Conditions: Alopecia
Keywords: growth factors, hair loss, females, regenerative therapy
Allocation: RANDOMIZED | Intervention model: CROSSOVER
Enrollment: 20
Healthy volunteers: yes

Brief summary:
The primary objective of this proposal is to conduct a study that assesses the safety, feasibility and efficacy of using PRP to treat this type of hair loss.

Detailed description:
Androgenic alopecia (AGA) is a common, progressive hair loss disorder affecting both sexes with significant negative impact on social and psychological well-being. The frequency and severity increases with age, and up to 80% of men, and 50% of women are affected by AGA over the course of their lives. While men are more frequently affected, the psychological impact is likely to be high for women where the social impact of hair loss is often devastating. Current medical therapies specifically approved by the US FDA are limited to minoxidil (for men and women) and finasteride (for men only) (Varothai), but variable responses and the need for indefinite use often result in patient fatigue and suboptimal compliance. Recently, there has been interest in treatments orientated to more biologically regenerative therapies, and consequently there have been numerous studies that have demonstrated successful use for platelet rich plasma (PRP) in treating AGA. PRP contains concentrated platelet cells derived from autologous whole blood that are believed to activate a cascade of growth factors when injected into an area of poor hair growth that stimulates hair growth. In this proposal we will examine whether PRP therapy provides similar or better hair growing capacity in women compared to the conventional topical application minoxidil.

Arms:
arm 1: Subjects will be randomized into the Platelet Rich Plasma group, treated for 12 weeks with a 2 month washout period in between treatments, then treated with Minoxidil Foam for 12 weeks. arm 2: Subjects will be randomized into the Minoxidil Foam group, treated for 12 weeks with a 2 month washout period in between treatments, then treated with Platelet Rich Plasma.

Interventions: intervention 1: Platelet Rich Plasma intervention 2: Minoxidil Foam
Intervention descriptions: intervention 1: Autologous Platelet Rich Plasma (PRP) will be isolated from blood collected from each subject at each treatment time point. 100 cc of PRP will be injected with a 30-gauge needle over the affected part of the scalp, approximately 1/10 cc/injection site. Treatments will take place every 4 weeks for a total of 3 treatments over 12 weeks. intervention 2: Minoxidil 5% topical foam will be applied once daily following manufacturer's instructions for a total of 12 weeks.

Protocol document:
Prot_SAP_000.pdf: 
 
 
A Randomized Controlled Trial Comparing Platelet Rich 
Plasma (PRP) to Minoxidil Foam for Treatment of 
Androgenic Alopecia in Women  
 
 
 NCT03488108 
 
06/01/2016 


1 
 
A Randomized Controlled Trial Comparing Platelet Rich Plasma 
(PRP) to Minoxidil Foam for Treatment of Androgenic Alopecia in 
Women  
 
 
 
 
Sponsor-Investigator: 
Alison Bruce, MD (Principal Investigator) 
Shane Shapiro, MD 
Funding Sponsor: 
 
Intramural Funding 
Mayo Center for Regenerative Medicine 
Study Product: 
Platelet Rich Plasma (PRP) 
Protocol Number: (IRBe) 
 
 
 
 
 
 
 
 
Initial version: 
 
 
6/1/2016 Version (1)  
 
 
 


2 
 
 
Table of Contents 
 
 
STUDY SUMMARY ................................................................................................................................................... 5 
1 
INTRODUCTION ............................................................................................................................................. 6 
1.1 
BACKGROUND ................................................................................................................................................. 6 
1.2 
INVESTIGATIONAL AGENT ............................................................................................................................... 6 
1.4 
CLINICAL DATA TO DATE ................................................................................................................................ 6 
1.5 
DOSE RATIONALE AND RISK/BENEFITS ........................................................................................................... 7 
2 
STUDY OBJECTIVES ..................................................................................................................................... 7 
3 
STUDY DESIGN ............................................................................................................................................... 8 
3.1 
GENERAL DESIGN ............................................................................................................................................ 8 
3.2 
PRIMARY STUDY ENDPOINTS .......................................................................................................................... 8 
3.3 
SECONDARY STUDY ENDPOINTS ..................................................................................................................... 8 
3.4 
PRIMARY SAFETY ENDPOINTS ......................................................................................................................... 8 
4 
SUBJECT SELECTION ENROLLMENT AND WITHDRAWAL ............................................................. 9 
4.1 
INCLUSION CRITERIA ....................................................................................................................................... 9 
4.2 
EXCLUSION CRITERIA...................................................................................................................................... 9 
4.3 
SUBJECT RECRUITMENT, ENROLLMENT AND SCREENING ................................................................................ 9 
4.4 
EARLY WITHDRAWAL OF SUBJECTS ................................................................................................................ 9 
4.4.1 
When and How to Withdraw Subjects ................................................................................................... 9 
4.4.2 
Data Collection and Follow-up for Withdrawn Subjects .................................................................... 10 
5 
STUDY DRUG ................................................................................................................................................. 10 
5.1 
DESCRIPTION ................................................................................................................................................. 10 
5.2 
TREATMENT REGIMEN .................................................................................................................................. 10 
5.3 
METHOD FOR ASSIGNING SUBJECTS TO TREATMENT GROUPS ...................................................................... 10 
5.4 
PREPARATION AND ADMINISTRATION OF STUDY DRUG ................................................................................ 10 
5.6 
PRIOR AND CONCOMITANT THERAPY ............................................................................................................ 11 
 
