Jorge U. Carmona

Autologous platelet concentrates as a treatment for musculoskeletal lesions in five horses

Two horses with acute tendinopathy of a superficial digital flexor tendon (sdft) and three horses with chronic proximal desmitis of the suspensory ligament (pdsl) were treated by injecting autologous concentrates of their platelets into the lesions. The lesions were monitored ultrasonographically and clinically. There were significant ultrasonographic and clinical improvements in the two horses with sdft, but no ultrasonographic improvements in the horses with pdsl; however, they improved clinically and became less lame. All the horses had returned to their pre-injury level of performance by six months after the completion of the treatment, and none of them had suffered a recurrence after 20 months.

Effects of the breed, sex and age on cellular content and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel

BackgroundThere is no information on the effects of the breed, gender and age on the cellular content and growth factor (GF) release from equine pure-platelet rich plasma (P-PRP) and pure-platelet rich gel (P-PRG). The objectives of this study were: 1) to compare the cellular composition of P-PRP with whole blood and platelet poor plasma (PPP); 2) to compare the concentration of transforming GF beta 1 (TGF-β1) and platelet derived GF isoform BB (PDGF-BB) between P-PRP treated with non-ionic detergent (P-PRP+NID), P-PRG (activated with calcium gluconate -CG-), PPP+NID, PPP gel (PPG), and plasma and; 3) to evaluate and to correlate the effect of the breed, gender and age on the cellular and GF concentration for each blood component. Forty adult horses, 20 Argentinean Creole Horses (ACH) and, 20 Colombian Creole Horses (CCH) were included. Data were analyzed by parametric (i.e.: t-test, one way ANOVA) and non parametric (Kruskal-Wallis test, Wilcoxon test) tests. Correlation analysis was also performed by using the Spearman and Pearson tests. A p ≤ 0.05 was set as significant for all tests. All the blood components were compared for platelet (PLT), leukocyte (WBC), TGF-β1 and PDGF-BB concentrations. The effect of the breed, gender and age on these variables was analyzed. A P ≤ 0.05 was accepted as significant for all the tests.ResultsPLT counts were 1.8 and 0.6 times higher in P-PRP than in whole blood and PPP, respectively; WBC counts were 0.5 and 0.1 times lower in P-PRP, in comparison with whole blood and PPP, respectively. TGF-β1 and PDGF-BB concentrations were 2.3 and 262 times higher, respectively, in P-PRG than in plasma, and 0.59 and 0.48 times higher, respectively, in P-PRG than in PPG. P-PRG derived from CCH females or young horses presented significantly (P < 0.001) higher PDGF-BB concentrations than P-PRG derived from ACH males or older horses.ConclusionsOur results indicated that P-PRP obtained by a manual method was affected by intrinsic factors such as the breed, gender and age. Equine practitioners should be aware that cellular and GF release from P-PRP/P-PRG could change according with the intrinsic variables associated with a patient in particular.

Intra-articular injections of autologous platelet concentrates in dogs with surgical reparation of cranial cruciate ligament rupture

OBJECTIVES The objective of this study was to evaluate by clinical, radiographic, and force plate gait analyses the effect of post-surgical intra-articular injections of autologous platelet concentrates (PC) in a small group of dogs with cranial cruciate ligament (CCL) rupture. METHODS The ten dogs used in this study were initially presented with CCL rupture and underwent ligament replacement surgery by fascia lata autograft guided by arthroscopy. Six dogs received three intra-articular injections of PC (PC group); one dose was injected immediately after surgery, and two additional doses were injected at two-week intervals. The remaining four dogs received only nutraceuticals (control group). All dogs were evaluated by clinical examination, serial radiography, and force plate gait analyses at monthly intervals up to 90 days. RESULTS The clinical follow-up of the PC-treated group indicated a better outcome than the control group. Radiographic evaluation was not conclusive. Values of peak vertical reaction force and vertical impulse of the affected limbs were only significantly larger on the 90th postoperative day in the PC group compared to the control group. CLINICAL SIGNIFICANCE Our results indicate that autologous PC might improve functional outcome after intra-articular cranial cruciate ligament repair. The effect of PC when using other repair procedures warrants additional studies.

In vitro bactericidal activity of equine platelet concentrates, platelet poor plasma, and plasma against methicillin-resistant Staphylococcus aureus

Los objetivos del estudio fueron: 1) evaluar el efecto antibacteriano de concentrados de plaquetas equinas (ePC) (activados o no con gluconato de calcio (CG)) frente a Staphylococcus aureus meticilino-resistente (MRSA) y 2) comparar su efecto antibacteriano contra plasma pobre en plaquetas (PPP) (activado con CG -PPP/GC-) y plasma (P). Los productos sanguineos fueron divididos en 4 grupos (ePC, ePC/CG, PPP/CG y P), mas un grupo control positivo (PCG) y otro control negativo. Los grupos se mezclaron con caldo Mueller-Hinton y MRSA. Las muestras fueron incubadas durante 1, 4, 8, 12 y 24 horas y se contaron las unidades formadoras de colonias. El crecimiento de las bacterias fue significativamente (P = 0,01) inhibido por el ePC, ePC/CG, PPP/CG y P en comparacion con el PCG durante las primeras 12 h. Solo a las 24 horas hubo una diferencia estadisticamente significativa (P = 0.01) y se observo un efecto antibacteriano para el ePC, ePC/CG y PPP/CG en comparacion con el PCG y P. Los ePCs y PPP equinos mostraron el mejor efecto antibacteriano in vitro contra el MRSA.

Evaluación del método del tubo para concentrar plaquetas felinas: estudio celular

SUMMARY The aim of this study was to evaluate a manual method for concentrating canine platelets and consequently produce autologous platelet concentrates (APCs) for clinical and experimental proposals. The APCs were obtained by venous blood collection in tubes with ACD solution and were spun at 191 g for 6 minutes. The cell counts of whole blood and APCs, were statistically different (P < 0.01) between the values for platelets, leukocytes, granulocytes, monocytes, but not for lymphocytes. A strong negative correlation (ρ = -0.702 P = 0.011) between the number of lymphocytes and platelets in the APCs was found. The collection efficiency of platelets was 29.9% and the concentration of platelets was 49.4% higher in APCs when compared to the whole blood samples. The results indicated that this simple centrifugation method allows concentrating canine platelets.

Uso de concentrados autólogos de plaquetas como terapia regenerativa de enfermedades crónicas del aparato musculoesquelético equino

SUMMARY Platelets are of pivotal importance for wound healing since they release growth factors that in turns produce chemotaxis, both cellular proliferation and differentiation, angiogenesis, and extracellular matrix deposition. The use of autologous platelet concentrates (APCs) has been proposed for accelerating wound healing, decreasing inflammation, to stimulate the regenerative capability of the injured tissues, to decrease the fibroblastic activity and to avoid the production of non functional scarring tissue. APCs could be obtained by several methods. Each method produces APCs of different both cellular and molecular quality. Recently, some basic and clinical information that justifies the usage of APCs for the treatment of degenerative musculoskeletal diseases in horses, such as osteoarthritis, tendinopathies and desmopathies has been published.

Niveles de factor de crecimiento transformante beta-3 y óxido nítrico en cuatro concentrados autólogos de plaquetas y plasma derivados de sangre equina

El objetivo de este estudio fue evaluar el metodo de centrifugado simple y doble en tubo para concentrar plaquetas, leucocitos, factor de crecimiento transformante beta-3 (TGF-s3) y oxido nitrico (NO). Se recogieron muestras de sangre entera de caballos clinicamente normales, las cuales fueron procesadas por medio del metodo de centrifugado simple y doble en tubo para obtener 4 concentrados de plaquetas (PCs): PC-A, PC-B, PC-C y PC-D. Los PCs y la sangre entera fueron analizados para hemograma mediante un sistema hematologico de citometria de flujo. Las concentraciones de TGF-s3 fueron analizadas mediante ELISA y el NO mediante la reaccion de Greiss. Las concentraciones de plaquetas y leucocitos fueron estadisticamente diferentes (P < 0,05) para cada PC y sangre entera. Los niveles de TGF-s3 y NO fueron similares para cada PC y sangre entera. En conclusion, el metodo de centrifugado simple y doble en tubo es un procedimiento de confianza para concentrar plaquetas en el caballo. Sin embargo, los niveles de TGF-s3 y NO no dependen del numero de plaquetas o de leucocitos concentrados con esta tecnica.

Effects of sodium citrate and acid citrate dextrose solutions on cell counts and growth factor release from equine pure-platelet rich plasma and pure-platelet rich gel

BackgroundThere is a lack information on the effects of the most commonly used anticoagulants for equine platelet rich plasmas (PRPs) elaboration on cell counts and growth factor release from platelet rich gels (PRGs). The aims of this study were 1) to compare the effects of the anticoagulants sodium citrate (SC), acid citrate dextrose solution A (ACD-A) and ACD-B on platelet (PLT), leukocyte (WBC) and on some parameters associated to platelet activation including mean platelet volume (MPV) and platelet distribution width (PDW) between whole blood, pure PRP (P-PRP) and platelet-poor plasma (PPP); 2) to compare transforming growth factor beta 1 (TGF-β1) and platelet-derived growth factor isoform BB (PDGF-BB) concentrations in supernatants from pure PRG (P-PRG), platelet-poor gel (PPG), P-PRP lysate (positive control) and plasma (negative control); 3) to establish the possible correlations between all the studied cellular and molecular parameters.ResultsIn all cases the three anticoagulants produced P-PRPs with significantly higher PLT counts compared with whole blood and PPP. The concentrations of WBCs were similar between P-PRP and whole blood, but significantly lower in PPP. The type of anticoagulant did not significantly affect the cell counts for each blood component. The anticoagulants also did not affect the MPV and PDW parameters. Independently of the anticoagulant used, all blood components presented significantly different concentrations of PDGF-BB and TGF-β1. The highest growth factor (GF) concentrations were observed from P-PRP lysates, followed by PRG supernatants, PPP lysates, PPG supernatants and plasma. Significant correlations were observed between PLT and WBC counts (ρ = 0.80), PLT count and TGF-β1 concentration (ρ = 0.85), PLT count and PDGF-BB concentration (ρ = 0.80) and PDGF-BB and TGF-β1 concentrations (ρ = 0.75). The type of anticoagulant was not correlated with any of the variables evaluated.ConclusionsThe anticoagulants did not significantly influence cell counts or GF concentrations in equine PRP. However, ACD-B was apparently the worst anticoagulant evaluated. It is necessary to perform additional research to determine the effect of anticoagulants on the kinetics of GF elution from P-PRG.

Concentrados autólogos de plaquetas como tratamiento de lesiones de tejidos blandos del aparato locomotor en caballos

The efficacy of an autologous platelet concentrate (APC) in 5 horses with soft tissue musculoskeletal injuries namely: superficial digital flexor tendon (SDF...

Evaluation of the anti-inflammatory effects of two platelet-rich gel supernatants in an in vitro system of cartilage inflammation

OBJECTIVES To study, in normal cartilage explants (CEs) challenged with lipopolysaccharide (LPS), the temporal effects (at 48 and 96h) of leukocyte- and platelet-rich gel (L-PRG) and pure platelet-rich gel (P-PRG) supernatants on the production and degradation of platelet-associated growth factors (GFs) (platelet-derived GF isoform BB [PDGF-BB] and transforming growth factor beta-1 [TGF-β1]), pro-inflammatory (tumour necrosis factor alpha [TNF-α]) and anti-inflammatory cytokines (interleukin 4 [IL-4] and IL-1 receptor antagonist [IL-1ra]). METHODS CEs from six horses were challenged with LPS and cultured for 96h with L-PRG and P-PRG supernatants at concentrations of 25% and 50%, respectively. The CE culture medium was changed every 48h and used for determination, by ELISA, of PDGF-BB, TGF-β1, TNF-α, IL-4 and IL-1ra. RESULTS Both the 25% and 50% PRG supernatants produced a different molecular profile in the culture media, unlike that of the CE challenged with LPS only. 50% L-PRG produced the most sustained release of growth factors and anti-inflammatory cytokines, although it produced the highest TNF-α release. PDGF-BB was significantly correlated with IL-1ra and TNF-α concentrations, whereas TNF-α was correlated with IL-4. CONCLUSIONS 50% L-PRG supernatant produced a more sustained concentration of growth factors and anti-inflammatory cytokines than the other hemoderivatives evaluated. This substance could be evaluated in animal models of arthritis or in patients with arthropathies.