Study of Effect of Docosahexaenoic Acid (DHA) and Eicosapentaenoic Acid (EPA) Supplementation on Biomarkers of Sub-Concussion Injuries in American Football
Phase: NA
Conditions: Traumatic Brain Injury (TBI), Chronic Traumatic Encephalopathy (CTE)
Sponsor: University of Arizona

Principal Investigator: Floyd H. Chilton, Ph.D.; Roberta D. Brinton, Ph.D. 
Study Title: Study of Effect of Docosahexaenoic Acid (DHA) and Eicosapentaenoic acid (EPA) on Biomarkers 
of Sub-concussion Injuries in American Football. 
IRB #: 1904553365 
 
1.​
Study Identification 
 
1.1.​
Unique Protocol Identification Number: 
1.2.​
Brief Title:  The effects of fish oil supplementation on the brain health of collegiate football 
athletes. 
1.2.1.​
Acronym: not required 
1.3.​
Official Title: Study of Effect of Docosahexaenoic Acid (DHA) and Eicosapentaenoic acid (EPA) 
Supplementation on Biomarkers of Sub-Concussion Injuries in American Football. 
1.4.​
Secondary ID: not required 
1.5.​
Study Type: Interventional (clinical trial) 
 
2.​
Study Status 
 
2.1.​
Record Verification Date:  Month:  Year:  
2.2.​
Overall Recruitment Status: Completed: The study has concluded normally; participants are 
no longer receiving an intervention or being examined (that is, last participant’s last visit has 
occurred) 
2.2.1.​
Why Study Stopped: NA 
2.3.​
Study Start date: May 29, 2019  
2.4.​
Primary Completion date: January 28, 2020 
2.5.​
Study Completion Date: January 28, 2020 
 
3.​
Sponsor/Collaborators 
 
3.1.​
Responsible Party, by Official Title: Principle Investigator 
3.1.1.​
Investigator Information: 
●​ Investigator Name: Floyd H. Chilton, Ph.D. 
●​ Investigator Official Title: Professor, Department of Nutritional Sciences; 
Associate; Director BIO5 Institute; Director, The Precision Wellness Initiative 
●​ Investigator Affiliation: University of Arizona 
3.2.​
Name of Sponsor: Center for Innovation in Brain Science, Roberta D. Brinton, Ph.D. 
3.3.​
Collaborators: NA 
 
4.​
Oversight 
 
4.1.​
Studies a U.S. FDA-regulated Drug Product: No  
4.2.​
Studies a U.S. FDA-regulated Drug Product: NA 
4.3.​
US FDA IND or IDE: No 
4.4.​
Product Manufactured in and Exported from U.S: Yes 
4.5.​
Human Subjects Review: Yes 
4.6.​
Human Subjects Protection Review Board Status: Submitted, approved 
1 
Version Date: 03/18/2020 

Principal Investigator: Floyd H. Chilton, Ph.D.; Roberta D. Brinton, Ph.D. 
Study Title: Study of Effect of Docosahexaenoic Acid (DHA) and Eicosapentaenoic acid (EPA) on Biomarkers 
of Sub-concussion Injuries in American Football. 
IRB #: 1904553365 
 
4.6.1.​
Board Approval Number: 1904553365 
4.6.2.​
Board Name: Internal Review Board 
4.6.3.​
Board Affiliation: University of Arizona 
4.6.4.​
Board Contact: Christine Melton-Lopez 
●​ Phone: (520) 626-8630 
●​ Extension: NA 
●​ Email: melton1@email.arizona.edu 
●​ Address: POB 210409, Tucson, AZ 85721 
4.7.​
Data Monitoring Committee: Yes 
4.8.​
FDA Regulated Intervention: No 
 
