Platelet-Rich Fibrin Can Neutralize Hydrogen Peroxide-Induced Cell Death in Gingival Fibroblasts

Kargarpour, Zahra; Nasirzade, Jila; Summa, Francesca Di; Panahipour, Layla; Miron, Richard J.; Gruber, Reinhard

doi:10.3390/antiox9060560

Cited by 18 publications

(18 citation statements)

References 40 publications

(71 reference statements)

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“…Conversely, lysates of heated PPP, similar to the red blood cell fraction, failed to provoke the expression of the TGF-β target genes and the activation of upstream signaling consisting of activation of the TGF-β receptor 1 kinase [32], as well as the phosphorylation and nuclear translocation of Smad2/3 [33]. This finding is in line with the catalase activity of PRF lysates that is abolished by heating [21]. There were variations among the blood and cell donors but the overall finding that PPP has a weaker TGF-β activity than buffy coat was consistent among all experiments.…”

Section: Discussionsupporting

confidence: 67%

“…Heating, however, not only inactivates plasminogen and coagulates albumin; it might also affect the activity of the catalase [21] and growth factors intrinsic to the PRF clot. We recently performed a proteomic analysis combined with a whole genome gene array of a traditional PRF membrane and identified that TGF-β was a major growth factor with respect to its activation of genes in oral fibroblasts [22].…”

Section: Introductionmentioning

confidence: 99%

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Liquid Platelet-Rich Fibrin and Heat-Coagulated Albumin Gel: Bioassays for TGF-β Activity

et al. 2020

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Liquid platelet-rich fibrin (PRF) can be prepared by high centrifugation forces separating the blood into a platelet-poor plasma (PPP) layer and a cell-rich buffy coat layer, termed concentrated PRF (C-PRF). Heating the liquid PPP was recently introduced to prepare an albumin gel (Alb-gel) that is later mixed back with the concentrated liquid C-PRF to generate Alb-PRF. PRF is a rich source of TGF-β activity; however, the overall TGF-β activity in the PPP and the impact of heating the upper plasma layer remains unknown. Here, we investigated for the first time the in vitro TGF-β activity of all fractions of Alb-PRF. We report that exposure of oral fibroblasts with lysates of PPP and the buffy coat layer, but not with heated PPP, provoked a robust increase in the TGF-β target genes interleukin 11 and NADPH oxidase 4 by RT-PCR, and for IL11 by immunoassay. Consistent with the activation of TGF-β signaling, expression changes were blocked in the presence of the TGF-β receptor type I kinase inhibitor SB431542. Immunofluorescence and Western blot further confirmed that lysates of PPP and the buffy coat layer, but not heated PPP, induced the nuclear translocation of Smad2/3 and increased phosphorylation of Smad3. The immunoassay further revealed that PPP and particularly BC are rich in active TGF-β compared to heated PPP. These results strengthen the evidence that not only the cell-rich C-PRF but also PPP comprise a TGF-β activity that is, however, heat sensitive. It thus seems relevant to mix the heated PPP with the buffy coat C-PRF layer to regain TGF-β activity, as proposed during the preparation of Alb-PRF.

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Section: Discussionsupporting

confidence: 67%

Section: Introductionmentioning

confidence: 99%

Liquid Platelet-Rich Fibrin and Heat-Coagulated Albumin Gel: Bioassays for TGF-β Activity

et al. 2020

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“…The 3T3-L1 pre-adipocyte mesenchymal cells are an established model for adipogenesis, also used for inflammation research [ 42 ]. Gingival fibroblasts are a widely used model for the investigation of inflammation of the oral cavity and oral disease [ 43 ] and have been utilized in PRF research [ 44 , 45 ]. The goal of the present study was to take advantage of the ST2 and 3T3-L1 cells and gingival fibroblasts to study the possible anti-inflammatory activity of the various PRF preparations in vitro.…”

