Can Brief Exposure to Hyperoxia Improve Function After Chronic Spinal Cord Injury?
Phase: EARLY_PHASE1
Conditions: Spinal Cord Injuries
Sponsor: University of Alberta

 
   
 
 
Protocol_Hyperoxia in SCI_v13 
 
Oct 1, 2021 
 
 
PROTOCOL COVER PAGE 
 
 
 
 
PROTOCOL NAME  
Can brief exposure to hyperoxia improve function after chronic spinal cord injury? 
 
 
PROTOCOL IDENTIFYING NUMBER 
Pro00061817 
 
 
PROTOCOL AMENDMENT VERSION 
Version 13 
 
PROTOCOL AMENDMENT DATE 
October 1, 2021 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
 
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GENERAL INFORMATION 
 
 
 
 
Name and address of the sponsor and monitor of the study 
 
University of Alberta, Edmonton, AB. 
 
 
Name and address of the person authorized to sign the protocol and amendments  
 
Jaynie Yang PT, PhD, 3-75 Corbett Hall, University of Alberta 
 
 
Name, title, address and telephone number(s) of the medical expert for the trial 
 
Eric Wong, MD, Associate Professor, Pulmonary Medicine, 3-127 Clinical Sciences Bldg, 
University of Alberta 
Tel: 780-492-3820 
 
 
Name and title of the investigator(s) and sub-investigators responsible for the trial with 
address and phone number(s)   
 
Jaynie Yang, Professor, 3-75 Corbett Hall, University of Alberta.  Tel: 780-492-2894 
David Bennett, Professor, 526 Heritage Medical Research Centre, University of Alberta. Tel: 
780-492-1516 
Karim Fouad, Professor, 3-87 Corbett Hall, University of Alberta. Tel: 780-492-5971 
Eric Wong, Associate Professor, 3-127 Clinical Sciences Bldg, University of Alberta. Tel: 780-
492-3820 
Chester Ho, Professor, 5-0520 G Katz Group Centre, University of Alberta. Tel: 780-492-4388 
 
 
Name, title and address of qualified physician who is responsible for trial-site related 
medical decisions 
 
Eric Wong, MD, Associate Professor, Pulmonary Medicine, 3-127 Clinical Sciences Bldg, 
University of Alberta 
Tel: 780-492-3820 
 
Trial site location and contact 
 
Location: Rm 2-151, Clinical Sciences Bldg., University of Alberta, Edmonton, AB 
Phone number: 780-492-5949 
Physical therapist: Michelle Barnes 
 
 

 
 
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Study Summary 
Title 
Can brief exposure to hyperoxia improve function after chronic spinal 
cord injury? 
Short Title 
N/A 
Protocol Number 
 Pro00061817, Version 13 
Phase 
Proof-of-principle study 
Methodology 
Prospective, double-blind, randomized crossover design  
Study Duration 
Projected end date March 31, 2026 
Study Center(s) 
Single centre 
Objectives 
Determine if breathing an increased proportion of oxygen above the 
concentration in normal room air results in changes in the sensory and 
motor function in people with severe spinal cord injury (SCI). 
 
Number of Subjects 
35 (25 with spinal cord injury, 10 uninjured controls) 
Diagnosis and Main 
Inclusion Criteria 
Chronic and subacute, traumatic spinal cord injury; uninjured healthy 
controls for ensuring experimental protocol without using oxygen 
Study Product, Dose, 
Route, Regimen 
Study products are: 1) oxygen (DIN# 02014408), and 2) compressed 
air (DIN# 02014483), delivered during different experiments through a 
face mask with flow rate of 10 litres/min, once per experiment, over 2 
experimental days, with experiments spaced by at least 2 weeks. 
Duration of 
administration 
2 minutes per experiment 
Reference therapy 
Placebo – compressed air (DIN# 02014483)  
Statistical 
Methodology 
Analysis will be guided by a statistician, Dr Ming Ye, consultant with 
the Faculty of Rehabilitation Medicine, Rehabilitation Research 
Centre. 
 
