petretta, M., Grigolo, B. Articular cartilage regeneration in osteoarthritis. Cells. 8 (11), 1305 (2019).
  2. Cui, A. Y., et al. Global, regional prevalence, incidence and risk factors of knee osteoarthritis in population-based studies. EClinicalMedicine. 29-30, 100587 (2020).
  3. Miyauchi, A., et al. Alleviation of murine osteoarthritis by deletion of the focal adhesion mechanosensitive adapter, Hic-5. Sci Rep. 9 (1), 15770 (2019).
  4. Wright, T. Biomechanical factors in osteoarthritis: the effects of joint instability. HSS J. 8 (1), 15-17 (2012).
  5. Patsika, G., Kellis, E., Kofotolis, N., Salonikidis, K., Amiridis, I. G. Synergetic and antagonist muscle strength and activity in women with knee osteoarthritis. J Geriatr Phys Ther. 37 (1), 17-23 (2014).
  6. Blalock, D., Miller, A., Tilley, M., Wang, J. X. Joint instability and osteoarthritis. Clin Med Insights. Arthritis and Musculoskelet Disord. 8, 15-23 (2015).
  7. Henriksen, M., Christensen, R., Danneskiold-Samsøe, B., Bliddal, H. Changes in lower extremity muscle mass and muscle strength after weight loss in obese patients with knee osteoarthritis: a prospective cohort study. Arthritis Rheum. 64 (2), 438-442 (2012).
  8. Schwartz, A. G., Lipner, J. H., Pasteris, J. D., Genin, G. M., Thomopoulos, S. Muscle loading is necessary for the formation of a functional tendon enthesis. Bone. 55 (1), 44-51 (2013).
  9. Felson, D. T. Osteoarthritis as a disease of mechanics. Osteoarthr Cartil. 21 (1), 10-15 (2013).
  10. Ma, S. N., et al. Effect of acupotomy on FAK-PI3K signaling pathways in KOA rabbit articular cartilages. Evid Based Complement Alternat Med. 2017, 4535326 (2017).
  11. Yu, P., et al. Research progress of experimental animal models of knee osteoarthritis. China Medical Herald. 16 (27), 41-44 (2019).
  12. Li, L. H., et al. Research progress of rabbit knee osteoarthritis model. Journal of Jiangxi University of Chinese Medicine. 31 (4), 108-113 (2019).
  13. Zhang, W., Gao, Y., Guo, C. Q., Khattab, I. Z. A., Mokhtari, F. Effect of acupotomy versus electroacupuncture on ethology and morphology in a rabbit model of knee osteoarthritis. J Tradit Chin Med. 39 (2), 229-236 (2019).
  14. An, X. Y., et al. Chondroprotective effects of combination therapy of acupotomy and human adipose mesenchymal stem cells in knee osteoarthritis rabbits via the GSK3 beta-cyclin D1-CDK4/CDK6 signaling pathway. Aging Dis. 11 (5), 1116-1132 (2020).
  15. Guo, C. Q., Liu, N. G. Analysis on the distribution features of Ashi points in Jingjin disorders. Journal of Basic Chinese Medicine. 17 (8), 899-900 (2011).
  16. Guo, C. Q., et al. Effects of acupotomy therapy on tenderness point around knee joint in patients with knee osteoarthritis: a randomized controlled clinical trial. Journal of Chengdu University of Traditional Chinese Medicine. 33 (3), 3-5 (2010).
  17. Zhao, Y., Dong, F. H., Zhang, K. Analysis of soft tissues mechanical changes and treatment of meridian tendon pain. Beijing Journal of Traditional Chinese Medicine. (9), 705-707 (2008).
  18. Zhang, L. P., Cheng, F., Liu, D. Y., Zhu, L. G. Application progress of biomechanics in knee osteoarthritis. Chinese Archives of Traditional Chinese Medicine. 34 (7), 1644-1647 (2016).
  19. Thomas, A. C., Sowers, M., Karvonen-Gutierrez, C., Palmieri-Smith, R. M. Lack of quadriceps dysfunction in women with early knee osteoarthritis. J Orthop Res. 28 (5), 595-599 (2010).
  20. Li, L., et al. Effects of unloaded muscle atrophy on contractile characteristics of hind-limb skeletal muscles in mice. Space Med Med Eng. 25 (5), 322-325 (2012).
check_url/kr/65584?article_type=t

Play Video

Cite This Article
LongFei, X., Yan, G., XiLin, C., TingYao, H., WenTing, Z., WeiWei, M., Mei, D., Yue, X., ChangQing, G. Application of Acupotomy in a Knee Osteoarthritis Model in Rabbit. J. Vis. Exp. (200), e65584, doi:10.3791/65584 (2023).

View Video