title: In vitro method for real-time, direct observation of cell-vascular graft interactions under simulated blood flow.
abstract: In the development of engineered vascular grafts, assessing the material's interactive properties with peripheral blood cells and its capacity to endothelialize are important for predicting in vivo graft behavior. Current in vitro techniques used for characterizing cell adhesion at the surface of engineered scaffolds under flow only facilitate a terminal quantification of cell/surface interactions. Here, we present the design of an innovative flow chamber for real-time analysis of blood-biomaterial interactions under controllable hemodynamic conditions. Decellularized human umbilical veins (dHUV) were used as model vascular allografts to characterize platelet, leukocyte, and endothelial cell (EC) adhesion dynamics. Confluent EC monolayers adhered to the lumenal surface of the grafting material were flow conditioned to resist arterial shear stress levels (up to 24 dynes/cm(2)) over a 48 h period, and shown to maintain viability over the 1 week assessment period. The basement membrane was imaged while whole blood/neutrophil suspensions were perfused across the HUV surface to quantify cell accumulation. This novel method facilitates live visualization of dynamic events, including cell adhesion, migration, and morphological adaptation at the blood-graft interface on opaque materials, and it can be used for preliminary assessment of clinically relevant biomaterials before implantation.
authors.affiliation: J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida , Gainesville, Florida.
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Uzarski
authors.forename: Aurore B
authors.initials: AB
authors.lastname: Van de Walle
authors.forename: Peter S
authors.initials: PS
authors.lastname: McFetridge
mesh_terms: D000328:Adult; D001775:Blood Circulation / Q000502:physiology*; D001807:Blood Vessel Prosthesis; D002450:Cell Communication; D048430:Cell Shape; D002478:Cells, Cultured; D003199:Computer Systems; D018922:HL-60 Cells; D018056:Hemorheology; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000648:ultrastructure; D006801:Humans; D036904:Leukocyte Rolling; D008027:Light; D009504:Neutrophils / Q000166:cytology; D010973:Platelet Adhesiveness; D010974:Platelet Aggregation; D033081:Shear Strength; D059008:Time-Lapse Imaging; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds