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].

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
<ol>
	<li>Raise 18, 6-month-old normal-grade male New Zealand rabbits (2.75 &#177; 0.25 kg) in standard single cages (12 h light/dark cycle, temperature 20-24 &#176;C, air humidity 40%-60%).</li>
</ol>
2. Grouping method
<ol>
	<li>Select 6 out of the 18 N...

Treating Knee Osteoarthritis with Tuina in a Rabbit Model

The authors declare no potential conflicts of interest.

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<sup class=...

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, reliable, and effective treatment method in Chinese medicine. This study uses Tuina combined with the knee joint rotary correction method to treat KOA. Tuina techniques such as the rotatory kneading and pressing method can balance muscle tissue, reduce pain, adjust inflammatory factor levels, improve tissue metabolism, and inhibit articular cartilage degeneration4,5. The knee joint rotary correction method can adjust the alignment of lower limb bones and joints, improve the knee joint gap, restore the normal force line, and balance the lower limb biomechanics6,7,8,9. Resistance exercises can enhance muscle mass and strength and promote cartilage tissue renewal10,11. A preliminary study found that this Tuina protocol is significantly more effective than oral glucosamine sulfate capsules in treating KOA, with a faster onset of action and significant inhibition of chondrocyte degeneration and repair of damaged cartilage tissue12. In the treatment of KOA, compared to Tuina therapy, non-steroidal anti-inflammatory drugs have adverse effects and unsatisfactory long-term efficacy, relatively high surgical risks and costs, and require certain indications for surgical treatment, with postoperative problems and periprosthetic complications13,14,15. When compared to drug therapy and surgery, Tuina treatment for KOA offers several advantages, including reduced side effects, lower risk, enhanced safety, cost-effectiveness, and longer-lasting efficacy. Additionally, it can effectively alleviate knee joint pain, swelling, popping, and restricted movement6,13,16,17.
However, the mechanism of Tuina for the treatment of KOA needs to be clarified, which limits the improvement and perfection of the treatment protocol for KOA. Therefore, studying the mechanism of Tuina intervention in KOA through animal experiments is an effective method. Rabbits, compared to rats, have a docile temperament and larger knee joints. The anatomical structure and cartilage biochemical indexes are similar to those of humans, so it is a suitable subject for studying the mechanism of knee joint disease by Tuina18. The KOA model established by injecting papain into the knee joint cavity of rabbits has the advantages of short modeling time, reduced trauma, high success rate, high survival rate, and similar pathological mechanism to KOA19. This study aims to establish a scientifically feasible animal experimental protocol for Tuina intervention in KOA and investigate the mechanism of Tuina.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9 % sodium chloride injection</td>
			<td>Sichuan Keren Pharmaceutical Co.</td>
			<td>Z22121903</td>
			<td></td>
		</tr>
		<tr>
			<td>-20&#176;C refrigerator</td>
			<td>Haier</td>
			<td>BD-328WL</td>
			<td></td>
		</tr>
		<tr>
			<td>4 % fixative solution</td>
			<td>Solarbio</td>
			<td>P1110</td>
			<td></td>
		</tr>
		<tr>
			<td>4&#176;C refrigerator</td>
			<td>Haier</td>
			<td>SC-315DS</td>
			<td></td>
		</tr>
		<tr>
			<td>Anhydrous ethanol</td>
			<td>Sinopharm</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Automatic tissue dewatering machine</td>
			<td>Dakowei (Shenzhen) Medical Equipment Co.</td>
			<td>HP30</td>
			<td></td>
		</tr>
		<tr>
			<td>Blast drying oven</td>
			<td>Shanghai Yiheng Scientific Instruments Co.</td>
			<td>DHG-9070A</td>
			<td></td>
		</tr>
		<tr>
			<td>Coverslip</td>
			<td>Biyuntian</td>
			<td>FCGF50</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric thermostat water bath</td>
			<td>Shanghai Yiheng Scientific Instruments Co.</td>
			<td>HWS-26</td>
			<td></td>
		</tr>
		<tr>
			<td>Embedding freezing table</td>
			<td>Changzhou Paishijie Medical Equipment Co.</td>
			<td>BM450</td>
			<td></td>
		</tr>
		<tr>
			<td>Embedding machine</td>
			<td>Changzhou Paishijie Medical Equipment Co.</td>
			<td>BM450A</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylenediaminetetraacetic acid decalcification solution</td>
			<td>Servicebio</td>
			<td>G1105-500ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescent inverted microscope</td>
			<td>Leica</td>
			<td>Leica DM IL LED</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin-eosin staining kit</td>
			<td>Cisco Jet</td>
			<td>EE0012</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrochloric acid</td>
			<td>Laiyang Economic and Technological Development Zone Fine Chemical Plant</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Medical joint goniometer</td>
			<td>KOSLO</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Neutral gum</td>
			<td>Cisco Jet</td>
			<td>EE0013</td>
			<td></td>
		</tr>
		<tr>
			<td>Normal-grade male New Zealand rabbit</td>
			<td>Jinan Xilingjiao Breeding and Breeding Center</td>
			<td>SCXK (Lu) 2020 0004</td>
			<td></td>
		</tr>
		<tr>
			<td>Papain(3000 U/mg)</td>
			<td>Bioss</td>
			<td>D10366</td>
			<td></td>
		</tr>
		<tr>
			<td>Pathological tissue bleaching and drying instrument</td>
			<td>Changzhou Paishijie Medical Equipment Co.</td>
			<td>PH60</td>
			<td></td>
		</tr>
		<tr>
			<td>Pet electric clippers</td>
			<td>Codos</td>
			<td>CP-3180</td>
			<td></td>
		</tr>
		<tr>
			<td>Rabbit fixing box</td>
			<td>any brand</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Rotating Slicer</td>
			<td>Leica</td>
			<td>531CM-Y43</td>
			<td></td>
		</tr>
		<tr>
			<td>Tuina technique parameter determination instrument</td>
			<td>Shanghai DuKang Instrument Equipment Co. Ltd.</td>
			<td>ZTC-<img alt="Equation 1" src="/files/ftp_upload/65763/65763eq01.jpg" style="margin: auto;" /></td>
			<td></td>
		</tr>
		<tr>
			<td>Ventilator</td>
			<td>TALY ELECTRIC</td>
			<td>C32</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>Fuyu Reagent</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

