Abstract Number: PB0487
Meeting: ISTH 2020 Congress
Background: Leukocyte-platelet rich fibrin (L-PRF) is extensively used in the dentistry field and other clinical scenarios due to its regeneration properties. Active platelets and leukocytes secrete growth factors and other proteins that contribute to wound healing.
Aims: The goal of the present study was to identify the proteins secreted by L-PRF membranes cultured in-vitro and differences in the secretome over time.
Methods: L-PRF membranes were obtained after centrifuging blood samples from healthy volunteers, in the absence of anticoagulants, following an established protocol. Membranes were cultured in DMEM and the secretome was collected at days 3, 7 and 21. Following precipitation, protein samples were separated by SDS-PAGE and analysed by LC-MS/MS. Initially, the secretome at day 3 of culture was analyzed. A SWATH-based quantitative differential proteomic analysis of the L-PRF secretome over time was also carried out.
Results: Overall, 705 proteins were identified in the secretome of L-PRF membranes after 3 days of culture, including growth factors and proteins related to platelet and neutrophil degranulation. Clathrin-mediated endocytosis was the top canonical pathway related to the proteins identified. Moreover, 202 differentially secreted proteins were quantified by SWATH when comparing secretomes at days 3, 7 and 21. The majority of them (187) were enriched at day 3. Some proteins that were also found in the initial qualitative analysis (CO3, TSP1 and MMP9) were validated by western blotting confirming an up-regulation at day 3. Other proteins such as fibrinogen and cathepsin S, which increased over time, were also validated corroborating the proteomic results. A quantitative growth factor analysis showed higher levels of PDGFA and EGF at day 3 versus day 7.
Conclusions: In summary, this is the most detailed proteome analysis of the L-PRF secretome to date. Proteins and growth factors identified, and their kinetics, provide novel information to further understand the wound healing properties of L-PRF.
To cite this abstract in AMA style:Hermida Nogueira L, Barrachina MN, Morán LA, Bravo S, Diz P, Blanco J, García Á. Deciphering the Protein Secretome of Leukocyte-platelet Rich Fibrin [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/deciphering-the-protein-secretome-of-leukocyte-platelet-rich-fibrin/. Accessed January 27, 2022.
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