Method to obtain platelet-rich plasma from rabbits (Oryctolagus cuniculus )

Abstract

RESUMO.-[Método de obtenção de plasma rico em plaquetas de coelhos (Oryctolagus cuniculus).] O plasma rico em plaquetas (PRP) é um produto de fácil obtenção a baixo custo, destacando-se pelos seus fatores de crescimento na reparação tecidual. Para obtenção do PRP, a centrifugação do sangue total é realizada com tempos e forças gravitacionais específicas. Assim, o presente trabalho teve por objetivo estudar o método da dupla centrifugação para obtenção do PRP, a fim de avaliar a eficácia de aumento daMethod to obtain platelet-rich plasma from rabbits (Oryctolagus cuniculus)¹ Platelet-rich plasma (PRP) is a product easy and inxpesnsive, and stands out to for its growth factors in tissue repair. To obtain PRP, centrifugation of whole blood is made with specific time and gravitational forces. Thus, the present work aimed to study a method of double centrifugation to obtain PRP in order to evaluate the effective increase of platelet concentration in the final product, the preparation of PRP gel, and to optimize preparation time of the final sample. Fifteen female White New Zealand rabbits underwent blood sampling for the preparation of PRP. Samples were separated in two sterile tubes containing sodium citrate. Tubes were submitted to the double centrifugation protocol, with lid closed and 1600 revolutions per minute (rpm) for 10 minutes, resulting in the separation of red blood cells, plasma with platelets and leucocytes. After were opened and plasma was pipetted and transferred into another sterile tube. Plasma was centrifuged again at 2000rpm for 10 minutes; as a result it was split into two parts: on the top, consisting of platelet-poor plasma (PPP) and at the bottom of the platelet button. Part of the PPP was discarded so that only 1ml remained in the tube along with the platelet button. This material was gently agitated to promote platelets resuspension and activated when added 0.3ml of calcium gluconate, resulting in PRP gel. Double centrifugation protocol was able to make platelet concentration 3 times higher in relation to the initial blood sample. The volume of calcium gluconate used for platelet activation was 0.3ml, and was sufficient to coagulate the sample. Coagulation time ranged from 8 to 20 minutes, with an average of 17.6 minutes. Therefore, time of blood centrifugation until to obtain PRP gel took only 40 minutes. It was concluded that PRP was successfully obtained by double centrifugation protocol, which is able to increase the platelet concentration in the sample compared with whole blood, allowing its use in surgical procedures. Furthermore, the preparation time is appropriate to obtain PRP in just 40 minutes, and calcium gluconate is able to promote the activation of platelets.INDEX TERMS: Platelet-rich plasma, platelet concentration, revascularization, rabbits.

Effect of Platelet Rich Plasma (PRP) Injection on the Endocrine Pancreas of the Experimentally Induced Diabetes in Male Albino Rats: A Histological and Immunohistochemical Study

“…In this study, the PRP preparation depended on double centrifugation method that resulted in a platelet concentration 3 times higher than the initial blood sample [17]. It was also depended on the endogenous activation of platelets to allow the platelets to make their action spontaneously.…”

“…The PRP preparation was carried out by adapting the protocol of the double centrifugation tube method [17]. In brief, rats were anesthetized with ether, 2 ml of blood was collected under aseptic technique from the retro-orbital plexus using capillary tube initially dipped in 3.2% sodium citrate, then collected into tubes containing 0.3 mL of the anticoagulant.…”

Biochemical and Molecular Mechanisms of Platelet-Rich Plasma in Ameliorating Liver Fibrosis Induced by Dimethylnitrosurea

“…Preparation of platelet-rich plasma PRP was prepared using the double centrifugation tube method [10]. Ten male Wistar rats were used as blood donors and anesthetized with ether, and 2 mL blood was collected using aseptic techniques from the retro-orbital plexus into tubes containing 0.3 mL (3.2% sodium citrate) anticoagulant.…”

Surgical sponge associated with platelets rich plasma in skin mesh grafts and layer in rabbits (Oryctolagus cuniculus)

“…The preparation of platelet-rich plasma was conducted as recommended by the literature (Pazzini et al, 2016b). The surgical procedure for reconstruction using graft was performed in a single moment, replacing the granulation tissue by platelet-rich plasma.…”