- ICH GCP
- US Clinical Trials Registry
- Clinical Trial NCT03711838
Redox Regulation of Satellite Cells and Skeletal Muscle Healing
December 15, 2022 updated by: Ioannis G. Fatouros, University of Thessaly
The Effect of Redox Potential on the Regulation of Satellite Cells and Skeletal Muscle Healing Following Exercise-Induced Muscle Damage
Skeletal muscle stem cells (Satellite cells) are indispensable for muscle growth and remodeling following myofibril damage.
Skeletal muscle trauma is present in numerous catabolic conditions, characterized by elevated proteolysis and muscle wasting such as, cancer cachexia and muscular dystrophy, which result in physical capacity impairment and a deteriorated quality of life.
Recent studies performed in animals and cell cultures indicate that the increased levels of inflammation and oxidative stress and the reduction of antioxidant defense may blunt the satellite cells response and myogenic programming during muscle healing.
However, evidence regarding the effects of redox status on satellite cells and muscle myogenic potential in humans is lacking.
Exercise-induced muscle damage bears striking similarities with the aforementioned conditions, which makes it a valuable tool to investigate the redox-dependent regulation of satellite cells during muscle healing.
Thus, the objectives of the present study are to examine the effects of redox status perturbation (via N-acetylcysteine administration) on intracellular pathways responsible for satellite cells responses at rest and following aseptic muscle trauma induced by damaging exercise.
Study Overview
Status
Completed
Intervention / Treatment
Detailed Description
A total number of 40-60 males, young individuals aged 18-30 years, will be initially enrolled in the study.
Then, participants will be allocated to either a 1) Low Respondents (LR) or a 2) High Respondents (HR) group based on the basal satellite cells content of their vastus lateralis muscle of their dominant leg.
In a double-blind, crossover, repeated measures design, participants will consume either Placebo (PLA) or N-acetylcysteine (NAC) before (7-day loading phase), on exercise day and for 8 consecutive days following a single bout of intense exercise (300 eccentric contractions at 30 deg/sec in an isokinetic dynamometer).
In both conditions, blood samples and muscle biopsies will be collected at baseline, before the exercise protocol and at 2- and 8-days post-exercise.
Muscle performance and soreness will also be assessed at the same time points.
Before the first trial, participants' dietary intake (via diet recalls) and physical activity (via accelerometry) will be analyzed and nutritional plans will be provided to participants in order to normalize their antioxidant and protein intake.
A 4-week washout period will be implemented between trials.
Blood samples will be analyzed for inflammation and oxidative stress markers.
Muscle samples will be analyzed for satellite cells responses and myogenic potential, protein levels of intracellular signaling proteins, muscle thiols and antioxidant enzymes activity.
Study Type
Interventional
Enrollment (Actual)
45
Phase
- Not Applicable
Contacts and Locations
This section provides the contact details for those conducting the study, and information on where this study is being conducted.
Study Locations
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Thessaly
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Trikala, Thessaly, Greece, 42100
- Laboratory of Exercise Biochemistry, Exercise Physiology,and Sports Nutrition, School of Physical Education and Sport Science, University of Thessaly
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Participation Criteria
Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.
Eligibility Criteria
Ages Eligible for Study
16 years to 28 years (Adult)
Accepts Healthy Volunteers
No
Genders Eligible for Study
Male
Description
Inclusion Criteria:
- No recent history of musculoskeletal injury
- Non-smokers.
- Abstain from any vigorous physical activity during the study
- Abstain from consumption of caffeine, alcohol, performance-enhancing or antioxidant supplements, NSAIDs and medications before (at least 6 months) and during the study.
Exclusion Criteria:
- A known NAC intolerance or allergy
- A recent febrile illness
- A recent history of muscle lesion and/or lower limb trauma
- Presence of metabolic diseases
- Use of anti-inflammatory medication.
- Use of medication interacting with muscle metabolism.
Study Plan
This section provides details of the study plan, including how the study is designed and what the study is measuring.
How is the study designed?
Design Details
- Primary Purpose: Basic Science
- Allocation: Randomized
- Interventional Model: Crossover Assignment
- Masking: Quadruple
Arms and Interventions
Participant Group / Arm |
Intervention / Treatment |
---|---|
Experimental: N-Acetylcysteine
N-Acetylcysteine supplementation: Orally, 40 mg/kg per day in 3 doses (250 ml each) for 7 consecutive days and immediately post-exercise.
The remaining 8 days, 40mg/kg per day in 3 doses (250 ml each).
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N-Acetylcysteine in a powder form diluted in a 250 ml drink containing 248 ml water and 2 ml of natural, non-caloric, flavoring-sweetener containing sucralose.
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Active Comparator: Placebo
Placebo administration: Orally 750 ml per day in 3 doses (250 ml each) for 7 consecutive days and immediately post-exercise.
The remaining 8 days, 750 ml per day in 3 doses (250 ml each).
