Figure 1

Decellularization efficiency and role of dextran: (A) Histological evaluation of human cornea decellularized by various concentrations of sodium deoxycholate (SD) with deoxyribonuclease I in comparison to the normal human cornea (NHC) using hematoxylin and eosin (H&E) staining. The higher magnification images of H&E staining showing the stromal cells in NHC (arrowheads) and cellular debris in 0.5% SD ( arrowheads). Scale bar = 100 µm. HLA-ABC staining (green) of NHC and DHC showing cellular expression of HLA in NHC and 0.5% SD treated DHC (white arrows); no HLA expression in 1% and 4% SD treated DHC cornea. DAPI staining reveals nuclei in NHC without remaining nuclei/nuclear debris in DHC. (B) The role of dextran during decellularization in both DM⁻ and DM⁺ cornea in comparison to respective controls. Graphs represent the thickness of DHC in various dextran conditions compared to NHC. Data are expressed as means ± S.E.M. (n = 5). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; (Wilcoxon signed-rank test). (C) AS-OCT images of DM⁻ and DM⁺ cornea in varying concentrations of dextran with respective controls. (D) Histological evaluation by staining with H&E on NHC and DHC showing no cell remnants in DHC but varying corneal thickness (in presence or absence of dextran) of both DM⁻ and DM⁺ cornea. Scale bar = 100 µm E) Confirmation of decellularization by quantification of residual DNA. The graph represents the quantity of DNA in both DM⁻ and DM⁺ cornea before and after decellularization (0 or 4% dextran). Data are expressed as means ± S.E.M. (n = 4). * p < 0.05; Mann–Whitney U test. DAPI 4′,6‐diamidino‐2‐phenylindole, DM⁻ cornea with the absence of Descemet membrane, DM⁺ cornea with the presence of Descemet membrane, AS-OCT anterior segment optical coherence tomography, HLA human leukocyte antigen.