Skin Prick Test in Healthy Volunteers of Any Sex, to Investigate the Potential Allergy to NeoMatriXTM Wound Matrix
Phase: NA
Conditions: Allergy Skin, Allergy, Dermatitis, Allergic Contact
Sponsor: NeXtGen Biologics, Inc.

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NeXtGen Biologics, Inc.  
12085 Research Drive, Box 17 
Alachua, FL 32615 
 
Jonelle Toothman 
800-209-7190 ext 2 
support@nextgenbiologics.com 
 
CONFIDENTIAL COMMUNICATION 
 
Skin Prick Test (SPT):   20 Patients 
 
ABBEVIATIONS 
AE  
–  
Adverse Event 
CFR 
- 
Code of Federal Regulations 
CRF 
- 
Case Report Form 
GCP 
- 
Good Clinical Practice 
ESS 
- 
Erythema Scoring Scale 
HIPAA - 
Health Insurance Portability and Accountability Act  
ICF 
- 
Informed Consent Form 
IRB  
- 
Institutional Review Board 
PI 
- 
Principal Investigator or designee 
RIPT 
- 
Repeated Insult Patch Test 
SAE 
- 
Serious Adverse Event 
SOP 
- 
Standard Operating Procedure 
SPT 
- 
Skin Prick Test 
 
DEFINITIONS 
Adverse Event (AE): Any negative experience that a volunteer has during the course of a clinical 
trial, including new or worsening symptoms or laboratory abnormalities. 
 
Belmont Report: A document entitled “Ethical Principles and Guidelines for the Protection of 
Human Subjects of Research” that was produced by the National Commission for the Protection 
of Human Subjects of Biomedical and Behavior Research in 1978.  The Belmont Report identifies 
three fundamental ethical principles for all human subject research: respect for persons, 
beneficence, and justice. 
 
Blinding:  When research subjects are unaware of the assigned “treatment.” In a single-blinded 
trial, the subjects do not know what treatment they are receiving.  In a double-blinded trial, the 
subjects and the investigators are unaware of the treatment assigned, as are the monitors and 
statisticians in some cases.   
 
Case Report Form (CRF): A record of information collected on each subject during the clinical 
trial. 
 
Clinical Investigation: A systematic trial designed to evaluate a test material in humans.  
 
Clinical Research:  A trial in human subjects. 
 
Clinical Trial:  Any investigation in humans meant to determine properties of a test material.  
 

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Ethics Committee:  An independent group of medical and nonmedical people who verify the 
integrity of a trial and ensure the safety, integrity, and human rights of the subjects. 
 
Exclusion Criteria:  Characteristic that would prevent a subject from being eligible to participate 
in a research trial, as specified in the protocol. 
 
Good Clinical Practices (GCP): International ethical and scientific quality standards for the 
design, conduct, monitoring, recording, auditing, analysis, and reporting of trials.  
 
Health Insurance Portability and Accountability Act (HIPAA): Legislation passed in 1996 that 
includes a privacy rule creating national standards to protect personal health information. 
 
Inclusion Criteria:  A list of requirements that a subject must meet to be eligible to participate in 
a research trial, as specified in the protocol.  
 
1 
PROTOCOL SUMMARY 
 
Title: 
Skin prick testing of healthy adult volunteers to investigate the potential of 
allergy to NeoMatriXTM Wound Matrix 
Design: 
Unblinded 
Test Articles: 
1. NeoMatriXTM Wound Matrix, an investigational device 
 
 
  
 
 
Duration: 
 
2-3 days 
Subjects: 
 
Sufficient healthy volunteer subjects so that 20 will complete the study 
Observations: 
Scoring of any skin reactions at 15 minutes, 6 hours and 1-2 days after 
prick test 
Location: 
 
Clinical Research Center, UB Clinical and Translational Research Center 
 
 
 
875 Ellicott Street, Buffalo, NY 
2 
KEY STUDY PERSONNEL AND RESPONSIBILITES 
 
Key Role 
Key Personnel 
General Responsibilities 
Principal Investigator 
(PI) 
Stanley Schwartz, MD, PhD 
Chief, Division of Allergy, 
Immunology & Rheumatology 
SUNY at Buffalo 
Buffalo General Medical Center 
 
The PI is responsible for ensuring 
sufficient resources are available to 
conduct the study, for investigating 
and reporting any serious adverse 
events to the Sponsor, the study 
design, compiling the results and 
writing the clinical report. 

