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Karen Chen, Gabriella Maria Fernandes Cunha, Sang Hoon Jung, Sei Kwang Hahn, Geon-Hui Lee, David Myung; Supramolecular guest-host hyaluronic acid hydrogels for epithelial cell delivery to the cornea. Invest. Ophthalmol. Vis. Sci. 2020;61(7):370.
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Restoration of a severely damaged corneal surface requires proper delivery and transplantation of cultured human epithelial cells. Here we investigated the feasibility of a supramolecular host-guest hydrogel assembly as a vehicle to deliver and engraft human epithelial cells to the cornea after total epithelial debridement. The goal was to determine whether the supramolecular gel provides a suitable environment for epithelial cell adhesion and proliferation on the cornea.
Hyaluronic acid (HA) was conjugated with cyclodextrin (CD) or adamantane (Ad). To form the supramolecular hydrogel, human corneal epithelial cells were first mixed with HA-CD and then mixed with HA-Ad in a 1:1 ratio of HA-CD to HA-Ad. To investigate hydrogel cytocompatibility, cells were seeded on the hydrogel and a live dead assay was performed at 4 and 10 days. Total epithelial debridement was performed on ex vivo rabbit corneas and then the gel and encapsulated cells were applied to the damaged corneas. At day 4, the corneas were fixed with PFA 4%, permeabilized and blocked with triton-x 0.5 % and goat serum 5 %, respectively. The tissue was incubated in anti-human nuclei antibody overnight to assess for the presence and adhesion of human epithelial cells to the cornea. Next, a secondary antibody anti-mouse Alexa 546 and DAPI were added and to the corneas and analyzed under confocal microscopy. Damaged corneas that received the encapsulated cells were compared to the corneas without encapsulated cells.
The supramolecular hydrogel was cytocompatible showing very few dead cells after 4 and 10 days. After 4 days, the presence of human epithelial cells on the damaged cornea was confirmed by the anti-human nuclei antibody, suggesting that the encapsulated cells were able to adhere and spread on the corneas that underwent total epithelial debridement, including the central cornea. There was no growth of host (rabbit) epithelial cells on the central cornea of the no treatment group.
Supramolecular hyaluronic acid host-guest hydrogels may be useful as a vehicle to delivered cultured epithelial cells to the cornea.
This is a 2020 ARVO Annual Meeting abstract.
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