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Placebo consisted of 248 ml water and 2 ml of natural, non-caloric, flavoring-sweetener containing sucralose.
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What is the study measuring?
Primary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
---|---|---|
Change in muscle satellite cells number (i.e. Pax7+ cells) and activation status (i.e. Pax7+/MyoD+ cells)
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Satellite cells number and activation status, will be assessed in muscle via immunohistochemistry.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Change in muscle myogenic mRNA expression
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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mRNA expression levels of Myogenic factor 5 (Myf5), myogenin and Myogenic factor 6 (Myf6/MRF4) and myostatin will be assessed in muscle using Real-Time Polymerase Chain Reaction (RT-PCR).
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Change in muscle inflammatory state
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Pro-inflammatory (M1+) and anti-inflammatory (M2+) macrophages will be measured in muscle using immunohistochemistry.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Change in intracellular antioxidant enzymes in muscle
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Protein levels of Glutathione peroxidase 3 (GPx3), Superoxide dismutase 1 (SOD1) and Thioredoxin (Trx1) will be measured using western blotting.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Change in muscle thiol content
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) will be measured spectophotometrically.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Change in intracellular signaling proteins in muscle
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Protein levels of IGF-1, Notch1 and Wnt3 will be measured using western blotting.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Change in skeletal muscle damage levels
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Skeletal muscle damage will be quantified via histochemistry using H&E staining.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Secondary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
---|---|---|
Resting metabolic rate (RMR)
Time Frame: At baseline
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RMR will be assessed after an overnight fast with participants in a supine position following a 15-min stabilization period by taking 30 consecutive 1-min VO2/CO2 measurements using a portable open-circuit indirect calorimeter with a ventilated hood system following a standard calibration protocol.
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At baseline
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Body composition
Time Frame: At baseline
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Body composition will be measured using a dual-energy x-ray absorptiometry scanner (DXA).
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At baseline
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Maximal oxygen consumption (VO2max)
Time Frame: At baseline
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VO2max will be assessed during continuous incremental running to volitional fatigue on a treadmill with a pulmonary gas exchange system (Oxycon Mobile; Sensor-Medics Corporation).
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At baseline
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Isokinetic strength
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Maximal knee extensor eccentric peak torque at 60 degrees will be assessed on an isokinetic dynamometer.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Change in delayed onset of muscle soreness (DOMS)
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Muscle soreness will assessed during palpation of the muscle belly and the distal region of relaxed vastus medialis, vastus lateralis and rectus femoris following three repetitions of of a full squat.
Subjects will rate their DOMS on a visual analogue scale (0-10).
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Physical activity
Time Frame: At baseline.
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Level of habitual physical activity will be assessed using accelerometry (ActiGraph GT3X-BT accelerometer).
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At baseline.
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Dietary intake
Time Frame: At baseline.
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Daily dietary intake will be assessed using 7-day diet recalls.
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At baseline.
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Interleukin-1β (IL-1β) in blood
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of IL-1β will be measured in plasma.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Interleukin-6 (IL-6) in blood
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of IL-6 will be measured in plasma.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Interleukin-8 (IL-8) in blood
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of IL-6 will be measured in plasma.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Interleukin-10 (IL-10) in blood
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of IL-10 will be measured in plasma.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Cortisol in blood
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of cortisol will be measured in serum.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Protein carbonyls (PC)
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of protein carbonyls will be measured in muscle.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Malondialdehyde (MDA)
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of malondialdehyde will be measured in muscle.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Total antioxidant capacity (TAC)
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of TAC will be measured in serum.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Catalase
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of catalase will be measured in red blood cell lysate.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Creatine Kinase (CK)
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Concentration of CK will be measured in serum.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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White blood cell count in blood
Time Frame: At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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White blood cell count will be measured in blood.
White blood cell count will be measured in blood.
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At baseline, before the exercise protocol and at days 2 and 8 following exercise.
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Collaborators and Investigators
This is where you will find people and organizations involved with this study.
Sponsor
Investigators
- Principal Investigator: Konstantinos Papanikolaou, PhDc, University of Thessaly, School of Physical Education & Sport Science
Publications and helpful links
The person responsible for entering information about the study voluntarily provides these publications. These may be about anything related to the study.
Study record dates
These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.
Study Major Dates
Study Start (Actual)
July 22, 2019
Primary Completion (Actual)
May 1, 2022
Study Completion (Actual)
August 1, 2022
Study Registration Dates
First Submitted
October 11, 2018
First Submitted That Met QC Criteria
October 16, 2018
First Posted (Actual)
October 19, 2018
Study Record Updates
Last Update Posted (Actual)
December 19, 2022
Last Update Submitted That Met QC Criteria
December 15, 2022
Last Verified
December 1, 2022
More Information
Terms related to this study
Additional Relevant MeSH Terms
Other Study ID Numbers
- Satellite Cells - UTH
Drug and device information, study documents
Studies a U.S. FDA-regulated drug product
No
Studies a U.S. FDA-regulated device product
No
This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.
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