Relative Centrifugal Force (RCF; G-Force) Affects the Distribution of TGF-β in PRF Membranes Produced Using Horizontal Centrifugation

Abstract

Solid platelet-rich fibrin (PRF) is produced with centrifugation tubes designed to accelerate clotting. Thus, activated platelets may accumulate within the fibrin-rich extracellular matrix even before centrifugation is initiated. It can thus be assumed that platelets and their growth factors such as transforming growth factor-β (TGF-β) are trapped within PRF independent of their relative centrifugal force (RCF), the gravitation or g-force. To test this assumption, we prepared PRF membranes with tubes where clotting is activated by a silicone-coated interior. Tubes underwent 210 g, 650 g and 1500 g for 12 min in a horizontal centrifuge. The respective PRF membranes, either in total or separated into a platelet-poor plasma and buffy coat fraction, were subjected to repeated freeze-thawing to prepare lysates. Gingival fibroblasts were exposed to the PRF lysates to provoke the expression of TGF-β target genes. We show here that the expression of interleukin 11 (IL11) and NADPH oxidase 4 (NOX4), and Smad2/3 signaling were similarly activated by all lysates when normalized to the size of the PRF membranes. Notably, platelet-poor plasma had significantly less TGF-β activity than the buffy coat fraction at both high-speed protocols. In contrast to our original assumption, the TGF-β activity in PRF lysates produced using horizontal centrifugation follows a gradient with increasing concentration from the platelet-poor plasma towards the buffy coat layer.

Evaluating the Effect of the Injectable PRF in comparison with the hyaluronic acid as an Intra-articular Medication Following Arthrocentesis in the Treatment of Internal Derangement

“…With regards to pain, it was reduced in both groups. These results complies with various other reports [7,14,21,[24][25][26][27][28] . At the third month, pain began to increase may be due to the need for treatment re-application.…”

“…Moreover, i-PRF achieved an improvement that was sustained all-over the follow up period. Pain reduction and better lubrication might allowed wider range of movement due to the enhanced mechanics and pain reduction which also might have decreased the incidence of trismus The better performance of i-PRF rather than HA go in agree with other studies [7,14,21,[24][25][26][27][28] . That might be the result of its ability of i-PRF to release higher concentrations of multiple growth factors, to induce higher fibroblast migration, and to express PDGF, TGF-β, and collagen1 which all could collaborate to provide more enhanced environment for the regeneration and repair of the defects.…”

“…Since Hyaluronic acid (HA) is the main constituent of the synovial fluid, it was widely utilized as an intra-articular injection therapy as a lubricant [21][22][23][24] . injectable PRF (i-PRF) is a very recently introduced regenerative material that contains higher concentrations of various growth factors and leukocytes rather than the ordinary PRF, and so it was introduced as a recent intra-articular injection material [25][26][27][28][29] . Therefore, the current study aimed at comparing the effect of the i-PRF with HA in cases of ID.…”

“…Therefore, a novel technique based on low speed centrifugation resulted in the introduction of the liquid PRF to enable the injection ability of that valuable material. This liquid PRF known as injectable PRF (i-PRF) has a concentration of leukocytes and various growth factors particularly vascular endothelial growth factor (VEGF) and transforming growth factor-beta (TGF-β) greater than that found in the ordinary PRF [25,26] . Leukocytes are well-known connector cells in the chain of cell-cell communication between the precursor cells and mesenchymal cells with regard to tissue regeneration and wound healing.…”

Platelet-Rich Fibrin Decreases the Inflammatory Response of Mesenchymal Cells

“…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.…”