Abstract
Blood concentrates are obtained by centrifugation of the patient's own peripheral blood. These concentrates revealed increasing importance in the regenerative medicine and tissue engineering. Many limitations are faced in preclinical research concerning the volume of blood needed to produce platelet-rich fibrin (PRF) matrices (10 mL). This study evaluated for the first time three different centrifugation protocols to obtain injectable PRF (i-PRF) matrices using a small blood volume of 3 mL. Small tubes were specially produced for this study. Blood was collected from 24 Wistar rats (ca. 200 g) for this study using the cardiac puncture technique. From each animal, two tubes were collected: the first was prepared according to the standard PRF protocol without anticoagulation and used for in vitro culturing and histology; the second was anticoagulated and used for automated cell count. Three different relative centrifugation forces (RCFs; high [710 g], medium [177 g], and low [44 g]) were evaluated in this study (n = 8 per RCF). The obtained PRF matrices were cultured over 10 days, and the supernatants were collected for quantification of growth factors (GFs), vascular endothelial growth factor (VEGF), transforming growth factor beta 1 (TGF-β1), and platelet-derived growth factor-BB (PDGF-BB). The results showed that it is possible to produce i-PRF using a small blood volume of 3 mL. Reducing the RCF led to a significant increase in the number of platelets and leukocytes, specifically lymphocytes, within the i-PRF matrices. GF release (VEGF, TGF-β1, and PDGF-BB) was continuously measurable in all evaluated i-PRF matrices over 10 days. The i-PRF matrices prepared using low RCF released significantly more GF compared to those prepared using a medium or high RCF over 10 days (p < 0.05). These findings are in accordance with the previously described low-speed centrifugation concept (LSCC), which was first established using human blood, and may serve as a proof of concept of the LSCC in another species, that is, rats. Reducing the initial blood volume provides many possibilities to implement PRF matrices in preclinical research as an autologous system. However, further studies are needed to evaluate the therapeutic effects of PRF matrices obtained using 10 mL blood compared to those obtained using 3 mL blood.
Impact Statement
This study evaluated for the first time the composition and bioactivity of platelet-rich fibrin (PRF) produced from small animal blood by reducing the initial blood volume needed for the preparation of PRF from 10 to 3 mL. The results showed that different preparation protocols of PRF produced using 3 mL of animal blood exhibit the same composition, properties, and bioactivity as PRF prepared using 10 mL human blood.
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