  
 
 

 
 
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Table of Contents 
1  
BACKGROUND ............................................................................................................................................... 1 
1.1 
INVESTIGATIONAL AGENT ........................................................................................................................... 1 
1.2 
PRECLINICAL DATA ..................................................................................................................................... 1 
1.3 
RISK/BENEFITS ............................................................................................................................................ 1 
1.4 
DOSE RATIONALE ........................................................................................................................................ 1 
1.5 
TRIAL CONDUCT ......................................................................................................................................... 1 
1.6 
POPULATION................................................................................................................................................ 1 
1.7 
LITERATURE ................................................................................................................................................ 1 
2 
TRIAL OBJECTIVES...................................................................................................................................... 2 
3 
TRIAL DESIGN ............................................................................................................................................... 2 
3.1 
PRIMARY STUDY ENDPOINTS/SECONDARY ENDPOINTS .............................................................................. 2 
3.2 
STUDY DESIGN/TYPE .................................................................................................................................. 2 
3.3 
RANDOMIZATION ........................................................................................................................................ 2 
3.4 
MAINTENANCE ............................................................................................................................................ 2 
3.5 
TRIAL TREATMENT ...................................................................................................................................... 2 
3.6 
DURATION ................................................................................................................................................... 2 
3.7 
DISCONTINUATION ...................................................................................................................................... 2 
3.8 
PRODUCT ACCOUNTABILITY ....................................................................................................................... 6 
3.9 
DATA IDENTIFICATION ................................................................................................................................ 6 
4 
SELECTION AND WITHDRAWAL OF SUBJECTS .................................................................................. 6 
4.1 
INCLUSION CRITERIA ................................................................................................................................... 6 
4.2 
EXCLUSION CRITERIA ................................................................................................................................. 7 
4.3 
SUBJECT WITHDRAWAL .............................................................................................................................. 7 
4.4 
TREATMENT OF SUBJECTS ........................................................................................................................... 8 
4.5 
MEDICATION ............................................................................................................................................... 8 
4.6 
MONITORING FOR SUBJECT COMPLIANCE .................................................................................................... 8 
5 
ASSESSMENT OF EFFICACY ...................................................................................................................... 4 
5.1 
EFFICACY PARAMETERS .............................................................................................................................. 4 
5.2 
METHOD AND TIMING ................................................................................................................................. 4 
6 
ASSESSMENT OF SAFETY ........................................................................................................................... 4 
6.1 
SAFETY PARAMETERS ................................................................................................................................. 4 
6.2 
METHOD AND TIMING ................................................................................................................................. 4 
6.3 
ADVERSE EVENT REPORTING ...................................................................................................................... 4 
6.4 
DEFINITIONS ................................................................................................................................................ 4 
6.5 
ADVERSE EVENT FOLLOW-UP ..................................................................................................................... 4 
7 
STATISTICAL PLAN ...................................................................................................................................... 5 
7.1 
STATISTICAL METHODS............................................................................................................................... 5 
7.2 
SUBJECT POPULATION(S) FOR ANALYSIS .................................................................................................... 5 
7.3 
SIGNIFICANCE ............................................................................................................................................. 5 
7.4 
TERMINATION CRITERIA ............................................................................................................................. 5 
7.5 
ACCOUNTABILITY PROCEDURE ................................................................................................................... 5 
7.6 
DEVIATION REPORTING ............................................................................................................................... 5 
8 
DIRECT ACCESS TO SOURCE DATA/DOCUMENTATION .................................................................. 5 
9 
QUALITY CONTROL AND QUALITY ASSURANCE .............................................................................. 5 
10 
ETHICAL CONSIDERATIONS ..................................................................................................................... 6 
11 
DATA HANDLING AND RECORD KEEPING ........................................................................................... 6 

 
 
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12 
FINANCE AND INSURANCE ........................................................................................................................ 6 
13 
PUBLICATION PLAN .................................................................................................................................... 6 
14 
SUPPLEMENTS ............................................................................................................................................... 6 
 

 
 
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List of Abbreviations 
 
 
ICH 
 
International Conference on Harmonisation 
 
CRF 
 
Case Report Form 
 
GCP  
Good Clinical Practice 
 
HREB  
Health Research Ethics Board 
 
 
 
 

 
Protocol_Hyperoxia in SCI_v13 
 
Oct 1, 2021 
 
1 Background 
 
1.1 Investigational Agent 
Pure oxygen (99%) from Praxair, Edmonton, DIN# 02014408, delivered through a face mask at 
a flow rate of 10 litres/min, for a period of 2 minutes.  Reference (placebo) agent is compressed 
air from Praxair, DIN# 02014483, delivered in the same way. 
 