6 
STUDY PROCEDURES ................................................................................................................................. 11 
6.1 
STUDY CALENDERS. ...................................................................................................................................... 12 
7 
STATISTICAL PLAN .................................................................................................................................... 13 
7.1 
SAMPLE SIZE DETERMINATION ..................................................................................................................... 13 
7.2 
STATISTICAL METHODS ................................................................................................................................. 13 
7.3 
SUBJECT POPULATION(S) FOR ANALYSIS ...................................................................................................... 14 
8 
SAFETY AND ADVERSE EVENTS ............................................................................................................. 14 
8.1 
DEFINITIONS .................................................................................................................................................. 14 
8.2 
RECORDING OF ADVERSE EVENTS ................................................................................................................. 16 
8.3 
REPORTING OF SERIOUS ADVERSE EVENTS AND UNANTICIPATED PROBLEMS .............................................. 16 
8.3.1 
Sponsor-Investigator reporting: notifying the Mayo IRB ................................................................... 16 
8.4 
STOPPING RULES ........................................................................................................................................... 16 
8.5 
MEDICAL MONITORING ................................................................................................................................. 16 
9 
DATA HANDLING AND RECORD KEEPING .......................................................................................... 17 
9.1 
DATA HANDLING .......................................................................................................................................... 17 
9.2 
CONFIDENTIALITY ......................................................................................................................................... 17 
9.3 
SOURCE DOCUMENTS .................................................................................................................................... 17 
9.4 
RECORDS RETENTION .................................................................................................................................... 17 
 


3 
 
10 
STUDY MONITORING, AUDITING, AND INSPECTING ...................................................................... 18 
10.1 
STUDY MONITORING PLAN ................................................................................................................... 18 
10.2 
AUDITING AND INSPECTING ...................................................................................................................... 18 
11 
ETHICAL CONSIDERATIONS ................................................................................................................... 18 
12 
STUDY FINANCES ........................................................................................................................................ 18 
12.1 
FUNDING SOURCE ..................................................................................................................................... 18 
12.2 
CONFLICT OF INTEREST ............................................................................................................................ 19 
12.3 
SUBJECT STIPENDS OR PAYMENTS ............................................................................................................ 19 
13 
PUBLICATION PLAN ................................................................................................................................... 19 
14 
REFERENCES ................................................................................................................................................ 19 
 
 
 
 
 
 


4 
 
 
List of Abbreviations 
 
 
LIST OF ABBREVIATIONS 
AGA 
Androgenic Alopecia 
 
 
AE 
Adverse Event/Adverse Experience 
 
 
CRF 
Case Report Form 
DSMB 
Data and Safety Monitoring Board 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practice 
HIPAA 
Health Insurance Portability and Accountability Act 
IB 
Investigator’s Brochure 
 
 
IRB 
MNX 
Institutional Review Board 
Minoxidil 
 
 
PHI 
Protected Health Information 
PI 
Principal Investigator 
PRP 
Platelet Rich Plasma 
SAE 
Serious Adverse Event/Serious Adverse Experience 
SOP 
Standard Operating Procedure 
 
 
 
 
 
 


5 
 
Study Summary 
Title 
Minoxidil Foam compared to PRP for Androgenic Alopecia in Women, 
A Randomized Controlled Pilot Trial 
Running Title 
PRP for Hair loss in Women 
Protocol Number 
1 
Phase 
Pilot 
Methodology 
Randomized, Controlled, Crossover 
Overall Study 
Duration 
12 Months 
Subject Participation 
Duration 
12 months 
Single or Multi-Site  
Single Site 
Objectives 
Determination of non-inferiority treatment of PRP compared to standard 
of care MNX in women 
Number of Subjects 
23 
Diagnosis and Main 
Inclusion Criteria 
Female, 18 years or older, Androgenic Alopecia grade I-II by Ludwig 
classification  
Study Product, Dose, 
Route, Regimen 
Platelet Rich Plasma 10cc, Injection by Nappage technique, 3 injections 
- 1 time every 4 weeks for 3 months 
Duration of 
Administration 
12 Weeks (1 time every 4 weeks for 3 months) 
Reference therapy 
Minoxidil 
Statistical 
Methodology 
Wilcoxon signed rank test for comparison of outcomes between 
treatments and McNemar’s exact test for categorical outcome variables  
 
 
 


6 
 
1 Introduction 
 
This document is a protocol for a human research study. This study will be carried out in accordance with the 
applicable United States government regulations and Mayo Clinic research policies and procedures.  
 
