Enhanced elastin synthesis and maturation in human vascular smooth muscle tissue derived from induced-pluripotent stem cells.

Obtaining vascular smooth muscle tissue with mature, functional elastic fibers is a key obstacle in tissue-engineered blood vessels. Poor elastin secretion and organization leads to a loss of specialization in contractile smooth muscle cells, resulting in over proliferation and graft failure. In this study, human induced-pluripotent stem cells (hiPSCs) were differentiated into early smooth muscle cells, seeded onto a hybrid poly(ethylene glycol) dimethacrylate/poly (l-lactide) (PEGdma-PLA) scaffold and cultured in a bioreactor while exposed to pulsatile flow, towards maturation into contractile smooth muscle tissue. We evaluated the effects of pulsatile flow on cellular organization as well as elastin expression and assembly in the engineered tissue compared to a static control through immunohistochemistry, gene expression and functionality assays. We show that culturing under pulsatile flow resulted in organized and functional hiPSC derived smooth muscle tissue. Immunohistochemistry analysis revealed hiPSC-smooth muscle tissue with robust, well-organized cells and elastic fibers and the supporting microfibril proteins necessary for elastic fiber assembly. Through qRT-PCR analysis, we found significantly increased expression of elastin, fibronectin, and collagen I, indicating the synthesis of necessary extracellular matrix components. Functionality assays revealed that hiPSC-smooth muscle tissue cultured in the bioreactor had an increased calcium signaling and contraction in response to a cholinergic agonist, significantly higher mature elastin content and improved mechanical properties in comparison to the static control. The findings presented here detail an effective approach to engineering elastic human vascular smooth muscle tissue with the functionality necessary for tissue engineering and regenerative medicine applications. STATEMENT OF SIGNIFICANCE: Obtaining robust, mature elastic fibers is a key obstacle in tissue-engineered blood vessels. Human induced-pluripotent stem cells have become of interest due to their ability to supplement tissue engineered scaffolds. Their ability to differentiate into cells of vascular lineages with defined phenotypes serves as a potential solution to a major cause of graft failure in which phenotypic shifts in smooth muscle cells lead to over proliferation and occlusion of the graft. Herein, we have differentiated human induced-pluripotent stem cells in a pulsatile flow bioreactor, resulting in vascular smooth muscle tissue with robust elastic fibers and enhanced functionality. This study highlights an effective approach to engineering elastic functional vascular smooth muscle tissue for tissue engineering and regenerative medicine applications.

Comparative analysis of in vitro oxidative degradation of poly(carbonate urethanes) for biostability screening.

The resistance to oxidation and environmental stress cracking of poly(carbonate urethanes) (PCUs) has generated significant interest as potential replacements of poly(ether urethanes) in medical devices. Several in vitro models have been developed to screen segmented polyurethanes for oxidative stability. High concentrations of reactive oxygen intermediates produced by combining hydrogen peroxide and dissolved cobalt ions has frequently been used to predict long-term oxidative degradation with short-term testing. Alternatively, a 3% H2O2 concentration without metal ions is suggested within the ISO 10993-13 standard to simulate physiological degradation rates. A comparative analysis which evaluates the predictive capabilities of each test method has yet to be completed. To this end, we have utilized both systems to test three commercially available PCUs with low and high soft segment content: Bionate PCU and Bionate II PCUs, two materials with different soft segment chemistries, and CarboSil TSPCU, a thermoplastic silicone PCU. Bulk properties of all PCUs were retained with minor changes in molecular weight and tensile properties indicating surface oxidative degradation in the accelerated system after 36 days. Soft segment loss and surface damage were comparable to previous in vivo data. The 3% H2O2 method exhibited virtually no changes on the surface or in bulk properties after 12 months of treatment despite previous in vivo results. These results indicate the accelerated test method more effectively characterized the oxidative degradation profiles than the 3% H2O2 treatment system. The lack of bulk degradation in the 12-month study also supports the hydrolytic stability of these PCUs.