5.​
Study Description 
 
5.1.​
Brief Summary: A short description of the clinical study, including a brief statement of the 
clinical study's hypothesis, written in language intended for the lay public. Limit: 5000 
characters.  
American football is one of the most popular sports in the U.S. Yet this sport is associated with 
increased risk of concussion (also known as mild traumatic brain injury, or mTBI) and 
subconcussive injury from repeated head impacts (RHI) due to the aggressive and highspeed 
nature of the game. Current protective equipment used by players are not sufficient to reduce 
concussion incidence and severity, nor are there any therapeutics available to prevent 
concussion. This study is a randomized, double-blind, placebo controlled trial, including 38 
research participants, to determine whether an omega-3 polyunsaturated fatty acids (PUFA) fish 
oil supplement containing 3.0 grams of docosahexaenoic acid (DHA; 22:6n-3) and 
eicosapentaenoic acid (EPA; 20:5n-3) can reduce blood biomarkers of sub-concussion injuries 
compared to placebo (high-oleic safflower oil) over a course of the American football season 
among collegiate football athletes. Secondary outcomes include 1) determining the effect of 
daily DHA and EPA supplementation on brain imaging markers of sub-concussion injuries, and 
2) a responder analysis to determine whether genotype differences in fatty acid desaturase 1 
(FADS1), FADS2, elongation of very long chain fatty acids 2 (ELOVL2), and ELOVL5 genes 
affect treatment outcome of DHA. An exploratory outcome is the discovery of novel and more 
sensitive metabolic or lipidomic biomarkers of sub-concussion injury. The dosage of DHA/EPA 
used in this study is generally safe, and procedures involved, monthly blood draws, surveys, 
and Magnetic Resonance Imaging (MRI), pose minimal risks to participants. While this study 
provides no direct benefit to participants, successful outcomes of this study can benefit the 
society by shedding light on development of potential preventative therapeutics for 
sports-induced mTBI and brain injury from RHI. The risk-benefit profile is appropriate for 
conducting this study. Based on preclinical studies and previous clinical study results, we expect 
that in comparison to placebo treatment, DHA and EPA treatment throughout the course of one 
American football season can maintain lower levels of sub-concussion associated biomarkers, 
2 
Version Date: 03/18/2020 

Principal Investigator: Floyd H. Chilton, Ph.D.; Roberta D. Brinton, Ph.D. 
Study Title: Study of Effect of Docosahexaenoic Acid (DHA) and Eicosapentaenoic acid (EPA) on Biomarkers 
of Sub-concussion Injuries in American Football. 
IRB #: 1904553365 
 
inflammatory cytokines, and cardiovascular risk markers. We also expect participants treated 
with DHA and EPA to have lower brain MRI imaging markers of sub concussion injury.  
 
5.2.​
Detailed Description: (not required) Definition: Extended description of the protocol, including 
more technical information (as compared to the Brief Summary), if desired. Do not include the 
entire protocol; do not duplicate information recorded in other data elements, such as Eligibility 
Criteria or outcome measures. Limit: 32,000 characters. 
 
6.​
Conditions and key words 
 
6.1.​
Primary Condition Being Studied: Sub-concussive brain injury as measured by biomarkers of 
axonal damage from repeated head impacts (RHI).  
6.2.​
Keywords: (not-required) concussion, mTBI, repeated head impacts, football, brain, Chronic 
traumatic encephalopathy, Magnetic resonance imaging, Neurofilament light chain, biomarker 
 
7.​
Study design 
 
7.1.​
Primary Purpose: Prevention 
7.2.​
Study Phase: NA 
7.3.​
Interventional Study Model: Parallel 
7.4.​
Model Description: Participants will be randomized to treatment (3 grams DHA and EPA (2:1 
weight ratio)) or placebo (3 grams high-oleic safflower oil) in a 1:1 allocation ratio based on their 
position on the football team and roster depth (starter vs. non-starter). Participants will be 
recruited as a single cohort. After screening, participants will be treated with DHA and EPA or 
matching placebo for 25 weeks (a full course of an American football season, including summer 
camp training sessions). A follow-up visit is scheduled 7 weeks after the final dosing at week 32 
for recovery and adverse event evaluation. 
7.5.​
Number of Arms: Two 
7.6.​
Masking Roles: Participant, investigator, care provider, outcomes assessor 
7.7.​
Masking Description: Study personnel will be kept blind to treatment assignments. The 
treatment assignment for each participant will be available to the investigator in a sealed 
envelope that may be opened only in the case of a serious adverse event which the investigator 
feels cannot be adequately treated without knowing the identity of the study medication. Code 
breakers will be collected and reviewed at the end of the study. If the blinding has been broken, 
then the investigator will provide documentation regarding the fact and indicate the other staff 
that received this information.  
7.8.​
Allocation: Randomized 
7.9.​
Enrollment Type: Actual 
7.10.​
Number of Subjects: 38 
 
