The impact of the centrifuge characteristics and centrifugation protocols on the cells, growth factors, and fibrin architecture of a leukocyte- and platelet-rich fibrin (L-PRF) clot and membrane

Abstract

L-PRF (leukocyte- and platelet-rich fibrin) is one of the four families of platelet concentrates for surgical use and is widely used in oral and maxillofacial regenerative therapies. The first objective of this article was to evaluate the mechanical vibrations appearing during centrifugation in four models of commercially available table-top centrifuges used to produce L-PRF and the impact of the centrifuge characteristics on the cell and fibrin architecture of a L-PRF clot and membrane. The second objective of this article was to evaluate how changing some parameters of the L-PRF protocol may influence its biological signature, independently from the characteristics of the centrifuge. In the first part, four different commercially available centrifuges were used to produce L-PRF, following the original L-PRF production method (glass-coated plastic tubes, 400 g force, 12 minutes). The tested systems were the original L-PRF centrifuge (Intra-Spin, Intra-Lock, the only CE and FDA cleared system for the preparation of L-PRF) and three other laboratory centrifuges (not CE/FDA cleared for L-PRF): A-PRF 12 (Advanced PRF, Process), LW-UPD8 (LW Scientific) and Salvin 1310 (Salvin Dental). Each centrifuge was opened for inspection, two accelerometers were installed (one radial, one vertical), and data were collected with a spectrum analyzer in two configurations (full-load or half load). All clots and membranes were collected into a sterile surgical box (Xpression kit, Intra-Lock). The exact macroscopic (weights, sizes) and microscopic (photonic and scanning electron microscopy SEM) characteristics of the L-PRF produced with these four different machines were evaluated. In the second part, venous blood was taken in two groups, respectively, Intra-Spin 9 ml glass-coated plastic tubes (Intra-Lock) and A-PRF 10 ml glass tubes (Process). Tubes were immediately centrifuged at 2700 rpm (around 400 g) during 12 minutes to produce L-PRF or at 1500 rpm during 14 minutes to produce A-PRF. All centrifugations were done using the original L-PRF centrifuge (Intra-Spin), as recommended by the two manufacturers. Half of the membranes were placed individually in culture media and transferred in a new tube at seven experimental times (up to 7 days). The releases of transforming growth factor β-1 (TGFβ-1), platelet derived growth factor AB (PDGF-AB), vascular endothelial growth factor (VEGF) and bone morphogenetic protein 2 (BMP-2) were quantified using ELISA kits at these seven experimental times. The remaining membranes were used to evaluate the initial quantity of growth factors of the L-PRF and A-PRF membranes, through forcible extraction. Very significant differences in the level of vibrations at each rotational speed were observed between the four tested centrifuges. The original L-PRF centrifuge (Intra-Spin) was by far the most stable machine in all configurations and always remained under the threshold of resonance, unlike the three other tested machines. At the classical speed of production of L-PRF, the level of undesirable vibra...

Standardized protocol proposed for clinical use of L-PRF and the use of L-PRF Wound Box®

“…No significant differences were found in the basic haematological confrontation, which had an average concentration of RBC 9. [12] and Dohan Ehrenfest et al [13,14] and those we observed in the horse model. In this comparison, it was verified that there are significant differences in the characteristics of the clot, but these differences are eliminated when the membranes derived from the compression were examined.…”

Thein vitrorelease of cytokines and growth factors from fibrin membranes produced through horizontal centrifugation

“…17 The ultrastructure of the fibrin membrane evidenced by SEM is a dense and intricate gauze-like mesh, very characteristic of the PRF elastic and resistant membranes. 14,19 It is important to notice that further studies are necessary to confirm that this apparent evidence of stability by direct observation reflects the maintenance on the integrity of the fibrin network, employing techniques such as SEM. 18 The absence of alterations in the size of these membranes (Fig.…”

Revisión Bibliográfica

“…Esta centrifugadora es la única hasta ahora aprobada por la FDA/CE del mercado. Se ha descrito que presenta el menor nivel de vibración (32) y que con ella se obtiene un coágulo de mejor calidad (33) cuando es comparada con otras tres centrifugadoras (A-PRF 12 (Advanced PRF, Process), LW -UPD8 (LW Scientific) y Salvin 1310 (Salvin Dental).…”

“…La Fibrina Rica en Plaquetas (PRF: Platelet Rich Fibrin) surgió en Francia en 2001 (3) como una segunda generación de concentrados plaquetarios con ciertas ventajas sobre el Plasma Rico en Plaquetas (PRP) (4,5) . Este biomaterial no requiere anticoagulantes en su preparación y muestra liberación sostenida de factores de crecimiento durante por lo menos 7 días (6,7) . La PRF contiene una matriz de fibrina con una gran cantidad de plaquetas y leucocitos, elementos clave que actúan en forma sinérgica promoviendo la cicatrización y la regeneración tisular (8) .…”