Introduction
Material and methods
Protocol development and eligibility criteria
Defining the focused question
Search strategy
Search terms
Criteria for study selection and inclusion
Exclusion criteria
Screening and selection of studies
Data extraction and analysis
Results
Selection of studies
Study (year) | Cell type | PRF preparation | Method | Main outcome induced by PRF |
---|---|---|---|---|
Beitzel et al. (2014) [17] | Human mesenchymal stem cells (MSCs) | 3000 rpm 10 min Hardware: NR | Cell viability by live and dead staining Cell adhesion assay to the PRF-Matrix Cell proliferation by incorporation of 3H-thymidine | Cell viability maintained Increased cell adhesion No effect in cell proliferation on collagen membranes |
Burnouf et al. (2012) [18] | Human embryonic kidney fibroblasts (HEK293), human osteoblastic cell line (MG-63), SIRC, NIH/3T3, periodontal ligament cells (PDL), gingival fibroblasts (GF) | 700 g 12 min Hardware:a | Cell proliferation by an automated cell counter | Increased proliferation of all cells except for NIH/3T3 after 7 days of stimulation |
Chang et al. (2010) [19] | Osteoblast cell line U2OS | 3000 rpm 12 min Hardware:a | Cell proliferation by MTT assay Western blot for ERK phosphorylation, RANKL and OPG expression | Increased proliferation after 1, 3 and 5 days Increased ERK phosphorylation during 5 days and OPG expression during 3 days |
Chang et al. (2011) [20] | Periodontal ligament fibroblasts (PDLFs) | 3000 rpm 12 min Hardware:a | Western blot for p-ERK and OPG ALP activity | Increased ERK phosphorylation and OPG protein expression up to 5 days Enhanced ALP activity up to 5 days |
Clipet et al. (2012) [21] | SaOS2 (osteoblasts), MRC5 (fibroblasts) KB (epithelial cells) | 400 g 12 min Hardware: NR | Cell proliferation by SRB assay Cytotoxic assay by SRB assay Cell cycle analysis by flow cytometry tests Gene expression of cbfa1, Col1, OC and OP by RT-PCR | Increased proliferation of SaOS2, MRC5 and KB Enhanced G2M phase in SAOS2 and MRC5 lineages Up-regulation of OP and OC on SAOS2 |
Dereli et al. (2018) [22] | Limbal epithelial cells | PRF gel: 1000 rpm/5 min PRF: 2700 rpm/12 min Hardware:b | Cell viability by live and dead staining | Cell viability maintained |
Dohle et al. (2018) [23] | Human outgrowth endothelial cells (OECs) primary osteoblasts (pOBs) | 700 rpm 3 min Hardware:c | Co-culture of OECs with pOBs Angiogenic activation by immunofluorescent staining ELISA for VEGF, PDGF-BB, E-selectin and ICAM-1 Gene expression of VEGFA, ICAM1, PDGF-BB, E-selectin. BMP2 and ALP by RT-PCR | Increased VEGF concentration on pOBs at 7 days Increased of PDGF and E-Sel levels in OECs and pOBs co-cultures at the mRNA level and protein level at 24 and 72 h Increased expression of ICAM1 and ALP at 24 h in OECs and pOBs co-cultures Upregulation of VEGF expression in PRF/co-culture at 1 and 7 days Increased BMP2 expression in PRF co-cultures cultivated at 1 and 7 days |
Ehrenfest et al. (2009) [24] | Osteoblasts, fibroblasts, preadipocytes, prekeratinocytes | 400 g 12 min Hardware:a | Proliferation assay Cytotoxicity by MTT assay Osteoblast differentiation by Von Kossa and ALP staining | Increased proliferation of fibroblasts and osteoblasts at 3, 7, 14 and 21 days in a dose-dependent manner on osteoblasts Absence of cytotoxicity in all cells Increased ALP activity after 3 days and up to 28 days and increased osteoblasts differentiation after 7 days and up to 28 days |
Ehrenfest et al. (2010) [25] | Human bone mesenchymal stem cells (BMSC) | 400 g 12 min Hardware:a | Proliferation and cytotoxicity by MTT assay Osteoblastic differentiation by ALP activity and quantification of the mineralization nodules | Increased proliferation in standard and osteogenic conditions in a dose-dependent manner Absence of cytotoxicity on BMSC in standard or osteogenic conditions Increased ALP activity in a dose-dependent manner in standard and osteogenic conditions |
