Glucuronoxylan-based quince seed hydrogel: A promising scaffold for tissue engineering applications

Highlights

• A novel tissue-engineering scaffold was fabricated by using glucuronoxylan-based quince seed hydrogel (QSH).

• QSH was characterized in terms of morphology, crosslinking and swelling capacity.

• QSH scaffold exhibits high biocompatibility and cell viability.

Abstract

Natural gums and mucilages from plant-derived polysaccharides are potential candidates for a tissue-engineering scaffold by their ability of gelation and biocompatibility. Herein, we utilized Glucuronoxylan-based quince seed hydrogel (QSH) as a scaffold for tissue engineering applications. Optimization of QSH gelation was conducted by varying QSH and crosslinker glutaraldehyde (GTA) concentrations. Structural characterization of QSH was done by Fourier Transform Infrared Spectroscopy (FTIR). Furthermore, morphological and mechanical investigation of QSH was performed by Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). The protein adsorption test revealed the suitability of QSH for cell attachment. Biocompatibility of QSH was confirmed by culturing NIH-3T3 mouse fibroblast cells on it. Cell viability and proliferation results revealed that optimum parameters for cell viability were 2 mg mL⁻¹ of QSH and 0.03 M GTA. SEM and DAPI staining results indicated the formation of spheroids with a diameter of approximately 300 μm. Furthermore, formation of extracellular matrix (ECM) microenvironment was confirmed with the Collagen Type-I staining. Here, it was demonstrated that the fabricated QSH is a promising scaffold for 3D cell culture and tissue engineering applications provided by its highly porous structure, remarkable swelling capacity and high biocompatibility.

Introduction

In recent years, hydrogels emerged as promising scaffold materials in 3D cell culture and tissue engineering field, since they are closely mimicking ECM structure and function [[1], [2], [3]]. Both natural and synthetic polymers have been developed and used in tissue engineering applications [1,[4], [5], [6], [7], [8], [9]]. However synthetic polymers often exhibit toxic and non-biocompatible properties, while natural polymers provide more suitable microenvironment for cellular studies [5,10]. Polysaccharides are one of the biggest clusters of natural polymers and an example of natural polymers with the hydrogel forming ability. They are widely used in food industry [[11], [12], [13]] as; gelling agents, stabilizers, emulsifiers, thickeners, and in medical/pharmaceutical industry [[14], [15], [16]] as; gelling agents, coatings, biofilms, and as a wound dressing. Their biocompatibility, biological activity, biodegradability, and hydrogel forming ability make them a valuable biomaterial for tissue engineering [5,[17], [18], [19]]. Alginate [20], cellulose [21], hyaluronic acid [22], chitosan [23], xanthan gum [15], guar gum [15], carrageenan [5,15], dextran [15], and gellan gum [15,17] are the most abundant polysaccharide types that are used as a scaffold in tissue engineering.

Plants seeds are one of the most common sources of plant-based polysaccharides. They contain high-molecular weight polysaccharides that form hydrocolloids [24,25], which are usually water-soluble. Gelation ability makes hydrocolloids suitable materials for varied applications in food, pharmaceutical, and medical industry. As biomaterials, hydrocolloids demonstrate excellent properties such as; high swelling capacity, bioactivity, biocompatibility, biodegradability, having antioxidant and anti-inflammatory features [15]. Quince (Rosaceae family) is a small, yellow fruit that is native to West Asia and Middle East regions, and it has been heavily used in traditional medicine as well as in pharmaceutical industry. Quince seed is an important source of hydrocolloid/hydrogel that is composed of mostly cellulose and hydrolysable polysaccharides such as glucuronoxylan [[26], [27], [28]], unsaturated fatty acids [29] and amino acids [29]. Quince seed hydrocolloid is richer in terms of polysaccharide content and has a higher molecular weight [11] than other commercial hydrocolloids like gellan gum [30], xanthan gum [31], guar gum [32], and locust bean gum [33]. Polysaccharide-rich QSH has outstanding mechanical and biological properties that make it a valuable and potential source as a biomaterial for medical applications. Although there are numerous studies on structural, physicochemical and mechanical properties of QSH [11,24,34], it's potential as a biomaterial has been under-evaluated and it has not been utilized in tissue engineering yet.

In this study, we aim to demonstrate the potential of QSH as a novel scaffold for tissue engineering application. For this aim, the gelation capacity of water-extracted QSH and its crosslinking ability with GTA was investigated. Chemical, structural and morphological characterization of QSH was done through ATR-FTIR, SEM and AFM analysis, respectively. Furthermore, cytotoxicity, cell viability, and proliferation were investigated through NIH-3T3 fibroblast cell culture to evaluate biocompatibility of QSH. We demonstrated the potential of QSH as a scaffold for 3D cell culture and tissue engineering applications.