Proteomic analysis of mesenchymal stromal cells secretome in comparison to leukocyte- and platelet-rich fibrin

Abstract

Abstract Secretomes of mesenchymal stromal cells (MSC) are emerging as a novel growth factor (GF)-based strategy for periodontal and bone regeneration. The objective of this study was to compare the secretome of human bone marrow MSC (BMSC) to that of leukocyte- and platelet-rich fibrin (L-PRF), an established GF-based therapy, in the context of wound healing and regeneration. Conditioned media from human BMSC (BMSC-CM) and L-PRF (LPRF-CM) were subjected to quantitative proteomic analysis using liquid chromatography with tandem mass spectrometry. Global profiles, gene ontology (GO) categories, differentially expressed proteins (DEPs), and gene set enrichment (GSEA) were identified using bioinformatic methods. Concentrations of selected proteins were determined using a multiplex immunoassay. Among the proteins identified in BMSC-CM (2157 proteins) and LPRF-CM (1420 proteins), 1283 proteins were common. GO analysis revealed similarities between the groups in terms of biological processes (cellular organization, protein metabolism) and molecular functions (cellular-/protein-binding). Notably more DEPs were identified in BMSC-CM (n=550) compared to LPRF-CM (n=118); these included several key GF, cytokines, and extracellular matrix (ECM) proteins involved in wound healing. GSEA revealed enrichment of ECM (especially bone-ECM) related processes in BMSC-CM and immune-related processes in LPRF-CM. Similar trends for inter-group differences in protein detection were observed in the multiplex analysis. Thus, while the secretomes of both BMSC and L-PRF are favourable for wound healing, BMSC-CM is additionally enriched for proteins/processes relevant for periodontal and bone regeneration.