1.1 
Background 
 
Androgenic alopecia (AGA) is a common, progressive hair loss disorder affecting both sexes with significant 
negative impact on social and psychological well-being. The frequency and severity increases with age, and up 
to 80% of men, and 50% of women are affected by AGA over the course of their lives. (Piraccini 2014) 
(Varothai) (Bologna - book)  While men are more frequently affected, the psychological impact is likely to be 
high for women where the social impact of hair loss is often devastating. Current medical therapies specifically 
approved by the US FDA are limited to minoxidil (for men and women) and finasteride (for men only) 
(Varothai), but variable responses and the need for indefinite use often result in patient fatigue and suboptimal 
compliance.  (Gupta)   
 
Recently, there has been interest in treatments orientated to more biologically regenerative therapies, and 
consequently there have been numerous studies that have demonstrated successful use for platelet rich plasma 
(PRP) in treating AGA. (Singhal) (Maria-Angeliki) (Gentile) (Trink) (Gkini) (Cerveli) (Uebel) (Greco) 
(Takikawa) (Kang) (Betsi) (Sclafani) (Khatu) (Schiavone) PRP contains concentrated platelet cells derived from 
autologous whole blood that are believed to activate a cascade of growth factors when injected into an area of 
poor hair growth that stimulates hair growth. (Jain)(Lubkowska) In this proposal we will examine whether PRP 
therapy provides similar or better hair growing capacity in women compared to the conventional topical 
application minoxidil. 
1.2 
Investigational Agent 
 
Platelet Rich Plasma (PRP)  
 
1.3 
Clinical Data to Date 
PRP is an autologous concentration of human platelets in a small volume of plasma that has a 4-7 times higher 
platelet concentration than baseline.(Jain) (Lubkowska ) (Parsley) Since activated platelets secrete a number of 
cytokines and growth factors, such as platelet-derived growth factor, epidermal growth factor, transforming 
growth factor, and vascular endothelial growth factor, (Jain) (Lubkowska ) (Parsley) PRP may provide an 
attractive therapy to promote hair growth in the declining hair follicle. 
 
Recent investigators have explored the use of PRP as a treatment modality for AGA, with most available studies 
to date showing positive results. (Singhal) (Maria-Angeliki) (Gentile ) (Trink) (Gkini) (Cerveli) PRP is 
attractive as a useful treatment for nonscarring alopecia’s such as AGA, because the treatment is safe and 
adverse effects appear to be negligible. (Singhal) (Maria-Angeliki) (Gentile ) (Trink) (Gkini) (Cerveli) 
However, the effect of PRP on hair growth has been studied primarily in men. Studies that include men and 
women usually do not analyze the data according to sex or comment on whether sex differences were observed. 
(Trink) (Takikawa) (Kang) (Betsi) (Sclafani) (Schiavone) (Gkini) Although sex differences are known to exist 
in the incidence, diagnosis and mechanisms leading to AGA, women are understudied (Farage) (Romanelli). 
The need to understand hair loss in women is underscored by the growing interest of cosmetic dermatologists 
and plastic surgeons to use PRP routinely in their aesthetic and rejuvenation practice to reduce/prevent hair loss. 


7 
 
 
A recent review of the available literature regarding PRP treatment for hair loss identified 14 articles describing 
the use of PRP, 12 of which were used for AGA. (Maria-Angeliki) This systematic review considered 7 studies 
to be of moderate quality, 4 of low quality and 3 of very low.  Of the 7 trials of moderate quality, all reported a 
statistically significant increase in the number of hairs and hair density following PRP therapy, indicating a 
positive response to treatment. The predominant criticisms of the trials to date are the lack of a consensus 
protocol regarding the exact concentration of PRP, dose, frequency, depth of injections, and number of required 
sessions. Variability in evaluation methods also limits accurate comparison between the studies (i.e., global 
photographs, phototrichograms, dermascopic images, hair pull test, and manual counting were all reported as 
measurement tools). In addition, these studies predominantly used men. Very little published data exists for 
PRP use in women with AGA. The studies that did include women examined very low numbers and/or did not 
analyze results according to sex so that it is not possible to evaluate whether PRP therapy is safe and efficacious 
in women. (Trink) (Takikawa) (Kang) (Betsi) (Sclafani) (Schiavone) (Gkini) Nonetheless, PRP appears to be a 
useful treatment for AGA, particularly in men, and there is an urgent need for randomized controlled studies 
with standardized measurement techniques and reproducible protocols to study clinical efficacy. Considering 
the widespread use of PRP in cosmetic clinics among women and the lack of data on the effect of PRP on hair 
regeneration in women there is a great need to perform a controlled trial in women. In future studies we would 
like to examine whether PRP is equally effective in men and women and to examine its mechanism of action. 
 
1.4 
Dose Rationale and Risk/Benefits 
 
We will dose PRP in a manner similar to previous trials. (Gentile, Trink, Cerveli) Fresh PRP will be harvested 
from the individual and their own PRP administered at three time points, once every 4 weeks for a totally 
treatment time of 12 weeks. Minoxidil will be used according to FDA and manufacturer instructions daily for 
12 weeks. 
 
Risks associated with PRP therapy include possible pain, redness, and/or irritation at the site of injections for up 
to 2-3 days post injection. Published human data indicate no significant adverse events related to injection of 
PRP into the skin and previous authors utilizing similar protocols reported no infection, fever, hematoma, tissue 
hypertrophy, or major adverse events among the subjects. (Singhal) (Cerveli) (Gentile) (Trink) (Gkini) Known 
possible risks for Minoxidil include chest pain, rapid heartbeat, faintness, dizziness, sudden or 
unexplained weight gain, swelling of the hands or feet, scalp irritation or redness, changes in 
hair color and texture, and unwanted facial hair growth. 
 
 
2 Study Objectives 
 
The overall goal of this study is to develop regenerative cell therapy for use in female patients with AGA. The 
primary objective of this proposal is to conduct a pilot study on the efficacy of using PRP to treat this type of 
hair loss in females. 
 
