Reduction of relative centrifugation force within injectable platelet-rich-fibrin (PRF) concentrates advances patients’ own inflammatory cells, platelets and growth factors: the first introduction to the low speed centrifugation concept

Abstract

PurposeThe aim of this study was to analyze systematically the influence of the relative centrifugation force (RCF) on leukocytes, platelets and growth factor release within fluid platelet-rich fibrin matrices (PRF).Materials and methodsSystematically using peripheral blood from six healthy volunteers, the RCF was reduced four times for each of the three experimental protocols (I–III) within the spectrum (710–44 g), while maintaining a constant centrifugation time. Flow cytometry was applied to determine the platelets and leukocyte number. The growth factor concentration was quantified 1 and 24 h after clotting using ELISA.ResultsReducing RCF in accordance with protocol-II (177 g) led to a significantly higher platelets and leukocytes numbers compared to protocol-I (710 g). Protocol-III (44 g) showed a highly significant increase of leukocytes and platelets number in comparison to -I and -II. The growth factors’ concentration of VEGF and TGF-β1 was significantly higher in protocol-II compared to -I, whereas protocol-III exhibited significantly higher growth factor concentration compared to protocols-I and -II. These findings were observed among 1 and 24 h after clotting, as well as the accumulated growth factor concentration over 24 h.DiscussionBased on the results, it has been demonstrated that it is possible to enrich PRF-based fluid matrices with leukocytes, platelets and growth factors by means of a single alteration of the centrifugation settings within the clinical routine.ConclusionsWe postulate that the so-called low speed centrifugation concept (LSCC) selectively enriches leukocytes, platelets and growth factors within fluid PRF-based matrices. Further studies are needed to evaluate the effect of cell and growth factor enrichment on wound healing and tissue regeneration while comparing blood concentrates gained by high and low RCF.

Reduction of relative centrifugal forces increases growth factor release within solid platelet-rich-fibrin (PRF)-based matrices: a proof of concept of LSCC (low speed centrifugation concept)

“…These observations highlight the influence of RCF reduction, i.e., from PRF (708 g) to A-PRF and A-PRF+ (208 g) on platelet distribution, thereby correlating with the previously demonstrated automated cell counting that indicated significantly more platelets in PRF matrices prepared with low RCF than with high RCF application [15]. A previous ex vivo immunohistochemical study demonstrated the distribution pattern in PRF and A-PRF, which included, in addition to platelets, a wide range of inflammatory cells that physiologically exist within the peripheral blood, such as leukocytes, including neutrophils and monocytes as well as lymphocytes [5].…”

“…Moreover, immunohistochemical evaluation indicated an equal distribution pattern of platelets in all clot regions in the case of A-PRF and A-PRF+, whereas in PRF, the majority of the platelets were located in the lower portion of the clot. These findings may be related to the LSCC (low speed centrifugation concept), indicating that reducing the applied RCF increases the number inflammatory cells and platelets as well as the growth factor release within the PRF-based matrices [15]. Because the centrifugation process depends on cell weight and density, a higher RCF may be the reason for the sedimentation of the majority of the platelets to the lower portion of the clot according to their density and size, as observed in PRF.…”

“…Further research on PRF-based matrices regarding their structure and composition indicates that adjusting the centrifugation time, i.e., reducing the spinning time and applying the same RCF as in the case of A-PRF, allows the introduction of a new PRF-based matrix, Advanced-PRF+ (A-PRF+). A previous systematic study demonstrated the influence of the RCF reduction on the leukocyte and platelet numbers as well as their role in growth factor release in fluid PRF-based matrices following the LSCC, which indicates that reducing the RCF enhances the cell number and growth factor release within PRF-based matrices [15]. Based on the LSCC, we examined modifications of the RCF and centrifugation times in solid PRF-based matricesand their influence on the growth factor release within the previously introduced PRF protocols with a solid structure; PRF, A-PRF and A-PRF+.…”

“…In addition, a modification of the preparation setting based on the previously LSCC (described low-speed centrifugation concept) is a first step in the reduction of the applied relative centrifugation force (RCF). This step was accompanied by a mild increase of centrifugation time, resulting in a so-called advanced PRF (A-PRF) [5,15]. Analysis of the structure and composition of A-PRF revealed a more porous structure compared to PRF [5].…”

Injectable platelet‐rich fibrin and microneedling for gingival augmentation in thin periodontal phenotype: A randomized controlled clinical trial

“…Injectable platelet‐rich fibrin (i‐PRF) has been produced by changing the type of the tube, centrifugation time and speed; specifically, the blood is centrifuged in plastic tubes at 700 rpm for 3 min (Miron & Choukroun, ). I‐PRF prepared according to low‐speed centrifugation concept can provide a significant advantage for the regeneration process, as it is rich in platelets, leucocytes and growth factors (Choukroun & Ghanaati, ). I‐PRF clots and forms a gel form after approximately 10–15 min and preserves its content in the tissue for sustained release (Miron & Choukroun, ).…”