A Comparative Study of The Effects of Anticoagulants on Pure Platelet-Rich Plasma Quality and Potency.

Hachidai Aizawa,Masayuki Nakamura,Hideo Masuki,Kazushige Isobe,Atsushi Sato,Tomoyuki Kawase,Tetsuhiro Tsujino,Koh Nakata,Taisuke Watanabe,Hideo Kawabata

doi:10.3390/biomedicines8030042

Abstract

It is generally accepted that citrate or the A-form of acid-citrate-dextrose (ACD-A) are suitable for preparing platelet-rich plasma (PRP) for regenerative therapy. However, this is based on evidence from blood transfusions and not from regenerative medicine. Thus, we examined the effects of anticoagulants, such as ACD-A, ethylenediaminetetraacetic acid (EDTA), and heparin, on the regenerative quality of PRP to address this gap. The blood samples were collected in the presence of anticoagulants and were processed to prepare pure-PRP. Platelet size, activation status, and intra-platelet free Ca2+ concentration were determined while using a hematology analyzer and flow cytometer. Platelet-derived growth factor-BB (PDGF-BB) was quantified while using an ELISA. In pure-PRP samples, EDTA caused platelet swelling and activation, but yielded the highest number of platelets. Heparin aggregated platelets and disturbed the overall counting of blood cells. However, no significant differences in PDGF-BB levels were observed among the anticoagulants tested. Moreover, when considering the easy preparation of platelet suspensions, without the need for high-level pipetting skills, these findings suggest the comparable potency of EDTA-derived pure-PRP in tissue regeneration and support the use of EDTA in the preparation of pure-PRP. Further in vivo studies are required in animal models to exclude the possible negative effects of including EDTA in pure-PRP preparations.

Highlights

The development and clinical use of platelet-rich plasma (PRP) is undoubtedly an outstanding innovation and it has markedly impacted regenerative medicine
We demonstrated that ACD-A preserved platelet morphology and functionality better than other anticoagulants that are used for PRP preparation
As ACD-A has a lower chelating ability compared with ethylenediaminetetraacetic acid (EDTA) [44] and has no detectable adverse effects on platelets, leukocytes, or other cells involved in tissue regeneration, it is a good choice for use as an anticoagulant

Summary

Introduction

The development and clinical use of platelet-rich plasma (PRP) is undoubtedly an outstanding innovation and it has markedly impacted regenerative medicine. PRP has been increasingly used in various fields of regenerative therapy to date since the first report appeared in 1998 [1]. Several in-vitro studies that were performed by our group and others have provided evidence to support its clinical application [2,3,4,5,6,7]. Its academic value has not been highly considered. A major reason for this paradoxical evaluation is due to the low predictable clinical outcomes of PRP. Several randomized controlled trials (RCT) have been performed while using PRP; all of the evaluations contradict each other, especially in bone regeneration.

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