 
 
 


8 
 
3 Study Design 
3.1 
General Design 
 
The study will be a randomized control trial following a crossover design to compare PRP to MNX in females.   
 
         0          1          2          3         4         5           6          7          8          9         10        11         12 
 
         Time (months) 
 
 
 
 
 
 
 
 
 
 
 
 
Subjects will be recruited from the clinical practice of the Department of Dermatology, Mayo Clinic Florida. 23 
females with AGA grades I-II based on the Ludwig Classification.  (Ludwig) 
  
 
Patients will be randomized into a PRP then MNX (Arm A) or MNX then PRP group (Arm B). Both 
treatments will be administered for 12 weeks with a 2 month washout period in between treatments. Hair 
density and adverse events will be assessed using Phototrichograms, Macroscopic Photos, physical exams and 
patient questionnaires. All patients will be followed for efficacy and adverse events at 3 month intervals until 1 
year after enrollment. 
 
3.2 
Primary Study Endpoints 
 
1) Hair growth related to each of the two treatment arms. 
2) Morbidity related to PRP scalp administration. 
3.3 
Secondary Study Endpoints 
 
1) To collect PRP samples to study the molecular mechanisms of PRP and hair growth 
3.4 
Primary Safety Endpoints 
 
1) Documentation will include levels of pain at injection sites, any evidence of adverse reaction in 
the skin (e.g., swelling, erythema, warmth) and any other concerns expressed by patients.  
4 Subject Selection Enrollment and Withdrawal 
4.1 
Inclusion Criteria 
1. Only female patients are eligible  
2. Patients must be 18 years of age or older  
3. Patients must have Androgenic Alopecia Grade I-II by Ludwig Classification 
-washout- 
Arm A (n=10) 
Arm B (n=10) 


9 
 
4. Patients must have been on stable birth control for the past  month if premenopausal. 
5. Patients are able and willing to provide written informed consent after the nature of the study is fully 
explained 
4.2 
Exclusion Criteria 
 
1. Patients with clinically abnormal hematology, serum chemistry, or screening laboratory results as 
reviewed by the Principal Investigator 
2. Patients who have undergone topical and systemic therapies for hair loss 3 months prior to the procedure 
3. Patients who have used any cosmetic product meant to address hair loss 3 months prior to enrollment 
4. Patients taking anti-rheumatic disease medication (including methotrexate or other anti-metabolites) 
within the 3 months prior to study entry.  
5. Patients previously having undergone hair transplant surgery prior to study entry  
6. Patients who are pregnant or currently breast-feeding children as Rogaine for Women is contra indicated 
for these women 
7. Patients who have taken spironolactone in the 3 months prior to study participation 
8. Patients with systemic, rheumatic or inflammatory disease or who are immunosuppressed 
9. Patients with ongoing infectious disease, including HIV and hepatitis  
10. Patients with clinically significant cardiovascular, renal, hepatic, endocrine disease, cancer, or diabetes 
11. Patients participating in a study of an experimental drug or medical device within 30 days of study entry 
12. Patients with history of platelet disorders, bone marrow aplasia 
13. Patients with a history of sepsis, or cancer must have been disease free for the past 5 years. 
14. Patients taking antiaggregating therapy 
15. Patients on anticoagulant therapy 
16. Patients with tendency to keloid formation 
17. Patients with uncompensated diabetes 
18. Patients with active skin disease or skin infection at intended treatment areas 
 
4.3 
Subject Recruitment, Enrollment and Screening 
 
Subjects will be recruited from the clinical practice of the Department of Dermatology, Mayo Clinic Florida. 
Patients seeking treatment for Androgenic Alopecia will be offered standard conservative medical therapy for 
their disease as part of routine dermatological treatment. As an alternative, patients will be informed of the 
study protocol, and offered the option to enroll if they meet the study eligibility criteria. 
 
4.4 
Early Withdrawal of Subjects 
4.4.1 When and How to Withdraw Subjects 
 
Subjects may withdraw from the study at any time for the following: 
• Subject safety issues 
• Subject decision to withdraw from the study (withdrawal of consent) 
• Subject’s decision to pursue other therapeutic regimens, either surgical or non-surgical 
 
4.4.2 Data Collection and Follow-up for Withdrawn Subjects 
 


10 
 
Even though a subject has withdrawn from the study, it may be important to collect some follow-up or survival 
data on subjects throughout the protocol defined follow-up period. Such data are important to the integrity of 
the final study analysis since early withdrawal could be related to the safety profile of the study drug. If a 
subject withdraws consent to participate in the study for subject safety reasons attempts will be made to obtain 
permission to collect follow up information whenever possible. Patients will be analyzed by intention to treat 
principles without some of the follow-up data as necessary. 
5 Study Drug  
5.1 
Description 
 
1. PRP – Autologous Platelet Rich Plasma isolated from blood collected from each patient at each 
treatment time point.   
 
2. Minoxidill 5% Topical Foam - Manufacturers Description: Indications: To regrow hair on top of the 
scalp.  Active Ingredient: Minoxidil 5% w/w (without propellant) Inactive Ingredients: butane, butylated 
hydroxytoluene, cetyl alcohol, citric acid, glycerin, isobutane, lactic acid, polysorbate 60, propane, 
purified water, SD alcohol 40-B, stearyl alcohol (Rogaine for Women). 
5.2 
Treatment Regimen 
 
