A small diameter elastic blood vessel wall prepared under pulsatile conditions from polyglycolic acid mesh and smooth muscle cells differentiated from adipose-derived stem cells

Abstract

Smooth muscle layer plays an important role in maintaining homeostasis of blood vessels, thus generating a functional smooth muscle layer is a prerequisite for successful construction of blood vessels via tissue-engineering approach. In this study, we investigated the feasibility of constructing an elastic vessel wall in small diameter (less than 6 mm) using smooth muscle cells (SMCs) differentiated from human adipose-derived stem cells (hASCs) under pulsatile stimulation in a bioreactor. With the induction of transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-4 (BMP4) in combination for 7 days, hASCs were found to acquire an SMC phenotype characterized by the expression of SMC-related markers including smooth muscle alpha actin (α-SMA), calponin, and smooth muscle myosin heavy chain (SM-MHC). The SMCs derived from hASCs were seeded in polyglycolic acid (PGA) unwoven mesh and the cell–scaffold complex were subjected to pulsatile stimulation in a bioreactor for 8 weeks. The vessel walls engineered under the dynamic stimulation for 8 weeks showed a dense and well-organized structure similar to that of native vessels. The differentiated hASCs with dynamic loading were found to maintain their SMC phenotype within 3-dimensional PGA scaffold with a high level of collagen deposition close to that of native ones. Vessels constructed in the static condition showed a loose histological structure with less expression of contractile proteins. More importantly, the engineered vessel under pulsatile stimulation exhibited significant improvement in biomechanical properties over that generated from static conditions. Our results demonstrated that hASCs can serve as a new cell source for SMCs in blood vessel engineering, and an elastic small-diameter vessel wall could be engineered by in vitro culture of SMC-differentiated hASCs on the PGA scaffold with matchable biomechanical strength to that of normal blood vessels under pulsatile stimulation.