tuina intervention in 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.

Author Spotlight: Using a Rabbit Model to Explore the Efficacy of Tuina in Treating Knee Osteoarthritis 

Tuina Intervention in Rabbit Model of Knee Osteoarthritis

When comparing the two types of therapy and the surgery, tuina treatment of full knee osteoarthritis offered several advantages, including reduced side effects, lower risk, enhanced safety, cost effectiveness, and longer-lasting efficacy. Additionally, it can effectively eliminate knee joint pain, swelling, popping, and restricted movement. This study offers scientifically sound, an affordable research particular, that will aid in exploring the mechanism underlying the effectiveness of tuina in the treatment of knee joint diseases.This study demonstrates the scientific feasibility of tuina treatment for knee osteoarthritis model rabit, highlighting its potential application in the study of knee osteoarthritis, and a similar knee joint-related conditions.

The degree of restricted knee motion and cartilage tissue damage reflects the severity of KOA. The knee joint&#39;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, ...

Watch this Scientific Journal Video about Using a Rabbit Model to Explore the Efficacy of Tuina in Treating Knee Osteoarthritis at JoVE.com

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-intervention-in-rabbit-model-of-knee-osteoarthritis

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JoVE Journal

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Training Prior to Executing Tuina on Knee Osteoarthritis Rabbit Model

Initiate training using the Tuina Technique Parameter Determination Tool prior to executing the Tuina manipulation. To do so, on the Tuina Manipulation Simulation Platform, perform the rotary kneading and pressing method using the thumb. At the same time, in the Tuina Manipulation Parameter Processing Software observe the magnitude, frequency, and duration of action of the applied force across the x, y, and z-axes.Standardize the training in an applied force of five newtons and a frequency of 60 times per minute. Maintain the standardized rotary kneading and pressing continuously for an operation time of 10 minutes.

Performing Tuina on Knee Osteoarthritis (KOA) Rabbit Model

To begin Tuina intervention on the model rabbit with osteoarthritis established in its left knee, place the animal in the rabbit fixation box, and sooth it by gently stroking it for about 10 seconds. Perform the rotary kneading method using the thumb on the rabbit's left patella and the left peri-knee having muscle stiffness and tendon knots. Maintaining a force of 5 Newtons, perform an up and down, round tip manipulation at a frequency of 60 times per minute for five minutes.Then proceed to perform Tuina on the acupoints listed on the screen. Using the thumb end, press onto each acupoint, and maintaining the same force and frequency as demonstrated previously, operate on each point for 30 seconds. To perform rotary correction on the rabbit knee joint, hold the femur with one hand while positioning the other hand behind the knee joint.Fix the lateral and medial tibial condyles using the thumb and ring fingers respectively. Secure the popliteal fossa with the index and middle fingers. Using the fingers as demonstrated, hold the skin in place, avoiding friction between the skin and fingers.Then keeping the direction of traction force parallel to the long axis of the tibia, apply traction and twisting force. Secure the femur again with one hand. Establish a grip on the medial and lateral edges of the patella using the thumb and little fingers of the other hand while securing the base of the patella with the index, middle, and ring fingers.Then, keeping the direction of the torsion force in line with the direction of the lower Xiyan, apply a torsion force. Estimating the left knee ROM in the different groups of rabbits revealed that the Tuina group showed significant improvement in knee joint mobility after receiving the Tuina intervention from week seven. However, the model group that did not receive Tuina intervention showed no significant increase in ROM.

1.1K Views.  Shandong University of Traditional Chinese Medicine.  The protocol describes a method for Tuina intervention in a rabbit model of knee osteoarthritis.

Treating Knee Osteoarthritis with Tuina in a Rabbit Model

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