RNAseq of Gingival Fibroblasts Exposed to PRF Membrane Lysates and PRF Serum

Overview

Journal Cells

Publisher MDPI

Specialties Biochemistry
Biophysics
Cell Biology
Molecular Biology

Date 2024 Aug 9

PMID 39120336

Authors

Atefe Imani

Layla Panahipour

Hannes Kuhtreiber

Michael Mildner

Reinhard Gruber

Affiliations

Soon will be listed here.

Abstract

Platelet-rich fibrin (PRF) is prepared by spontaneous coagulation of fractionated blood. When squeezed between two plates, PRF is separated into solid PRF membranes and a liquid exudate, the PRF serum. The question arises regarding how much the overall activity remains in the PRF membranes and what is discarded into the PRF serum. To this end, we have exposed gingival fibroblasts to lysates prepared from PRF membranes and PRF serum, followed by bulk RNA sequencing. A total of 268 up- and 136 down-regulated genes in gingival fibroblasts exposed to PRF membrane lysates were significantly regulated under the premise of a minimum log2 with 2.5-fold change and a minus log10 significance level of two, respectively. PRF serum only caused 62 up- and 32 down-regulated genes under these conditions. Among the 46 commonly up-regulated genes were CXCL1, CXCL5, CXCL6, CXCL8, IL33, IL6, and PTGS2/COX2, stanniocalcin-1-all linked to an inflammatory response. PRF membrane lysates further increased chemokines CCL2, CCL7, CXCL2, CXCL3, and IL1R1, IL1RL1, and IL1RN, as well as the paracrine factors IL11, LIF, IGF1, BMP2, BMP6, FGF2, and CCN2/CTGF, and all hyaluronan synthases. On the other hand, PRF serum increased DKK1. The genes commonly down-regulated by PRF membrane lysates and PRF serum included interferon-induced protein with tetratricopeptide repeats (IFIT1, IFIT2, IFIT3) and odd-skipped-related transcription factors (OSR1 and OSR2), as well as FGF18 and GDF15, respectively. Taken together, PRF membrane lysates, compared to PRF serum, cause a more complex response in gingival fibroblasts, but each increased chemokine expression in gingival fibroblasts.

Citing Articles

RNA Sequencing Revealed a Weak Response of Gingival Fibroblasts Exposed to Hyaluronic Acid.

Panahipour L, Imani A, Dos Santos Sanches N, Kuhtreiber H, Mildner M, Gruber R Bioengineering (Basel). 2025; 11(12.

PMID: 39768125 PMC: 11726844. DOI: 10.3390/bioengineering11121307.

References

Wetzler M, Talpaz M, Lowe D, Baiocchi G, Gutterman J, Kurzrock R . Constitutive expression of leukemia inhibitory factor RNA by human bone marrow stromal cells and modulation by IL-1, TNF-alpha, and TGF-beta. Exp Hematol. 1991; 19(5):347-51. View

Heberle H, Meirelles G, da Silva F, Telles G, Minghim R . InteractiVenn: a web-based tool for the analysis of sets through Venn diagrams. BMC Bioinformatics. 2015; 16:169. PMC: 4455604. DOI: 10.1186/s12859-015-0611-3. View

Miron R, Moraschini V, Fujioka-Kobayashi M, Zhang Y, Kawase T, Cosgarea R . Use of platelet-rich fibrin for the treatment of periodontal intrabony defects: a systematic review and meta-analysis. Clin Oral Investig. 2021; 25(5):2461-2478. PMC: 8060184. DOI: 10.1007/s00784-021-03825-8. View

de Witt S, Verdoold R, Cosemans J, Heemskerk J . Insights into platelet-based control of coagulation. Thromb Res. 2014; 133 Suppl 2:S139-48. DOI: 10.1016/S0049-3848(14)50024-2. View

Koenen R . The prowess of platelets in immunity and inflammation. Thromb Haemost. 2016; 116(4):605-12. DOI: 10.1160/TH16-04-0300. View

Subbarao K, Nattuthurai G, Sundararajan S, Sujith I, Joseph J, Syedshah Y . Gingival Crevicular Fluid: An Overview. J Pharm Bioallied Sci. 2019; 11(Suppl 2):S135-S139. PMC: 6555362. DOI: 10.4103/JPBS.JPBS_56_19. View