Gassling et al. (2013) [26] | Human osteoblasts | 400 g 12 min Hardware: NR | Cell viability by live and dead staining Biocompatibility and cell proliferation by lactate dehydrogenase, BrdDU, MTT and WST-1 Alkaline phosphatase activity by ELISA | Cell viability maintained Increased ALP activity |
Gassling et al. (2010) [27] | Human periosteal cells | 400 g 12 min 2700 rpm Hardware:d | Cell viability by live and dead staining Biocompatibility test by LDH test and MTT assay Cell Proliferation by BrdU | Cells seeded on PRF membranes maintained their viability PRF membranes were biocompatible No effect on proliferation |
He et al. (2009)[28] | Rat calvaria osteoblasts | 400 g 10 min Hardware: NR | Cell Proliferation ALP activity Mineralization assay by alizarin red staining | Increased proliferation at 5 days and enhanced mineralization at 14 days No effect on ALP activity |
He et al. (2016) [29] | Human dental pulp cell (hDPC) | 400 g 10 min Hardware: NR | Proliferation assay by CCK-8 Gene expression of ALP and DSPP by RT-PCR | Increased proliferation after 5 and 7 days Up-regulation of ALP and DSPP expression |
Hong et al. (2018) [30] | Human Stem Cells of the Apical Papilla (SCAPs) | 400 g 10 min Hardware: NR | Cell proliferation assay Cell migration assay Osteogenic differentiation by alizarin red staining Gene expression of ALP, BSP, DMP1, and DSPP by RT-PCR | Increased proliferation at 1, 3, 5 and 7 days Enhanced migration at 12 and 24 h Increased mineralization at 7 and 14 days Down-regulation of ALP, BSP, DMP1 expression but not DSPP after 7 days Up-regulation of ALP, BSP, DMP1 and DSPP expression after 14 days |
Huang et al. (2010) [31] | Dental pulp cells (DPCs) | 3000 rpm 10 min Hardware:a | Cell proliferation by MTT assay Western blot for OPG ALP activity assay | Increased DPC proliferation at 1, 3 and 5 days Up-regulation of OPG expression in a time-dependent manner Increased ALP activity |
Kang et al. (2011) [32] | Human alveolar bone marrow stem cells (hABMSCs) | 400 g/10 min + 230 g/10 min Hardware: NR | Cell proliferation by BrdU assay Cell migration by wound-healing assay Mineralization by alizarin red staining | Increased proliferation Reduced migration at 1 and 2 days Enhanced mineralization |
Khurana et al. (2017) [33] | Dental pulp stem cells (DPSCs) periodontal ligament stem cells (PDLSCs) | 3000 rpm 10 min Hardware: NR | Cell viability by trypan blue | Absence of cytotoxicity at 7 days |
Kim et al. (2017) [34] | Human derived osteoblasts | 400 g 10 min Hardware:e | Cell proliferation by MTT assay Osteoblast differentiation by Sulforhodamine B assay ALP activity | Increased proliferation at 1, 2 and 3 days Enhanced osteoblast differentiation Increased ALP activity at 2 and 3 days |
Kim et al. (2017) [34] | Human dental pulp cells (HDPCs) | 400 g 10 min Hardware: NR | Cell viability by MTT assay ELISA for IL-1β, IL-6, and IL-8 Western blot for VCAM1 and ICAM1 and odontoblastic differentiation markers DSP and DMP1 ALP Activity Mineralization by alizarin red staining | Absence of cytotoxicity Reduced LPS-induced pro-inflammatory cytokines IL-1b, IL-6, and IL-8 at 1 and 3 days and LPS-induced adhesion molecules VCAM1 and ICAM1 at 1 day Enhanced LPS-induced up-regulation of odontoblastic differentiation markers DSP and DMP1 at 3 days Increased ALP activity and mineralization at day 7 in LPS treated cells |
Kobayashi et al. (2015) [35] | Human umbilical vein endothelial cells (HUVECs) | 600 g/2 min + 500 g/4 min + 800 g/3 min Hardware:f | Scratch assay Western blot for VEGFR2 New blood vessel formation by CAM assay | Increased migration Enhanced phosphorylation of VEGFR2 in a dose-dependent manner Increased number of blood capillaries |
Fujioka-Kobayashi et al. (2017)[36] | Human gingival fibroblasts | L-PRF: 708 g/12 min A-PRF: 200 g/14 min A-PRF+: 200 g/8 min Hardware:c | Cell viability by live-dead staining Cell migration assay Cell proliferation by MTS assay Gene expression of TGFβ, PDGF, and COL1a2 by RT-PCR | Cell viability maintained Increased migration Increased proliferation at 3 and 5 days Up-regulation of PDGF and COL1a2 expression and TGFβ, PDGF and COL1a2 at day 7 A-PRF+ produced the highest expression of TGFβ and COL1a2 |