Patients will be randomized into a PRP then MNX (Arm A) or MNX then PRP group (Arm B). Patients 
randomized to Arm A will receive injections of PRP using the nappage technique (Sivagnanam) every 4 weeks 
for a total of 3 treatments over 12 weeks. Patients will then have a 2 month (8 week) washout period lasting 
until the end of month 4. Starting on month 5 they will receive crossover treatment with Minoxidil by applying 
the 5% foam once daily following manufacture instructions (Rogaine For Women) for a total of 3 months after 
which they will be followed until 1 year after enrollment. Patients randomized to Arm B will apply Minoxidil 
topical foam once daily following manufacture instructions for a total of 3 months followed by a two month (8 
week) washout period. At the 5 month mark, the patients in Arm B will receive crossover treatment with PRP 
harvested and administered in the same manner as the patients in Arm A.  At the end of the PRP treatment 
patients in Arm B will be followed until 1 year after enrollment.  
5.3 
Method for Assigning Subjects to Treatment Groups 
 
Assignment of treatments will be randomized using dynamic allocation.  
5.4 
Preparation and Administration of Study Drug 
 
PRP will be harvested and prepared in the Mayo Jacksonville Dermatology practice.  20 mLs of blood will be 
collected from patients using standard venipuncture technique.  Collected blood will be processed using a 
standard dual spin centrifugation protocol resulting in approximately 10.5 mLs of PRP.   The 10 cc of PRP will 
be injected with a 30-gauge needle using the nappage technique (Sivagnanam) over the affected part of the 
scalp, approximately 1/10 cc / injection site.  At the investigators discretion scalp may be numbed with 
Gebauer’s Pain Ease Nonflammable Medium Stream Spray Instant Topical Anesthetic or Impax Lisocaine and 
Prilocaine Cream, USP 2.5%.  The remaining 0.5 mL of PRP will be collected for future research purposes.  
  
5.5 
Prior and Concomitant Therapy 
 
Subjects will not be allowed any concomitant therapy for hair loss treatment. 


11 
 
No prior hair replacement / treatment surgeries.  
No topical and systemic therapies for hair loss 3 months prior to study enrollment. 
6 Study Procedures 
 
a. At baseline all patients will undergo a clinical exam, assessments of hair density using 
phototrichograms, macroscopic photographs, patient questionnaires and labs including CBC, ferritin, 
thyroid function, testosterone, TSH and pregnancy test for premenopausal women. All patients will also 
have a small area shaved at the center of the treatment area, as well as a small tattoo injection to allow 
for follicle counting via the computerized software. 
b. At 3 months, 5 months, 8 months and 12 months, patients will have repeat clinical exams and 
phototrichograms, macroscopic photographs and complete patient questionnaires. 
c. At 12 months, repeat serum tests (CBC, ferritin, thyroid function, testosterone, TSH) will be performed. 
d. Adverse events will be assessed at each visit (3, 5, 8 and 12 months post enrollment) as well as 1 week 
after each injection of PRP. 
e. Shampoo and conditioner will be standardized with the study supplying Free & Clear brand products to 
the patients free of charge for the duration of the study. 
f. Pre-menopausal patients will undergo a second pregnancy test before beginning the cross-over 
treatment. 
 


12 
 
6.1 PRP vs Minoxidil Study Calendars 
 
Arm A – PRP then MNX 
 
 
Procedure 
Baseline 
 
 
Week 1 
 
 
Week 4 
 
 
Week 5 
 
 
Week 8 
 
 
Week 9 
 
 
Month 3/ 
Week 12 
 
Month 5/ 
Week 20 
 
Month 8/ 
Week 32 
 
Month 12/ 
Week 48 
 
Consent 
X 
  
  
  
 
 
  
  
  
  
Clinical Exam 
X 
  
  
  
 
 
X 
X 
X 
X 
Labs 
X 
  
  
  
 
 
  
 
X*   
  
X 
Phototrichograms &   
Macroscopic Photos 
X 
  
  
  
 
 
X 
X 
X 
X 
Questionnaire 
X 
  
  
  
 
 
X 
X 
X 
X 
PRP Injections 
X 
  
X 
  
X 
 
  
  
  
  
Minoxidil Treatment Daily 
 
  
  
  
 
 
 
X 
 
 
Adverse Events 
  
X 
X 
X 
X 
X 
X 
X 
X 
X 
Clinical Assessment of 
 Injection Sites 
  
X 
  
X 
 
X 
 
 
 
 
Collection of Research 
Samples 
X 
 
X 
 
X 
 
 
 
 
 
*- pre-menopausal patients will have a second pregnancy test at the end of the washout period before beginning the cross-over treatment. 
Arm B – MNX then PRP 
 
 
Procedure 
Baseline 
 
 
Month 3/ 
Week 12 
 
Month 5/ 
Week 20 
 
Week 21 
 
 
Week 24 
 
 
Week 25 
 
 
Week 28 
 
 
Week 29 
 
 
Month 8/ 
Week 32 
 
Month 12/ 
Week 48 
 
Consent 
X 
  
  
  
  
  
  
  
  
  
Clinical Exam 
X 
 X 
X 
  
  
  