1.2 Preclinical Data 
    
An abstract from the preclinical trial in rodents is included below.  This study details the 
mechanisms underlying chronic hypoxia in the spinal cord distal to a complete spinal cord 
transection in a rodent model, and how brief exposure to breathing high levels of oxygen 
modifies the motor behaviour of the rodent.  Because there is no way to measure the oxygenation 
in the human spinal cord non-invasively, we wish to determine if brief exposure to pure oxygen 
in a human with severe spinal cord injury modifies the sensory and motor behaviour in a way 
similar to that in rodents.  If so, it would suggest that the human spinal cord distal to a severe 
spinal cord injury is also chronically hypoxic, and interferes with sensory and motor function.  
Ways to restore oxygenation to the spinal cord could then be pursued in future studies. 
 
Pericytes impair capillary blood flow and motor function after chronic spinal 
cord injury 
 
Yaqing Li1, Ana M. Lucas-Osma1, Sophie Black1, Mischa V. Bandet2, Marilee J. Stephens1, 
Romana Vavrek1, Leo Sanelli1, Keith K. Fenrich1, Antonio Dinarzo3,  
Stella Dracheva3, Ian R. Winship2, Karim Fouad1 and David J. Bennett1 
1Neuroscience and Mental Health Institute and Faculty of Rehabilitation Medicine, University of 
Alberta, Canada; 2Neuroscience and Mental Health Institute and Department of Psychiatry, 
University of Alberta, Canada; 3Department of Genetics and Genomic Sciences, Icahn School of 
Medicine at Mount Sinai, New York, NY, USA 
 
Nature Medicine (2017) 23(6):733 doi:10.1038/nm.43311 
 
ABSTRACT 
Blood vessels in the CNS are controlled by neuronal activity, including widespread vessel 
constrictions (tone) induced by brainstem neurons that release the monoamines serotonin and 
noradrenaline, and local vessel dilation mediated by glutamatergic neuron activity. We examine 
how vessel tone adapts to the loss of these monoamines after spinal cord injury (SCI) in rats. We 
find that, months after SCI, the cord below the injury is in a chronic state of hypoxia, produced by a 
paradoxical excess activity in monoamine receptors (5-HT1), despite the absence of monoamines. 
These receptors activate pericytes that locally constrict capillaries, reducing blood flow to ischemic 
levels. The paradoxical receptor activity results from trace amines (tryptamine) produced by 
pericytes that ectopically express the enzyme aromatic-L-amino-acid-decarboxylase (AADC), 

 
 
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which synthesizes trace amines directly from dietary amino acids (tryptophan).  However, blocking 
monoamine receptors or AADC, or briefly inhaling increased oxygen, produces long-term relief 
from hypoxia and improves locomotor function. 
 
1.3 Risk/Benefits 
Oxygen therapy with up to 100% oxygen for much longer than we propose to use (which is 2 
mins) is used in the management of pulmonary conditions and emergency medicine (Vogelsinger 
et al 2017; Branson 2018; Brugnliaux et al 2018), and is also used to enhance physical 
performance in athletics (Brugnliaux et al 2018). Symptoms of oxygen toxicity does not start to 
develop in humans breathing pure oxygen at normobaric (i.e., 1 atmospheric pressure, which is 
our proposed condition) until after 4 - 24 hours of exposure (Klein 1990). People with chronic, 
spinal cord injury have been exposed to much longer duration of oxygen (up to 4 hours) with no 
adverse effects reported (Gui et al 2014).  
 
