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Tuina Intervention in Rabbit Model of Knee Osteoarthritis

Published: August 25th, 2023



1Shandong University of Traditional Chinese Medicine, 2Liaoning University of Traditional Chinese Medicine, 3Shandong University of Traditional Chinese Medicine Affiliated Hospital
* These authors contributed equally

The protocol describes a method for Tuina intervention in a rabbit model of knee osteoarthritis.

Knee osteoarthritis (KOA) is mainly characterized by degenerative changes in the knee joint's cartilage and surrounding soft tissues. The efficacy of Tuina in treating KOA has been confirmed, but the underlying mechanism needs to be investigated. This study aims to establish a scientifically feasible KOA rabbit model treated with Tuina to reveal the underlying mechanisms. For this, 18, 6-month-old normal-grade male New Zealand rabbits were randomly divided into sham, model, and Tuina groups, with 6 rabbits in each group. The KOA model was established by injecting 4% papain solution into the knee joint cavity. The Tuina group was intervened with Tuina combined with the knee joint rotary correction method for 4 weeks. Only the standard grasping and fixation were performed in sham and model groups. At the end of the 1-week intervention, the knee joint range of motion (ROM) was observed, and cartilage hematoxylin-eosin (HE) staining was done. The study shows that Tuina could inhibit chondrocyte apoptosis, repair cartilage tissue, and restore knee joint ROM. In conclusion, this study demonstrates the scientific feasibility of Tuina treatment for KOA model rabbits, highlighting its potential application in the study of KOA and similar knee joint-related conditions.

Knee osteoarthritis (KOA) is a degenerative disease of the knee joint, mainly manifested by knee pain, swelling, deformation, and restricted movement, with a high disability rate and a higher prevalence in women, with 527.81 million patients with osteoarthritis worldwide in 2019 and its global prevalence accounting for 60.6% of the total global prevalence of OA1. Clinically, the treatment of KOA is usually divided into non-surgical and surgical therapies. Non-surgical therapies include physiotherapy, pharmacotherapy, and platelet-rich plasma injection therapy2,3. Tuina is a common, safe....

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The study was approved by the ethics committee of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine (approval number: 2020-29).

1. Experimental animals

  1. Raise 18, 6-month-old normal-grade male New Zealand rabbits (2.75 ± 0.25 kg) in standard single cages (12 h light/dark cycle, temperature 20-24 °C, air humidity 40%-60%).

2. Grouping method

  1. Select 6 out of the 18 N.......

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The degree of restricted knee motion and cartilage tissue damage reflects the severity of KOA. The knee joint's ROM reflects the degree of restriction of knee joint motion. The smaller the knee joint ROM, the more serious the limitation of knee joint motion. On the contrary, the bigger the knee joint ROM, the more normal the degree of knee motion. HE staining to observe the morphology and structure of cartilage tissue reflects the degree of cartilage tissue damage. The more irregular the surface of cartilage tissue, .......

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The design of the experimental protocol is particularly important to investigate the mechanism of Tuina in treating KOA. KOA modeling was performed on rabbits by injection of papain at Waixiyan. Waixiyan is located in the lateral crypt of the patellar ligament, which is easy to locate, and the joint space between the femur and tibia is large here during knee flexion, which makes it easy to inject into the knee joint cavity and prevents damage to the surrounding tissues, so it is easy to establish the KOA model

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This work was supported by the Shandong Provincial Traditional Chinese Medicine Science and Technology Project (2021Q080) and the Qilu School of Traditional Chinese Medicine Academic School Inheritance Project [Lu-Wei-Letter (2022) 93].


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Name Company Catalog Number Comments
0.9 % sodium chloride injection Sichuan Keren Pharmaceutical Co. Z22121903
-20°C refrigerator Haier BD-328WL
4 % fixative solution Solarbio P1110
4°C refrigerator Haier SC-315DS
Anhydrous ethanol Sinopharm
Automatic tissue dewatering machine Dakowei (Shenzhen) Medical Equipment Co. HP30
Blast drying oven Shanghai Yiheng Scientific Instruments Co. DHG-9070A
Coverslip Biyuntian FCGF50
Electric thermostat water bath Shanghai Yiheng Scientific Instruments Co. HWS-26
Embedding freezing table Changzhou Paishijie Medical Equipment Co. BM450
Embedding machine Changzhou Paishijie Medical Equipment Co. BM450A
Ethylenediaminetetraacetic acid decalcification solution Servicebio G1105-500ML
Fluorescent inverted microscope Leica Leica DM IL LED
Hematoxylin-eosin staining kit Cisco Jet EE0012
Hydrochloric acid Laiyang Economic and Technological Development Zone Fine Chemical Plant
Medical joint goniometer KOSLO
Neutral gum Cisco Jet EE0013
Normal-grade male New Zealand rabbit Jinan Xilingjiao Breeding and Breeding Center SCXK (Lu) 2020 0004
Papain(3000 U/mg) Bioss D10366
Pathological tissue bleaching and drying instrument Changzhou Paishijie Medical Equipment Co. PH60
Pet electric clippers Codos CP-3180
Rabbit fixing box any brand
Rotating Slicer Leica 531CM-Y43
Tuina technique parameter determination instrument Shanghai DuKang Instrument Equipment Co. Ltd. ZTC-Equation 1
Ventilator TALY ELECTRIC C32
Xylene Fuyu Reagent

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