Platelet-Rich Plasma Obtained with Different Anticoagulants and Their Effect on Platelet Numbers and Mesenchymal Stromal Cells Behavior In Vitro

28.7.2016
Stem Cells International

Platelet-Rich Plasma Obtained with Different Anticoagulants and Their Effect on Platelet Numbers and Mesenchymal Stromal Cells Behavior In Vitro

Ronaldo José Farias Corrêa do Amaral,1,2 Nemias Pereira da Silva,1 Natália Ferreira Haddad,1 Luana Siqueira Lopes,1 Fábio Dias Ferreira,1 Ricardo Bastos Filho,3 Paola Alejandra Cappelletti,1 Wallace de Mello,1,4 Eric Cordeiro-Spinetti,2 and Alex Balduino1,2

1Excellion Serviços Biomédicos, Amil/UnitedHealth Group, 25651-000 Petrópolis, RJ, Brazil
2Laboratório de Biologia e Tecnologia Celular, Universidade Veiga de Almeida, 20270-150 Rio de Janeiro, RJ, Brazil
3Universidade Federal Fluminense, 24033-900 Niterói, RJ, Brazil
4Centro Universitário Celso Lisboa, 20950-091 Rio de Janeiro, RJ, Brazil

Received 8 January 2016; Revised 9 April 2016; Accepted 27 April 2016

Academic Editor: Jiabing Fan

Copyright © 2016 Ronaldo José Farias Corrêa do Amaral et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Abstract

There are promising results in the use of platelet-rich plasma (PRP) for musculoskeletal tissue repair. However, the variability in the methodology for its obtaining may cause different and opposing findings in the literature. Particularly, the choice of the anticoagulant is the first definition to be made. In this work, blood was collected with sodium citrate (SC), ethylenediaminetetraacetic acid (EDTA), or anticoagulant citrate dextrose (ACD) solution A, as anticoagulants, prior to PRP obtaining. Hematological analysis and growth factors release quantification were performed, and the effects on mesenchymal stromal cell (MSC) culture, such as cytotoxicity and cell proliferation (evaluated by MTT method) and gene expression, were evaluated. The use of EDTA resulted in higher platelet yield in whole blood; however, it induced an increase in the mean platelet volume (MPV) following the blood centrifugation steps for PRP obtaining. The use of SC and ACD resulted in higher induction of MSC proliferation. On the other hand, PRP obtained in SC presented the higher platelet recovery after the blood first centrifugation step and a minimal change in MSC gene expression. Therefore, we suggest the use of SC as the anticoagulant for PRP obtaining.

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