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

Abstract: 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

Osseointegração clínica-radiológica do compósito hidroxiapatita-lignina entre implante metálico e tecido ósseo em coelho Clinical-radiographic osseointegration of hydroxyapatite-lignin composite between metallic implant and bone tissue in rabbit

Para avaliar o compósito hidroxiapatita-lignina na osseointegração entre implante metálico e o tecido ósseo, foram utilizados 20 coelhos adultos, nos quais foi realizada uma falha óssea na face lateral proximal de ambas as tíbias. Na tíbia esquerda, introduziu-se, no canal medular, um pino intramedular de Schanz revestido com o compósito em sua parte rosqueada, após o preenchimento daquele com 1000mg do compósito. A falha cortical foi preenchida com o compósito. O mesmo foi feito na tíbia direita, porém sem a utilização do compósito, servindo como controle. A avaliação clínica baseou-se na deiscência, claudicação, sensibilidade dolorosa e circunferência tibial. Foram realizadas radiografias imediatamente após a cirurgia e aos oito, 30, 60, 90 e 120 dias do pós-operatório. A maioria dos animais apresentou evolução clínica normal. Nas radiografias do grupo tratado, houve decréscimo da radiopacidade no defeito e no espaço medular até tornar-se semelhante à do osso circunvizinho, quadro inverso ao do grupo-controle. Conclui-se que o compósito hidroxiapatita-lignina não mostrou indícios clínicos de rejeição e que o tecido visualizado na altura da falha óssea e ao redor do pino intramedular tinha radiopacidade semelhante à do osso circunvizinho, o que sugere que o material promoveu a integração com o tecido ósseo.This study evaluated a sinthetic hydroxyapatite-lignin composite for osseoingration between metallic implant and bone tissue. Twenty New Zealand rabbits were used. A defect was made a the proximal region of lateral surfaces of both tibias. An intramedullary pin was inserted in the medullary cavity of the left tibia after filling the cavity with 1,000mg of the composite and covering the thread part of the pin with the composite. The same procedure was made in control tibias without the composite. Clinical evaluation was based on inflammatory reaction, dehiscence, lameness, pain, and tibial circumference. Radiographs were made immediately after surgery and then on days eight, 30, 60, 90, and 120. Most of the animals presented normal clinical progression. Radiographies of treated group showed decrease in the defect and medullary cavity radiopacity to the same pattern of the surrounding bone, while the contrary happened in control group; but, in this group, the medullary cavity remained radioluscent at the last observation date. It can be concluded that the hydroxyapatite-lignin composite did not show clinical signs of rejection and that radiopacity of the tissue in the bone defect and surrounding the pin was similar to bone radiopacity, suggesting that the material promoted osseointegration