Hyaluronan Hydrogels for a Biomimetic Spongiosa Layer of Tissue Engineered Heart Valve Scaffolds.

Advanced tissue engineered heart valves must be constructed from multiple materials to better mimic the heterogeneity found in the native valve. The trilayered structure of aortic valves provides the ability to open and close consistently over a full human lifetime, with each layer performing specific mechanical functions. The middle spongiosa layer consists primarily of proteoglycans and glycosaminoglycans, providing lubrication and dampening functions as the valve leaflet flexes open and closed. In this study, hyaluronan hydrogels were tuned to perform the mechanical functions of the spongiosa layer, provide a biomimetic scaffold in which valve cells were encapsulated in 3D for tissue engineering applications, and gain insight into how valve cells maintain hyaluronan homeostasis within heart valves. Expression of the HAS1 isoform of hyaluronan synthase was significantly higher in hyaluronan hydrogels compared to blank-slate poly(ethylene glycol) diacrylate (PEGDA) hydrogels. Hyaluronidase and matrix metalloproteinase enzyme activity was similar between hyaluronan and PEGDA hydrogels, even though these scaffold materials were each specifically susceptible to degradation by different enzyme types. KIAA1199 was expressed by valve cells and may play a role in the regulation of hyaluronan in heart valves. Cross-linked hyaluronan hydrogels maintained healthy phenotype of valve cells in 3D culture and were tuned to approximate the mechanical properties of the valve spongiosa layer. Therefore, hyaluronan can be used as an appropriate material for the spongiosa layer of a proposed laminate tissue engineered heart valve scaffold.

Electrospun Polyurethane and Hydrogel Composite Scaffolds as Biomechanical Mimics for Aortic Valve Tissue Engineering.

In this study, a composite scaffold consisting of an electrospun polyurethane and poly(ethylene glycol) hydrogel was investigated for aortic valve tissue engineering. This multilayered approach permitted the fabrication of a scaffold that met the desired mechanical requirements while enabling the 3D culture of cells. The scaffold was tuned to mimic the tensile strength, anisotropy, and extensibility of the natural aortic valve through design of the electrospun polyurethane mesh layer. Valve interstitial cells were encapsulated inside the hydrogel portion of the scaffold around the electrospun mesh, creating a composite scaffold approximately 200 µm thick. The stiffness of the electrospun fibers caused the encapsulated cells to exhibit an activated phenotype that resulted in fibrotic remodeling of the scaffold in a heterogeneous manner. Remodeling was further explored by culturing the scaffolds in both a mechanically constrained state and in a bent state. The constrained scaffolds demonstrated strong fibrotic remodeling with cells aligning in the direction of the mechanical constraint. Bent scaffolds demonstrated that applied mechanical forces could influence cell behavior. Cells seeded on the outside curve of the bend exhibited an activated, fibrotic response, while cells seeded on the inside curve of the bend were a quiescent phenotype, demonstrating potential control over the fibrotic behavior of cells. Overall, these results indicate that this polyurethane/hydrogel scaffold mimics the structural and functional heterogeneity of native valves and warrants further investigation to be used as a model for understanding fibrotic valve disease.