Abstract
Objective: The purpose of this study was to test the hypothesis that explanted perfused arteries can serve as the initial endothelial cell culture source to evaluate the onset of angiogenesis in a cellulose acetate electrospun scaffold.
Methods: Electrospun scaffolds with fiber diameters roughly controlled in three broad ranges: 0.01 to 0.2, 0.2 to 1, and 1 to 5 μ m (Nanomed Nanotechnol Biol Med 2:37–41, 2006), were used in cell culture to determine which provides the best culture topology. This scaffold was then tested in a bioassay chamber whose cellular source was an explanted abdominal aorta from donated euthanized mice. Scaffolds were draped over a cannulated vessel perfused for 24 h. Cell viability, density, and morphology were quantified.
Results: The largest fiber diameter group provided the best culture topology for human umbilical vein endothelial cells, showing high cell viability and density, and enhanced elongated cell morphology. Addition of single-walled carbon nanotubes decreased cell density significantly but chitosan heightened cell density and promoted spontaneous capillary tube like structure. Viability of endothelial cells increased with higher flow in the bioassay chamber.
Conclusions: Endothelial cells showed a growth preference towards larger diameter fibers. Addition of chitosan improved culture conditions. Thus, this study provides a proof of principle for the possibility of co-culturing tissue engineered vascular networks from a perfused explant.
This work was supported by the following grants: NIH HL55492 (to M.F.), NIGMS R25-GM62492 (NIH MARC to S.G.), and T34-GM08655 (to S.G.). The authors thank Dr. Martha Furie (Department of Pathology, Stony Brook University) for the generous gift of HUVECs.