If there is any sign of discomfort during an experiment suggestive of early stage oxygen toxicity, 
such as tracheal irritation, abnormal rise in blood pressure or other symptoms (Klein 1990), 
oxygen will be discontinued. Dr Wong (co-investigator and medical expert on this trial) will be 
available by phone during all experiments with participants with spinal cord injury, and a 
licensed Physiotherapist (PT) will be present during these experiments. 
 
If a participant has any signs of distress (related or unrelated to the study), the PT will evaluate 
the situation, and contact Dr Wong for consultation, or arrange for transportation to the 
emergency department if warranted. The laboratory where all the experiments will take place is 
located in the Clinical Sciences Building at the University of Alberta. This Building is physically 
connected with the University of Alberta Hospital. 
 
1.4 Dose Rationale  
Oxygen is delivered in the same way as has been tested in rodents.  The percentage of oxygen is 
also similar to that used in the animal study (Li et al 2017).  The period of exposure is chosen to 
be slightly longer than that used in rodents (1 min in rodents vs 2 min in humans) to allow 
delivery of oxygen in a much larger organism, but still well below the time frame when 
complications have been reported (≥4 hours) (Klein 1990). 
 
1.5 Trial Conduct 
This study will be conducted in compliance with the protocol approved by the University of 
Alberta Health Research Ethics Board (HREB), and according to Good Clinical Practice (GCP) 
standards.  No deviation from the protocol will be implemented without the prior review and 
approval of the HREB, except where it may be necessary to eliminate an immediate hazard to a 
research subject.  In such case, the deviation will be reported to the HREB as soon as possible. 

 
 
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1.6 Population 
Subacute or chronic (≥3 months post-injury), traumatic spinal cord injury. See inclusion and 
exclusion criteria (Sections 4.1 and 4.2, respectively, below). 
A small number of uninjured participants will be included to test the experimental set-up using 
compressed air only.  These participants will not be included in the testing of the experimental 
agents, but rather are used to ensure that the protocol runs smoothly prior to testing participants 
with spinal cord injury. 
1.7 Literature 
Ways to promote recovery from severe spinal cord injury has been extremely limited in 
humans.  Individuals with spinal cord injury, diagnosed with ASIA Impairment Scale A and B, 
have historically tended not to improve a great deal over time, or even progressively decrease in 
function (Steeves et al 2011; Zariffa et al 2011; Aimetti et al 2019). Recently, epidural 
stimulators implanted over the spinal cord have resulted in some return of function in such 
individuals (Harkema et al 2011; Angeli et al 2014; Angeli et al 2018), but it requires surgical 
implantation of a stimulator, which comes with all the potential complications of such a 
procedure. 
Recent work in rats with complete and incomplete spinal cord injuries (Li et al 2017) suggests 
that the spinal cord distal to the lesion site suffers from chronic hypoxia, resulting from excessive 
activity of monoamine receptors in pericytes, which surround capillaries and constrict blood 
flow.  Brief (1 min) inhalation of pure oxygen in these rats reduces the hypoxia for ~20 min, 
resulting in improved motor and sensory function. We will test this idea in humans with chronic, 
severe, spinal cord injury. 
 
2 Trial Objectives 
Determine if breathing an increased proportion of oxygen above the concentration in normal 
room air results in changes in the sensory and motor function in people with sub-acute and 
chronic, severe spinal cord injury (SCI). 
 
3 Trial Design 
The scientific integrity of the trial and the credibility of the data from the trial depend 
substantially on the trial design.  The trial design is as follows (shown in schematic form in 
Figure 1).  
 

 
 
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Figure 1. Crossover experimental design for participants with spinal cord injury.   Participants will be 
randomized into Group 1 or Group 2.  Group 1 will receive oxygen during Phase 1, and compressed air 
in Phase 2.  Group 2 will receive the same agents in opposite order.  Measures will be taken before 
and after the exposure.  Experiments will be spaced at least 1 week apart. 
 