  
  
X 
X 
Labs 
X 
  
X* 
  
  
  
  
  
 
X  
Phototrichograms &  
Macroscopic Photos 
X 
X 
X 
  
  
  
  
  
X 
X 
PRP Injections 
 
  
X 
  
X 
  
X 
  
  
  
Minoxidil Treatment Daily 
X 
 
 
 
 
 
 
 
 
 
Questionnaire 
X 
X 
X 
  
  
  
  
  
X 
X 
Adverse Events 
  
X 
X 
X 
X 
X 
X 
X 
X 
X 
Clinical Assessment of 
 Injection Sites 
  
 
 
X 
  
X 
  
X 
 
 
Collection 
of 
Research 
Samples 
 
 
X 
 
X 
 
X 
 
 
 
*- pre-menopausal patients will have a second pregnancy test at the end of the washout period before beginning the cross-over treatment. 


13 
 
7 Statistical Plan 
7.1 
Sample Size Determination 
 
A total of 23 patients will be enrolled in this study. This sample size will be sufficient in order to gain valuable 
data to be used in the design of a future, larger study. Given the pilot/feasibility nature of the study, no formal 
power calculations were performed. 
 
7.2 
Statistical Methods 
 
Data Analysis – Given the design, all statistical analysis will be focused on the difference in outcomes, and 
differences in outcomes compared to baseline, for each treatment within individual patients. For continuous or 
ordinal outcome variables, we will use a Wilcoxon signed rank test for comparison of outcomes between PRP 
and Minoxidil, while for categorical outcome variables we will use McNemar’s exact test for these. Continuous 
outcome measures will be summarized using the sample mean, standard deviation, median, minimum, 25th and 
75th percentiles, and maximum. Categorical variables will be summarized with number and percentage. 95% 
confidence intervals (CIs) will be estimated where appropriate. P-values of 0.05 or less will be considered as 
statistically significant, however the results of these tests will be interpreted with the low power of this pilot 
study in mind; more emphasis will be placed on graphical and descriptive summaries than on the results of 
statistical tests. No adjustment for multiple testing will be made owing to the exploratory, pilot nature of the 
study. Given the prospective nature of the study, we expect to encounter little if any missing data. However in 
the event of missing data, no attempt to impute this missing data will be made in this pilot study; missing data 
will be treated as missing. 
 
Descriptive Statistics 
 
We will summarize continuous variables using the sample mean, standard deviation, median, minimum, 25th 
and 75th percentiles, and maximum. We will summarize categorical variables with number and percentages. 
Ordinal variables will be summarized using the sample median, minimum, 25th and 75th percentiles, and 
maximum, as well as with number and percentage.  
 
Handling of Missing Data 
 
Given the prospective nature of the study, we expect to encounter little if any missing data. However in the 
event of missing data, no attempt to impute this missing data will be made in this pilot study; missing data will 
be treated as missing. 
 
Multiplicity 
 
No adjustment for multiple testing will be made owing to the exploratory, pilot nature of the study. P-value of 
0.05 or less will be considered as statistically significant. 
 
Primary Hypothesis: 
 
We hypothesize that PRP administration for AGA will be safe and without significant complications for the 
patient.  AGA will be comparable in efficacy or superior to conventional treatment with MNX. 
 


14 
 
The primary endpoints of the study, occurrence of adverse reactions and morbidity, are both dichotomous 
categorical variables. As such, the proportion for which these outcomes occur will be estimated along with 95% 
CIs. These proportions will be compared between treatments using McNemar’s exact test 
 
 
Interim Analysis 
 
No interim analysis will be performed in this pilot study. 
 
7.3 
Subject Population(s) for Analysis 
All-treated population:  Any subject randomized into the study that received study drug.  
 
 
8 Safety and Adverse Events 
8.1 
Definitions 
 
Unanticipated Problems Involving Risk to Subjects or Others (UPIRTSO) 
Any unanticipated problem or adverse event that meets the following three criteria:  
• Serious: Serious problems or events that results in significant harm, (which may be physical, 
psychological, financial, social, economic, or legal) or increased risk for the subject or others (including 
individuals who are not research subjects). These include: (1) death; (2) life threatening adverse 
experience; (3) hospitalization - inpatient, new, or prolonged; (4) disability/incapacity - persistent or 
significant; (5) birth defect/anomaly; (6) breach of confidentiality and (7) other problems, events, or new 
information (i.e. publications, DSMB reports, interim findings, product labeling change) that in the 
opinion of the local investigator may adversely affect the rights, safety, or welfare of the subjects or 
others, or substantially compromise the research data, AND 
• Unanticipated: (i.e., unexpected) problems or events are those that are not already described as potential 
risks in the protocol, consent document, not listed in the Investigator’s Brochure, or not part of an 
underlying disease. A problem or event is "unanticipated" when it was unforeseeable at the time of its 
occurrence. A problem or event is "unanticipated" when it occurs at an increased frequency or at an 
increased severity than expected, AND 
• Related: A problem or event is "related" if it is possibly related to the research procedures. 
 
 
Adverse Event 
 
An untoward or undesirable experience associated with the use of a medical product (i.e. drug, device, biologic) 
in a patient or research subject. 
 
