Authors’ Disclosure Statement: The authors report that Arthrex donated syringes for generating platelet-rich plasma. Dr. Fortier reports that she is a paid consultant for Arthrex. Dr. Cole reports that he receives intellectual property royalties from, is a paid consultant, and provides research support to Arthrex. This article was supported by the National Institute of Health and the Harry M. Zweig Memorial Fund for Equine Research. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Acknowledgment: The authors would like to thank Paula Sharp for her generous technical assistance in generating the article.
Dr. Wilson is a Veterinarian, Susitna Holistic Veterinary Services, Wasilla, Alaska. Dr. Fortier is James Law Professor of Surgery, Department of Clinical Sciences, and Ms. Goodale is a Veterinary Student, Cornell University, Ithaca, New York. Dr. Cole is an Orthopedic Surgeon and Professor, Department of Orthopedics, Rush University Medical Center, Chicago, Illinois.
Address correspondence to: Lisa A. Fortier, DVM, PhD, Department of Clinical Sciences, Cornell University, 930 Campus Road, Ithaca, NY 14853 (email, laf4@cornell.edu).
Am J Orthop. 2018;47(4). Copyright Frontline Medical Communications Inc. 2018. All rights reserved.
Brooke H. Wilson, DVM, MS Brian J. Cole, MD Margaret B. Goodale, BS Lisa A. Fortier, DVM, PhD . Short-Term Storage of Platelet-Rich Plasma at Room Temperature Does Not Affect Growth Factor or Catabolic Cytokine Concentration. Am J Orthop.
April 13, 2018
TAKE-HOME POINTS
Blood can be stored at room temperature for up to 4 hours before making PRP without loss in activity.
PRP can be stored at room temperature for up to 4 hours before administration to a patient without loss in activity.
Manual, direct smear analysis is as accurate as automated counting for measuring platelet concentration in PRP.
References
ABSTRACT
The aim of this study was to provide clinical recommendations about the use of platelet-rich plasma (PRP) that was subjected to short-term storage at room temperature. We determined bioactive growth factor and cytokine concentrations as indicators of platelet and white blood cell degranulation in blood and PRP. Additionally, this study sought to validate the use of manual, direct smear analysis as an alternative to automated methods for platelet quantification in PRP.
Blood was used to generate low-leukocyte PRP (Llo PRP) or high-leukocyte PRP (Lhi PRP). Blood was either processed immediately or kept at room temperature for 2 or 4 hours prior to generation of PRP, which was then held at room temperature for 0, 1, 2, or 4 hours. Subsequently, bioactive transforming growth factor beta-1 and matrix metalloproteinase-9 were measured by ELISA (enzyme-linked immunosorbent assay). Manual and automated platelet counts were performed on all blood and PRP samples.
There were no differences in growth factor or cytokine concentration when blood or Llo PRP or Lhi PRP was retained at room temperature for up to 4 hours. Manual, direct smear analysis for platelet quantification was not different from the use of automated machine counting for PRP samples, but in the starting blood samples, manual platelet counts were significantly higher than those generated using automated technology.
When there is a delay of up to 4 hours in the generation of PRP from blood or in the application of PRP to the patient, bioactive growth factor and cytokine concentrations remain stable in both blood and PRP. A manual direct counting method is a simple, cost-effective, and valid method to measure the contents of the PRP product being delivered to the patient.
Platelet-rich plasma (PRP) is used to promote healing in many areas of medicine, such as dental surgery,1,2 soft-tissue injury,3,4 orthopedic surgery,5,6 wound healing,7 and veterinary medicine.8,9 Despite its extensive use, there are still questions about the clinical efficacy of PRP.10-12 Due to biological heterogeneity between patients13,14 and differences between available manufacturing kits,13,15 PRP can be highly variable between patients. There are classification schemes to categorize the various types of PRP,16-18 which can be divided broadly into low-leukocyte PRP (Llo PRP) and high-leukocyte PRP (Lhi PRP). PRP can be used as a point of care therapy, prepared and used immediately, or it can be used during a surgical procedure. In some institutions, blood is drawn by a phlebotomist, processed in the hospital laboratory, and then delivered to the operating room. In other instances, PRP is generated patient-side by the primary attending physician’s team, who draws the blood and processes it for immediate use.5,19 Delays at any step in these various scenarios could lead to the blood or the resultant PRP remaining at room temperature from minutes to several hours prior to administration to the patient. This variability in PRP protocols between clinical and surgical settings adds to concerns regarding the stability and efficacy of the biologic.
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