8.​
Arms, Groups, and Interventions 
3 
Version Date: 03/18/2020 

Principal Investigator: Floyd H. Chilton, Ph.D.; Roberta D. Brinton, Ph.D. 
Study Title: Study of Effect of Docosahexaenoic Acid (DHA) and Eicosapentaenoic acid (EPA) on Biomarkers 
of Sub-concussion Injuries in American Football. 
IRB #: 1904553365 
 
 
8.1.​
Arm 1 Type: Active comparator 
8.2.​
Arm 1 Title: Treatment 
8.3.​
Arm 2 Type: Placebo Comparator 
8.4.​
Arm 2 Title: Placebo 
8.5.​
Intervention 1: 
8.5.1.​
Intervention 1 Type: Dietary Supplement 
8.5.2.​
Intervention 1 Name: Fish oil supplement containing docosahexaenoic acid (DHA; 
22:6n-3) and eicosapentaenoic acid (EPA; 20:5n-3) 
8.5.3.​
Intervention 1 Description: Participants consumed a dietary supplement consisting of 3 
grams refined fish oils as DHA and EPA ethyl esters in 2:1 ratio 5 days a week for 25 
weeks. The product is stabilized with identity preserved mixed natural tocopherols and 
maintains stability for 24 months when stored in original, unopened, light-resistant 
containers at a temperature below 30° Celsius. Refined fish oil was encapsulated so six 
capsules provide 3 gram per dose. Certificate of analysis confirming manufacture date, 
active ingredient, fill weight, and microbiology safety, and disintegration time was 
provided. A dose of 3 grams DHA and EPA, 5 days per week, was based on a previous 
clinical study in a similar population using the same compounds5. This dose is 
considered generally safe, and can provide sufficient bioavailability to significantly 
reduce plasma NfL level (a biomarker of brain injury and neurodegeneration)5. The dose 
remained constant for each participant during the study period. If a participant 
experiences an adverse event (AE) considered to be possibly related to the study dietary 
supplement, the investigator has the option to split up the dose throughout the day, 
recommend the participant take the supplement with food or to discontinue the 
participant. 
8.6.​
Intervention 2: 
8.6.1.​
Intervention 2 Type: Placebo 
8.6.2.​
Intervention 2 Name:  
8.6.3.​
Intervention 2 Description: Participants consumed a placebo consisting of 
encapsulated high-oleic safflower oil. Each placebo capsule contains 866.9 mg/capsule 
active ingredient. The placebo intervention included consumption of 6 capsules 5 days a 
week for 25 weeks.  A certificate of analysis confirming manufacture date (06/03/2019), 
active ingredient (safflower oil), fill weight (866.9 milligrams), and microbiology safety (all 
negative), and disintegration time (3 minutes) was provided. 
8.7.​
Arm/Intervention Cross-Reference: Arm 1 = Intervention 1; Arm 2 = Intervention 2 
 
9.​
Outcome Measures 
9.1.​
Outcome 1: Primary Outcome Measure  
9.1.1.​
Title 1: The effect of DHA and EPA Omega-3 PUFA fish oil supplementation on blood 
biomarkers of brain injury compared to placebo (high-oleic safflower oil) over a course of 
4 
Version Date: 03/18/2020 

