Gelatin-Based Electrospun Fibers Insolubilized by Horseradish Peroxidase-Catalyzed Cross-Linking for Biomedical Applications

Abstract

Gelatin-based water-insoluble nanofibers with a diameter of 160 nm were obtained from electrospinning aqueous solutions containing gelatin with phenolic hydroxyl (Ph) moieties (Gelatin-Ph) and horseradish peroxidase (HRP). The water insolubility of the nanofibers was accomplished through HRP-catalyzed cross-linking of the Ph moieties by exposing the electrospun nanofibers to air containing hydrogen peroxide. The HRP activity in the electrospun nanofibers was 65% that of native HRP. The cytocompatibility necessary for tissue engineering applications of the water-insoluble Gelatin-Ph nanofibers was confirmed by the adhesion and viability of human embryonic kidney-derived HEK293 cells.

Layered dermal reconstitution through epigallocatechin 3-gallate loaded chitosan nanoparticle within enzymatically crosslinked polyvinyl alcohol/collagen fibrous mat

“…The H 2 O 2 molecules from saturated air could be mildly diffused into fibrous structure and provide uniform crosslinking without specific concerns for shape deformation and liquefaction of electrospun fibers before proceeding of covalent crosslinking of Ph moieties to each other and stable solidification of fibers. 28 The usefulness of HRP-catalyzed crosslinking of polymers with Ph moieties (polymer-Ph) in an aqueous solution for obtaining stable 3D construct in bulk gels is well recognized. 26,29,30 Polymers possessing Ph moieties enzymatically crosslinked and hydrogelated in presence of HRP& H 2 O 2 .…”

Effectiveness of insolubilized polyvinyl alcohol-based electrospun fibers loaded methylprednisolone by enzyme-catalyzed cross-linking in spinal cord regeneration

“…The covalent crosslinking using chemicals such as glutaraldehyde and carbodiimide crosslinking are not safe, and the rate of crosslinking is too low [9,11,21,44]. Recently, Sakai et al reported the feasibility of brous mat fabrication through HRP-mediated crosslinking for GelaPh within minutes, that proved viability of corresponding approach for other biopolymer development and application for biomedical elds [31]. Therefore, here we have attempted to synthesize Ph conjugated PVA and Gela as an electrospinning media and then crosslinked their molecules through HRP-mediated crosslinking using vaporized H 2 O 2 as an electron donor.…”

“…Meanwhile, addition of GelaPh in higher concentration improved tensile strength in lower degree due to reduction of PVAPh crosslinking and correspondingly higher GelaPh crosslinking in the hydrogel structure since Gela inherently possessed a weak mechanical strength [11,23,31]. The PVAPh brous mat extended without breaking up to 183% strain, which was highest among brous mats (Fig.…”

“…Such hydrogels were obtained by adding an H2O2 aqueous solution to that of polymer-Phs and HRP [24,29,30]. However, to the best of our knowledge fabrication of brous mats through HRP-mediated reaction is not developed well for most of biopolymers [31][32][33].…”

Phenolated alginate-collagen hydrogel induced chondrogenic capacity of human amniotic mesenchymal stem cells

“…34 Among the wide range of crosslinking enzymes, Horseradish peroxidase (HRP) has been widely used to catalyze polymers with phenolic hydroxyl (Ph) moieties using hydrogen peroxide (H 2 O 2 ) as an oxidant under mild conditions. 35–38 The advantages of using HRP for biological applications include fast gelation rate, having moderate substrate specificity, manageable gelation rate, and crosslinking density. 39–41 Besides carbodiimide activation chemistry which is used for alginate phenolation has been shown to have a variety of advantages, such as enhancement of osteo-compatibility, 42 cellular attachment, 43 and muco-adhesiveness.…”