Copenhagen Wound Healing Center, Bispebjerg Hospital, University of Copenhagen, Copenhagen, Denmark; Digestive Disease Center, Bispebjerg Hospital, University of Copenhagen, Copenhagen, Denmark; Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Vox Sang. 2014 Jul;107(1):37-43. doi: 10.1111/vox.12120. Epub 2013 Dec 10.
Autologous platelet-rich fibrin (PRF(®)) is prepared by the automatic Vivostat(®) system. Conflicting results with Vivostat PRF in acute wound healing prompted us to examine its cellular and biomolecular composition. Specifically, platelets, selected growth factors and matrix metalloproteinase (MMP)-9 were quantified using novel analytical methods.
Ten healthy non-thrombocytopenic volunteers donated blood for generation of intermediate fibrin-I and final PRF. Anticoagulated whole blood and serum procured in parallel served as baseline controls. Leucocyte, erythrocyte and platelet counts in whole blood and fibrin-I were determined by automated haematology analyser. Platelet concentration in PRF was quantified manually by stereologic analysis of Giemsa-stained tissue sections, and the total content of five growth factors and MMP-9 by enzyme-linked immunosorbent assays.
The number of leucocytes and erythrocytes was reduced (P < 0·001), whereas platelets increased (P < 0·001) in fibrin-I versus whole blood. PRF contained 982 ± 206 × 10(9) platelets/l representing 3·9-fold (P < 0·001) enrichment relative to whole blood. Growth factor abundance in Vivostat PRF and serum was in descending order: transforming growth factor-β1 [5·1-fold higher in PRF than serum, P < 0·001] > platelet-derived growth factor (PDGF)-AB [2·5-fold, P < 0·01] > PDGF-BB [1·6-fold, P < 0·05] > vascular endothelial growth factor > basic fibroblast growth factor [75-fold, P < 0·001]. MMP-9 was reduced 139-fold (P < 0·001) compared with serum, reflecting leucocyte depletion in PRF.
The gained knowledge on platelet enrichment and biomolecular constituents may guide clinicians in their optimal use of Vivostat PRF for tissue regenerative applications.
自体富血小板纤维蛋白(PRF(®))由自动 Vivostat(®)系统制备。 Vivostat PRF 在急性伤口愈合中的结果存在冲突,这促使我们研究其细胞和生物分子组成。具体而言,使用新型分析方法定量了血小板、选定的生长因子和基质金属蛋白酶(MMP)-9。
10 名健康非血小板减少症志愿者捐献血液以生成中间纤维蛋白-I 和最终 PRF。同时采集抗凝全血和血清作为基线对照。使用自动血液学分析仪测定全血和纤维蛋白-I 中的白细胞、红细胞和血小板计数。通过 Giemsa 染色组织切片的体视学分析手动定量 PRF 中的血小板浓度,并通过酶联免疫吸附试验定量五种生长因子和 MMP-9 的总量。
与全血相比,纤维蛋白-I 中的白细胞和红细胞数量减少(P < 0·001),而血小板数量增加(P < 0·001)。PRF 中含有 982 ± 206 × 10(9)个血小板/L,相对于全血富集了 3.9 倍(P < 0·001)。Vivostat PRF 和血清中生长因子的丰度依次为:转化生长因子-β1 [PRF 中比血清高 5.1 倍,P < 0·001] >血小板衍生生长因子(PDGF)-AB [2.5 倍,P < 0·01] > PDGF-BB [1.6 倍,P < 0·05] > 血管内皮生长因子 >碱性成纤维细胞生长因子[75 倍,P < 0·001]。与血清相比,MMP-9 减少了 139 倍(P < 0·001),反映了 PRF 中白细胞的耗竭。
关于血小板富集和生物分子成分的知识可指导临床医生优化 Vivostat PRF 在组织再生应用中的使用。