Evaluation of platelet indices and complete blood count in canine mammary tumors

Malignant mammary tumors in humans and bitches cause hematological disorders such as anemia, erythrocytosis, thrombocytosis, hyperproteinemia, and leucopenia. Novel studies have been conducted on the predictive and prognostic values of platelet (PLT) indices in human breast cancer (HBC). However, there is little information about the alterations in hematological parameters in canine mammary tumors (CMTs). The aims of this study were to evaluate the platelet indices and complete blood count (CBC) parameters in bitches with and without mammary tumor and to assess the above mentioned parameters with regard to histological tumor types and grades. A total of 71 bitches were enrolled in this study. The bitches in the study group were divided into 2 groups which consisted of malignant epithelial mammary tumors (group EMT; n = 43) and malignant mixed mammary tumors (group MMT; n = 12). Control group (group C) consisted of clinically and gynaecologically healthy 16 bitches. Blood samples were obtained to perform the CBC and PLT indices analysis. Histopathological examinations were carried out under a light microscope. Histological tumor types and malignancy grades were classified. The bitches with mammary tumor showed significantly increased PLT values and decreased hematocrit (HCT), hemoglobin (HGB) and mean corpuscular hemoglobin (MCH) values versus the healthy ones, regardless of the tumor type. However, in comparisons with the group C, mean platelet volume (MPV) and mean corpuscular hemoglobin concentration (MCHC) values were different only in the group MMT, while plateletcrit (PCT) and mean corpuscular volume (MCV) values were different only in the group EMT. Also white blood cell (WBC), PLT and PCT values were higher than the referenced laboratory ranges in grade 3 tumors. In the presented study, MPV was considerably correlated with PLT, platelet distribution width (PDW) and PCT. Also, PCT and PLT had high sensitivity and specificity to distinct EMT and MMT from the healthy bitches. Microcytic and hypochromic anemia occurs due to the decrease in the amount of HGB. Levels of MCV, MCH, and MCHC in the HBC group were reported to be significantly lower than in humans without breast cancer. Although anemia did not occur in EMT and MMT groups, obtained significances in the HCT, HGB, MCV, MCH, and MCHC levels between the bitches with and without mammary tumor were in line with the previous reports. In this study, WBC levels in grade 3 tumors were significantly higher than grade1 tumors (P < 0.05). Whereas levels of WBC in grade 1 and grade 2 tumors were in referenced laboratory ranges, it was higher in grade 3. Increased level of WBC in grade 3 was supposed to be due to the rise in malignancy as previously reported. Thrombocytosis was detected in 48.83% and 41.66% of the bitches in EMT and MMT groups, respectively. The higher percentage of CMTs with thrombocytosis in this study might be due to the difference in referenced upper limit of PLT in previous studies. The elapsed time between tumor formation and clinical presentation could be another influencing factor. Although PLT and PCT values were not significant according to the histological grading in this study, both parameters were found to be higher in grade 3 than the normal reference values. Further studies conducted with higher populations may lead the differences in these parameters to significance. With the support of further studies, alterations in the above mentioned parameters in bitches may contribute in the diagnosis process, management of treatment and may constitute an easy way to have an idea about the prognosis of mammary tumors.(AU)

Protocols for preparation of platelet rich plasma (PRP) in Quarter Horses / Protocolos para o preparo de plasma rico em plaquetas (PRP) em cavalos Quarto de Milha

This study compared two protocols for preparation of platelet rich plasma (PRP) and evaluated the association between manual and automated methods for platelet count using a prospective study design. Eight clinically healthy Quarter Horses had venous blood samples collected at rest. After collection, blood samples were centrifuged twice, using two different protocols including a period of sample resting, either at the start or at the end of the protocol. Platelet counting at the start of the protocol, during, and after obtaining PRP was conducted manually or with an automated counter, followed by comparison of the two methods. In order to investigate platelet degranulation during the protocol, vascular endothelial growth factor (VEGF) was measured at each preparation stage. The protocol with sample resting before centrifugation yielded a more concentrated PRP, and the study verified that both manual and automated methods are comparable and can be used interchangeably for platelet counting. VEGF concentration did not differ significantly between protocols, or among protocol stages. The results indicate that choice of protocol for PRP preparation will affect the quantity of platelets in the final product, although platelet degranulation was not observed as evidenced by the stable VEGF concentrations measured. A larger yield of non-degranulated platelets in PRP is desirable since more α-granules will be present, therefore Protocol II is recommended. Both manual and automated counts reliably allow clinicians to obtain platelet counts and the choice of utilizing a manual or automated method is unlikely to interfere with evaluation of the final PRP product.(AU)

Este estudo comparou dois protocolos de preparo de plasma rico em plaquetas (PRP) e avaliou a associação entre dois métodos de contagem plaquetária - um manual e o outro automático através de um estudo prospectivo. Sangue venoso de oito equinos da raça Quarto de Milha foi coletado e em seguida foi centrifugado duas vezes utilizando-se dois protocolos distintos: um com descanso antes da primeira centrifugação e outro após a segunda centrifugação. A contagem plaquetária ao início, no meio e ao final dos protocolos foi realizada manualmente e pelo método automatizado, seguida de comparação entre os dois métodos. Para investigar a degranulação plaquetária ocorrida durante o preparo do PRP, o fator de crescimento vascular endotelial (VEGF) foi mensurado em cada estágio dos protocolos. O método utilizando o descanso da amostra antes da primeira centrifugação proporcionou a obtenção de um PRP mais concentrado, além de o estudo verificar que ambos os métodos de contagem plaquetária (manual e automatizado) são comparáveis e podem ser usados indiferentemente. A concentração de VEGF não foi significativamente diferente entre os estágios de preparo do PRP. Os resultados indicam que o método de preparo afeta a quantidade de plaquetas obtidas no PRP, apesar da degranulação plaquetária não ter sido observada, como evidenciado pela concentração estável de VEGF. Uma maior concentração de plaquetas no PRP é desejável, pois indica que um maior número de α-grânulos estará presente na amostra, portanto, conclui-se que o Protocolo II é mais recomendável. Tanto o método manual, quanto o automatizado, pode ser usado de maneira confiável para a contagem plaquetária, não interferindo com a avaliação do produto final (PRP).(AU)

Criopreservação do plasma rico em plaquetas de equinos / Equine platelet-rich plasma cryopreservation

Avaliou-se o congelamento do plasma rico em plaquetas (PRP) de equinos, a -196ºC em nitrogênio líquido, utilizando-se como crioprotetor o DMSO em duas concentrações (3% e 6%), e, como ponto final, a avaliação da morfologia e da agregometria plaquetária. Foram utilizadas 12 amostras de PRP em duas repetições. Previamente ao congelamento, as amostras foram submetidas a um resfriamento lento (-0,07ºC/minuto) até a temperatura final de 4-5ºC. A criopreservação do PRP equino, incluindo um resfriamento lento a 4-5ºC, previamente ao congelamento a -197ºC em nitrogênio líquido, foi similar para as concentrações do crioprotetor DMSO a 3% ou 6%, quando avaliado o percentual de ativação e de agregação plaquetária.

Equine platelet-rich plasma (PRP) frozen at -196°C in liquid nitrogen using DMSO as a cryoprotectant in two different concentrations (3% and 6%) was evaluated, using platelet morphology and aggregometry as the final parameters. Twelve PRP samples were used in two repetitions. The samples were submitted to slow cooling prior to frozen (-0.07°C/minute) until they reached the temperature of 4-5°C. Platelet cryopreserved in 3% or 6% DMSO, presented similar efficacy when the percentage of activation and platelet aggregation was evaluated.