Fish-Scale Collagen Membrane Seeded with Corneal Endothelial Cells as Alternative Graft for Endothelial Keratoplasty Transplantation

Abstract

The human corneal endothelium has limited regeneration capacity. Several methods have been developed in an attempt to repair it. Descemet stripping automated endothelial keratoplasty (DSAEK) is commonly performed on patients with endothelial dysfunction. However, donor demand far exceeds donor supply. Here, we prepared fish-scale collagen membrane (FSCM) and seeded it with CECs in preparation for corneal endothelial transplantation. The fish scales were decellularized, decalcified, and curved. The FSCM was inspected by fluorescence microscopy, SEM, and TGA to validate decellularization, microstructure, and decalcification, respectively. The cytotoxicity of FSCM and the viability of the cells in contact with it were evaluated by LDH and WST-1, respectively. CEC tight junctions and ZO-1 structure were observed by SEM and confocal microscopy. FSCM seeded with CECs were implanted to rabbit anterior chambers to evaluate host tissue reactions to it. FSCM biocompatibility and durability were also assessed. The results showed that FSCM has excellent transparency, adequate water content, and good biocompatibility. The cultivated CECs mounted on the FSCM were similar to normal CECs in vivo. The FSCM plus CECs developed here have high potential efficacy for endothelial keratoplasty transplantation.

Characteristics of Marine Biomaterials and Their Applications in Biomedicine

“…This approach can directly produce neurovascular structure in the infarct cavity and modulate inflammatory, scar, and neural stem cell responses in the adjacent brain. Li, KY et al seeded corneal en-dothelial cells (CEC) on fish scale collagen membrane (FSCM) and transplanted them [164]. After FSCM was implanted into the anterior chamber of rabbit cornea, it was found that the corneal transparency and thickness were normal at all time points, and no edema or turbidity was observed [164].…”

“…Li, KY et al seeded corneal en-dothelial cells (CEC) on fish scale collagen membrane (FSCM) and transplanted them [164]. After FSCM was implanted into the anterior chamber of rabbit cornea, it was found that the corneal transparency and thickness were normal at all time points, and no edema or turbidity was observed [164]. FSCM was suitable as a cell carrier for corneal transplantation.…”

Limbal Stem Cells on Bacterial Nanocellulose Carriers for Ocular Surface Regeneration

“…to enhance hESC‐derived retinal pigment epithelial cells attachment to synthetic electrospun membranes drawing similar conclusions. [ 51 ] Hence, the requirement of surface modifications to use BNC as a cell culture substrate seems to be cell type‐dependent. This cell‐specificity has also been described by Pertile and colleagues [ 52 ] who reported that nitrogen plasma activation improved the affinity of microvascular cells and neuroblasts for BNC but had no effect on fibroblasts.…”

Special Issue: Leaders in Biomedical Engineering

“…Li and coworkers applied the decellularization technique to develop decellularized fish-scale collagen membranes with adequate strength and transparency. 1 These membranes allowed corneal endothelial cells to attach and grow with tight cell conjugation and normal morphology. After the fish-scale collagen membranes were implanted into rabbit anterior chambers, corneal clarity and thickness remained normal throughout the course of examination.…”

“…A good example of natural materials is fish scales, which are multilayered and well-oriented. Li and co-workers applied the decellularization technique to develop decellularized fish-scale collagen membranes with adequate strength and transparency . These membranes allowed corneal endothelial cells to attach and grow with tight cell conjugation and normal morphology.…”