Basic Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Orthop. Jun 18, 2022; 13(6): 587-602
Published online Jun 18, 2022. doi: 10.5312/wjo.v13.i6.587
Content of blood cell components, inflammatory cytokines and growth factors in autologous platelet-rich plasma obtained by various methods
Maciej Dejnek, Jarosław Witkowski, Helena Moreira, Sylwia Płaczkowska, Piotr Morasiewicz, Paweł Reichert, Aleksandra Królikowska
Maciej Dejnek, Jarosław Witkowski, Paweł Reichert, Department of Trauma Surgery, Wroclaw Medical University, Wroclaw 50-556, Poland
Helena Moreira, Department of Medical Science Foundation, Wroclaw Medical University, Wroclaw 50-556, Poland
Sylwia Płaczkowska, Teaching and Research Diagnostic Laboratory, Department of Laboratory Diagnostics, Wroclaw Medical University, Wroclaw 50-556, Poland
Piotr Morasiewicz, Institute of Medical Sciences, Department of Orthopaedic and Trauma Surgery, University of Opole, Opole 45-052, Poland
Aleksandra Królikowska, Ergonomics and Biomedical Monitoring Laboratory, Department of Physiotherapy, Wroclaw Medical University, Wroclaw 50-355, Poland
Author contributions: Dejnek M, Reichert P, and Królikowska A designed and coordinated the study; Dejnek M, Witkowski J, Moreira H, and Płaczkowska S performed the experiments, acquired and analyzed data; Dejnek M, Reichert P, Królikowska A, and Morasiewicz P interpreted the data; Dejnek M wrote the manuscript; all authors approved the final version of the article.
Supported by the Wroclaw Medical University as a Regional Center of Excellence in the field of medical sciences and health sciences implemented under the funds of the Ministry of Science and Higher Education (Republic of Poland) in the program "Regional Initiative of Excellence" in the years 2019-2022, No. RID.Z501.20.008.
Institutional review board statement: The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Institutional Ethics Committee of Wroclaw Medical University (KB - 163/2020, 30.03.2020).
Informed consent statement: All study participants provided informed written consent prior to study enrollment.
Conflict-of-interest statement: The authors declare no conflict of interest. The founders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.
Data sharing statement: The datasets used during the current study are available from the corresponding author on reasonable request.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Maciej Dejnek, MD, Research Scientist, Surgeon, Department of Trauma Surgery, Wroclaw Medical University, ul. Borowska 213, Wroclaw 50-556, Poland. maciej.dejnek@student.umed.wroc.pl
Received: January 18, 2022
Peer-review started: January 18, 2022
First decision: March 24, 2022
Revised: April 4, 2022
Accepted: May 7, 2022
Article in press: May 7, 2022
Published online: June 18, 2022
Processing time: 149 Days and 7.4 Hours
ARTICLE HIGHLIGHTS
Research background

Autologous platelet-rich plasma (PRP) therapy is a method used to treat a variety of diseases related to soft tissue degeneration. The main idea behind this is to improve local healing and stimulate regeneration by administering large amounts of platelet-derived growth factors and cytokines. There are many commercial kits available to assist in obtaining PRP in an outpatient setting.

Research motivation

Due to the wide variety of PRP preparation systems, there are justified doubts about the quality of the obtained samples. Differences in the content of biologically active compounds between some PRP systems have already been demonstrated. However, only a small number of available systems and a limited number of cytokines and growth factors have been investigated.

Research objectives

To compare PRP obtained using four different commercial preparation systems in terms of the content of biologically active components, correlations between those components and their repeatability in each method.

Research methods

After obtaining informed consent from participants, whole blood was collected from 12 young healthy male volunteers, and 4 different PRP samples were prepared from each of them in a single-donor model. PRP samples were prepared using different commercial kits: Arthrex Autologous Conditioned Plasma (ACP) Double Syringe System (Arthrex Inc., United States), the Mini GPS III Platelet Concentration System (Biomet Inc., United States), the Xerthra PRP kit (Biovico Sp. z o.o., Poland) and Dr. PRP (Rmedica, Republic of Korea). The content of cellular components in each sample was assessed using an automatic laboratory analyzer Mindray BC-5150 (Shenzhen Mindray Bio-Medical Electronics Co., PRC). To quantify the content of seven selected growth factors (Epidermal growth factor (EGF), Fibroblast Growth Factor- basic, Hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), Transforming Growth Factor-β1, Platelet-Derived Growth Factor-AA, Platelet-Derived Growth Factor-BB and thirteen inflammatory cytokines [(Interferon-α2, Interleukin-10, Interleukin-12p70, Interleukin-17A, Interleukin-18 (IL-18), Interleukin-23 (IL-23), Interleukin-33, Interleukin-6, Interleukin-8, Interferon-γ, Monocyte Chemoattractant Protein-1 (MCP-1), Tumor Necrosis Factor α (TNF-α)], bead-based multiplex immunoassays LEGENDplexTM (BioLegend, United States) that use fluorescence-encoded beads and flow cytometer measurements were performed.

Research results

Differences between PRPs obtained with various preparation systems were found in terms of cellular composition, repeatability, platelet capture efficiency, concentrations of growth factors and inflammatory cytokines. The highest ability to concentrate platelets (PLT) above the baseline was obtained with Mini GPS III (5.05 x) and the lowest with Arthrex ACP (1.47 x). Those two systems had the best repeatability of platelet concentrations assessed as the coefficient of variation of 13.25% and 12.18%, respectively. The highest concentrations of Epidermal growth factor, hepatocyte growth factor, vascular endothelial growth factor, platelet-derived growth factor-AA, platelet-derived growth factor-BB, IL-18, Interleukin-1β, Interleukin-8, MCP-1 and TNF-α were found in PRP with the highest PLT, white blood cells and red blood cells concentrations (obtained with Mini GPS III), and positive significant (P < 0.05) correlations between cell components and these paracrine factors (except TNF-α) were revealed.

Research conclusions

The study provided new data on the differences between PRP obtained with the various commercial systems. The range of analyzed cytokines far exceeded the ranges investigated in earlier publications. The presented findings should help researchers and clinicians choose the system that best meets their expectations.

Research perspectives

Further research should be focused on the comparison of PRPs obtained using different techniques in the context of their biological effect on soft tissues in vitro and their clinical efficacy in various diseases.