Quantitative Near-Infrared Imaging of Platelets in Platelet-Rich Fibrin (PRF) Matrices: Comparative Analysis of Bio-PRF, Leukocyte-Rich PRF, Advanced-PRF and Concentrated Growth Factors.

Hachidai Aizawa,Sadahiro Yamaguchi,Atsushi Sato,Taisuke Watanabe,Tetsuhiro Tsujino,Tomoyuki Kawase,Kazushige Isobe,Kazuhiro Okuda,Hajime Okudera,Yutaka Kitamura

doi:10.3390/ijms21124426

Hachidai Aizawa, Sadahiro Yamaguchi + Show 8 more

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https://doi.org/10.3390/ijms21124426

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Abstract

Platelet-rich fibrin (PRF) is a fibrin matrix enriched with platelets. The PRF matrix is thought to form a steep gradient of platelet density around the region corresponding to the buffy coat in anticoagulated blood samples. However, this phenomenon has not yet been proven. To visualize platelet distribution in PRF in a non-invasive manner, we utilized near-infrared (NIR) imaging technology. In this study, four types of PRF matrices, bio-PRF, advanced-PRF (A-PRF), leukocyte-rich PRF (L-PRF), and concentrated growth factors (CGF) were compared. Blood samples collected from healthy, non-smoking volunteers were immediately centrifuged using four different protocols in glass tubes. The fixed PRF matrices were sagittally divided into two equal parts, and subjected to modified immunohistochemical examination. After probing with NIR dye-conjugated secondary antibody, the CD41+ platelets were visualized using an NIR imager. In L-PRF and CGF, platelets were distributed mainly on and below the distal surface, while in bio-PRF and A-PRF, platelet distribution was widespread and homogenous. Among three regions of the PRF matrices (upper, middle, and lower), no significant differences were observed. These findings suggest that platelets aggregate on polymerizing fibrin fibers and float up as a PRF matrix into the plasma fraction, amending the current “gradient” theory of platelet distribution.

Highlights

Platelet-rich fibrin (PRF), as its name implies, is a fibrin matrix enriched with platelets and their growth factors [1]
In the A-PRF matrix prepared by slow spin, the platelets were distributed nearly homogeneously
To confirm the validity of the imaging data, we examined the platelet distribution using conventional immunohistochemical methods using paraffin sections

Summary

Introduction

Platelet-rich fibrin (PRF), as its name implies, is a fibrin matrix enriched with platelets and their growth factors [1]. This characteristic constitution has led PRF to be widely used in regenerative medicine. Micrographic examination methods using scanning electron microscopy (SEM) and immunohistochemical techniques have been the only practical options for evaluating platelet distribution [4,5,6,7]. The data obtained using these methods are often too fragmented to adequately evaluate platelet distribution in three-dimensional (3D) structures by reconstruction

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