Design of modular gellan gum hydrogel functionalized with avidin and biotinylated adhesive ligands for cell culture applications

Abstract

This article proposes the coupling of the recombinant protein avidin to the polysaccharide gellan gum to create a modular hydrogel substrate for 3D cell culture and tissue engineering. Avidin is capable of binding biotin, and thus biotinylated compounds can be tethered to the polymer network to improve cell response. The avidin is successfully conjugated to gellan gum and remains functional as shown with fluorescence titration and electrophoresis (SDS-PAGE). Self-standing hydrogels were formed using bioamines and calcium chloride, yielding long-term stability and adequate stiffness for 3D cell culture, as confirmed with compression testing. Human fibroblasts were successfully cultured within the hydrogel treated with biotinylated RGD or biotinylated fibronectin. Moreover, human bone marrow stromal cells were cultured with hydrogel treated with biotinylated RGD over 3 weeks. We demonstrate a modular and inexpensive hydrogel scaffold for cell encapsulation that can be equipped with any desired biotinylated cell ligand to accommodate a wide range of cell types.

3D printing of gellan‐dextran methacrylate IPNs in glycerol and their bioadhesion by RGD derivatives

“…Despite its interesting properties, unmodified Ge is a relatively bioinert material, showing a weak ability to support cell adhesion that limits its applications to anchorage‐independent cells 39,50 . To address this restriction and make it an optimal substitute for ECM for anchorage‐dependent cells, Ge must be modified 51,52 . Recently, Ge derivatization with RGD‐containing peptides has emerged as a highly effective strategy to enhance cell‐biomaterial interactions, by exploiting the affinity of integrins for the arginine‐glycine‐aspartic acid sequence 53–55 .…”

“…39,50 To address this restriction and make it an optimal substitute for ECM for anchorage-dependent cells, Ge must be modified. 51,52 Recently, Ge derivatization with RGD-containing peptides has emerged as a highly effective strategy to enhance cellbiomaterial interactions, by exploiting the affinity of integrins for the arginine-glycine-aspartic acid sequence. [53][54][55] In this work, a dextran methacylate (DexMa)/gellan-RGD (GeRGD)-based IPN system was developed using glycerol as a non-conventional solvent, which allows both to speed up the kinetic of crosslinking, thanks to its capability to act as a co-initiator in the radical process 32 and to provide the polymeric mixture with enhanced viscoelastic properties that make it suitable as bioink to print different complex scaffolds.…”

Avidin-Conjugated Nanofibrillar Cellulose Hydrogel Functionalized with Biotinylated Fibronectin and Vitronectin Promotes 3D Culture of Fibroblasts

“…36 Also, avidinconjugated gellan gum hydrogel has been prepared and functionalized by biotinylated adhesive ligands such as B-FN and biotinylated cyclic RGD; this was demonstrated to be a promising tool for 3D cell culture studies. 37 Recently, Fernandes et al used avidin and biotinylated multi-arm PEGs to design affinity-triggered hydrogels. The authors showed that avidin and PEG-[biotin] 4 were capable to selfassemble instantaneously, and these hydrogels were used to encapsulate pluripotent stem cells and to induce neural lineage.…”

“…Avidin retained its biotin-binding activity during the 3D printing process . Also, avidin-conjugated gellan gum hydrogel has been prepared and functionalized by biotinylated adhesive ligands such as B-FN and biotinylated cyclic RGD; this was demonstrated to be a promising tool for 3D cell culture studies . Recently, Fernandes et al used avidin and biotinylated multi-arm PEGs to design affinity-triggered hydrogels.…”