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Protein Supplementation and Skeletal Muscle Healing Process

Primary Purpose

Skeletal Muscle Damage, Exercise-induced Aseptic Inflammation, Skeletal Muscle Performance

Status
Completed
Phase
Not Applicable
Locations
Greece
Study Type
Interventional
Intervention
Milk protein isolate
Placebo
Sponsored by
University of Thessaly
About
Eligibility
Locations
Arms
Outcomes
Full info

About this trial

This is an interventional basic science trial for Skeletal Muscle Damage

Eligibility Criteria

18 Years - 30 Years (Adult)MaleAccepts Healthy Volunteers

Inclusion Criteria:

  • a) recreationally trained as indicated by the maximal oxygen consumption levels (VO2max > 45 ml/kg/min), b) engaged in systematic exercise at least three times per week for > 12 months, c) non-smokers, d) abstained from any vigorous physical activity during the study, e) abstained from consumption of caffeine, alcohol, performance-enhancing or antioxidant supplements, and medications during the study.

Exclusion Criteria:

  • a) a recent febrile illness, b) history of muscle lesion, c) lower limb trauma

Sites / Locations

  • Exercise Biochemistry Laboratory, School of Physical Education & Sports Sciences, University of Thessaly

Arms of the Study

Arm 1

Arm 2

Arm Type

Experimental

Active Comparator

Arm Label

Protein

Placebo

Arm Description

Milk protein isolate supplementation: orally, 20g of protein immediately post-exercise and then 20g every 3h on 3 occasions (+3, +6, +9), on the exercise day. The remaining 8 days, 20g daily with breakfast.

Placebo administration: orally 500 ml, immediately post-exercise as well as at +3, +6 and +9 hours, on the exercise day. The remaining 8 days, 500 ml daily with breakfast.

Outcomes

Primary Outcome Measures

Change in reduced glutathione in blood
Concentration of reduced glutathione in red blood cells
Change in protein carbonyls in serum
Concentration of protein carbonyls
Change in protein carbonyls in muscle
Protein carbonyl concentration in quadriceps skeletal muscle group
Change in thiobarbituric acid and reactive substances in serum
Thiobarbituric acid reactive substances concentration in serum
Change in oxidized glutathione in blood
Concentration of oxidized glutathione in red blood cells
Change in total antioxidant capacity in serum
Total antioxidant capacity in serum
Change in catalase activity in serum
Catalase activity in serum
Change in creatine kinase activity in plasma
Change in C-reactive protein in plasma
Change in white blood cell count in blood
Changes in volume and morphological complexity of immune cells
Change in neutrophil count in blood
Change in glucose concentration in blood
Change in insulin concentration in blood
Change in testosterone concentration in plasma
Change in cytokine concentration in plasma
Measurement of IL-1β, IL-4, IL-6, IL-8, IL-10, TNF-α
Change in adhesion molecule concentration in blood
Change in intracellular signalling proteins in muscle
Measurement of phosphorylation levels of mammalian target of rapamycin (mTOR), ribosomal protein S6 (rpS6) and nuclear factor kB (NFkB), and protein expression levels of forkhead box protein O1 (FOXO1), HSP70, and parkin.
Change in proteasome activities in muscle
Measurement of LLVY, LSTR and LLE
Change in protein expression level of proteasome subunits
Measurement of B1i, B2i, B5i, B5, B1, B2 and α7

Secondary Outcome Measures

Change in muscle function of knee extensor and flexor muscle
Assessment of muscle peak and mean torque of knee extensors and flexors on an isokinetic dynamometer at 0, 90 and 180 degrees/sec
Body composition
Assessment of percent (%) body mass
Maximal aerobic capacity
Assessment of maximal oxygen consumption
Change in dietary intake profile
Assessment of dietary intake with emphasis on protein consumption

Full Information

First Posted
June 24, 2016
Last Updated
October 3, 2016
Sponsor
University of Thessaly
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1. Study Identification

Unique Protocol Identification Number
NCT02816411
Brief Title
Protein Supplementation and Skeletal Muscle Healing Process
Official Title
Effects of Protein Supplementation on Skeletal Muscle Regeneration and Healing Process Following Exercise-induced Aseptic Injury
Study Type
Interventional

2. Study Status

Record Verification Date
July 2016
Overall Recruitment Status
Completed
Study Start Date
May 2014 (undefined)
Primary Completion Date
February 2016 (Actual)
Study Completion Date
July 2016 (Actual)

3. Sponsor/Collaborators

Responsible Party, by Official Title
Principal Investigator
Name of the Sponsor
University of Thessaly

4. Oversight

Data Monitoring Committee
No

5. Study Description

Brief Summary
In this study the investigators utilized protein supplementation over an 8-day period following eccentric exercise-induced muscle damage in order to test the initial hypotheses : i) protein supplementation after exercise-induced muscle injury affects exercise-induced aseptic inflammation and muscle performance.
Detailed Description
The objective was to examine weather protein supplementation is able to affect the inflammatory response as well as recovery of muscle performance following an intense eccentric exercise protocol. In a double-blind, counterbalanced design, 14 men received either Placebo (PLA) or milk protein isolate (PRO) for 8 consecutive days following a single bout of exercise (300 eccentric contractions at 30 deg/sec). In both conditions, performance was assessed at baseline, immediately post-exercise, 2h post-exercise and daily for 8 consecutive days. Blood samples were collected at baseline, 2h post-exercise and daily for the remaining 8 days. Muscle biopsies from vastus lateralis were collected at baseline as well as at day 2 and day 8 of the post-exercise period.

