Investigating the Efficacy of Low or High Dose Tart Cherry on Recovery From Exercise Induced Muscle Damage and Establishing Its Molecular Mechanisms of Action

Sponsor

PepsiCo Global R&D (Industry)

Overall Status

Recruiting

CT.gov ID

NCT04725149

Collaborator

(none)

Enrollment

Location

Arms

28.5

Anticipated Duration (Months)

1.2

Patients Per Site Per Month

Study Details

Study Description

Brief Summary

To determine the dose-response effects of 10-day tart cherry product consumption (0, 30 ml, and 60 ml) on knee extensor isometric strength 24-h and 48-h after muscle damaging exercise and to elucidate the mechanisms of action for TC supplementation.

Condition or Disease	Intervention/Treatment	Phase
Muscle Recovery	Other: 500 ml base solution Other: 500 ml containing 30 ml tart cherry concentrate diluted with water and base solution Other: 500 ml containing 60 ml tart cherry concentrate diluted with water only	N/A

Detailed Description

Montmorency cherry supplementation has been shown to exert antioxidant and anti-inflammatory effects that can be beneficial for improving recovery from exercise. However, the investigator is currently not aware of how tart cherry polyphenol supplementation produces these effects. This study aims to determine the dose-response effects of 10-day tart cherry product consumption (placebo, 30 ml, and 60 ml Montmorency tart cherry concentrate in a 500 ml beverage) on knee extensor isometric strength 24-h and 48-h after muscle damaging exercise on supplementation Day 8; on enhancing recovery of other measures of muscle function (single leg eccentric and concentric force development and single leg jump height) and muscle soreness; on reducing markers of oxidative stress and inflammation in plasma, muscle and urine; on inducing signaling in muscle via the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway to upregulate endogenous antioxidant enzymes in muscle; on inhibiting muscle cyclo-oxygenase (COX-1 and COX-2).

This study also aims to identify and establish the molecular mechanisms of action through which tart cherry polyphenols exert antioxidant and anti-inflammatory effects. Oxidative stress and inflammatory signaling in primary human myogenic cells will be assessed by incubating primary human myogenic cells (commercial cell line) in sera derived from 6 participants consuming 8-day placebo vs. 60 ml/day Montmorency tart cherry concentrate supplement. Subjects are permitted to participate in both the molecular mechanism of action part of the study (Part A, no exercise component) and the damaging exercise muscle recovery part of the study (Part B).

Study Design

Study Type:

Interventional

Anticipated Enrollment :

34 participants

Allocation:

Randomized

Intervention Model:

Crossover Assignment

Intervention Model Description:

Repeated measures, double-blind, counter-balancedRepeated measures, double-blind, counter-balanced

Masking:

Quadruple (Participant, Care Provider, Investigator, Outcomes Assessor)

Masking Description:

Taste-matched placebo

Primary Purpose:

Basic Science

Official Title:

Investigating the Efficacy of Low or High Dose Tart Cherry on Recovery From Exercise Induced Muscle Damage and Establishing Its Molecular Mechanisms of Action

Actual Study Start Date :

Dec 14, 2020

Anticipated Primary Completion Date :

Apr 30, 2023

Anticipated Study Completion Date :

Apr 30, 2023

Arms and Interventions

Arm	Intervention/Treatment
Placebo Comparator: 0 ml tart cherry concentrate Still cherry-flavored beverage, similar in appearance, taste, aroma and calories to the 30 ml and 60 ml tart cherry concentrate beverages	Other: 500 ml base solution Color, taste, aroma, and calorie matched water base
Experimental: 30 ml tart cherry concentrate "Low dose" tart cherry concentrate beverage	Other: 500 ml containing 30 ml tart cherry concentrate diluted with water and base solution Montmorency cherry concentrate
Experimental: 60 ml tart cherry concentrate "High dose" tart cherry concentrate beverage	Other: 500 ml containing 60 ml tart cherry concentrate diluted with water only Montmorency cherry concentrate

Arm

Intervention/Treatment

Placebo Comparator: 0 ml tart cherry concentrate

Still cherry-flavored beverage, similar in appearance, taste, aroma and calories to the 30 ml and 60 ml tart cherry concentrate beverages

Other: 500 ml base solution

Color, taste, aroma, and calorie matched water base

Experimental: 30 ml tart cherry concentrate

"Low dose" tart cherry concentrate beverage

Other: 500 ml containing 30 ml tart cherry concentrate diluted with water and base solution

Montmorency cherry concentrate

Experimental: 60 ml tart cherry concentrate

"High dose" tart cherry concentrate beverage

Other: 500 ml containing 60 ml tart cherry concentrate diluted with water only

Montmorency cherry concentrate

Outcome Measures

Primary Outcome Measures

Isometric knee extensor strength [Changes from immediately before and immediately after muscle damaging exercise on day 8 of tart cherry supplementation to 24 hours and 48 hours after muscle damaging exercise (days 9 and 10 of supplementation).]
Maximum voluntary contraction (MVC) as a measure of functional recovery from muscle damage. Unit of measurement is Newtons. Stronger is better.