Principal Investigator: Floyd H. Chilton, Ph.D.; Roberta D. Brinton, Ph.D. 
Study Title: Study of Effect of Docosahexaenoic Acid (DHA) and Eicosapentaenoic acid (EPA) on Biomarkers 
of Sub-concussion Injuries in American Football. 
IRB #: 1904553365 
 
the American football season among collegiate football athletes exposed to repeated 
subconcussive head impacts. 
9.1.2.​
Time Frame 1: 32 weeks from baseline to one month postseason/treatment 
9.1.3.​
Description 1: (not required) Blood-based biomarkers for sub-concussion injury 
(neurofilament light (NFL), Tau protein (Tau), ubiquinone C-terminal hydrolase  L1 
(UCH-L1)) and can lower inflammatory cytokines (C-reactive protein (CRP), tumor 
necrosis factor 1 (TNF-α), interleukin 6 (IL6)) will be measured at baseline, 
mid-treatment, post treatment, and one month (recovery) after the end of treatment and 
the football season. 
9.2.​
Outcome 2: Primary Outcome Measure 
9.2.1.​
Title 2: The effect of DHA and EPA Omega-3 PUFA fish oil supplementation on blood 
cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), and triglyceride 
levels in collegiate football athletes. 
9.2.2.​
Time Frame 2: 25 weeks from baseline to end of treatment 
9.2.3.​
Description 2: (not required) A lipid panel will be assessed at baseline and end of 
treatment. 
9.3.​
Outcome 3: Secondary Outcome Measure 
9.3.1.​
Title 3: The effect of DHA and EPA Omega-3 PUFA fish oil supplementation on brain 
imaging markers of sub-concussion injury collegiate football athletes over the course of 
the American football season. 
9.3.2.​
Time Frame 3: 25 weeks from baseline to end of treatment 
9.3.3.​
Description 3: (not required) MRI scans were obtained at baseline and end of 
treatment. 
9.4.​
Outcome 4: Secondary Outcome Measure 
9.4.1.​
Title 4: The effect of genotype differences on DHA and EPA treatment outcomes on 
blood based biomarker or brain MRI imaging biomarker as described in specific aim 1. 
9.4.2.​
Time Frame 4: 32 weeks from baseline to one month post treatment and post end of 
football season. 
9.4.3.​
Description 4: (not required)  
9.5.​
Outcome 5: Other Pre-Specified Outcome Measures 
9.5.1.​
Title 5: Conduct non-targeted metabolomic analysis to potentially identify additional 
potential biomarkers for sub-concussion injuries.  
9.5.2.​
Time Frame 4: 32 weeks from baseline to one month post treatment and post end of 
football season. 
9.5.3.​
Description 5: (not required)  
 
10.​
Eligibility 
10.1.​
Sex: Male 
10.1.1.​
Gender-Based: (not required) No 
10.2.​
Age Limits:  
10.2.1.​
Minimum Age: 18 
5 
Version Date: 03/18/2020 

Principal Investigator: Floyd H. Chilton, Ph.D.; Roberta D. Brinton, Ph.D. 
Study Title: Study of Effect of Docosahexaenoic Acid (DHA) and Eicosapentaenoic acid (EPA) on Biomarkers 
of Sub-concussion Injuries in American Football. 
IRB #: 1904553365 
 
10.2.2.​
Unit of Time: Years 
10.2.3.​
Maximum Age: NA 
10.2.4.​
Unit of Time: NA 
10.3.​
Accepts healthy Volunteers: Yes 
10.4.​
Eligibility Criteria:  
10.4.1.​
Inclusion Criteria 
●​ University of Arizona  
●​ National Collegiate Athletic Association (NCAA)  
●​ Collegiate football players  
●​ Cleared to participate in university athletics as determined by the team physician.  
●​ >18 years. 
10.4.2.​
Exclusion Criteria 
●​ Chronic daily anti-inflammatory medication (>20 d). 
●​ Medications for blood lipids.  
●​ Active fish oil or omega-3 fatty acid supplementation. 
●​ Consumption of more than two servings of fish per week. 
●​ Injured and unable to participate in regular schedule conditioning or 
competitions. 
●​ Prior diagnosis acute concussion. 
●​ Allergic to fish or shellfish. 
 
 
 
 
 
 
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