Abstract
BackgroundThe clinical use of autologous platelet concentrates (also known as platelet-rich plasma) on the field of regenerative therapy, in the last decade has been the subject of several studies especially in equine medicine and surgery. The objectives of this study was: 1) to describe and compare the cellular population in whole blood, lower fraction (A) and upper fraction (B) of platelet concentrates, 2) to measure and compare the transforming growth factor beta 1 (TGF-β1) concentration in plasma and both platelet concentrates after be activated with calcium gluconate or batroxobin plus calcium gluconate and, 3) to determine correlations between cell counts in platelet concentrates and concentrations of TGF-β1. Blood samples were taken from 16 dogs for complete blood count, plasma collection and platelet concentrates preparation. The platelet concentrates (PC) were arbitrarily divided into two fractions, specifically, PC-A (lower fraction) and PC-B (upper fraction). The Platelet concentrates were analyzed by hemogram. After activated with calcium gluconate or batroxobin plus calcium gluconate, TGF-β1 concentration was determined in supernatants of platelet concentrates and plasma.ResultsThere were differences statistically significant (P < 0.05) for the platelet count and leukocyte count and TGF-β1 concentration between whole blood, plasma and both platelet concentrates. A significant correlation was found between the number of platelets in both platelet concentrates and TGF-β1 concentration. Platelet collection efficiency was 46.34% and 28.16% for PC-A and PC-B, respectively. TGF-β1 concentration efficiency for PC activated with calcium gluconate was 47.75% and 31.77%, for PC-A and PC-B, respectively. PC activated with batroxobin plus CG showed 46.87% and 32.24% for PC-A and PC-B, respectively.ConclusionsThe methodology used in this study allows the concentration of a number of platelets and TGF-β1 that might be acceptable for a biological effect for clinical or experimental use as a regenerative therapy in dogs.
Highlights
The clinical use of autologous platelet concentrates on the field of regenerative therapy, in the last decade has been the subject of several studies especially in equine medicine and surgery
This situation is different for platelet concentrates (PC) derived from equine blood [24]. Both PC obtained in this study permitted concentrate two (PC-B) and three (PC-A) fold the concentration of TGF-β1 respect to the basal concentration of this protein in plasma. These findings suggest that plasma platelets were scarcely activated at the moment of blood extraction, since the MPV remained lower in whole blood in comparison with the same parameter in PC arbitrarily classified as lower fraction (PC-A)
This protein was measured in the present study with a specific canine antibody for TGF-β1, whereas the studies mentioned [29,30] used a human TGF-β1 antibody. This is only an assumption and further studies are necessary to validate the actual utility of human or canine ELISA kits for canine TGF-β1 measurement. Plasma and both PC of this study presented higher concentrations of TGF-β1 in comparison with the results of the same protein from autologous conditioned plasma (ACP) and plasma obtained with ACD-A from blood of dogs [31]
Summary
The clinical use of autologous platelet concentrates ( known as platelet-rich plasma) on the field of regenerative therapy, in the last decade has been the subject of several studies especially in equine medicine and surgery. The objectives of this study was: 1) to describe and compare the cellular population in whole blood, lower fraction (A) and upper fraction (B) of platelet concentrates, 2) to measure and compare the transforming growth factor beta 1 (TGF-β1) concentration in plasma and both platelet concentrates after be activated with calcium gluconate or batroxobin plus calcium gluconate and, 3) to determine correlations between cell counts in platelet concentrates and concentrations of TGF-β1. Platelet alpha granules [4,6] contain at least seven GF directly involved in the healing process Of these proteins, transforming growth factor beta isoform 1 (TGF-β1) is of pivotal importance for its actions on cell proliferation, angiogenesis and extracellular matrix deposition [7]. Combination of both includes batroxobin plus calcium gluconate [10]
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