 
 
This is a prospective, double-blind, interventional, placebo controlled trial.  Participants will 
be randomly allocated to Group 1 or Group 2.  Group 1 will receive oxygen (99%) in Phase 
1, and room air (placebo) in Phase 2.  Group 2 will receive the agents in opposite order to 
Group 1.  Measures of sensory function will be taken before and after each exposure. Two 
experimental days are planned for each phase, so that von Frey hairs and reflexes are tested 
on separate occasions to avoid cross-contamination.  Both high oxygen and room air will be 
delivered through a loose fitting face mask approved by Health Canada (license #81363, 
supplied by Glenwood Labs, Canada, part # 1041). The flow rate will be 10 litre/min for a 2-
min period. 
 
The participants with SCI will be blinded.  The experiments will be spaced by at least 2 
weeks. 
 
Primary outcome measure: skin sensation will be collected at regular intervals using 
calibrated Von Frey Hairs. Reflex and skin sensation will be recorded throughout the 30 min 
prior to and 30 min after exposure to high oxygen or room air at regular intervals.  
Secondary outcome measure: leg reflexes will be elicited by electrical stimulation of the 
skin, and responses measured by surface electromyography.  
 
The experimenter collecting the sensory data and the experimenter analyzing the reflex data 
will be blinded to the oxygen/air exposure, to avoid bias.  The blinding ends for both the 
participant and the experimenter once the both experiments are completed for that 
participant. 
 
Stopping rules/discontinuation criteria: if any symptoms of early oxygen toxicity emerge, 
including acute tracheobronchitis, which could be a mild tickling sensation, later followed 
by inspirational pain, cough, or a constant retrosternal burning sensation (Klein 1990), the 
experiment will be stopped immediately. Normally, these types of symptoms are not 

 
 
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experienced until after 3 or more hours of exposure to high oxygen.  If such an incident 
occurs with any participant, the study team will consider terminating the study. 
Uninjured participants (n=10) will be included to test the study protocol for reflexes and von 
Frey hairs, but using compressed air only.  This is to ensure the smooth running of experiments 
for the participants with spinal cord injury, and to provide reliability information on our testing 
procedures.  Four experiments will be included to test the von Frey hairs and the reflexes in 
separate experiments, each repeated once to examine reliability of the testing (see Figure 2). 
 
 
 
 
Figure 2. Experimental design for 
uninjured participants.  Four 
experiments are planned for each 
participant.  Only compressed air will be 
used in these experiments.  Von Frey Hairs 
will be used to test sensation on 2 separate 
occasions, and reflexes will be used in the 
other 2 occasions. 
 
 
3.1 Primary Study Endpoints/Secondary Endpoints 
The primary endpoint is the skin sensation as quantified by von Frey Hairs at specific skin 
locations above and below the level of injury.  The exact skin location will vary, since different 
participants will have different levels of injury. The secondary endpoint is the reflex excitability 
as measured by surface electromyography (EMG) in the leg muscles, and induced by electrical 
stimulation of the surface of the skin on the foot arch.   
 
3.2 Study Design/Type 
Prospective, double-blind, placebo controlled cross-over trial, with randomized order of 
presentation of high concentration of oxygen (99%) vs room air (placebo).  All participants will 
be exposed twice to both conditions over the course of the 4 experiments, and measures will be 
taken before and after each exposure. See Figure 1 and Point 3 Study Design for details. 
 
3.3 Randomization 
The allocation of participants into Group 1 or Group 2 will be block randomized (block size 2-4) 
for each participant using a random sequence generator.  Only one researcher present in the 
experiment will know the order of presentation for ease of administration and for safety, but all 
other experimenters doing the measurements and analyzing the data will be blinded. 
 

