Harnessing topographical & biochemical cues to enhance elastogenesis by paediatric cells for cardiovascular tissue engineering applications

Abstract

The development of tissue-engineered vascular grafts (TEVGs) with a biomimetic extracellular matrix (ECM) structure, including a mature elastic network, remains a key challenge for the production of grafts with long-term functionality. The aim of this study was to investigate the influence of aligned nanofiber substrates on ECM protein synthesis by neonatal smooth muscle cells (SMCs), and to examine the combined effects of this topographical cue in conjunction with transforming growth factor beta-1 (TGF-β1) - a biochemical elastogenic promoter. Glass coverslips were coated in electrospun fibrinogen nanofibers (average diameter < 500 nm) with either a randomly-orientated or aligned topography. Human umbilical artery smooth muscle cells (hUASMCs) were cultured on the electrospun substrates for 7 and 14 days, with or without a 2 ng/ml TGF-β1 supplement. The ECM structure was analysed using immunohistochemistry and the quantity of secreted elastin in the cell layer was measured using a dye-binding assay.Aligned fiber substrates induced a directed orientation of both the seeded cells and cell-synthesized ECM fibers. Cells cultured on aligned fibers exhibited a significant increase in the expression of phenotypic contractile proteins, as well as increases in the secreted elastin content of the cell layer, compared to cells cultured on randomly-orientated substrates. TGF-β1 supplementation was shown to synergistically increase secreted elastin from cells cultured on aligned fiber substrates (p < 0.05). Aligned nanofiber scaffolds can be used to direct cellular orientation, elastin-related protein synthesis and cell phenotype, and consequently there is potential for their application in the development of TEVGs as part of a multi-pronged strategy to promote elastic fiber formation.