Caterino Chiara, Della Valle Giovanni, Aragosa Federica, De Biase Davide, Ferrara Gianmarco, Lamagna Francesco, Fatone Gerardo
Department of Veterinary Medicine and Animal Production, University of Naples "Federico II", Naples, Italy.
Department of Pharmacy/DIFARMA, University of Salerno, Fisciano, Italy.
Front Vet Sci. 2022 Apr 15;9:861255. doi: 10.3389/fvets.2022.861255. eCollection 2022.
Leukocyte-Platelet-Rich Fibrin (L-PRF) is a second generation of platelet concentrates; it was widely used, as an autologous platelet-based wound sealant and hemostatic agent in surgical wound healing. L-PRF clot or membrane is a solid fibrin-based biomaterial, with a specific 3D distribution of the leukocytes and platelet aggregates. This biological scaffold releases growth factors (i.e., TGF- β1, PDGF-AB, VEGF) and matrix proteins (fibronectin, vitronectin and thrombospondin-1) during the healing process after the application. To the Authors' knowledge both in human and veterinary medicine a single standardised protocol was not reported. This prospective study aimed to apply Crisci's L-PRF protocol (which is characterised by 30" of acceleration, 2' at 2,700 rpm, 4' at 2,400 rpm, 3' at 3,000 rpm, and 36" of deceleration and arrest) sin canine species, evaluate macroscopically and histologically the L-PRF membranes obtained by using Wound Box to standardise the L-PRF protocol in dogs and to evaluate the clinical feasibility of using L-PRF membranes by quantitative analysis of growth factors over 7 days. One hundred twenty-eight dogs in good general condition with no history of recent NSAIDs intake (15 days of washout) and/or any medication or disease related to coagulation process met inclusion criteria and therefore were enrolled. We obtained 172 membrane L-PRF membranes by 86 dogs: half of them underwent macroscopic and histological analysis, the other 86 underwent ELISA analysis. The Wound Box gave a membrane of mean (±SD) length (cm), width (cm) and weight (g) of 1.97 (±0.89), 0.95 (±0.36), 0.46 (±0.20) respectively. Histology analysis confirmed a well-defined histoarchitecture with five layers reproducing density and distribution of blood cells in this biomaterial. Finally, the ELISA assay performed with 22 L-PRF membranes showed a peak in growth factors at 6 h after membrane production, followed by a decrease in release at 24 and 72 h and a second peak in release at 168 h after production. Statistical analysis of demographic variables (age, sex, and body condition score BCS) and the average of growth factors determined by the ELISA assay did not reveal statistical significance, except for the BCS factor compared with the production of VEGF. Our data confirm the effectiveness of this protocol and of Wound Box to produce L-PRF membranes in dogs.
富白细胞血小板纤维蛋白（L-PRF）是第二代血小板浓缩物；它作为一种基于自体血小板的伤口密封剂和止血剂，在外科伤口愈合中被广泛应用。L-PRF凝块或膜是一种基于纤维蛋白的固体生物材料，白细胞和血小板聚集体具有特定的三维分布。这种生物支架在应用后的愈合过程中释放生长因子（即转化生长因子-β1、血小板衍生生长因子-AB、血管内皮生长因子）和基质蛋白（纤连蛋白、玻连蛋白和血小板反应蛋白-1）。据作者所知，在人类和兽医学中均未报道单一的标准化方案。这项前瞻性研究旨在将克里斯奇的L-PRF方案（其特点是加速30秒、2700转/分钟持续2分钟、2400转/分钟持续4分钟、3000转/分钟持续3分钟、减速和停止36秒）应用于犬类，通过宏观和组织学评估使用伤口盒获得的L-PRF膜，以标准化犬类的L-PRF方案，并通过对生长因子进行7天的定量分析来评估使用L-PRF膜的临床可行性。128只一般状况良好、近期无非甾体抗炎药摄入史（洗脱15天）和/或无任何与凝血过程相关的药物或疾病的犬符合纳入标准，因此被纳入研究。我们从86只犬身上获得了172片L-PRF膜：其中一半进行宏观和组织学分析，另外86片进行酶联免疫吸附测定（ELISA）分析。伤口盒制备的膜的平均（±标准差）长度（厘米）、宽度（厘米）和重量（克）分别为1.97（±0.89）、0.95（±0.36）、0.46（±0.20）。组织学分析证实了其具有明确的组织结构，有五层结构，再现了这种生物材料中血细胞的密度和分布。最后，对22片L-PRF膜进行的ELISA检测显示，生长因子在膜制备后6小时达到峰值，随后在24小时和72小时释放量下降，在制备后168小时释放量再次达到峰值。除了身体状况评分（BCS）因子与血管内皮生长因子（VEGF）的产生相关外，对人口统计学变量（年龄、性别和身体状况评分BCS）以及ELISA检测确定的生长因子平均值进行的统计分析未显示出统计学意义。我们的数据证实了该方案和伤口盒在犬类中制备L-PRF膜的有效性。