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Study Coordinator 
Robin Stein, RN, BSN 
The Study Coordinator will be 
responsible for conducting the 
study on a daily basis. 
Monitor 
NeXtGen Appointee 
 
Project Coordinator 
Jonelle Toothman 
Sponsor representative who is the 
primary point of contact and 
represents the Sponsor (NeXtGen 
Biologics) 
 
3 
STUDY VISIT CHART 
Day 
01 
02-03 
Prick Test Performed 
ü 
 
Clinically Evaluate Prick Site 
15 mins       6 hours 
ü                         ü 
 
     ü 
 
4 
INTRODUCTION, BACKGROUND AND OBJECTIVE 
Skin prick tests for immediate hypersensitivity have been the gold standard as assays for allergies 
since the 1860s and are currently used by allergists identify allergic reactions to specific 
allergens. These tests are minimally invasive and are used on patients of all ages from infants to 
seniors. They have the advantage of being highly sensitive and specific and produce rapid 
results. 
 
Therapies for facilitating wound healing currently are limited especially with regard to chronic 
wounds and remain a significant unmet need. Chronic wounds including non-healing skin ulcers 
such as pressure ulcers, diabetic neuropathic ulcers and vascular insufficiency ulcers, are a 
source of substantial morbidity and mortality for hospitalized patients and residents of nursing 
homes. Non-healing and chronic wounds increase the risk of infection and can result in limb 
amputation and death.  
 
Numerous topical treatments for non-healing wounds have produced mixed results and an ideal 
treatment remains to be achieved. The test product of this proposal, NeoMatriXTM Wound Matrix, 
is a wound dressing for the management of wounds, acute and chronic. NeoMatriXTM Wound 
Matrix has the features of a biocompatible biomaterial. Despite its stringent preparation 
protocols and having completed a range of biocompatibility testing, prior to utilization in human 
clinical trials, it remains to be tested in a small pilot study to determine if it can elicit any allergic 
responses. While it is highly unlikely that any study subject will have de novo, immediate allergic 
reactions to NeoMatriXTM Wound Matrix, the current protocol is designed to document this and 
determine whether study subjects may show any delayed allergic reactions. 
 
The objective of this study is to investigate the potential of NeoMatriXTM Wound Matrix to cause 
an allergic response in healthy volunteers. The biomaterial is applied to the forearm of study 
subjects, a drop of saline solution is placed on the membrane and using a standard skin prick 
test device the underlying epidermis is superficially pricked through the overlying membrane.  
After the test, the area of the skin is observed after 15 minutes and 6 hours to see if a reaction 
develops. The test site will again be evaluated at 1-2 days after the initial prick test.  The skin will 
be examined for a “wheal”—a raised, itchy bump and surrounding erythematous “flare”. 

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and 
flare whose diameter can be measured and exceed a wheal at the negative control by 4 mm. 
Pricking through a drop of saline serves as a negative control that is not expected to produce a 
significant wheal and flare.
 
, to reiterate, this is 
completely unanticipated but remains to be proven. 
5 
STUDY DESIGN 
The prick test is not blinded. 
6 
SELECTION OF SUBJECTS  
6.1 
Screenings 
A sufficient number of subjects will be recruited to ensure a minimum of 20 subjects complete 
the study. Subjects must satisfy the inclusion and exclusion criteria, must be prepared to accept 
the prohibitions and restrictions and must give written informed consent. 
The suitability of each potential subject will be confirmed before their acceptance by review of a 
study specific pre-treatment questionnaire. 
 
6.2 
Inclusion criteria  
6.2.1 Healthy volunteers, of either sex, aged at least 18 years. 
6.2.2 Completed written informed consent and receive a copy of their executed ICF.  
6.2.3 Volunteers must be capable of understanding and following directions in English. 
 
6.3 
Exclusion criteria  
6.3.1 Pregnancy or lactation.  
6.3.2 Inadequate or non-existent contraception (women of child bearing potential only).  
6.3.3 A current skin disease of any type apart from mild facial acne (e.g. eczema, psoriasis). 
6.3.4 Heavy alcohol consumption. 
6.3.5 Current use or history of repeated use of recreational drugs. 
6.3.6 A febrile illness lasting more than 24 hours in the six days prior to test. 
6.3.7 Significant past medical history of hepatic, renal, cardiac, pulmonary, digestive, 
hematological, neurological, locomotor or psychiatric disease.  
6.3.8 
 
 
 
 