Secondary Outcome Measures

Single leg eccentric and concentric isokinetic force production [Changes from immediately before and immediately after muscle damaging exercise on day 8 of tart cherry supplementation to 24 hours and 48 hours after muscle damaging exercise (days 9 and 10 of supplementation).]
Dynamic muscle strength measured by isokinetic contraction (IKC, 60 deg.s-1). More forceful is better.
Single leg vertical jump height (SLVJ) [Changes from immediately before and immediately after muscle damaging exercise on day 8 of tart cherry supplementation to 24 hours and 48 hours after muscle damaging exercise (days 9 and 10 of supplementation).]
Dynamic muscle strength measured by jump height (cm). Higher is better.
Leg muscle soreness by Pressure Pain tolerance (PPT) [Changes from immediately before and immediately after muscle damaging exercise on day 8 of tart cherry supplementation to 24 hours and 48 hours after muscle damaging exercise (days 9 and 10 of supplementation).]
Pressure pain tolerance (PPT) measured by algometer. Higher tolerance is better.
Leg muscle soreness by Visual Analogue Scale (VAS) [Changes from immediately before and immediately after muscle damaging exercise on day 8 of tart cherry supplementation to 24 hours and 48 hours after muscle damaging exercise (days 9 and 10 of supplementation).]
Measured using a visual analogue pain scale from 0 mm (no pain) to 100 mm (extremely painful). Lower score is better.
Leg muscle biopsy for nuclear factor k-B (NFκB) [Changes from pre and post exercise on day 8 of tart cherry supplementation to pre-exercise on days 9 and 10 of supplementation. Biopsies are done after blood samples and before muscle soreness measures.]
Inflammation marker expression activity of the NFκB pathway will be measured by immunoblotting. Lower is better.
Leg muscle biopsy for cyclo-oxygenase COX1 protein [Changes from pre and post exercise on day 8 of tart cherry supplementation to pre-exercise on days 9 and 10 of supplementation. Biopsies are done after blood samples and before muscle soreness measures.]
Inflammation marker of cyclo-oxygenase measured by total protein carbonylation (immunoblotting). A reduction (inhibiting) is better.
Leg muscle biopsy for cyclo-oxygenase COX1 activity [Changes from pre and post exercise on day 8 of tart cherry supplementation to pre-exercise on days 9 and 10 of supplementation. Biopsies are done after blood samples and before muscle soreness measures.]
Inflammation marker of cyclo-oxygenase activity measured by commercially available colorimetric assay. A reduction (inhibiting) is better.
Leg muscle biopsy for nuclear factor erythroid 2-related factor 2 (Nrf2), absolute expression, nuclear translocation [Changes from pre and post muscle damaging exercise on day 8 of tart cherry supplementation to pre-exercise on days 9 and 10 of supplementation. Biopsies are done after blood samples and before muscle soreness measures.]
Oxidative damage marker Nrf2 pathway intended to upregulate endogenous antioxidant enzymes in muscle. Induction of signaling is better.
Leg muscle biopsy for endogenous antioxidant enzyme expression [Changes from pre and post muscle damaging exercise on day 8 of tart cherry supplementation to pre-exercise on days 9 and 10 of supplementation. Biopsies are done after blood samples and before muscle soreness measures.]
Enzymes (superoxide dismutase 1, superoxide dismutase 2, glutathione peroxidase 1 and catalase) protein expression quantified by immunoblotting. Higher values (upregulation by Nrf2) is better.
Leg muscle biopsy for immune cell infiltration [Changes from pre and post muscle damaging exercise on day 8 of tart cherry supplementation to pre-exercise on days 9 and 10 of supplementation. Biopsies are done after blood samples and before muscle soreness measures.]
Inflammation marker response measured by immunohistochemistry. Less infiltration is better.
Plasma interleukin-6 (IL-6) [Changes from pre and post muscle damaging exercise on day 8 of tart cherry supplementation to pre-exercise on days 9 and 10 of supplementation. Pre exercise blood sampling is done after urine collection. Post exercise blood sampling done before biopsy.]
Plasma inflammatory stress response measured by ELISA. A reduction is better.
Plasma protein carbonyls [Changes from pre and post muscle damaging exercise on day 8 of tart cherry supplementation to pre-exercise on days 9 and 10 of supplementation. Pre exercise blood sampling is done after urine collection. Post exercise blood sampling done before biopsy.]
Oxidative stress response and antioxidant status measured by ELISA. A reduction is better.
Plasma phenolic concentration [Changes from pre and post muscle damaging exercise on day 8 of tart cherry supplementation to pre-exercise on days 9 and 10 of supplementation. Pre exercise blood sampling is done after urine collection. Post exercise blood sampling done before biopsy.]
LC-MS-MS metabolite analysis to evaluate plasma levels associated with tart cherry supplementation at 0, 30 and 60 ml/day.
Urine oxidation-reduction potential (ORP) [Change from pre-exercise value day 8 to days 9 and 10 of tart cherry supplementation. Collected upon arrival to the study site.]
Urinary oxidation-reduction potential to measure oxidative damage and antioxidant status measured by probe. Less oxidative stress/free radical production is better.

