The following research examines the creation of a new small-diameter bypass graft. It is clinically relevant to patients who need distal arterial bypass, coronary artery bypass, or hemodialysis access, but who do not have adequate autologous vein for their surgeries. Future investigations will involve further tissue engineering of this vascular scaffold (eg, autologous endothelial seeding of its lumen) and testing the clinical usefulness of the completed graft.

No abstract

Physical and biochemical forces exist that are necessary for the persistent attachment and function of ECs on native and prosthetic blood vessels. The optimization of conditions that permit regeneration of these attachment forces may allow rapid establishment of a durable, biocompatible EC monolayer. We examined the effects of three major factors, protein substrate, EC incubation time, and shear stress, on the attachment kinetics of human adult ECs to two different polymers. ECs were incubated up to 30 minutes on polymers (PS or PET) coated with extracellular matrix proteins: collagen I/III, fibronectin, collagen IV/V, laminin, gelatin, or saline control. After incubation, continued attachment in the presence of shear stress (created in a rotating disc device) between zero and 90 dynes/cm2 for 30 minutes was evaluated. Maximal adherence was observed on all substrates by 30 minutes. Therefore, after a 30-minute incubation, the percentage of cells attached (postshear ECs/preshear ECs/preshear ECs X 100) was measured as a function of shear stress. ECs attached to a matrix of fibronectin or collagen I/III demonstrated shear-resistant adherence after as little as 5 minutes of static incubation before initial shear exposure. By 30 minutes, more than 90% of the ECs on both matrices demonstrated the ability to remain attached in the presence of 90 dynes/cm2 of shear stress. We conclude that forces that attach ECs to surfaces are affected by temporal factors (incubation time) and substrate composition and may be quantified with a defined shear stress detachment assay. Understanding and manipulating these temporal physiochemical parameters should allow one to re-create an optimal EC monolayer on a blood-contacting surface.

No abstract

The purpose of the present study was to culture human adult endothelial cells (HAECs) on a long-term basis in the laboratory. Previous inability to accomplish this has been the major impediment to the in vitro study of endothelialization of prosthetic grafts with human cells, a problem of significant clinical relevance. We have been successful in developing a technique that allows HAECs from human adult arteries, veins, and capillaries to proliferate vigorously in culture for up to 80 population doublings. HAECs are grown on a gelatin surface (medium 199 containing 20% fetal calf serum). Heparin and endothelial cell growth factor (ECGF) are required for optimal growth. With this technique, which will be described in detail, over 10(23) HAECs can be produced from each 1 cm2 of vascular tissue. This makes large numbers of HAECs available for high-density seeding on prosthetic grafts prior to implantation. It also permits for the first time with human cells the in vitro study of prosthetic grafts--HAEC interactions and the factors that enhance optimal growth and adherence to prosthetic materials. It is hoped that identification of the factors promoting graft endothelialization in combination with high-density seeding will lower graft thrombogenicity and therefore result in greater graft longevity than has been possible heretofore.