 
 
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Maintenance: the order of presentation can be disclosed to the participant and experimenters 
after all data has been collected on a participant. 
3.4 Trial Treatment 
The trial is an exposure to high oxygen (99%), delivered to the participant through a Health 
Canada approved (lic. #81363) face mask (supplied by Glenwood Labs Canada, Part #1041).  
The oxygen and room air are stored in high-pressure cylinders, which will be labelled with the 
investigational product name, study title, lot number and expiry date.  The pressure is stepped 
down with Health Canada approved regulators, supplied by Western/Scott Fetzer Enterprise: 
WES M1-346-15FM for room air, and WES M1-540-15FM for high oxygen.  The exposure to 
high oxygen and room air will both be for 2 min, and will occur on different experimental days. 
3.5 Duration 
The participant will be involved in four sessions of testing at a minimum of 2-week intervals 
between every testing session, each of which will take approximately 1.5 to 2 hours.  A total 
duration of involvement in the study is a minimum of 7 weeks, with some possibly longer to fit 
in the 4 experiments according to participant and experimenter availability. 
3.6 Discontinuation 
Early signs of oxygen toxicity include acute tracheobronchitis, including a mild tickling 
sensation, later followed by inspirational pain, cough, and if more severe, a constant retrosternal 
burning sensation.  If any of these symptoms are reported, we will immediately stop the delivery 
of high concentration of oxygen.  Normally, these types of symptoms are not experienced until 
after 3 or more hours of exposure to high oxygen (Klein 1990). 
3.7 Product Accountability 
We will test oxygen exposure against room air (placebo). 
3.8 Data Identification 
The data will be recorded on RedCap with the assistance of the office of Quality Management 
for Clinical Research at the University of Alberta.  RedCap data will include all relevant Case 
Report Forms, Informed Consent Forms, data from the von Frey hairs, and analyzed data from 
the reflexes. 
4 Selection and Withdrawal of Subjects 
4.1 Inclusion Criteria 
SCI participants:  
i) 
traumatic SCI with onset ≥3 months prior,  
ii) 
between 18 - 65 yr old,  
iii) 
ASIA Impairment Scale at discharge classified as A, B or C,  
iv) 
injury level between C5 and T10,  
v) 
able to give informed, written consent. 

 
 
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Uninjured controls:  
i) 
no previous spinal cord injury or health condition that would confound the 
experiment or make it unsafe for the participant (see exclusion criteria),  
ii) 
between 18-65 yr old,  
iii) 
able to give informed, written consent. 
4.2 Exclusion Criteria 
 
SCI participants:  
i) 
frequent uncontrolled autonomic dysreflexia,  
ii) 
uncontrolled high blood pressure,  
iii) 
cardiac or cardiovascular disease,  
iv) 
cancer,  
v) 
active urinary tract infection,  
vi) 
active pressure sores,  
vii) 
signs of deep vein thrombosis in the legs,  
viii) 
severe swelling of the feet and/or legs,  
ix) 
severe cognitive impairment,  
x) 
pulmonary dysfunction such as chronic obstructive pulmonary disease or acute 
respiratory infection,  
xi) 
any condition which would be exacerbated by sitting or lying in one position for 2 
hours, such as low back pain,  
xii) 
pregnancy. 
 
Uninjured controls:  
i) 
any active medical or neurological disorders,  
ii) 
cardiac or cardiovascular disease,  
iii) 
cancer,  
iv) 
signs of deep vein thrombosis in the legs,  
v) 
severe swelling of the feet and/or legs,  
vi) 
severe cognitive impairment,  
vii) 
pulmonary dysfunction such as chronic obstructive pulmonary disease or acute 
respiratory infection,  
viii) 
any condition which would make it difficult to remain sitting or lying in one position 
for over 2 hours, such as low back pain,  
ix) 
pregnancy. 
4.3 Subject Withdrawal 
Participants can withdraw at any time without consequence to them. They simply have to tell us 
they do not wish to continue.  There will be no follow-up, and no consequence to the 
participants.  There will be no need to replace participants, as this is not a clinical trial.  We have 
factored in 10%-15% attrition.   
 

 
 
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4.4 Treatment of Subjects 
During an experiment, participants will transfer to a wide, physical therapy plinth, and positioned 
in sitting, with the torso about 45 degrees from the vertical.  The agent, high oxygen or room air, 
will be administered just once for a short duration.  No other treatment is involved. On each 
testing day, participants will be exposed to one condition, either high oxygen or room air, and 
outcome measures will be taken before and after each exposure. On 2 of the testing days, the 
participants will be exposed to high oxygen, and on the other 2 days they will be exposed to 
room air.  Testing sessions are spaced at least 2 weeks apart from each other. 
4.5 Medication 
 
 
As we have excluded all people with respiratory dysfunction, none of the participants will be on 
respiratory medications.  We will document all other medications the participants are taking. 
 
