ABSTRACT
A method for the creation of a type IV collagen (CNIV) scaffold on polytetrafluoroethylene (PTFE) for endothelial cell attachment is described. This mimic for the basal lamina can be used in the seeding and retention of endothelial cells for blood contacting devices. The CNIV-PTFE production technique can be defined as three processes: (i) creation of a reactive superacidic/ionic PTFE surface with retained hydrophobic characteristics; (ii) activation of this surface via covalent attachment of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC); and (iii) conjugation of the EDC with human CNIV resulting in the covalent binding of protein to the PTFE surface. This article demonstrates exciting new results showing that a reaction of CNIV to this particular surface results in a unique matrix assembly of CNIV scaffolds similar to those found in the basal lamina. This assembly is concentration-dependant, occurring in a narrow window around 0.435 microM. Following the fabrication of the CNIV matrix assembly, porcine aortic endothelial cells (PAEC) were seeded onto this material. Results described in this article demonstrate that the PAEC subsequently aligned with the direction of shear, filled voids created by dead or detached cells, and divided during the 6 h of experimentation. Under static conditions, cells remained viable for 1 week of testing. This was not observed with PAEC attached to glass with adsorbed Vitrogen. In summary, this article describes a novel biotechnological breakthrough that enables the creation of stable endothelial cell monolayers useful for fabricating blood contacting devices.