 
 
 
Serious Adverse Event 


15 
 
 
Adverse events are classified as serious or non-serious.  Serious problems/events can be well defined and 
include; 
• death 
• life threatening adverse experience 
• hospitalization 
• inpatient, new, or prolonged; disability/incapacity 
• persistent or significant birth defect/anomaly 
 
or other events that in the opinion of the sponsor-investigator may have adversely affected the rights, 
safety, or welfare of the subjects or others, or substantially compromised the research data. 
 
All adverse events that do not meet any of the criteria for serious, should be regarded as non-serious adverse 
events.  
 
Adverse Event Reporting Period 
For this study, the study treatment follow-up period is 12 months after enrollment. The adverse event 
monitoring period will end at the end of the follow-up period. If an event is reported after this time it would not 
be considered study related. 
 
Preexisting Condition 
A preexisting condition is one that is present at the start of the study.  A preexisting condition should be 
recorded as an adverse event if the frequency, intensity, or the character of the condition worsens during the 
study period. 
 
General Physical Examination Findings 
At screening, any clinically significant abnormality should be recorded as a preexisting condition.  At the end of 
the study, any new clinically significant findings/abnormalities that meet the definition of an adverse event must 
also be recorded and documented as an adverse event.  
 
Post-study Adverse Event 
All unresolved adverse events should be followed by the sponsor-investigator until the events are resolved, the 
subject is lost to follow-up, or the adverse event is otherwise explained.  At the last scheduled visit, the sponsor-
investigator should instruct each subject to report, to the sponsor-investigator, any subsequent event(s) that the 
subject, or the subject’s personal physician, believes might reasonably be related to participation in this study.  
 
Abnormal Laboratory Values 
A clinical laboratory abnormality should be documented as an adverse event if it was exhibited as a severe 
reaction i.e. swelling, that required hospitalization or additional clinical intervention.  
 
 
Hospitalization, Prolonged Hospitalization or Surgery 
Any adverse event that results in hospitalization or prolonged hospitalization should be documented and 
reported as a serious adverse event unless specifically instructed otherwise in this protocol.  Any condition 
responsible for surgery should be documented as an adverse event if the condition meets the criteria for an 
adverse event.  
 
 
Neither the condition, hospitalization, prolonged hospitalization, nor surgery are reported as an adverse event in 
the following circumstances:  


16 
 
• Hospitalization or prolonged hospitalization for diagnostic or elective surgical procedures for a 
preexisting condition.  Surgery should not be reported as an outcome of an adverse event if the purpose 
of the surgery was elective or diagnostic and the outcome was uneventful. 
• Hospitalization or prolonged hospitalization required to allow efficacy measurement for the study. 
• Hospitalization or prolonged hospitalization for therapy of the target disease of the study, unless it is a 
worsening or increase in frequency of hospital admissions as judged by the clinical investigator. 
 
8.2 
Recording of Adverse Events 
All adverse events will be documented as required under 21 CFR part 1271. Depending on the nature and 
seriousness of the adverse reaction, the IRB will be notified according to the Mayo Clinic IRB policy. If An 
SAE occurs it will also be reported to the FDA as per regulations 
 
At each contact with the subject, the study team must seek information on adverse events by specific 
questioning and, as appropriate, by examination.  Information on all adverse events should be recorded 
immediately in the source document, and also in the appropriate adverse event section of the case report form 
(CRF) or separate worksheet.  All clearly related signs, symptoms, and abnormal diagnostic, laboratory or 
procedure results should recorded in the source document. 
 
All adverse events occurring during the study period must be recorded.  The clinical course of each event 
should be followed until resolution, stabilization, or until it has been ultimately determined that the study 
treatment or participation is not the probable cause.  Serious adverse events that are still ongoing at the end of 
the study period must be followed up, to determine the final outcome. 
 
8.3 
Reporting of Serious Adverse Events and Unanticipated Problems 
When an adverse event has been identified, the study team will take appropriated action necessary to protect the 
study participant and then complete the Study Adverse Event Worksheet and/or log.  The sponsor-investigator 
will evaluate the event and determine the necessary follow-up and reporting required. 
 
8.3.1 Sponsor-Investigator reporting: notifying the Mayo IRB 
The sponsor-investigator will report to the Mayo IRB any UPIRTSOs and Non-UPIRTSOs according to the 
Mayo IRB Policy and Procedures. 
8.4 
Stopping Rules  
Study enrollment and treatment procedures will be suspended in the event that 3 successive subjects experience 
a Serious Adverse Event within the first 3 months after treatment.  The study would only be resumed after a 
thorough review of the incidents and any corrective and preventative actions have been put in place along with 
consultation between the study team and the IRB. 
8.5 
Medical Monitoring 
It is the responsibility of the Principal Investigator to oversee the safety of the study at his/her site.  This safety 
monitoring will include careful assessment and appropriate reporting of adverse events as noted above, as well 
as the construction and implementation of a site data and safety-monitoring plan (see section 10  “Study 
Monitoring, Auditing, and Inspecting”).  Medical monitoring will include a regular assessment of the number 
and type of serious adverse events. 
 


