Post-graduate program in Public Health, Department of Biophysics and Pharmacology, Federal University of Rio Grande Norte (UFRN), Natal, RN, 59072-970, Brazil.
Center for MicroElectroMechanical Systems (CMEMS-UMINHO), University of Minho, Guimarães, 4800-058, Portugal.
Clin Oral Investig. 2019 Mar;23(3):1309-1318. doi: 10.1007/s00784-018-2555-2. Epub 2018 Jul 12.
The aim of the present study was to evaluate the blood cell content, morphological aspects, gene expression of type I collagen, and release of growth factors on an injectable platelet rich fibrin (i-PRF).
Blood samples were collected from 15 volunteers to prepare i-PRF samples. Peripheral blood was used as a control group. Blood clot and i-PRF samples were cultured for 10 days. The supernatant of the samples was collected for ELISA immunoassay quantification of PDGF and VEGF growth factors over periods of 1, 8, 24, 72, and 240 h. I-PRF and blood clot samples were biologically characterized using histological and immunohistochemistry analysis for IL-10, osteocalcin, and TGF-β. Scanning electron microscopy (SEM) was used to inspect the fibrin network and distribution of blood platelets and leukocytes. Reverse transcriptase polymerase chain reaction (RT-PCR) method was used to evaluate gene expression for type I collagen.
A higher concentration of platelets and lymphocytes was recorded in i-PRF than in peripheral blood (p < 0.05). The release of VEGF was higher in blood clot samples (1933 ± 704) than that for i-PRF (852 ± 376; p < 0.001). Immunohistochemistry showed upregulation of TGF-B, IL-10, and osteocalcin in the i-PRF group. RT-PCR showed increased type I collagen gene expression in i-PRF (p < 0.05). SEM images revealed agglomeration of platelets in some regions, while a fibrin networking was noticeable in the entire i-PRF sample.
Injectable platelet rich fibrin becomes a good approach for soft and mineralized tissue healing considering the formation of a three-dimensional fibrin network embedding platelets, leukocytes, type I collagen, osteocalcin, and growth factors. Indeed, the injectable platelet rich fibrin can be indicated in several medical applications regarding bioactivity, simplied technique, and flowable mixing with other biomaterials.
Morphological, cell, and protein characterization of platelet rich fibrin provides a better understanding of the clinical effects and improvement of clinical guidelines for several medical applications. Once well physicochemical and biologically characterized, the use of an injectable platelet rich fibrin can be extended to other applications in the field of orthopedics, periodontics, and implant dentistry on the repairing process of both soft and mineralized tissues.
本研究旨在评估可注射富血小板纤维蛋白（i-PRF）的细胞含量、形态学特征、I 型胶原的基因表达和生长因子的释放。
从 15 名志愿者中采集血液样本以制备 i-PRF 样本。外周血作为对照组。将血液凝块和 i-PRF 样本培养 10 天。收集样本的上清液，使用 ELISA 免疫分析法在 1、8、24、72 和 240 小时时对 PDGF 和 VEGF 生长因子进行定量。使用组织学和免疫组织化学分析，对 i-PRF 和血凝块样本中的白细胞介素 10（IL-10）、骨钙素和转化生长因子-β（TGF-β）进行生物学特征分析。扫描电子显微镜（SEM）用于检查纤维蛋白网络和血小板及白细胞的分布。逆转录聚合酶链反应（RT-PCR）方法用于评估 I 型胶原的基因表达。
i-PRF 中的血小板和淋巴细胞浓度明显高于外周血（p<0.05）。i-PRF 中的 VEGF 释放量高于血凝块样本（1933±704 比 852±376；p<0.001）。免疫组织化学显示 i-PRF 组 TGF-B、IL-10 和骨钙素表达上调。RT-PCR 显示 i-PRF 中 I 型胶原基因表达增加（p<0.05）。SEM 图像显示在某些区域血小板聚集，而整个 i-PRF 样本中可见纤维蛋白网络。
考虑到形成三维纤维蛋白网络嵌入血小板、白细胞、I 型胶原、骨钙素和生长因子，可注射富血小板纤维蛋白是一种治疗软组织和矿化组织的良好方法。事实上，可注射富血小板纤维蛋白具有生物活性、简化技术和与其他生物材料混合的流动性等特点，可应用于多种医疗领域。
富血小板纤维蛋白的形态学、细胞和蛋白质特征有助于更好地了解其临床效果，并为多种医疗应用提供临床指导的改进。一旦充分理化和生物学特性得到表征，可注射富血小板纤维蛋白的应用可以扩展到骨科、牙周科和种植牙科领域中软组织和矿化组织修复过程中的其他应用。
