Architectural organization and functional features of early endothelial progenitor cells cultured in a hyaluronan-based polymer scaffold.
Neovascularization can be improved using polymer scaffolds supporting endothelial progenitor cells (EPCs). The aim of the present study was to investigate whether human early EPCs (eEPCs) could be efficiently cultured in a hyaluronan-based non-woven mesh (HYAFF-11). eEPCs were seeded on HYAFF-11 at the density of 1 x 10(6)/cm(2) and cultured with endothelial differentiating factors for 3 weeks. After 24 h, nearly 90% of EPCs were adherent. Cell viability, evaluated by methyltetrazolium test, was greater in HYAFF-11 than on the most commonly used fibronectin-coated dishes, even if a progressive decline in viability was observed starting from approximately the second week of culture. eEPCs easily migrated to and aggregated on the scaffold. Evidence of active protein synthesis and features of endothelial differentiation, including cellular transcytotic channels and micropinocytotic vesicles, was revealed using electron microscopy, immunofluorescence, and reverse transcriptase polymerase chain reaction analysis. eEPCs cultured in the scaffold also showed a certain angiogenic activity, as demonstrated by hepatocyte growth factor transcription and vascular endothelial growth factor secretion. In conclusion, eEPCs can migrate and adhere inside HYAFF-11, maintain their pre-endothelial phenotype, and express angiogenic factors, especially within the first week of growth. These results indicate that non-woven HYAFF-11 could be a promising candidate as a vehicle for eEPCs for regenerative medicine applications.
Division of Clinical Pathology, Department of Radiological and Histocytopathological Sciences, University of Bologna, Bologna, Italy.
Gianandrea
G
Pasquinelli
Maria Cristina
MC
Vinci
Chiara
C
Gamberini
Catia
C
Orrico
Laura
L
Foroni
Carlo
C
Guarnieri
Astrid
A
Parenti
Mauro
M
Gargiulo
Fabrizio
F
Ledda
Claudio Marcello
CM
Caldarera
Claudio
C
Muscari
D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006801:Humans; D006820:Hyaluronic Acid / Q000494:pharmacology*; D008855:Microscopy, Electron, Scanning; D011108:Polymers / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000648:ultrastructure; D054457:Tissue Scaffolds / Q000737:chemistry*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D015415:Biomarkers; D011108:Polymers; D042461:Vascular Endothelial Growth Factor A; D006820:Hyaluronic Acid