Zhao Y, Zhang Y, Lu W, Sun R, Guo R, Cao X . The diagnostic/prognostic roles and biological function of the IFIT family members in acute myeloid leukemia. BMC Med Genomics. 2023; 16(1):296. PMC: 10657597. DOI: 10.1186/s12920-023-01735-0. View

Miron R, Fujioka-Kobayashi M, Sculean A, Zhang Y . Optimization of platelet-rich fibrin. Periodontol 2000. 2023; 94(1):79-91. DOI: 10.1111/prd.12521. View

Kargarpour Z, Nasirzade J, Panahipour L, Mitulovic G, Miron R, Gruber R . Platelet-Rich Fibrin Increases BMP2 Expression in Oral Fibroblasts via Activation of TGF-β Signaling. Int J Mol Sci. 2021; 22(15). PMC: 8347014. DOI: 10.3390/ijms22157935. View

10.

Wu Y, Xing J, Li X, Yang Y, Shao H, Li J . Roles of interferon induced protein with tetratricopeptide repeats (IFIT) family in autoimmune disease. Autoimmun Rev. 2023; 22(11):103453. DOI: 10.1016/j.autrev.2023.103453. View

11.

Goedhart J, Luijsterburg M . VolcaNoseR is a web app for creating, exploring, labeling and sharing volcano plots. Sci Rep. 2020; 10(1):20560. PMC: 7689420. DOI: 10.1038/s41598-020-76603-3. View

12.

Erickson P, Cleves P, Ellis N, Schwalbach K, Hart J, Miller C . A 190 base pair, TGF-β responsive tooth and fin enhancer is required for stickleback Bmp6 expression. Dev Biol. 2015; 401(2):310-23. PMC: 4424077. DOI: 10.1016/j.ydbio.2015.02.006. View

13.

You J, Hodge C, Hoque M, Petsoglou C, Sutton G . Human Platelets and Derived Products in Treating Ocular Surface Diseases - A Systematic Review. Clin Ophthalmol. 2020; 14:3195-3210. PMC: 7567560. DOI: 10.2147/OPTH.S265701. View

14.

Lindemann S, Tolley N, Dixon D, McIntyre T, Prescott S, Zimmerman G . Activated platelets mediate inflammatory signaling by regulated interleukin 1beta synthesis. J Cell Biol. 2001; 154(3):485-90. PMC: 2196422. DOI: 10.1083/jcb.200105058. View

15.

Kargarpour Z, Nasirzade J, Panahipour L, Miron R, Gruber R . Platelet-Rich Fibrin Decreases the Inflammatory Response of Mesenchymal Cells. Int J Mol Sci. 2021; 22(21). PMC: 8583104. DOI: 10.3390/ijms222111333. View

16.

Kargarpour Z, Panahipour L, Mildner M, Miron R, Gruber R . Lipids of Platelet-Rich Fibrin Reduce the Inflammatory Response in Mesenchymal Cells and Macrophages. Cells. 2023; 12(4). PMC: 9954017. DOI: 10.3390/cells12040634. View

17.

Nasirzade J, Kargarpour Z, Hasannia S, Strauss F, Gruber R . Platelet-rich fibrin elicits an anti-inflammatory response in macrophages in vitro. J Periodontol. 2019; 91(2):244-252. PMC: 7065136. DOI: 10.1002/JPER.19-0216. View

18.

Love M, Huber W, Anders S . Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014; 15(12):550. PMC: 4302049. DOI: 10.1186/s13059-014-0550-8. View

19.

Siamwala J, Pagano F, Dubielecka P, Ivey M, Guirao-Abad J, Zhao A . IL-1β-mediated adaptive reprogramming of endogenous human cardiac fibroblasts to cells with immune features during fibrotic remodeling. Commun Biol. 2023; 6(1):1200. PMC: 10673909. DOI: 10.1038/s42003-023-05463-0. View

20.

Kolberg L, Raudvere U, Kuzmin I, Adler P, Vilo J, Peterson H . g:Profiler-interoperable web service for functional enrichment analysis and gene identifier mapping (2023 update). Nucleic Acids Res. 2023; 51(W1):W207-W212. PMC: 10320099. DOI: 10.1093/nar/gkad347. View