Encapsulation of Corneal Stromal Stem Cells within Supramolecular Host-Guest Hyaluronic Acid Gels

Abstract

Purpose : Corneal stromal stem cells (CSSCs) have shown promise in the treatment of corneal injury and scarring. However, the optimum way to deliver them to the cornea has yet to be determined, and their phenotypic expression in vitro remains an important area of study. Here, we investigate whether a supramolecular host-guest hyaluronic acid (HA) gel may serve as a suitable microenvironment for the culture and delivery of CSSCs based on their growth factor secretions and wound healing effects.

Methods : Live-dead analysis was used to track cell the viability and proliferation of CSSCs within the HA host-guest gel after the passage through a 23g needle. The presence of TGF-beta, TSG-6, FGF-2, keratinocyte growth factor (KGF), and hepatocyte growth factor (HGF) were analyzed using reverse transcription polymerase chain reaction.
To track the effects of the HA host-guest gel with and without CSSCs on corneal wound healing, a rabbit cornea organ culture keratectomy model was performed, where deep stromal wounds were filled with the HA-host guest gel with and without encapsulated CSSCs.

Results : CSSC viability within the HA host-guest gel was found to be over 90%. Growth factor expression was significantly higher in CSSCs grown in the HA host-guest matrix compared to CSSCs grown in a commercial medium on standard 2D culture plates. Human CSSCs within the HA gels remained viable with keratectomy wound sites and enhanced epithelial healing in organ culture compared to either HA host-guest gels alone or no treatment controls. HA host-guest gels alone also improved the rate of epithelialization compared to no treatment controls.

Conclusions : Supramolecular host-guest HA gels show promise as a material to promote corneal wound healing on its own, and as a cytocompatible microenvironment for the culture and delivery of CSSCs that stimulates the production of key growth factors and enhances epithelial wound healing in an organ culture model of deep stromal injury.

This is a 2020 ARVO Annual Meeting abstract.