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

Abstract

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 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.

Dual-Crosslinked Oxidized Pectin/N-Succinyl Chitosan Hydrogel Containing Graphene Oxide Nanosheets for Tissue Engineering Application

“…Pectin (PE), which is also called polygalacturonic acid, is an anionic polysaccharide extracted from primary cell plants. Pectin has been widely used in a variety of biomedical applications, due to its important properties such as biocompatibility, biodegradability, capability to inhibiting the activity of macrophages and neutrophils, anti-in ammatory effects, and the capability to restrain cells, drugs, and genes (Li et al, 2019, Kaushik et al, 2020. On the backbone chain, there are hydroxyl, carboxyl and carboxymethyl groups which makes it readily susceptible to functionalization and modi cation.…”

Fabrication of multilayer tubular scaffolds with aligned nanofibers to guide the growth of endothelial cells

“…Electrospinning has been widely used to fabricate scaffolds, because it is easier to blend or mix various materials and build non-delamination layers to develop multi-layered scaffolds. 20–22 This method enables the fabrication of fine polymer fibrous mats composed of fibers whose diameters range from several microns to 100 nm or less. 23 Furthermore, electrospinning is a simple and versatile method to fabricate a 3 D nanofiber scaffold that mimics the nanofibrous native ECM structure.…”

Microfibrous Extracellular Matrix Changes the Liver Hepatocyte Energy Metabolism via Integrins

“…Electrospinning is another technology to fabricate 3D fibrous ECM, which applies a high voltage (kV range) to turn a polymer solution to micro-/nano-fibers with tunable fiber and pore sizes 24 . A myriad of cell types including hepatocytes 25 have been successfully 3D cultured on electrospun fibers showing improved functions [26][27][28][29] .…”