Source Paper
Feng-Sheng Wang, Chung-Wen Kuo, Jih-Yang Ko, Yu-Shan Chen, Shao-Yu Wang et al.
Antioxidants • 2020
Compromised autophagy and mitochondrial dysfunction downregulate chondrocytic activity, accelerating the development of osteoarthritis (OA). Irisin, a cleaved form of fibronectin type III domain containing 5 (FNDC5), regulates bone turnover and muscle homeostasis. Little is known about the effect of Irisin on chondrocytes and the development of osteoarthritis. This study revealed that human osteoarthritic articular chondrocytes express decreased level of FNDC5 and autophagosome marker LC3-II but upregulated levels of oxidative DNA damage marker 8-hydroxydeoxyguanosine (8-OHdG) and apoptosis. Intra-articular administration of Irisin further alleviated symptoms of medial meniscus destabilization, like cartilage erosion and synovitis, while improved the gait profiles of the injured legs. Irisin treatment upregulated autophagy, 8-OHdG and apoptosis in chondrocytes of the injured cartilage. In vitro, Irisin improved IL-1β-mediated growth inhibition, loss of specific cartilage markers and glycosaminoglycan production by chondrocytes. Irisin also reversed Sirt3 and UCP-1 pathways, thereby improving mitochondrial membrane potential, ATP production, and catalase to attenuated IL-1β-mediated reactive oxygen radical production, mitochondrial fusion, mitophagy, and autophagosome formation. Taken together, FNDC5 loss in chondrocytes is correlated with human knee OA. Irisin repressed inflammation-mediated oxidative stress and extracellular matrix underproduction through retaining mitochondrial biogenesis, dynamics and autophagic program. Our analyses shed new light on the chondroprotective actions of this myokine, and highlight the remedial effects of Irisin on OA development.
Objective: Analysis of walking patterns and gait characteristics of injured legs using the CatWalk system to measure footprint area, contact area, swing speed, and duty cycle
This is a Gait Profile Analysis protocol using mouse as the model organism. The procedure involves 8 procedural steps, 8 equipment items, 3 materials. Extracted from a 2020 paper published in Antioxidants.
Model and subjects
mouse • C57L/B6 • male • 12-week-old • 15
Study window
~8 week study window
Core workflow
Animal anesthesia • Destabilized medial meniscus (DMM) surgery • Irisin preparation
Primary readouts
Key equipment and reagents
Verified items
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12-week-old male C57L/B6 mice were anesthetized using inhaled isoflurane delivered via Vetflo anesthetics vaporizer
Note: Anesthesia used for surgical procedures
“12-week-old male C57L/B6 mice were anesthetized by inhaled isoflurane using a Vetflo anesthetics vaporizer (Kent Scientific Corporation, Torrington, CT, USA)”
Medial meniscus of the left knee was removed using aseptic surgical procedures
Note: Surgical procedure performed on anesthetized mice to induce osteoarthritis model
“Medial meniscus of the left knee was removed using aseptic surgical procedures”
Irisin recombinant protein (20 mg/mL) was dissolved in normal saline and filtered through 0.22 µm filters
Note: Preparation performed 1 week post-operatively, prior to injection
“20 mg/mL of Irisin recombinant protein were dissolved in normal saline and filtered through 0.22 µm filters”
A total of 10 µL Irisin mixture was intra-articularly injected into DMM-injured knees using an insulin needle under ultrasound guidance (10-22 MHz LOGIO system)
Note: Performed at 1 week post-operatively. Ultrasound guidance used to ensure accurate injection into knee joint
“At 1 week postoperatively, a total of 10 µL Irisin mixtures were intra-articularly injected into DMM-injured knees using an insulin needle under the guidance of sonography (10–22 MHz, LOGIO, GE Healthcare, Princeton, NJ, USA)”
Walking patterns and gait characteristics of injured legs were investigated using the CatWalk analysis system. Mice walked across the illuminated glass walkway and footprint patterns were recorded
Note: Testing performed at 8 weeks post-operatively on sham (n=5), DMM (n=5), and DMM+Irisin (n=5) groups
“The walking patterns and gait characteristics of injured legs were investigated using the Catwalk analysis system (Noldus Information Technology, Leesburg, VA, USA)”
Footprint histograms were computed from CatWalk recordings using CatWalk software 9.1 according to manufacturer's instructions
Note: Software automatically processes video recordings of paw prints
“Footprint histograms were computed using CatWalk software 9.1 (Noldus Information Technology, Leesburg, VA, USA), according to the maker's instructions”
Footprint area (cm²), maximum contact area (%), swing speed (cm/s), and duty cycle (%) were calculated using CatWalk XT's Automatic Footprint Classification software
Note: Four primary gait parameters were extracted from the footprint data
“Footprint area (cm2), maximum contact area (%), swing speed (cm/s) and duty cycle (%) were calculated using CatWalk XT's Automatic Footprint Classification software”
At 8 weeks post-operatively, mice were euthanized using an overdose of anesthetics. Knee joints were dissected for histological examination
Note: Performed after gait analysis testing was complete
“At the end of the experiment, the mice were euthanatized using an overdose of anesthetics. Knee joints were dissected for histological examination”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
Analysis of walking patterns and gait characteristics of injured legs using the CatWalk system to measure footprint area, contact area, swing speed, and duty cycle
Objective
Analysis of walking patterns and gait characteristics of injured legs using the CatWalk system to measure footprint area, contact area, swing speed, and duty cycle
Subjects
From papermouse • C57L/B6 • male • 12-week-old
Sample count
From paper15
Cohort notes
From paperThree experimental groups: sham (n=5), DMM (n=5), DMM+Irisin (n=5).
