Hydrazone crosslinked hyaluronan-based hydrogels for therapeutic delivery of adipose stem cells to treat corneal defects

Abstract

Corneal blindness is a worldwide problem, plagued by insufficient amount of high-quality donor tissue. Cell therapy using human adipose stem cells (hASCs) has risen as an alternative to regenerate damaged corneal stromal tissue, the main structural and refractive layer of the cornea. Herein we propose a method to deliver hASCs into corneal defects in hyaluronan (HA)-based hydrogels, which form rapidly in situ by hydrazone crosslinking. We fabricated two different HA-based hydrazone-crosslinked hydrogels (HALD1-HACDH and HALD2-HAADH), and characterized their swelling, degradation, mechanical, rheological and optical properties and their ability to support hASC survival. To promote hASC attachment and survival, we incorporated collagen I (col I) to the more stable HALD1-HACDH hydrogel, since the HALD2-HAADH hydrogel suffered swift degradation in culture conditions. We then used an organ culture model with excised porcine corneas to study the delivery of hASCs in these three hydrogels for stromal defect repair. Although all hydrogels showed good hASC survival directly after encapsulation, only the collagen-containing HALD1-HACDH-col I hydrogel showed cells with elongated morphology, and significantly higher cell metabolic activity than the HALD1-HACDH gel. The addition of col I also increased the stiffness and reduced the swelling ratio of the resulting hydrogel. Most importantly, the corneal organ culture model demonstrated these hydrogels as clinically feasible cell delivery vehicles to corneal defects, allowing efficient hASC integration to the corneal stroma and overgrowth of corneal epithelial cells.

Designing Scaffolds for Corneal Regeneration

“…Koivusalo et al demonstrated using a porcine organ culture model that modified hyaluronic acid based hydrogels seeded with adipose derived stem cells could be used to repair damaged cornea without the need for sutures. [151] Hyaluronic acid has also been suggested as a suitable xeno-free substrate for culturing corneal epithelial cells [149] and as a potential carrier for endothelial cells. [163,164] Despite some promising results, one potential limiting factor with using hyaluronic acid for corneal tissue regeneration is that the material has been associated with lymphangiogenesis in the limbus, although this requires further study.…”

Screening of Hydrogels for Human Pluripotent Stem Cell–Derived Neural Cells: Hyaluronan‐Polyvinyl Alcohol‐Collagen‐Based Interpenetrating Polymer Network Provides an Improved Hydrogel Scaffold

“…Aldehyde groups were introduced to HA either by using periodate oxidation (HA1) or selective oxidation of diol‐modified HA (HA2). The syntheses and determinations of the degree of substitution (DS%) of modified HA1 and HA2 components were carried out according to previously reported procedures . Hydrazide groups were introduced to PVA using glycine ethyl ester and hydrazine as a source of the hydrazide unit.…”

Bioengineering Approaches for Corneal Regenerative Medicine

“…Delivered stem cells were integrated into the cornea within 7 days, and the repopulation of epithelial cells was reported within this period. Longer preclinical observations are required to investigate the efficiency of hASCs delivery for treating corneal stromal defects [134].…”