Liang et al. (2018) [37] | Nanofat-derived stem cells (NFSCs) | 2700 rpm 12 min Hardware: NR | Cell proliferation by CCK-8 Gene expression of VEGF, bFGF, PDGF and TGFβ by RT-PCR Protein levels by Western blot | Increased proliferation Increased expression and protein levels of VEGF, bFGF, PDGF and TGFβ Enhanced osteogenic, adipogenic and chondrogenic differentiation |
Miron et al. (2017) [38] | Human gingival fibroblasts (HGF) | I-PRF:700 rpm 60 g 3 min Hardware:c | Cell viability by live and dead assay Cell migration by Boyden chamber Cell proliferation by MTS assay Gene expression of TGFβ, PDGF and COL1a2 by RT-PCR | Absence of cytotoxicity Increased cell migration Increased proliferation after 5 days Upregulation of COL1a2 and PDGF expression after 3 days, and TGFβ and COL1a2 after 7 days |
Moradian et al. (2017) [39] | Human bone marrow mesenchymal stem cells (BMMSCs) | 400 g 10 min Hardware: NR | Cell proliferation by MTT assay | Increased proliferation at 7 days |
Schär et al. (2015) [40] | Human bone marrow-derived MSC and HUVEC | 400 g 12 min Hardware:b | Cell migration assay by Boyden chambers | Increased migration of MSC and HUVEC cells at 7 days and at 8 h, respectively |
Park et al. (2018) [41] | Human umbilical vein endothelial cell (HUVEC) | 400 g 12 min Hardware:a | Cell proliferation by MTT assay Cytotoxicity assay by adenylate kinase (AK) release from dead cells Cell migration by Boyden chamber assay Cell attachment by Green Nucleic Stain-Kit | Increased proliferation, migration and attachment of cells by coating porcine-matrices with PRF Increased cytotoxicity at day 1 |
Passaretti et al. (2014) [42] | HUVEC, skin fibroblasts | 400 g 12 min Hardware:a | Cell proliferation by Bürker chamber counting and automated cell counter | Increased cell proliferation at 24 h |
Saeed et al. (2017) [43] | Dental pulp stem cells (DPSCs) | 2700 rpm 12 min + 1800 rpm/5 min Hardware: NR | Cell proliferation by MTS assay Osteogenic differentiation by alizarin red assay | Reduced cell viability at 1 and 3 days by most PRF concentrations Enhanced osteogenic differentiation at 7 days |
Vahabi et al. (2015) [44] | Human gingival fibroblasts cell line (HGF) | 400 g 12 min Hardware: NR | Viability and proliferation by MTT assay | Cell viability maintained during the first 24 h, at 48 and 72 h cell viability is reduced Increased proliferation only the first 24 h |
Wang et al. (2018) [45] | Human primary osteoblasts | I-PRF:700 rpm 60 g 3 min Hardware:c | Cell viability by live and dead staining Cell migration by polyethylene terephthalate cell culture inserts Cell adhesion assay by staining cells with 4′,6-diamidino-2-phenylindole Cell proliferation by CCK-8 ALP activity assay Mineralization by alizarin red staining Gene expression of COL1A, Runx2, ALP, OCN by RT-PCR Immunofluorescence staining for OC expression | Cell viability maintained Increased migration at 1 day No effect on cell adhesion Increased proliferation at 3 and 5 days Enhanced ALP activity and mineralized nodule formation Up-regulation of ALP expression at 3 days and OCN, Runx2 and COL1A at 14 days Increased staining intensity of OCN |
Wang et al. (2017) [46] | Human gingival fibroblasts | 700 rpm 60 g 3 min Hardware:c | Cell viability by live-dead staining assay Cell migration assay by polyethylene terephthalate cell culture inserts Cell adhesion assay Cell proliferation by CCK-8 Gene expression of PDGF, TGFβ and COL1a1 and FN1 Collagen Type I Staining Immunostaining | Cell viability maintained irrespective of the titanium surface Increased migration in tissue culture plates and titanium surfaces No effect on cell adhesion Increased cell proliferation after 3 and 5 days Upregulation of PDGF, TGF-b, COL1 and FN1 expression levels on all surfaces Increased fluorescence intensity of collagen type 1 on all surfaces. |