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6.3.10 
 
 
 
 
erials.  
6.3.12 Current treatment by a physician for allergy unless physician consulted by Investigator 
and participation approved. 
6.3.13 Participation in a repeat insult patch test (RIPT), skin prick test (SPT) or follow-up work 
within the last month. 
6.3.14 Sensitization or questionable sensitization in a RIPT or SPT. 
6.3.15 Recent immunization (less than 10 days prior to prick test). 
6.3.16 A medical history indicating atopy or severe dematographism. 
6.4 
Prohibitions and restrictions for the duration of the study 
6.4.1 No use of aspirin or non-steroidal anti-inflammatory or corticosteroid drugs for the 
duration of the study. 
6.4.2 
 
6.4.3 
 
 
 
METHOD 
7.1 
Test Articles – NeoMatriXTM, extracellular matrix xenograft, 8mm disc 
7.2 
Negative Skin Test Control– sterile normal saline 
7.3 
Positive Skin Test Control – Sterile histamine 
     
 
7.4 
Materials 
• 
Test article NEOMATRIXTM 8mm disc 
• 
Positive control solution 
• 
Negative control solution 
• 
 
• 
Sharps container for disposal of picks  
• 
Marker pen for the skin 
• 
Ruler for measuring reactions 
• 
Sterile cotton balls for wiping skin 
• 
Gloves (latex-free) 
• 
Recording sheets, Case Report Forms 
• 
Syringes and needles 
• 
Cetirizine 10 mg tablets 
• 
Hydrocortisone cream 1% 
 
 

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7.5 
Devices used for skin prick testing 
• 
 
7.6 
Procedure 
The subject needs is placed in a comfortable position with volar surface of the forearm 
 
 
The room should be private and at a comfortable temperature. It is 
advisable to provide the patient with a magazine or something to occupy themselves for the 15-
20 minutes that is required for the test (and to distract them from any discomfort).  
 
The test area should be clean, free from moisturizers and area for skin pricks should be more than 
5 cm from the wrist and 3 cm from the antecubital fossa and marked by numbers on the skin to 
identify the test article, negative control and positive control. Pricks should be made 
immediately adjacent to the numbers to avoid confusion between allergens. Prick tests should 
be at least 2 cm apart to avoid overlapping reactions and false-positive results.  
 
 
 
  
  
 
Positive and negative controls will be applied directly with the Picks.   
 
A separate Pick is used to prick each site.  For example, there should be 3 Picks used per subject, 
one for each test site. Prick each site separately. 
 
  
 
 
 
7.7 
Grading of Test Sites - Reading Results 
• 
Initial reading after 15 minutes looking for the presence or absence of a wheal and flare 
at all three sites and measuring the diameter of both the wheal and flare if present. The 
wheal at the positive control must exceed a wheal at the negative control by 4 mm.  
• 
A second reading is conducted about 6 hours after the SPT administration and similar 
measurements are taken.  
• 
The subjects return to the office 1-2 days after the initial administration for a third test 
reading and measurements.  
• 
A positive reaction is present when there is a measurable wheal of >3 mm . Small wheals 
are confirmed by palpation. A flare alone is measured but is not of any clinical 
significance.   
 
7.8 
Characteristics of Wheal and Flare 
Wheals are determined by palpation or oblique lighting and the diameter is measured in mm. If 
the wheal is not asymmetrical two diameters are measured (smallest and largest) and an 
average value is calculated. Flares, areas of erythema, are similarly measured but are not 
considered clinically significant. 

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8 
SAFETY - ADVERSE EVENTS 
8.1 
Relationship to Test Material 
The relationship or association of the AE to the test material will be characterized as unlikely, 
possible or probably.  Assessments shall be recorded on the CRF.  
 
Causality 
Term 
 
Assessment Criteria 
Probable 
Event or laboratory test abnormality, with plausible time relationship 
to test material exposure 
Unlikely to be attributed to condition (or disease) or other products in 
use by subject 
Response to withdrawal clinically reasonable 
Rechallenge satisfactory or not required 
Possible 
Event or laboratory test abnormality, with reasonable time relationship 
to test material intake 
Could also be explained by condition (or disease) or other products in 
use by subject 
 
Unlikely 
Event or laboratory test abnormality, with a time to test material 
intake that makes a relationship improbable (but not impossible) 
Condition (or disease) or other products provide plausible explanations 
 
For safety analyses, AEs that are classified as a possible or probable association to a test 
material shall be considered test material-related AEs. 
 
Follow-up of the AE, after the date of discontinuation of exposure to test material is 
required if the AE or its sequelae persist.  Follow-up is required until the event or its 
sequelae resolve or stabilize at a level acceptable to the PI   and to the Medical Monitor. 
 
