Department of Oral Biology, University Clinic of Dentistry, Medical University of Vienna, 1090 Vienna, Austria.
Department of Periodontology, School of Dental Medicine, University of Bern, 3012 Bern, Switzerland.
Int J Mol Sci. 2022 May 24;23(11):5897. doi: 10.3390/ijms23115897.
The preparation of platelet-rich fibrin (PRF) requires blood centrifugation to separate the yellow plasma from the red erythrocyte fraction. PRF membranes prepared from coagulated yellow plasma are then transferred to the defect sites to support tissue regeneration. During natural wound healing, however, it is the unfractionated blood clot (UBC) that fills the defect site. It is unclear whether centrifugation is necessary to prepare a blood-derived matrix that supports tissue regeneration. The aim of the present study was to compare lysates prepared from PRF and UBC based on bioassays and degradation of the respective membranes. We report here that lysates prepared from PRF and UBC membranes similarly activate TGF-β signaling, as indicated by the expression of interleukin 11 (IL-11), NADPH oxidase 4 (NOX-4) and proteoglycan 4 (PRG4) in gingival fibroblasts. Consistently, PRF and UBC lysates stimulated the phosphorylation and nuclear translocation of Smad3 in gingival fibroblasts. We further observed that PRF and UBC lysates have comparable anti-inflammatory activity, as shown by the reduction in lipopolysaccharide (LPS)-induced IL-6, inducible nitric oxidase synthase (iNOS) and cyclooxygenase 2 (COX-2) expression in RAW264.7 cells. Moreover, inflammation induced by Poly (1:C) HMW and FSL-1, which are agonists of Toll-like receptor (TLR) 3 and 2/6, respectively, was reduced by both PRF and UBC. PRF and UBC lysates reduced the nuclear translocation of p65 in LPS-induced RAW264.7 cells. In contrast to the similar activity observed in the bioassays, UBC membranes lack the structural integrity of PRF membranes, as indicated by the rapid and spontaneous disintegration of UBC membranes. We show here that the lysates prepared from PRF and UBC possess robust TGF-β and anti-inflammatory activity. However, visual inspection of the PRF and UBC membranes confirmed the negative impact of erythrocytes on the structural integrity of membranes prepared from whole blood. The data from the present study suggest that although both UBC and PRF have potent TGF-β and anti-inflammatory activity, UBC does not have the strength properties required to be used clinically to prepare applicable membranes. Thus, centrifugation is necessary to generate durable and clinically applicable blood-derived membranes.
富血小板纤维蛋白（PRF）的制备需要血液离心，以将黄色血浆与红色红细胞部分分离。然后将从凝固的黄色血浆中制备的 PRF 膜转移到缺陷部位以支持组织再生。然而，在自然伤口愈合过程中，充满缺陷部位的是未分馏的血凝块（UBC）。目前尚不清楚是否需要离心来制备支持组织再生的血液衍生基质。本研究的目的是通过生物测定和各自膜的降解来比较从 PRF 和 UBC 制备的裂解物。我们在这里报告，从 PRF 和 UBC 膜制备的裂解物通过白细胞介素 11（IL-11）、NADPH 氧化酶 4（NOX-4）和蛋白聚糖 4（PRG4）的表达，类似地激活 TGF-β信号，在牙龈成纤维细胞中。一致地，PRF 和 UBC 裂解物刺激牙龈成纤维细胞中 Smad3 的磷酸化和核易位。我们进一步观察到，PRF 和 UBC 裂解物具有类似的抗炎活性，如脂多糖（LPS）诱导的 IL-6、诱导型一氧化氮合酶（iNOS）和环氧化酶 2（COX-2）在 RAW264.7 细胞中的表达降低所示。此外，由多聚（1：C）HMW 和 FSL-1 诱导的炎症，分别是 Toll 样受体（TLR）3 和 2/6 的激动剂，也被 PRF 和 UBC 降低。PRF 和 UBC 裂解物减少了 LPS 诱导的 RAW264.7 细胞中 p65 的核易位。与生物测定中观察到的相似活性相反，UBC 膜缺乏 PRF 膜的结构完整性，如 UBC 膜的快速和自发崩解所示。我们在这里表明，从 PRF 和 UBC 制备的裂解物具有强大的 TGF-β和抗炎活性。然而，对 PRF 和 UBC 膜的目视检查证实了红细胞对全血制备的膜的结构完整性的负面影响。本研究的数据表明，尽管 UBC 和 PRF 都具有强大的 TGF-β和抗炎活性，但 UBC 不具备临床应用所需的强度特性，无法制备适用的膜。因此，离心是产生耐用且临床适用的血液衍生膜所必需的。
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