Wei et al. (2017) [47] | Nanofat-derived Stem Cells (NFSCs) | 2700 rpm 12 min Hardware: NR | Proliferation assay by CCK-8 Gene expression of adipogenic differentiation markers PPARγ2, C/EBPα, and ADD1 by RT-PCR | Increased proliferation after 3 days Enhanced adipogenic differentiation at 14 days in a dose dependent-manner Up-regulation of PPARγ2, C/EBPα, and ADD1 expression |
Wirohadidjojo et al. (2016)[48] | Ultraviolet-A (UVA)-irradiated human dermal fibroblasts (HDFs) | 400 g 12 min Hardware: NR | Proliferation assay by MTT assay Collagen deposition assay Cell migration rate assay | Increased proliferation, migration and collagen deposition in UVA-irradiated HDFs |
Woo et al. (2016) [49] | Human dental pulp cells (HDPCs) | 400 g 10 min Hardware: NR | Cell viability by MTT and MTA assay ALP activity assay ALP staining Alizarin red staining for mineralization formation Western Blot for DSP,DMP1, BMP 2/4, phospho-smad1/5/8 | Cell viability maintained at 48 h Increased protein levels of DSP and DMP1 and enhanced ALP activity. Increased mineralization Activation of BMP 2/4 signaling and phosphorylation of SMAD1/5/8 |
Wu et al. (2012) [50] | Human osteoblast cell line (U2OS) | 3000 rpm 12 min Hardware:c | Cell attachment assay by WST-1 assay Cell proliferation assay Western blot analysis for p-Akt, HSP 47 and LOX | Increased cell attachment the first 3 h Increased proliferation at 1, 3 and 5 days. Enhanced Akt phosphorylation, HSP 47 expression, and LOX expression in U2OS cells |
Xu et al. (2016) [51] | Human breast adipose-derived stem cells (HBASCs) | 2700 rpm 12 min Hardware: NR | Attachment of HBASCs on scaffolds in the presence of PRF, Rg1 or both by fluorescence imaging | Increased proliferation and attachment on scaffolds |
Zhao et al. (2013) [52] | Periodontal ligament stem cells (PDLSCs) | 400 g 10 min Hardware: NR | Cell viability and proliferation assay by MTT assay ALP activity Gene expression of BSP, OCN, ColI, and CP23 by RT-PCR | Cell viability maintained Increased proliferation during 7 days Reduced ALP activity at 7 days Down-regulation of BSP and OCN expression at 7, 14 and 21 days Up-regulation PDL-related genes ColI and CP23 |
Study (year) | Cell type | PRF preparation | Method | Main outcome induced by PRF |
---|---|---|---|---|
Bucur et al. (2019) [53] | Fibroblast cell line L929 | 200 g 14 min Hardware: NR | Cell proliferation and migration using RCTA Scratch assay | No effect on proliferation neither on migration |
Elgamal et al. (2019) [47]. | Adipose mesenchymal stem cells (MSC) | 1500 rpm 14 min Hardware: NR | Cell proliferation by MTT assay | Increased proliferation |
Gervois et al. (2019) [54] | Human dental pulp stem cells (hDPSCs), neural stem cell (NSC) | 400 g 12 min Hardware:c | Metabolic activity assay by MTT assay Cell proliferation by PI assay Cell migration by transwell migration assay | Decreased and increased metabolic activity, dependent on the PRF concentration Increased proliferation of hDPSCs but no effect on NSC Increased migration of NSC |
Gomez et al. (2019) [55] | Endothelial cells (EC) | 2000 rpm 7 min Hardware: NR | Counting of the cells viewed under phase-contrast microscope | Increased cell growth |
Herrera-Vizcaíno et al. (2019) [56] | Human dermal vascular endothelial cells (HDMECs) Human fibroblasts (HF) | 44 g or 710 g 8 min Hardware:a | Immunostaining of endothelial cells marker CD31 Immunohistochemical detection of CD31 Protein levels of VEGF, PDGF-BB and TGFβ by ELISA | Increased CD31-positive cells Increased concentration of PDGF-BB of TGFβ at day 4 in HDMECs and HF No effect on VEGF |
