Fabrication and Characterization of Pectin Hydrogel Nanofiber Scaffolds for Differentiation of Mesenchymal Stem Cells into Vascular Cells

Despite significant progress over the past few decades, creating a tissue-engineered vascular graft with replicated functions of native blood vessels remains a challenge due to the mismatch in mechanical properties, low solution biological function, and rapid occlusion caused by restenosis of small diameter vessel grafts (<6 mm diameter). A scaffold with similar mechanical properties and biocompatibility to the host tissue is ideally needed for the attachment and proliferation of cells to support the building of engineered tissue. In this study, pectin hydrogel nanofiber scaffolds with two different oxidation degrees (25 and 50%) were prepared by a multistep methodology including periodate oxidation, electrospinning, and adipic acid dihydrazide crosslinking. Scanning electron microscopy (SEM) images showed that the obtained pectin nanofiber mats have a nano-sized fibrous structure with 300-400 nm fiber diameter. Physicochemical property testing using Fourier transform infrared (FTIR) spectra, atomic force microscopy (AFM) nanoindentations, and contact angle measurements demonstrated that the stiffness and hydrophobicity of the fiber mat could be manipulated by adjusting the oxidation and crosslinking levels of the pectin hydrogels. Live/Dead staining showed high viability of the mesenchymal stem cells (MSCs) cultured on the pectin hydrogel fiber scaffold for 14 days. In addition, the potential application of pectin hydrogel nanofiber scaffolds of different stiffness in stem cell differentiation into vascular cells was assessed by gene expression analysis. Real-time polymerase chain reaction (RT-PCR) results showed that the stiffer scaffold facilitated the differentiation of MSCs into vascular smooth muscle cells, while the softer fiber mat promoted MSC differentiation into endothelial cells. Altogether, our results indicate that the pectin hydrogel nanofibers have the capability of providing mechanical cues that induce MSC differentiation into vascular cells and can be potentially applied in stem cell-based tissue engineering.