TY - JOUR
T1 - Exosomes
T2 - Novel effectors of human platelet lysate activity
AU - Torreggiani, E.
AU - Perut, F.
AU - Roncuzzi, L.
AU - Zini, N.
AU - Baglìo, S. R.
AU - Baldini, N.
PY - 2014
Y1 - 2014
N2 - Despite the popularity of platelet-rich plasma (PRP) and platelet lysate (PL) in orthopaedic practice, the mechanism of action and the effectiveness of these therapeutic tools are still controversial. So far, the activity of PRP and PL has been associated with different growth factors (GF) released during platelet degranulation. This study, for the first time, identifies exosomes, nanosized vesicles released in the extracellular compartment by a number of elements, including platelets, as one of the effectors of PL activity. Exosomes were isolated from human PL by differential ultracentrifugation, and analysed by electron microscopy and Western blotting. Bone marrow stromal cells (MSC) treated with three different exosome concentrations (0.6 μg, 5 μg and 50 μg) showed a significant, dose-dependent increase in cell proliferation and migration compared to the control. In addition, osteogenic differentiation assays demonstrated that exosome concentration differently affected the ability of MSC to deposit mineralised matrix. Finally, the analysis of exosome protein content revealed a higher amount of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), plateletderived growth factor (PDGF-BB) and transforming growth factor beta 1 (TGF-β1) as compared to PL. In regards to RNA content, an enrichment of small RNAs in exosomes as compared to donor platelets has been found. These results suggest that exosomes consistently contribute to PL activity and could represent an advantageous nanodelivery system for cell-free regeneration therapies.
AB - Despite the popularity of platelet-rich plasma (PRP) and platelet lysate (PL) in orthopaedic practice, the mechanism of action and the effectiveness of these therapeutic tools are still controversial. So far, the activity of PRP and PL has been associated with different growth factors (GF) released during platelet degranulation. This study, for the first time, identifies exosomes, nanosized vesicles released in the extracellular compartment by a number of elements, including platelets, as one of the effectors of PL activity. Exosomes were isolated from human PL by differential ultracentrifugation, and analysed by electron microscopy and Western blotting. Bone marrow stromal cells (MSC) treated with three different exosome concentrations (0.6 μg, 5 μg and 50 μg) showed a significant, dose-dependent increase in cell proliferation and migration compared to the control. In addition, osteogenic differentiation assays demonstrated that exosome concentration differently affected the ability of MSC to deposit mineralised matrix. Finally, the analysis of exosome protein content revealed a higher amount of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), plateletderived growth factor (PDGF-BB) and transforming growth factor beta 1 (TGF-β1) as compared to PL. In regards to RNA content, an enrichment of small RNAs in exosomes as compared to donor platelets has been found. These results suggest that exosomes consistently contribute to PL activity and could represent an advantageous nanodelivery system for cell-free regeneration therapies.
KW - Bone marrow stromal cells
KW - Cell-free regeneration therapies
KW - Exosomes
KW - Growth factors
KW - Nanodelivery system
KW - Platelet lysate
KW - Platelet richplasma
KW - RNA
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84907818281&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/25241964
UR - http://www.scopus.com/inward/record.url?scp=84907818281&partnerID=8YFLogxK
U2 - https://doi.org/10.22203/eCM.v028a11
DO - https://doi.org/10.22203/eCM.v028a11
M3 - Article
C2 - 25241964
SN - 1473-2262
VL - 28
SP - 137
EP - 151
JO - European Cells and Materials
JF - European Cells and Materials
ER -