Method Article

A Visualized Sensitized Acupoint Procedure for Knee Osteoarthritis: Behavioral, Thermal, and Molecular Assessments

DOI:

10.3791/68967

⸱

August 29th, 2025

 ,  ,  ,  ,  ,  ,  , 
1School of Acupuncture, Moxibustion and Tuina, Beijing University of Chinese Medicine, 2Department of Traditional Chinese Medicine, Air Force Medical Center

* These authors contributed equally

In This Article

Summary

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This study details a rat fixation method suitable for photographing acupuncture and infrared thermography. Furthermore, it demonstrates that electroacupuncture at sensitized acupoints inhibits local inflammatory pain.

Abstract

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Knee osteoarthritis (KOA), a leading cause of musculoskeletal pain and functional disability worldwide, remains challenging to manage due to the limited efficacy of current pharmacological and surgical interventions in achieving sustained pain relief. Although acupuncture has demonstrated clinical promise in alleviating KOA-related pain, its underlying mechanisms require further elucidation. Application of acupuncture to sensitized acupoints can markedly enhance the effectiveness of therapy. This protocol establishes a rat model of KOA through intra-articular injection of sodium monoiodoacetate (MIA) and systematically describes an acupuncture protocol targeting sensitized acupoints, including restraint methods, acupoint localization, and acupuncture parameters. Therapeutic efficacy was comprehensively evaluated through pain behavioral assessments, infrared thermographic analysis of the knee joint, inflammatory cytokine detection in serum, and histopathological examination of cartilage tissue. Results indicate that acupuncture at sensitized acupoints significantly alleviates pain phenotypes in MIA-induced model rats by inhibiting the release of pro-inflammatory mediators, blocking nociceptive signal transmission, improving blood microcirculation, regulating local joint temperature, and delaying joint degeneration.

Introduction

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Knee osteoarthritis (KOA), a prevalent degenerative joint disorder, is characterized by chronic inflammation and whole-joint pathology, including cartilage erosion, subchondral bone remodeling, osteophyte development, and synovial inflammation1,2. Clinically, patients typically present with pain, muscle weakness, joint instability, transient morning stiffness, and progressive functional limitations. KOA imposes a substantial global disease burden, affecting approximately 365 million individuals with functional disability, with epidemiological models predicting a 74.9% escalation in case numbers from 2020 basel....

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Protocol

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All experimental protocols were approved and rigorously supervised by the Animal Ethics Committee of Beijing University of Chinese Medicine (ID: bucm-2024091102-3197), and all procedures strictly adhered to the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals. Thirty male Sprague-Dawley rats (weight range: 210-250 g) were used in this study. The reagents and equipment used are listed in the Table of Materials.

1. Experimental animals and groups

  1. House the animals in the dedicated animal facility.
  2. Maintain ambient temperature at 22 °C ± 2 ....

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Results

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No animals died during the entire experiment. Results of the operation procedure duration and needle detachment count for each rat in both the NF and EA groups indicated that the EA group, following restriction with nylon cable ties, experienced reduced operation time and fewer needle detachments (P < 0.001; Figure 1H,I). Analysis of acupoint pain thresholds revealed no statistically significant differences in mechanical pain sensitivity between GB34, ST36, and n.......

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Discussion

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MIA, a metabolic inhibitor, selectively disrupts cellular aerobic glycolysis, thereby inducing chondrocyte apoptosis17,18,19. Intra-articular MIA administration results in chondrocyte depletion and subsequent articular cartilage degeneration, characterized by morphological irregularities and histopathological alterations that closely resemble human OA pathology20. Thus, it is widely utilized in research o.......

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Disclosures

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All authors have declared no potential conflicts of interest.

Acknowledgements

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This research was supported by the National Key R&D Program of China (grant 2023YFC3502700) and the Fundamental Research Funds for the Central Universities (2025-JYB-XJSJJ014)

....

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
0-10ul Adjustable-Volume PipettorServicebioIC012395160823
0-200ul Adjustable-Volume PipettorServicebioIC021029160323
1.5 ml eppendorf tubesServicebioEP-150-M
10ul micropipette tipServicebioTP-10
1ml syringeShanghai Zhiyu Medical Equipment Co., LTDZSQ-1
200ul micropipette tipServicebioTP-200
22-gauge needle Kruuse112416c
2ml eppendorf tubesServicebioEP-150-M
5ml red blood tubeShanghai Xinle Biotechnology Co. LTDCXG-5R
75% alcoholShandong Anjie Gaoke Disinfection Technology Co. LTD75-500ml
Adherent slideCITOTEST188105
Automatic Plate WasherRaytoRT-3100C
Blood collection needleJiangxi Ruibang Industrial Group Co. LTDCXZ-07
Bone tissue safranin and solid green staining solutionShanghai Yuan Ye Biotechnology Co., Ltd.R32730
Cover glassCitotest Labware Manufacturing Co.,Ltd10212432C
DehydratorDIAPATHDonatello
Electronic balanceMettler-ToledoME203E/02
Electrothermal blast drying ovenLaboteryGEL-70
Embedding machineWuhan Junjie Electronics Co., LtdJB-P5
Environmental Friendly Dewaxing Transparent LiquidServicebioG1128-1L
Enzyme label detectorBioTeKEpoch
EthanolSCRC100092683
Ethylenediamine tetraacetic acid Macklin60-00-4
Fat-free cotton ballCaoxian Hualu Sanitary Material Co. LTDTZMQ-500
High speed refrigerated centrifugeDragonD3024R
High Speed Tissue GrinderServicebioKZ-III-F
Hot Plate AnalgesiometerKew basisKW-CT-1
Imaging systemNikonNIKON DS-U3
Infrared thermal imagerTesto AGTesto 865
IodophorShandong Anjie Gaoke Disinfection Technology Co. LTDDF-500
IR SoftTesto AG
IsofluraneRWD life scienceR510-22-10
Latex glovesBeijing Ruijing Latex Products Co. LTDRJJC-S
Mechanical Paw Poking Test DeviceKew basisKW-RB
MIASigma206-165-7
Neutral gumSCRC10004160
Normal butanolSCRC100052190
Normal saline solutionSHIMENSLYS-500ml
Paraffin liquidMacklin8012-95-1
Paraformaldehyde Fixative (Neutral)ServicebioG1101
Pathology slicerShanghai Leica Instrument Co., LtdRM2016
Rat IL-1 beta  ELISA Kit GER0002-96tServicebioAC30253754
Rat TNF-alpha ELISA Kit GER0004-96tServicebioAC40253711
SD ratsBeijing Sibefei Biotechnology Co., LtdSCXK (jing) 2024-0001 
Small Animal Anesthesia Machine YuYAN ABM
Surgical maskwinnerWJKZ-1
Tissue spreaderZhejiang Kehua Instrument Co., Ltd KD-P
Ultrapure Water Polishing SystemAiken water ElAK-RO-C2
Upright optical microscopeNikonNIKON ECLIPSE E100
UrethaneMacklin2621-79-6
Vortex MixerServicebioMV-100
XyleneSCRC10023418

References

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  1. Giorgino, R., et al. Knee osteoarthritis: Epidemiology, pathogenesis, and mesenchymal stem cells: What else is new? An update. Int J Mol Sci. 24 (7), 6405(2023).
  2. Li, J., et al. MRI-based texture ....

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Tags

Sensitized AcupointsAcupuncture ProtocolPain Behavioral AssessmentInfrared ThermographyInflammatory CytokinesCartilage HistopathologyRat KOA ModelJoint MicrocirculationNociceptive Signal Transmission

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