Eligibility Criteria

Criteria

Ages Eligible for Study:

18 Years to 40 Years

Sexes Eligible for Study:

Male

Accepts Healthy Volunteers:

Yes

Inclusion Criteria:

Male in good health
18-40y
BMI ≤ 29.9
Recreationally active but not highly trained (Participating in sport or physical activity at least once per week for the last 6 months and lower limb resistance training ≤ 1/week) (Part B only)
No lower limb musculoskeletal injury that prevented exercise in the last 3 months (Part B only)
Understanding of the procedures to be undertaken as part of the study
Willing to participate in the exercise protocols (Part B only) and follow supplementation guidelines and other instructions provided by the experimenter
Willing to abstain from caffeine, alcohol and strenuous exercise for two 5 day periods (from 48h prior to muscle damaging exercise and throughout the follow up period; days 6-10 of the trial) for Part B, and abstain from caffeine and alcohol for two 3-day periods for Part A
Willing to keep food diaries for the duration of each arm of the trial and avoid multiple servings per day of foods that have a high polyphenol content from a list provided.
Willing to construct their diet for the first arm of the trial from a list of recipes provided, and to then replicate this diet during the second arm of the trial.
Willing to wear a triaxial accelerometer throughout each arm of the trial, and during the second arm of the trial to replicate their physical activity pattern completed in the first arm of the trial.
Willing to abstain from consumption of anti-inflammatory, analgesic medications or those with anti-coagulant effects especially NSAIDs and aspirin for 48h prior to and throughout each arm of the trial. (Part B only)
Willing to abstain from icing or applying compression bandages to the exercised/biopsied leg (Part B only)
Ability to provide voluntary written informed consent to participate in the study.

Exclusion Criteria:

Female
<18y or >40y
BMI ≥ 30
Inactive or well-trained (see inclusion criteria) (Part B only)
Lower limb musculoskeletal injury in the last 3 months (Part B only)
Under the care of a physician
Known health condition that could interfere with any element of the data collection protocol. Any known pulmonary, gastrointestinal (e.g., irritable bowel syndrome, Crohn's disease), kidney, cardiovascular, or metabolic diseases, blood-borne infections, or coagulopathies including deep vein thrombosis (DVT)
Current regular consumption of nutritional/dietary supplements or previous use of any dietary supplements in the past 6 months known to have a lasting effect.
Current use of prescription medication; including the use of an inhaler
Non-steroidal anti-inflammatory drugs, aspirin or other analgesic medication or any anti-coagulants are prohibited during the study and for 48h prior to commencing supplementation
Allergy/sensitivity/intolerance to cherries, any cherry derived product, or any of the additional ingredients found within the TC test products, including sensitivity to sorbitol.
Allergy to lidocaine or other local anaesthetics. (Part B only)
Active drug or alcohol dependency
Current or active usage of nicotine or tobacco related products
Participation in a clinical trial within the past 30 days or concurrently during this trial
Participation in any PepsiCo trial within past 6 months, except that Part A participants may take part in Part B after a >=14 day washout from Part A
Inability to provide written informed consent