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      Cellular components and TGF-β1 content of a closed Tube system for Platelet Rich Plasma acquisition in horse] Translated title: [Componentes celulares e TGF-β1 do plasma rico em plaquetas obtido por sistema fechado em equinos]

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          Abstract

          ABSTRACT Platelet-rich plasma (PRP) has been proposed as an agent to accelerate the healing process and stimulate the regenerative capacity of tissues due to its abundance of growth factors. A large variety of kits and protocols are available to obtain PRP by different cell-separation systems. However, the lack of standardization may lead to inconsistent results. The aim of this study was to characterize cellular composition, platelet parameters using the ADVIA 120 flow cytometer, and TGF-β1 concentration from the PRP product obtained through a closed system, using simple centrifugation. Six clinically healthy horses were used in this study. The protocol in the closed system resulted in approximately 1.6-fold higher platelet and approximately 2.0-fold lower white blood cell concentrations in comparison with whole blood values. The evaluated system was efficient in concentrating platelets and in retrieving a small number of leukocytes, using a protocol of single centrifugation at low speed.

          Translated abstract

          RESUMO O plasma rico em plaquetas (PRP) tem sido proposto como um agente para acelerar o processo de cicatrização e estimular a capacidade regenerativa dos tecidos, devido à sua abundância de fatores de crescimento. Uma grande variedade de kits e de protocolos está disponível para obtenção de PRP, utilizando-se diferentes sistemas de separação de células. No entanto, a falta de padronização pode levar a resultados inconsistentes. O objetivo deste estudo foi caracterizar a composição celular, os parâmetros plaquetários pelo citômetro de fluxo ADVIA 120 e a concentração de TGF-β1 do PRP obtido em sistema fechado, por centrifugação simples. Seis cavalos clinicamente saudáveis foram usados neste estudo. O protocolo no sistema fechado resultou em concentrações de plaquetas aproximadamente 1,6 vez maior e concentrações de leucócitos aproximadamente 2,0 vezes menores em comparação com os valores do sangue total. O sistema avaliado foi eficiente na concentração de plaquetas e na recuperação de um pequeno número de leucócitos, utilizando um protocolo de centrifugação única em baixa velocidade.

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          Most cited references36

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          Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF).

          The topical use of platelet concentrates is recent and its efficiency remains controversial. Several techniques for platelet concentrates are available; however, their applications have been confusing because each method leads to a different product with different biology and potential uses. Here, we present classification of the different platelet concentrates into four categories, depending on their leucocyte and fibrin content: pure platelet-rich plasma (P-PRP), such as cell separator PRP, Vivostat PRF or Anitua's PRGF; leucocyte- and platelet-rich plasma (L-PRP), such as Curasan, Regen, Plateltex, SmartPReP, PCCS, Magellan or GPS PRP; pure plaletet-rich fibrin (P-PRF), such as Fibrinet; and leucocyte- and platelet-rich fibrin (L-PRF), such as Choukroun's PRF. This classification should help to elucidate successes and failures that have occurred so far, as well as providing an objective approach for the further development of these techniques.
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            Platelet-rich plasma: evidence to support its use.

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              Growth factor and catabolic cytokine concentrations are influenced by the cellular composition of platelet-rich plasma.

              Previous studies of bioactive molecules in platelet-rich plasma (PRP) have documented growth factor concentrations that promote tissue healing. However, the effects of leukocytes and inflammatory molecules in PRP have not been defined. The hypothesis for this study was that the concentration of growth factors and catabolic cytokines would be dependent on the cellular composition of PRP. Controlled laboratory study. Platelet-rich plasma was made from 11 human volunteers using 2 commercial systems: Arthrex ACP (Autologous Conditioned Plasma) Double Syringe System (PRP-1), which concentrates platelets and minimizes leukocytes, and Biomet GPS III Mini Platelet Concentrate System (PRP-2), which concentrates both platelets and leukocytes. Transforming growth factor-β1 (TGF-β1), platelet-derived growth factor-AB (PDGF-AB), matrix metalloproteinase-9 (MMP-9), and interleukin-1β (IL-1β) were measured with enzyme-linked immunosorbent assay (ELISA). The PRP-1 system consisted of concentrated platelets (1.99×) and diminished leukocytes (0.13×) compared with blood, while PRP-2 contained concentrated platelets (4.69×) and leukocytes (4.26×) compared with blood. Growth factors were significantly increased in PRP-2 compared with PRP-1 (TGF-β1: PRP-2 = 89 ng/mL, PRP-1 = 20 ng/mL, P < .05; PDGF-AB: PRP-2 = 22 ng/mL, PRP-1 = 6.4 ng/mL, P < .05). The PRP-1 system did not have a higher concentration of PDGF-AB compared with whole blood. Catabolic cytokines were significantly increased in PRP-2 compared with PRP-1 (MMP-9: PRP-2 = 222 ng/mL, PRP-1 = 40 ng/mL, P < .05; IL-1β: PRP-2 = 3.67 pg/mL, PRP-1 = 0.31 pg/mL, P < .05). Significant, positive correlations were found between TGF-β1 and platelets (r(2) = .75, P < .001), PDGF-AB and platelets (r(2) = .60, P < .001), MMP-9 and neutrophils (r(2) = .37, P < .001), IL-1β and neutrophils (r(2) = .73, P < .001), and IL-1β and monocytes (r(2) = .75, P < .001). Growth factor and catabolic cytokine concentrations were influenced by the cellular composition of PRP. Platelets increased anabolic signaling and, in contrast, leukocytes increased catabolic signaling molecules. Platelet-rich plasma products should be analyzed for content of platelets and leukocytes as both can influence the biologic effects of PRP. Depending on the clinical application, preparations of PRP should be considered based on their ability to concentrate platelets and leukocytes with sensitivity to pathologic conditions that will benefit most from increased platelet or reduced leukocyte concentration.
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                Author and article information

                Journal
                abmvz
                Arquivo Brasileiro de Medicina Veterinária e Zootecnia
                Arq. Bras. Med. Vet. Zootec.
                Universidade Federal de Minas Gerais, Escola de Veterinária (Belo Horizonte, MG, Brazil )
                0102-0935
                1678-4162
                February 2022
                : 74
                : 1
                : 93-100
                Affiliations
                [4] Belo Horizonte Minas Gerais orgnameUniversidade Federal de Minas Gerais orgdiv1Escola de Veterinária Brazil
                [2] Cerdanyola del Vallès Cataluña orgnameUniversitat Autónoma de Barcelona orgdiv1Department of Animal Medicine and Surgery Spain
                [5] Brasilia Distrito Federal orgnameMinistério da Educação orgdiv1CAPES Foundation Brazil
                [3] Contagem Minas Gerais orgnameCentro Universitário UNA Brazil
                [1] Bom Despacho Minas Gerais orgnameCentro Universitário UNA Brazil
                Article
                S0102-09352022000100093 S0102-0935(22)07400100093
                10.1590/1678-4162-12257
                d34e271a-3602-4e67-9c37-682480ac1902

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 15 March 2021
                : 28 September 2021
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 36, Pages: 8
                Product

                SciELO Brazil

                Categories
                Veterinary Medicine

                platelet concentrate,growth factors,PRP,protocolo de centrifugação,concentrado plaquetário,fatores de crescimento,concentration protocol

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