In House Double Centrifugation Method for Preparation of Homologous Platelet-Rich Plasma (Prp) in Dogs

The production of inexpensive autologous canine platelet rich plasma (PRP) with no requirement for specific equipment may provide a relatively easy source of growth factors and cell-rich fractions for stimulating healing processes and regeneration. In dogs many different manual or semi-automated methods for PRP production are described achieving wide ranges of platelet concentration, but few studies provide information about platelet activation and platelet-derived-growth-factor (PDGF) concentration. The aim of this study was to evaluate the in-house manual double centrifugation method that would produce small amounts of autologous canine PRP from a relatively small volume of whole blood. Thus making it a practical technique for autologous PRP production in the clinic. A double centrifugation protocol was followed for all samples. Platelet counts (PLT/μL) leucocyte counts (WBC/μL) and PCV (packed cell volume), were calculated on the WB and PRP Blood smears were performed from all whole blood and PRP samples to evaluate the percentage of activated platelets. PDGF were measured on PRP samples by ELISA. The mean platelet concentration was 906900 platelets/μL ± 166259 in a mean of 1.01 ml PRP obtained from 8 ml of whole blood. There was a significant difference (p < 0.0001) between the percentage of activated platelets in WB (22.9 ± 14.2) and in PRP (39.05 ± 19.4). The mean concentration of PDGF in PRP was 1562.23 ± 684.72 ng/L. Poor correlation was found between PDGF concentration and activated platelet percentages in PRP. This method appears to produce adequate concentration of platelets and PDGF and may be appropriate for clinical use.