4.6 Monitoring for subject compliance 
Not applicable.  Participants will be in our lab the entire time. 
 
5 Assessment of Efficacy 
This is a one-time exposure to high oxygen during two experimental sessions.  It is not meant to 
be efficacious as a treatment at this time.  The experiment is a scientific study to determine how 
high oxygen, delivered for a short period, affects people with and without spinal cord injury. If 
exposure to high oxygen has a similar effect as it does in other animals with spinal cord injury, 
then new studies will be designed in the future to explore ways to improve oxygenation in people 
with spinal cord injury.   
5.1 Efficacy Parameters 
Not applicable.  The study is a proof-of-principle study that the high oxygen results in motor and 
sensory changes below the level the spinal cord injury in the short term.  It is not meant to be a 
therapeutic agent. 
 
5.2 Method and Timing 
N/A 
 
6 Assessment of Safety 
 
6.1 Safety Parameters 
Screening: Participants will be screened in person by a study team member and will be required 
to sign a release to allow the study team to examine their medical record, to ensure there are no 
contraindications for the study. An approval form will be signed by their family physician, or Dr. 

 
 
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Ho (co-investigator) in the case where a potential participant does not have a family physician. 
The approval indicates they see no medical reason why the individual should not participate. 
 
COVID-19 precautions: During the Covid-19 pandemic, additional measures have been taken to 
protect both the participants and the experimenters.  These measures are as follows: 
 All participants and experimenters must be fully vaccinated with a Health Canada 
Approved vaccine at least 2 weeks prior to the initial visit.  This is in keeping with the 
University of Alberta regulations for all individuals attending campus. 
 A screening questionnaire will be administered prior to admission to the lab at every visit 
to ensure an absence of symptoms and close contact with Covid-19 positive individuals. 
 All participants and experimenters must wear medical grade masks provided by the 
laboratory. 
 During administration of the agent, a medical grade mask will be placed over the oxygen 
mask to protect both the experimenters and participants. 
 Documentation of people in and out of the lab with dates and times will be enforced. 
 Reporting of a Covid-19 positive test subsequent to a visit is required of all people 
attending the experiment.  Should such an incident occur, all people in the lab who could 
have been exposed to the virus will be notified by phone. 
6.2 Method and Timing 
Participants will be instrumented with a non-invasive heart rate (HR) and tissue oxygen 
saturation monitor, clipped to the finger (Pulsox-300). HR will be constantly monitored through 
the experiment, including 30 min after the termination of the exposure. We expect HR could 
decrease slightly (~10 beats/min) (Waring et al 2003) during exposure.  If these changes exceed 
10 bpm during the intervention exposure, the intervention will be stopped. The participants will 
be monitored for 30 min after the exposure only, because there are no known subsequent effects 
from the short, 2-min exposure to oxygen.  To ensure that there are no lingering effects, they will 
be contacted by phone by a physical therapist the following day to check for any issues.  The 
phone check will be done for exposure to both oxygen and compressed air to ensure blinding. 
 
6.3 Adverse Event Reporting 
We will comply with HREB requirements to report any adverse events.  The event will be 
reported in writing to the HREB, using the approved Local Serious Adverse Event Report form.  
Further, all serious unexpected adverse drug reactions (SUSARs) will be reported in writing to 
Health Canada. 
6.4 Definitions 
Unanticipated events are any events during the exposure or through the 30 min following the 
exposure that exceed the HR changes described above or other health issues. 
 
Serious events are any events during the same period of time (i.e., exposure to oxygen and 30 
min period following exposure) that led to the need to seek immediate medical attention at a 
hospital emergency and requires inpatient hospitalization. 
 

 
 
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Related events will be defined as events that occurred during or after exposure (within 30 min) to 
the agent (99% oxygen) which were not present prior to exposure. 
   
6.5 Adverse Event Follow-up 
We will follow-up with the participant daily, either through phone, email or in-person until the 
condition has been resolved. The HREB will be notified. 
 