Animal anesthesia
Destabilized medial meniscus (DMM) surgery
Irisin preparation
Intra-articular Irisin injection
Footprint area (cm²)
From paperFootprint histograms computed using CatWalk software 9.1.
Artifact type
Longitudinal gait metrics and per-animal performance tables
Comparison focus
Compare recovery trajectory across post-injury timepoints and treatment conditions
Maximum contact area (%)
From paperFootprint histograms computed using CatWalk software 9.1.
Artifact type
Longitudinal gait metrics and per-animal performance tables
Comparison focus
Compare recovery trajectory across post-injury timepoints and treatment conditions
Swing speed (cm/s)
From paperFootprint histograms computed using CatWalk software 9.1.
Artifact type
Longitudinal gait metrics and per-animal performance tables
Comparison focus
Compare recovery trajectory across post-injury timepoints and treatment conditions
Duty cycle (%)
From paperFootprint histograms computed using CatWalk software 9.1.
Artifact type
Longitudinal gait metrics and per-animal performance tables
Comparison focus
Compare recovery trajectory across post-injury timepoints and treatment conditions
Footprint area (cm²)
From paperRaw artifact
Per-run gait capture with paw placement, timing, and stride features for each animal
Processed artifact
Cleaned gait metrics table and recovery trend summary across timepoints
Final reported form
Group comparisons of gait indices, stride metrics, or recovery curves
Maximum contact area (%)
From paperRaw artifact
Per-run gait capture with paw placement, timing, and stride features for each animal
Processed artifact
Cleaned gait metrics table and recovery trend summary across timepoints
Final reported form
Group comparisons of gait indices, stride metrics, or recovery curves
Swing speed (cm/s)
From paperRaw artifact
Per-run gait capture with paw placement, timing, and stride features for each animal
Processed artifact
Cleaned gait metrics table and recovery trend summary across timepoints
Final reported form
Group comparisons of gait indices, stride metrics, or recovery curves
Duty cycle (%)
From paperRaw artifact
Per-run gait capture with paw placement, timing, and stride features for each animal
Processed artifact
Cleaned gait metrics table and recovery trend summary across timepoints
Final reported form
Group comparisons of gait indices, stride metrics, or recovery curves
Acquisition
Capture run-level gait data for each animal and preserve the timepoint or treatment labeling.
Preprocessing / cleaning
Footprint histograms computed using CatWalk software 9.1.
Scoring or quantification
Quantify the primary readouts for this experiment: Footprint area (cm²); Maximum contact area (%); Swing speed (cm/s); Duty cycle (%).
Normalization
Normalize image-derived measurements against the matched acquisition or segmentation rules before comparing groups.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Footprint area (cm²), Maximum contact area (%), Swing speed (cm/s), Duty cycle (%).
Source links and direct wording from the methods section for validation and deeper review.
Citation
Feng-Sheng Wang et al. (2020). Irisin Mitigates Oxidative Stress, Chondrocyte Dysfunction and Osteoarthritis Development through Regulating Mitochondrial Integrity and Autophagy. Antioxidants
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Kent Scientific Corporation • Vetflo
Noldus Information Technology
GE Healthcare • LOGIO
Carl Zeiss
Olympus
Applied Biosystems • ABI 7900
Noldus Information Technology
Noldus Information Technology
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Evidence Quotes
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