6. Conditions and Keywords

Primary Disease or Condition Being Studied in the Trial, or the Focus of the Study
Skeletal Muscle Damage, Exercise-induced Aseptic Inflammation, Skeletal Muscle Performance, Intracellular Signaling in Skeletal Muscle, Proteasome Activation

7. Study Design

Primary Purpose
Basic Science
Study Phase
Not Applicable
Interventional Study Model
Crossover Assignment
Masking
ParticipantInvestigator
Allocation
Randomized
Enrollment
14 (Actual)

8. Arms, Groups, and Interventions

Arm Title
Protein
Arm Type
Experimental
Arm Description
Milk protein isolate supplementation: orally, 20g of protein immediately post-exercise and then 20g every 3h on 3 occasions (+3, +6, +9), on the exercise day. The remaining 8 days, 20g daily with breakfast.
Arm Title
Placebo
Arm Type
Active Comparator
Arm Description
Placebo administration: orally 500 ml, immediately post-exercise as well as at +3, +6 and +9 hours, on the exercise day. The remaining 8 days, 500 ml daily with breakfast.
Intervention Type
Dietary Supplement
Intervention Name(s)
Milk protein isolate
Intervention Description
Milk protein isolate in a powder form consisted of 80% casein and 20% whey protein. 20g were diluted into 500 ml water. Placebo consisted of 365 ml water, 125 ml sugar-free cordial and 2g of low-calorie glucose/dextrose powder.
Intervention Type
Dietary Supplement
Intervention Name(s)
Placebo
Intervention Description
500 mL drink that contained water (375 mL), sugar-free cordial (125 mL) and 2 g of low-calorie glucose/dextrose powder.
Primary Outcome Measure Information:
Title
Change in reduced glutathione in blood
Description
Concentration of reduced glutathione in red blood cells
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in protein carbonyls in serum
Description
Concentration of protein carbonyls
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in protein carbonyls in muscle
Description
Protein carbonyl concentration in quadriceps skeletal muscle group
Time Frame
1h before exercise, 2 days post-exercise, 8 days post-exercise
Title
Change in thiobarbituric acid and reactive substances in serum
Description
Thiobarbituric acid reactive substances concentration in serum
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in oxidized glutathione in blood
Description
Concentration of oxidized glutathione in red blood cells
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in total antioxidant capacity in serum
Description
Total antioxidant capacity in serum
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in catalase activity in serum
Description
Catalase activity in serum
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in creatine kinase activity in plasma
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in C-reactive protein in plasma
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in white blood cell count in blood
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Changes in volume and morphological complexity of immune cells
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in neutrophil count in blood
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in glucose concentration in blood
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in insulin concentration in blood
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in testosterone concentration in plasma
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in cytokine concentration in plasma
Description
Measurement of IL-1β, IL-4, IL-6, IL-8, IL-10, TNF-α
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in adhesion molecule concentration in blood
Time Frame
1h before exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Change in intracellular signalling proteins in muscle
Description
Measurement of phosphorylation levels of mammalian target of rapamycin (mTOR), ribosomal protein S6 (rpS6) and nuclear factor kB (NFkB), and protein expression levels of forkhead box protein O1 (FOXO1), HSP70, and parkin.
Time Frame
1h before exercise, 2 days post-exercise, 8 days post-exercise
Title
Change in proteasome activities in muscle
Description
Measurement of LLVY, LSTR and LLE
Time Frame
1h before exercise, 2 days post-exercise, 8 days post-exercise
Title
Change in protein expression level of proteasome subunits
Description
Measurement of B1i, B2i, B5i, B5, B1, B2 and α7
Time Frame
1h before exercise, 2 days post-exercise, 8 days post-exercise
Secondary Outcome Measure Information:
Title
Change in muscle function of knee extensor and flexor muscle
Description
Assessment of muscle peak and mean torque of knee extensors and flexors on an isokinetic dynamometer at 0, 90 and 180 degrees/sec
Time Frame
1h before exercise, 5 min post-exercise, 2h post-exercise, daily for 8 days post-exercise
Title
Body composition
Description
Assessment of percent (%) body mass
Time Frame
One day before exercise
Title
Maximal aerobic capacity
Description
Assessment of maximal oxygen consumption
Time Frame
One day before exercise
Title
Change in dietary intake profile
Description
Assessment of dietary intake with emphasis on protein consumption
Time Frame
1h before exercise, daily for 8 days post-exercise

10. Eligibility

Sex
Male
Minimum Age & Unit of Time
18 Years
Maximum Age & Unit of Time
30 Years
Accepts Healthy Volunteers
Accepts Healthy Volunteers
Eligibility Criteria
Inclusion Criteria: a) recreationally trained as indicated by the maximal oxygen consumption levels (VO2max > 45 ml/kg/min), b) engaged in systematic exercise at least three times per week for > 12 months, c) non-smokers, d) abstained from any vigorous physical activity during the study, e) abstained from consumption of caffeine, alcohol, performance-enhancing or antioxidant supplements, and medications during the study. Exclusion Criteria: a) a recent febrile illness, b) history of muscle lesion, c) lower limb trauma
Overall Study Officials:
First Name & Middle Initial & Last Name & Degree
IOANNIS G. FATOUROS, Ph.D.
Organizational Affiliation
UNIVERSITY OF THESSALY, SCHOOL OF PHYSICAL EDUCATION & SPORTS SCIENCES
Official's Role
Principal Investigator
Facility Information:
Facility Name
Exercise Biochemistry Laboratory, School of Physical Education & Sports Sciences, University of Thessaly
City
Karies, Trikala
ZIP/Postal Code
42100
Country
Greece

12. IPD Sharing Statement

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Protein Supplementation and Skeletal Muscle Healing Process

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