Preparation and characterization of leukocyte‐ and platelet‐rich fibrin membrane derived from cats' blood

Abstract

Autologous platelet concentrates have been used in regenerative medicine in humans due to the abundance of growth factors, but there are only a few reports in small animals. This study aimed to prepare and characterize a leukocyte and platelet-rich fibrin membrane (L-PRF) produced with blood obtained from cats. Thirteen client-owned healthy adult Maine Coon cats were enrolled. The blood samples were collected and centrifuged at 650g for 12 min using a centrifuge specifically designed for this application. The L-PRF clot was removed from the tube and red blood cell base layer was separated, leaving buffy coat intact. After this, L-PRF clot was compressed by specialized metal plate for 30-60 s, and L-PRF membrane was obtained. Light microscopy examination of the membranes showed three distinct layers: white part, buffy coat, and red part. Immunohistochemical analysis demonstrated expression of vascular endothelial growth factor and platelet derived growth factor. The scanning electron microscopy showed that three-dimensional architecture of fibrin network was more compact in the area near the buffy coat. In conclusion, the method used allowed the characterization of the L-PRF membrane composition, which presented cell types and fibrin network architecture similar to those described in the human species.

Evaluation of a Standardized Protocol for Plasma Rich in Growth Factors Obtention in Cats: A Prospective Study

“…Moreover, two recent articles describe the use of a platelet-rich fibrin matrix in the resolution of an oronasal fistula ( 36 ) and the application of PRP in a cutaneous wound secondary to mammary gland tumor removal with satisfactory results ( 37 ). In addition, plasma-rich fibrin (PRF), a second-generation PC used to regenerate tissues and promote early healing without risk of infection, has been identified and proposed to treat different conditions in cats ( 38 ). Compared to PRP and PGRF, PRF are preparations with a high-density fibrin network.…”

Histological, Immunohistochemical, Biomechanical, and Wettability Evaluations of the Leukocyte- and Platelet-Rich Fibrin Membranes Derived from Canine Blood

“…1,4,5 However, in a study in cats using the same centrifuge as the present study, producing a membrane required centrifugation at 650 g for 12 min. 11 Thus, the standardization of protocol for species must be considered. The centrifuge choice is also very important since a study comparing four commercially available centrifuges used to produce L-PRF demonstrated that the type of vibration affects the quality of the platelet concentrate and can cause significant damage to the cell content and, consequently, affect the quality of the platelet concentrate.…”

“…8,10 In a study of the L-PRF membrane in cats, VEGF - positive was scored as 4 and 3, respectively, in 69.23% and 30.77% of the samples; and PDGF-positive was scored as 3 and 2, respectively, in 53.85% and 46.15% of the samples. 11 Lower scores were observed in the present study since 50% of the membranes had a score of 3% and 30% a score of 2 for VEGF, and 50% of the membranes had a score of 1% and 40% a score of 2 for PDGF. VEGF and PDGF are considered inducers of angiogenesis, 25 and the intensity of expression of these growth factors may influence the purpose of the L-PRF membrane application.…”

“…Then, the samples were stained with Harris hematoxylin. Immunohistochemistry was assessed by semi - quantitative score, 11 where 0 – no staining, 1 – 1%–25% of cells staining, 2 – 26%–50% of cells staining, 3 – 51%–75% of cells staining, and 4 – above 75% of cells staining.…”

“…In addition, a positive expression of PDGF-BB, VEGF, interleukins (IL-1, IL-4), tumor necrosis factor (TNF), and TGF-β 1 in immunohistological staining has been shown. 7–11…”