7 Statistical Plan 
 
7.1 Statistical Methods 
The data will be analyzed promptly, typically within a week of data collection.  Statistical 
analysis will be done later after all data is collected and a statistician will be consulted to assist 
with analyses.  The staff analyzing the reflex data off-line will be blinded. 
7.2 Subject Population(s) for Analysis 
This is a pilot study.  Participant numbers are very small, as there is no prior data to allow a 
calculation of sample size.  Data on leg reflexes, skin sensation and oxygen saturation levels over 
time will be analyzed pre- and post-exposure and compared with the control condition (i.e., , 
room air).   
7.3 Significance 
P<0.05 
7.4 Termination Criteria 
If serious adverse events (defined in 6.4) related to the exposure to high oxygen occurs in a 
participant, the study will be re-evaluated, and termination of the study considered. 
7.5 Accountability Procedure 
Missing data could occur if a participant drops out of the study prior to completion of all 
measurements.  In that case, we will use whatever data is collected, as it will still be useful to 
understand any changes that were attributable to the intervention/placebo. 
7.6 Deviation Reporting 
Any deviation(s) from the original statistical plan will be described and justified in the protocol 
and/or in the final report. 
 
8 Direct Access to Source Data/Documentation 
We will provide direct access to trial-related monitoring, audits, IHREB review and regulatory 
inspection(s), in full compliance with the HREB regulations.  All source data will be retained for 
a minimum period of 25 years, in accordance with regulations for clinical trials. 

 
 
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9 Quality Control and Quality Assurance 
The study will be in compliance with the Good Clinical Practice (GCP) guidelines, as 
documented in the ICH Harmonised Tripartite Guideline. The investigators welcome audits and 
allow access to source data from the Institutional Health Research Ethics Board and the Office of 
Quality Management in Clinical Research at the University of Alberta. 
10 Ethical Considerations 
This study will be conducted according to Canadian and international standards of GCP.  
Applicable government regulations and University of Alberta research policies and procedures 
will also be followed. 
 
This protocol and any amendments will be submitted to the University of Alberta HREB for 
formal approval to conduct the study.  The decision of the HREB concerning the conduct of the 
study will be made in writing to the investigator.   
 
All participants for this study will be provided a consent form describing this study and 
providing sufficient information for participants to make an informed decision about their 
participation in this study.  This consent form will be submitted with the protocol for review and 
approval by the HREB.  The formal consent of a participant, using the HREB-approved consent 
form, will be obtained before that participant is submitted to any study procedure.  This consent 
form must be signed by the participant, and the investigator-designated research professional 
obtaining the consent.  
11 Data Handling and Record Keeping 
Only the researchers and their trainees and laboratory staff will handle the data.  The majority of 
data collected will be in electronic form.  All electronic data will be coded by a participant 
number, so there will be no identifiable information stored with the data.  The data will be stored 
on the computer servers in Corbett Hall, and in RedCap.  Identifiable information in hardcopy 
will be stored in a locked room within a locked cabinet.  If there is any electronic data that 
contains identifiable information, such as a master copy of the participants and their codes, this 
will be encrypted on the servers. Hardcopy, identifiable information will include the participants’ 
full name, contact information (email and/or phone number), AHS number, physician’s name 
and number, and medical information extracted from the AHS files.  We need contact 
information to schedule the experiment, and medical information to confirm suitability for the 
experiment. 
 
12 Finance and Insurance 
There will be no cost to the participant.  Parking or public transit costs will be covered by the 
researchers. Insurance is covered by the University, as this is a research project carried out on the 
University campus, by University employees. 
 

 
 
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13 Publication Plan 
The results of this study will be published in a scientific journal.  The results may also be 
presented at national and international conferences. 
14 Supplements 
 Safety Data Sheet for compressed air 
 Safety Data Sheet for pure oxygen 
 
 
 
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Angeli CA, Boakye M, Morton RA, et al. (2018) Recovery of over ground walking after chronic 
motor complete spinal cord injury.  N Engl J Med 379(13):1244-1250.  
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