8.2 
Severity of Adverse Events 
The severity of each AE will be graded using the following criteria: 
 
.  
Grade 
Description 
1 
Mild – Minor AEs requiring no specific medical intervention including 
asymptomatic laboratory findings only, or finding of marginal clinical relevance 
2 
Moderate – AEs including urticaria requiring minimal, local, and/or noninvasive 
intervention 
3 
Severe – Severe and undesirable AEs involving significant symptoms requiring 
hospitalization or invasive intervention; transfusion; elective interventional 
radiological procedure; or therapeutic endoscopy or operation 

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4 
Life-threatening or debilitating – AEs complicated by acute, life-threatening 
metabolic or cardiovascular complications such as circulatory  
failure, 
hemorrhage, or sepsis.  Also, life-threatening physiologic consequences; need 
for intensive   care   or   emergent    invasive   procedure;    emergent    
interventional radiological procedure, therapeutic  endoscopy or operation. 
5 
Death 
 
If the Grade changes within a day, the maximum Grade shall be recorded. 
 
 
If the Grade changes over a longer period of time, the changes shall be recorded as 
separate events (having separate onset and stop dates for each grade). 
 
8.3 
Serious Adverse Events 
An SAE is defined as any AE that suggests a significant clinical hazard, contraindication, side 
effect, or precaution. This includes any event which: 
 
• Results in death 
• Is life-threatening 
• Results in persistent or significant disability/incapacity 
• Results in or prolongs an existing inpatient hospitalization 
• Results in a congenital anomaly/birth defect 
• Is an  important medi ca l event based on medical judgment, which  jeopardizes the 
subject and requires medical or surgical intervention to prevent one of the outcomes 
listed above. 
 
The PI will evaluate all serious AEs as to their intensity, relation to test material, outcome, and 
action taken.   
 
M a n a g e m e n t  o f  A d v e r s e  E v e n t s  
No adverse reaction is anticipated.  The patient will be evaluated at 15 minutes after SPT and 
then again at 6 hours after SPT.  Minor adverse events (e.g. itching at the skin test site) will be 
examined by the nursing staff of the CRC; the PI will be contacted as needed.  
• 
Over-the-counter (OTC) antihistamines will be given, which relieve minor symptoms.  
Antihistamines prevent symptoms such as hives by blocking histamine receptors. 
• 
Persistent itching, swelling or redness, will be reduced with ice and at the test site we shall 
apply 1% hydrocortisone cream one time. 
• 
Should the subject develop urticaria they will be provided with a 5 day course of 
antihistamine (10 mg cetirizine orally once daily)  
While serious adverse events are not anticipated the most likely serious event would be a 
systemic allergic reaction or anaphylaxis which would most likely occur within 20 minutes while 
the subject is under close evaluation at the CRC.  
• 
The subject will be assisted to lie on their back with feet raised about 12 inches, clothing 
loosened to ease breathing, covered with a blanket and coached to stay calm.  They 
will be turned on their side if vomiting or bleeding.  
• 
Epinephrine (adrenaline) may be used, depending on the severity. 

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• 
Should anaphylaxis occur Epinephrine 1:1000 will be administered immediately by 
intramuscular injection and repeated every 10 min. as needed.  The PI will be summoned 
stat and the subject will be transported to the Emergency Department of the Buffalo 
General Medical Center located on the ground floor of the same building housing the 
Clinical Research Center.  
9 
REPORTING 
9.1 
Evaluation of Results – this simple study design, consists of only one physiological measure 
(that of wheal/flare) measured at 3 time points: T1-15 minutes after SPT, T2=6hr after SPT and 
T3=24-48 hrs. after SPT.  Our power analysis calculation indicates at a confidence level of 99% 
and confidence interval of 1.35, we will need a sample size of n=20.  No statistical analysis will be 
performed. No significant effect or interaction effects are anticipated hence the small sample 
size.  
9.2 
Interim Reports – unexpected findings will be reported to the Sponsor. At the conclusion of 
the study, a draft report will be sent to the Sponsor and contain all of the information to be 
included on the final report.  Comments made by the Sponsor may be incorporated into the 
draft report after which it will be issued as the final report and signed by the PI.  
9.3 
Correction or Additions to the Final Report – the sponsor will review the draft final report 
and make necessary edits prior to the final report being issued. 
 
TEST ARTICLE PRECAUTIONS: 
 
 
 
 
 
 
 
TEST ARTICLE POTENTIAL COMPLICATIONS: 
 
 
 
 
 
 
 
 
 
 
 
 
 
TEST ARTICLE STORAGE: