Investigation of the Viability, Adhesion, and Migration of Human Fibroblasts in a Hyaluronic Acid/Gelatin Microgel‐Reinforced Composite Hydrogel for Vocal Fold Tissue Regeneration

Abstract

The potential use of a novel scaffold biomaterial consisting of crosslinked hyaluronic acid (HA)–gelatin (Ge) composite microgels was investigated for use in treating vocal fold injury and scarring. Cell adhesion integrins and kinematics of cell motion were investigated in two- and three-dimensional culture conditions, respectively. Human vocal fold fibroblast (hVFF) cells were seeded on HA–Ge microgels attached to a HA hydrogel thin film. The results showed that hVFF cells established effective adhesion to HA–Ge microgels through the ubiquitous expression of β1 integrin in the cell membrane. The microgels were then encapsulated in a three-dimensional HA hydrogel for the study of cell migration. The cells within the HA–Ge microgel-reinforced composite hydrogel (MRCH) scaffold had an average motility speed of 0.24±0.08 μm/minute. The recorded microscopic images revealed features that are presumably associated with lobopodial and lamellipodial cell migration modes within the MRCH scaffold. Average cell speed during lobopodial migration was greater than that during lamellipodial migration. The cells moved faster in the MRCH than in the HA–Ge gel without microgels. These findings support the hypothesis that HA–Ge MRCH promote cell adhesion and migration; thereby they constitute a promising biomaterial for vocal fold repair.

Gelatin‐polysaccharide composite scaffolds for 3D cell culture and tissue engineering: Towards natural therapeutics

“…Thiolated HA (3,3‐dithiobis‐[propanoic dihydrazide]) or thiol‐carboxymethyl HA can be crosslinked to form a hydrogel by the addition of PEG diacrylate. These composites have been used as injectable scar‐free 3D cell scaffolds for wound healing or as 3D cell culture scaffolds . Cyto‐adhesiveness of the material can be increased by co‐crosslinking with gelatin, modified with thiol groups, yielding a gel with tripeptide Arg‐Gly‐Asp (RGD) motifs for binding integrins on cell surfaces .…”

“…These composites have been used as injectable scar-free 3D cell scaffolds for wound healing or as 3D cell culture scaffolds. [247][248][249] Cyto-adhesiveness of the material can be increased by co-crosslinking with gelatin, modified with thiol groups, yielding a gel with tripeptide Arg-Gly-Asp (RGD) motifs for binding integrins on cell surfaces. 250 Moreover, HA can be chemically modified to acquire hydrophobic properties, which may be homogeneously mixed with gelatin to form hydrophobic-hydrophilic mixed gels used as 3D scaffolds for the chondrogenic differentiation of MSCs.…”

Tissue engineering-based therapeutic strategies for vocal fold repair and regeneration

“…[123] These microparticle-reinforced hydrogels promoted hVFF adhesion, spreading, proliferation and exerted enhanced cell migration (average motility speed: 0.24 μm/min) in the external HA network as a result of controlling cellular adhesion at the interface of densely-crosslinked microgels, a phenomenon not observed in HA-Gtn composites lacking microgels. [124]…”

A Review of Hyaluronic Acid and Hyaluronic Acid-based Hydrogels for Vocal Fold Tissue Engineering

“…Microgels made with thiolated HA-gelatin and reinforced to a composite hydrogel 76 show better adhesion and migration of vocal fold fibroblasts on these scaffolds in comparison with HA-gelatin scaffolds without the reinforced microgels. Carbylan-GSX, along with carboxymethylated HA (Extracel), has also been used as a delivery vehicle for combined therapy with bone marrow-mesenchymal stem cells embedded in the hydrogel.…”