Endothelial and smooth muscle cell seeding onto processed ex vivo arterial scaffolds using 3D vascular bioreactors

Biomaterials derived from ex vivo tissues offer a viable alternative to synthetic materials for organ replacement therapies. In this study, we describe the use of a tissue engineering scaffold derived from ex vivo arterial tissue to assess vascular cell adhesion within a three-dimensional perfusion bioreactor. With the aim of maximizing seeding efficiency, five methods for endothelial cell (EC) and three independent methods for vascular smooth muscle cell (VSMC) adhesion were explored. Seeded constructs were maintained in vascular bioreactors under pulsatile flow conditions, culminating at 165 ml/min at 1.33 Hz to validate cell attachment and retention over time. Progressive modification of the seeding and flow regime protocols resulted in an increased of EC retention from 5.1 to 634 cells/mm2. Seeding VSMCs as sheets rather than cell suspensions bound and stabilized surface EC matrix fibers, resulting in multiple cell layers adhered to the scaffold with cells migrating to the medial/adventitial boundary. In conjunction with the bioscaffold, the vascular perfusion system serves as a useful tool to analyze cell adhesion and retention by allowing controlled manipulation of seeding and perfusion conditions.