Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering

Abstract

The idea of transplanting a sheet of laboratory‐grown corneal endothelium dates back to 1978; however, the ideal scaffold is still lacking. We hypothesized that human crystalline lens capsules (LCs) could qualify as a scaffold and aimed to characterize the properties of this material for endothelial tissue engineering. LCs were isolated from donor eyes, stored at −80 °C, and decellularized with water and trypsin‐EDTA. The decellularization was investigated by nuclear staining and counting and the capsule thickness was determined by optical coherence tomography and compared with Descemet's membrane (DM). Transparency was examined by spectrometry, and collagenase degradation was performed to evaluate its resistance to degradation. Cell‐scaffold interaction was assessed by measuring focal adhesions surface area on LC and plastic. Finally, primary corneal endothelial cells were grown on LCs to validate the phenotype. Trypsin‐EDTA decellularized most effectively, removing 99% of cells. The mean LC thickness was 35.76 ± 0.43 μm, whereas DM measured 25.93 ± 0.26 μm (p < .0001). Light transmission was 90% for both LC and DM. On a collagenase challenge, LC and amniotic membrane were digested after 13 hr, whereas DM was digested after 17 hr. The surface area of focal adhesions for cells grown on coated LCs was at least double that compared with other conditions, whereas tight junctions, ion pumps, and hexagonal morphology were well maintained when endothelial cells were cultured on LCs. In conclusion, LCs demonstrate excellent scaffolding properties for tissue engineering and sustain the cell phenotype and can be considered a suitable substrate for ocular tissue engineering or as a template for future scaffolds.

Decellularization methods for developing porcine corneal xenografts and future perspectives

“…However, after transplantation into rabbits, the corneas gradually regained transparency and the angiogenesis that developed immediately after transplantation started to regress after the first 2 weeks . Van den Bogerd et al have successfully used the freeze‐thaw method on anterior capsule of the lens to make a useful scaffold for corneal endothelial cells (CECs).…”

“…A comparative study found that the acellular human lens capsule is a better scaffold to create ES when compared with human Descemet's membranes and human amniotic membranes. 50 In animal studies, freeze-thaw, N 2 gas, HHP, and SDS-treated DPCXs displayed repopulation of recipient keratocytes in graft stroma after lamellar keratoplasty. 10,22,48,51 However, the recellularization activity of PLA and glycerol combined with chemical crosslinking seems to be a preferred alternative.…”

“…A comparative study found that the acellular human lens capsule is a better scaffold to create ES when compared with human Descemet's membranes and human amniotic membranes . Lee et al demonstrated a porcine model of corneal endothelialization and successfully obtained an ES on DPCXs with porcine CECs.…”

Exploring the Mesenchymal Stem Cell Secretome for Corneal Endothelial Proliferation

“…Endo: corneal endothelium medium; Exofree: BM MSC medium with exosome depleted FBS; WT FBS: BM MSC medium with wild-type FBS. 4 Stem Cells International identified growth factors, there was neither a simulation of growth, even more, there was an increase in PDT, i.e., a slower cell growth, for cell supplemented with a 10 ng/mL combination of EGF-IGFBP2-IGFBP6 (Figure 7(g)).…”

“…One possible strategy to overcome these issues is to tissue engineer an endothelial layer in the lab. This product would be composed of ex vivo grown HCEnCs on a suitable cell scaffold for transplantation [1,4,5]. Although the scaffold approach is the most commonly explored, cell suspension therapies have also been trialed in 11 patients [6,7].…”