Department of Veterinary Surgery and Animal Reproduction, School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.
Material Science and Engineering Laboratory - IBTEC, São Paulo State University (UNESP), Botucatu, Brazil.
J Vet Dent. 2023 Sep;40(3):212-219. doi: 10.1177/08987564231152594. Epub 2023 Feb 1.
This study aimed to perform histological, immunohistochemical, biomechanical, and wettability assessments of leukocyte- and platelet-rich fibrin (L-PRF) membranes obtained from the blood of healthy dogs. Ten client-owned Labrador Retriever dogs were enrolled. Blood samples were obtained from the external jugular vein using a vacuum tube without anticoagulant, which was immediately centrifuged at 400 for 12 min in a dedicated centrifuge. The L-PRF clot was removed from the tube, and the red clot was released from the buffy coat using a spatula. The membrane was produced using a PRF box. Histological examination identified the three portions of the L-PRF membranes. The first portion was composed mainly of red blood cells with the presence of a low number of leukocytes among them. The second portion was composed of white blood cells, mainly neutrophils. The third portion was composed of the fibrin network which was characterized by acidophilic staining. The immunohistochemical analysis showed that vascular endothelial growth factor and platelet-derived growth factor were expressed in all samples at different intensities, both in cellular components and fibrin mesh. The tensile test and wettability assessments were measured in membranes 30 min and 3 h after production. The 30 min L-PRF membranes supported twice the ultimate tensile strength compared to 3 h L-PRF membranes. The wettability of the 30 min sample membranes was statistically higher than the 3 h sample membranes. In conclusion, the centrifugation protocol allowed production of the L-PRF membrane using canine blood and this was confirmed by histological and immunohistochemical analysis. The mechanical resistance and wettability of the L-PRF membrane were significantly reduced over time.
本研究旨在对从健康犬血液中获得的白细胞和血小板丰富纤维蛋白（L-PRF）膜进行组织学、免疫组织化学、生物力学和润湿性评估。招募了 10 只客户拥有的拉布拉多猎犬。使用无抗凝剂的真空采血管从颈外静脉采集血液，立即在专用离心机中以 400 转/分离心 12 分钟。从管中取出 L-PRF 凝块，并用刮刀从白细胞层中释放出红色凝块。使用 PRF 盒生产膜。组织学检查确定了 L-PRF 膜的三个部分。第一部分主要由红细胞组成，其中有少量白细胞。第二部分由白细胞组成，主要是中性粒细胞。第三部分由纤维蛋白网络组成，其特征是嗜酸性染色。免疫组织化学分析显示，血管内皮生长因子和血小板衍生生长因子在所有样本中均以不同强度表达，细胞成分和纤维蛋白网均有表达。在生产后 30 分钟和 3 小时测量拉伸试验和润湿性评估。30 分钟的 L-PRF 膜的极限拉伸强度是 3 小时 L-PRF 膜的两倍。30 分钟样本膜的润湿性明显高于 3 小时样本膜。总之，离心方案允许使用犬血生产 L-PRF 膜，这通过组织学和免疫组织化学分析得到了证实。随着时间的推移，L-PRF 膜的机械阻力和润湿性显著降低。