Kargarpour et al. (2019) [57] | Murine primary macrophages, RAW264.7 cells | 1570 rpm 12 min Hardware:d | Cell viability by MTT assay and live-dead staining Cell proliferation by BrdU Caspase 3 activity assay Western blot for cleaved caspase 3 Gene expression of osteoclast marker genes TRAP, Cathepsin K, DCSTAMP, NFATc1, OSCAR and pro and anti-apoptotic marker genes, caspase-3, Bax and Bcl2L1 by RT-PCR TRAP staining Pit formation assay | Maintenance of cell viability and enhanced cell proliferation Reduced caspase 3 activity and suppression of cleaved caspase-3 Supression of osteoclastogenesis Suppression of the expression of TRAP, Cathepsin K, DCSTAMP, NFATc1, OSCAR Reduced number of multinucleated TRAP positive cells Reduced pit formation on dentine slices |
Kasnak et al. (2019) [58] | Human oral keratinocyte (HMK) cells | 2700 rpm 15 min Hardware:e | Cell proliferation by CellTiter 96 assay Protein levels of IL1β, IL1Ra, IL8, MCP1, and VEGF by ELISA | Increased proliferation on titanium and hydroxyapatite discs Increased concentration of IL1β, IL1Ra, IL8, MCP1, and VEGF on titanium and hydroxyapatite discs |
Li et al. (2018) [59] | Human periodontal ligament cells (hPDLCs) | 750 g 12 min + 500 g 5 min Hardware: NR | Cell proliferation using CCK-8 ALP activity assay Mineralization assay by alizarin red staining Gene expression of RUNX2, Osterix and Osteocalcin by RT-PCR Protein expression of RUNX2 by Western blot | Increased proliferation at day 1, 2 and 3 Increased ALP activity at 7 and 14 days Increased mineralization at 14 days Increased expression of RUNX2, Osterix at 5 and 7 days and Osteocalcin at 7 days Increased protein expression of RUNX2 at day 5 |
Mahendran et al. (2019)[60] | Fibroblast cell line L929 | 3000 rpm 10 min Hardware: NR | Cell viability by MTT assay | Maintenance of cell viability |
Mudalal et al. (2019) [61] | Human gingival fibroblast | 3000 rpm 12 min Hardware:b | Gene expression of IL1β, IL6 and TNFα by RT-PCR | Decreased expression of IL1β, IL6 and TNFα |
Nasirzade et al. (2019) [62] | Murine primary macrophages, Raw 264.7 cells | 1570 rpm 12 min Hardware:d | Gene expression of M1 marker genes IL1β, IL6; M2 genes Arg1, Ym1 and lipoxygenases, ALOX12, and ALOX15 by RT-PCR IL6 levels by ELISA NF-휅B intracellular translocation by Immunofluorescent | Decreased expression of IL1β and IL6 Increased expression of Arg1, Ym1 and lipoxygenases Increased IL6 protein level Reduced intracellular translocation of NF-휅B |
Ratajczak et al. (2018) [63] | HUVEC | 400 g 12 min Hardware:c | Cell proliferation by MTT assay and propidium iodide assay Cell migration by transwell migration assay Angiogenic potential by Tube Formation assay | Increased proliferation Enhanced migration Increased tube formation |
Steller et al. (2019) [64] | Primary human osteoblasts (OB) | 400 g 10 min Hardware:f | Viability by MTT assay Cell adhesion to titanium surface by RTCA and cell wash assay | Maintenance of cell viability No effect on adhesion to titanium surface |
Steller et al. (2019) [64] | Human gingival fibroblasts (GF), human osteoblasts (hOB) | 400 g 10 min Hardware:f | Cell viability by MTT assay Cell migration by scratch assay | Cell viability maintained at 24 and 72 h in OB and at 72 h in GF Increased migration of GF and OB at 24, 48 and 72 h Increased proliferation |
Verboket et al. (2019)[40] | Bone marrow monuclear cells (BMC) | 60 g 3 min 208 g 8 min Hardware:a | Metabolic activity by MTS assay Gene expression of VEGFA, ICAM1, MMP2, MMP7, MMP9, TGF-β1, BCL2, BAX, ALP, COL1a1, FGF23, and OPN by RT-PCR Determination of apoptosis using Annexin-V-staining | Increased metabolic activity at day 14 Increased expression of SPPI at day 2 and 7, TGFβ, MMP2, at day 7 and MMP9 at day 14 No effect on ICAM, ALP, COL1a1, FGF23 and other genes. PRF did not induce apoptosis |
Wang et al. (2019) [65] | Dermal skin fibroblast cell | 60 g 3 min Hardware: NR | Cell viability using live and dead staining Cell migration assay Cell proliferation using CCK-8 Gene expression of PDGF, TGFβ, COL1a1, and FN1 by RT-PCR Immunofluorescent staining of collagen type I | Maintenance of cell viability Increased the migration Increased proliferation at day 3 and 5 Increased expression of PDGF, TGFβ, COL1a1, and FN1 at 3 and 7 days. Increased collagen type I staining |