The Role of Platelet Homeostasis in a Novel Topical PRP Formulation

December 2020 | Volume 19 | Issue 12 | Original Article | 1215 | Copyright © December 2020


Published online November 25, 2020

Shaun Wootten BSEa, Zoe Diana Draelos MDb, Robert S Kellar PhDc,d, Lawrence Rheins PhDa

aDepartment of Research and Development, Aesthetics Biomedical Inc., Phoenix, AZ
bDermatology Consulting Services, PLLC, High Point, NC
cDevelopment Engineering Sciences, LLC, Flagstaff, AZ
dNorthern Arizona University, Center for Materials Interfaces in Research & Applications, Flagstaff, AZ

Abstract
Background: Topical platelet-rich plasma (PRP) must demonstrate stability to insure biologic activity in aesthetic medicine.
Objective: The objective of this research was to evaluate the role of platelet homeostasis in a novel PRP topical cosmetic formulation to provide facial appearance improvement.
Methods: The stability of the topical PRP formulation was evaluated in vitro followed by clinical in vivo testing. The in vitro evaluation examined platelet stability and morphology over a 90-day period within the preservative cosmetic base utilizing ELISA and light microscopy (LM)/scanning electron microscopy (SEM). The in vivo clinical study enrolled 20 subjects in a 120-day double blind split face study to evaluate the effect of 5–7x concentrated PRP compared to 2–3x concentrated PRP on facial photoaging. Cosmetic effect was evaluated by the subject and the dermatologist investigator on a 5-point ordinal scale at baseline, week 8, and week 16.
Results: 90-day stability for the topical PRP formulation was verified via ELISA and LM/SEM. ELISA showed the PRP was more inactive than control conditions via analyte concentration curves (PDGF-AB, EGF, and P-Selectin). LM/SEM demonstrated the PRP had less aggregation/activation over time within the cosmetic base and that refrigeration is superior to room-temperature storage thus delaying full platelet degranulation. The in vivo clinical study demonstrated parity between 20ml and 60ml PRP in terms of clinical efficacy.
Conclusion: Platelets remain viable for up to 90 days in a refrigerated cosmetic vehicle with demonstrated topical clinical PRP facial benefits. PRP kits of 20ml and 60ml volumes for topical PRP are equally efficacious.

J Drugs Dermatol. 2020;19(12):1215-1218. doi:10.36849/JDD.2020.5495

INTRODUCTION

Platelet-rich plasma (PRP) originally was used in the orthopedic and dental arenas for its regenerative effects in wound healing.1-2 PRP is gaining popularity in aesthetic medicine as an injectable and topical treatment for improving the appearance of photoaged skin.3-5 Efficacious topical PRP for long term usage was not previously possible due to shelf life and stability limitations.

Platelets are anucleaic cytoplasmic fragments derived from megakaryocytes found in bone marrow. The platelet contains mitochondria, microtubules, and α, δ, λ-granules.6 There may be 50 to 80 granules in a platelet containing biologically active substances including adhesive proteins, plasma proteins, cellular mitogens, coagulation factors, protease inhibitors, micro vesicles, and exosomes.7 During platelet activation, degranulation occurs leading to the secretion of biologically active molecules. Degranulation of PRP must be inhibited until the topical preparation is applied to the skin; thus, creating long-term stability challenges.

PRP stability can be achieved by 3 key mechanisms: (1) reduce platelet activation during blood collection, preparation, centrifugation processing, and storage; (2) reduce glucose consumption and lactate production; (3) provide ample glucose during storage.8 These concepts were incorporated into the PRP topical vehicle utilized in this research containing electrolytes, an electrolytic resistant chassis, glucose, and a preservative system with anti-fungal/anti-bacterial property that also functions as a preservative for the platelet's active biomolecules. This research examined the ability of the topical vehicle to maintain platelet granule stability while delivering an aesthetically pleasing PRP-containing anti-aging formulation to the face.

METHODS

In Vitro PRP Stability Assessment
Four (4) healthy female or male subjects who met all inclusion criteria and none of the exclusion criteria and signed informed consent (IntegReview Institutional Review Board; Austin, TX) were enrolled in this multi-site study. Subjects underwent phlebotomy to obtain 200mL of blood for PRP preparation. During phlebotomy, 50mL of whole blood specimen was harvested per subject in a 60mL syringe containing 10mL of sodium citrate anti-coagulant. The anti-coagulant/whole blood