Leukocyte-and Platelet-Rich Fibrin for enhanced tissue repair: an in vitro study characterizing cellular composition, growth factor kinetics and transcriptomic insights

富含白细胞和血小板的纤维蛋白可增强组织修复:一项表征细胞组成、生长因子动力学和转录组学见解的体外研究

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作者:Birgit Coucke, Ellen Dilissen, Jonathan Cremer, Rik Schrijvers, Tom Theys, Laura Van Gerven

Background

Leukocyte- and platelet-rich fibrin (L-PRF) is an autologous platelet concentrate, prepared by centrifugation of blood and consisting of a dense fibrin network with incorporated leukocytes and platelets. This study aims to perform an in-depth analysis of the cells, growth factors, and transcriptome of L-PRF.

Conclusion

We characterized L-PRF using in vitro experiments, to obtain an insight in the composition of the material including a possible mechanistic role for tissue healing. This was the first study characterizing L-PRF at a combined cellular, proteomic, and transcriptional level.

Results

Fresh, 1 week and 2 weeks cultured human L-PRF membranes and liquid L-PRF glue were characterized on cellular and transcriptional level using flow cytometry (n = 4), single-cell RNA sequencing (n = 5) and RT-qPCR. Growth factor kinetics were investigated using ELISA (EGF, VEGF, PDGF-AB, TGF-β1, bFGF). L-PRF contained a large number of viable cells (fresh 97.14 ± 1.09%, 1 week cultured 93.57 ± 1.68%), mainly granulocytes in fresh samples (53.9 ± 19.86%) and T cells in cultured samples (84.7 ± 6.1%), confirmed with scRNA-seq. Monocytes differentiate to macrophages during 1 week incubation. Specifically arterial L-PRF membranes were found to release significant amounts of VEGF, EGF, PDGF-AB and TGF-β1.

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