17 
 
9 Data Handling and Record Keeping 
9.1 
Data Handling 
Data will be collected and entered by the study coordinator assigned to the project, and stored into a REDCap 
database. 
9.2 
Confidentiality 
Information about study subjects will be kept confidential and managed according to the requirements of the 
Health Insurance Portability and Accountability Act of 1996 (HIPAA).  Those regulations require a signed 
subject authorization informing the subject of the following:  
• What protected health information (PHI) will be collected from subjects in this study 
• Who will have access to that information and why 
• Who will use or disclose that information 
• The rights of a research subject to revoke their authorization for use of their PHI.  
In the event that a subject revokes authorization to collect or use PHI, the investigator, by regulation, retains the 
ability to use all information collected prior to the revocation of subject authorization.  For subjects that have 
revoked authorization to collect or use PHI, attempts should be made to obtain permission to collect at least 
vital status (long term survival status that the subject is alive) at the end of their scheduled study period. 
 
Hard copy data such as consent forms will be stored in locked file cabinets; electronic data will be stored in 
secure web-based database (REDCap) that will be designed with the help of the statistician.  
9.3 
Source Documents 
Source data are all information, original records of clinical findings, observations, or other activities in a clinical 
trial necessary for the reconstruction and evaluation of the trial.  Source data are contained in source documents.  
Examples of these original documents, and data records include: hospital records, clinical and office charts, 
laboratory notes, memoranda, subjects’ diaries or evaluation checklists, pharmacy dispensing records, recorded 
data from automated instruments, copies or transcriptions certified after verification as being accurate and 
complete, microfiches, photographic negatives, microfilm or magnetic media, x-rays, subject files, and records 
kept at the pharmacy, at the laboratories, and at medico-technical departments involved in the clinical trial. 
 
Data Management 
Study data will be managed in a study specific REDCap system 
 
Data Security and Confidentiality 
 
The REDCap system has built in systems for control of access, data integrity and audit trails. Access and 
confidentiality are controlled in a manner similar to other institutional systems. 
9.4 
Records Retention 
The sponsor-investigator will maintain records and essential documents related to the conduct of the study.  
These will include subject case histories and regulatory documents. 
 
The sponsor-investigator will retain the specified records and reports for whichever is longer; 
1. Up to 2 years after the marketing application is approved for the drug; or, if a marketing application is 
not submitted or approved for the drug, until 2 years after shipment and delivery of the drug for 
investigational use is discontinued and the FDA has been so notified. OR 
 
2. As outlined in the Mayo Clinic Research Policy Manual “Access to and Retention of Research Data 
Policy” http://mayocontent.mayo.edu/research-policy/MSS_669717. 


18 
 
10 Study Monitoring, Auditing, and Inspecting 
10.1 Study Monitoring Plan 
The investigator will allocate adequate time for such monitoring activities.  The Investigator will also ensure 
that the monitor or other compliance or quality assurance reviewer is given access to all the study-related 
documents and study related facilities (e.g. pharmacy, diagnostic laboratory, etc.), and has adequate space to 
conduct the monitoring visit. 
 
The study specific DSMP will be submitted to the IRB as a separate document in the IRB application. 
 
This study will be monitored on a routine basis during the conduct of the trial.  The Mayo Clinic Office of 
Research Regulatory Support will assist in coordination of the clinical monitoring for the trial as a service for 
the sponsor-investigator.  Clinical trial monitoring requires review of the study data generated throughout the 
duration of the study to ensure the validity and integrity of the data along with the protection of human research 
subjects.  This will assist sponsor-investigators in complying with Food and Drug Administration regulations. 
10.2 Auditing and Inspecting 
The investigator will permit study-related monitoring, audits, and inspections by the IRB, the sponsor, and 
government regulatory agencies, of all study related documents (e.g. source documents, regulatory documents, 
data collection instruments, study data etc.).  The investigator will ensure the capability for inspections of 
applicable study-related facilities (e.g. pharmacy, diagnostic laboratory, etc.). 
 
Participation as an investigator in this study implies acceptance of potential inspection by government 
regulatory authorities and applicable compliance offices. 
11 Ethical Considerations 
This study is to be conducted according to United States government regulations and Institutional research 
policies and procedures. 
 
This protocol and any amendments will be submitted to a properly constituted local Institutional Review Board 
(IRB), in agreement with local legal prescriptions, for formal approval of the study.  The decision of the IRB 
concerning the conduct of the study will be made in writing to the sponsor-investigator before commencement 
of this study. 
 
All subjects for this study will be provided a consent form describing this study and providing sufficient 
information for subjects 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 IRB for the study.  The formal consent of a 
subject, using the Approved IRB consent form, must be obtained before that subject undergoes any study 
procedure.  The consent form must be signed by the subject or the subject’s legally authorized representative, 
and the individual obtaining the informed consent. 
 
12 Study Finances 
12.1 Funding Source 
Mayo Center for Regenerative Medicine accelerated research Intramural funding. 
 


19 
 
12.2 Conflict of Interest 
Any study team member who has a conflict of interest with this study (patent ownership, royalties, or financial 
gain greater than the minimum allowable by their institution, etc.) must have the conflict reviewed by a properly 
constituted Conflict of Interest Committee with a Committee-sanctioned conflict management plan that has 
been reviewed and approved by the study sponsor-investigator prior to participation in this study. 
12.3 Subject Stipends or Payments 
No subject payments or reimbursement will be offered. 
 
13 Publication Plan 
The Principal Investigator will be responsible for manuscript preparation and submission to a suitable 
orthopedic journal for publication of results. 
 
14 References 
 
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