Section: Introductionmentioning

confidence: 99%

Platelet-Rich Fibrin Decreases the Inflammatory Response of Mesenchymal Cells

Kargarpour

Nasirzade

Panahipour

et al. 2021

IJMS

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Chronic inflammation is a pathological process where cells of the mesenchymal lineage become a major source of inflammatory mediators. Platelet-rich fibrin (PRF) has been shown to possess potent anti-inflammatory activity in macrophages, but its impact on mesenchymal cells has not been investigated. The aim of this study was, therefore, to expose mesenchymal cells to inflammatory cytokines together with lysates generated from liquid platelet-poor plasma (PPP), the cell-rich buffy coat layer (BC; concentrated-PRF or C-PRF), and the remaining red clot layer (RC), following centrifugation of blood. Heating PPP generates an albumin gel (Alb-gel) that when mixed back with C-PRF produces Alb-PRF. Membranes prepared from solid PRF were also subjected to lysis. We report here that lysates of PPP, BC, and PRF decreased the cytokine-induced expression of interleukin 6 (IL6) and nitric oxide synthase (iNOS) in the bone marrow-derived ST2 cells. Consistently, PPP, BC, and PRF greatly decreased the phosphorylation and nuclear translocation of p65 in ST2 cells. The inflammatory response caused by Pam3CSK4 was reduced accordingly. Moreover, PPP, BC, and PRF reduced the enhanced expression of inflammatory mediators IL6 and iNOS in 3T3-L1 pre-adipocyte mesenchymal cells, and iNOS and CCL5 in murine calvarial cells. Surprisingly, PRF lysates were not effective in reducing the inflammatory response of human gingival fibroblasts and HSC2 epithelial cells. The data from the present study suggest that both liquid PRF and solid PRF exert potent anti-inflammatory activity in murine mesenchymal cells.

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“…After removing the serum, we obtained a white clot. This procedure is similar to our previously protocol used to prepare PRF membranes [ 18 , 19 , 20 ]. The clotted fraction before and after removing the serum by a gauze was used to measure the wet weight and dry weight, respectively.…”

Section: Methodsmentioning

confidence: 99%

Fibrinogen Concentrations in Liquid PRF Using Various Centrifugation Protocols

et al. 2022

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Liquid platelet-rich fibrin (PRF) is produced by fractionation of blood without additives that initiate coagulation. Even though liquid PRF is frequently utilized as a natural source of fibrinogen to prepare sticky bone, the concentration of fibrinogen and the overall amount of “clottable PRF” components have not been evaluated. To this aim, we prepared liquid PRF at 300, 700, and 2000 relative centrifugal force (RCF), for 8 min and quantified the fibrinogen levels by immunoassay. We report here that, independent of the RCF, the fibrinogen concentration is higher in the platelet-poor plasma (PPP) compared to the buffy coat (BC) fraction of liquid PRF and further decreases in the remaining red fraction. We then determined the weight of the clotted PRF fractions before and after removing the serum. The PPP and BC fractions consist of 10.2% and 25.3% clottable matrix suggesting that more than half of the weight of clottable BC is caused by cellular components. Our data provide insights into the distribution of fibrinogen in the different fractions of liquid PRF. These findings suggest that PPP is the main source of clottable fibrinogen, while the BC is more a cell source when it comes to the preparation of sticky bone.

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Platelet-Rich Fibrin Can Neutralize Hydrogen Peroxide-Induced Cell Death in Gingival Fibroblasts

Cited by 18 publications

References 40 publications

Liquid Platelet-Rich Fibrin and Heat-Coagulated Albumin Gel: Bioassays for TGF-β Activity

Liquid Platelet-Rich Fibrin and Heat-Coagulated Albumin Gel: Bioassays for TGF-β Activity

Platelet-Rich Fibrin Decreases the Inflammatory Response of Mesenchymal Cells

Fibrinogen Concentrations in Liquid PRF Using Various Centrifugation Protocols

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