Abstract 233: Quantification of Histological and Biomechanical Properties of Tissue-Engineered Vascular Graft in Inferior Vena Cava

Abstract

Background: We developed the first tissue engineered vascular graft (TEVG) for use in congenital heart surgery and confirmed its significant potential via an initial clinical trial (Shinoka et al., JTCVS 2005). The primary complication of TEVG at medium term was stenosis attributed to suboptimal neovessel remodeling, which necessitated an investigation of the mechanics and mechanobiology to predict diverse aspects of the neovessel formation. Here we present the novel tools to investigate the evolving biomechanical properties of TEVG in a mouse inferior vena cava (IVC) replacement model. Hypothesis: The biomechanical properties of a TEVG evolve nonlinearly in time from a construct-dominated stiffness to a neovessel dominated stiffness. Novel biaxial biomechanical tests are required to quantify such changes rigorously. Method: Thirty-six CB17 SCID/beige mice were implanted with TEVG (PGA + P[CL/LA]) as IVC interposition graft. Twelve tissue engineered neovessels were harvested at 2, 6, and 12 weeks after implantation. Neovessels were characterized biomechanically using a custom, computer-controlled biaxial testing device, and compared with native veins. Extracellular matrix (ECM) remodeling of the neovessel was characterized histologically, biochemically, and molecular biologically. Result: The biaxial data revealed an improved compliance of the neovessel over time. Similarly, the axial stretch response of the TEVG became more like the native vein at 12 weeks. This response was quantified as a ratio of neo-vessel to native vein and improved to reach 52% at 12 weeks from 11% at 2 weeks. Scaffold mass in vivo, estimated by remaining scaffold on histology, showed a significant decline at 6 and 12 weeks. Gene expression of both type I and III collagen peaked at 2 weeks, and total collagen mass quantified by a Sircol™ assay revealed that total collagen peaked at 2 weeks but decreased gradually to the level of native vein. Gene expression of both tropoelastin and fibrillin-1 peaked at 2 weeks, whereas elastin mass quantified by Fastin™ assay showed a delayed peak level of elastin at 6 weeks. Gene expression of MMP-2 and 9 peaked at 6 and 2 weeks, respectively, implicating robust ECM remodeling at early time points. Conclusion: Our novel histo-mechanical approach is the first to show that neovessel formation is a dynamic process characterized by progressive degradation of the scaffold and increased ECM remodeling, which yields biomechanical properties of the TEVG similar to native vessel within 12 weeks in vivo.