The Effect of Pulsatile Loading and Scaffold Structure for the Generation of a Medial Equivalent Tissue Engineered Vascular Graft

摘要

A reliable and cost-effective scaffold for tissue-engineered vascular graft that would not only support cell proliferation and growth but also maintain cell phenotype has been a long-term challenge. In this study, we propose a biodegradable and biomimetic copolymer of gelatin with vinyl acetate synthesized via a graft copolymerization technique to generate tubular scaffolds for vascular tissue engineering. Two fabrication techniques, freeze drying and electrospinning, were used to generate the differing architectures for the scaffolds and characterized. The electrospun scaffolds were found to have a faster rate of mass loss in physiological saline of 81.72% within 4 months compared with 60% mass loss for the freeze-dried samples, though the materials were more crystalline. Vascular (v) smooth muscle cells (SMCs) were seeded on these tubes, which were then subjected to dynamic pulsatile stimulation on a vascular bioreactor for a week. Gross examination of the tissue-engineered constructs revealed that the cells secreted extensive extracellular matrix, with the dynamically conditioned samples exhibiting well-orientated SMCs and collagenous fibers in comparison with growth in static conditions. In addition, the alignment of cells in the direction of strain was greater in the electrospun constructs. The electrospun scaffolds maintained the characteristic contractile phenotype of SMCs, which was confirmed by higher gene expression rates of contractile protein markers like SM22α and calponin. A significant increase in the total matrix components (collagen and elastin) in the electrospun constructs compared with the freeze-dried samples was confirmed by biochemical analysis. The results of this study indicate that a combination approach involving a biomimetic scaffold with the nanofibrillar architecture and good mechanical strength conducive to the growth of SMCs and the use of the pulsatile forces to modulate the cell morphology and phenotypic plasticity of vSMCs helps in the successful engineering of a medial layer of blood vessel.