Supplemented media used in the culture of human corneal endothelial cells.

<p>Supplemented media used in the culture of human corneal endothelial cells.</p

Donor information.

<p>COD: cause of death. Donor age ranged from 10 year-old to 42 year old with a median age of 26 year old. Days taken from death of donor to the initiation of corneal endothelial cell culture ranged from 3 days to 13 days with a median of 8 days. Experiment A: morphological assessment/growth profile - P0 to P1; Experiment B: morphological assessment/growth profile - P1 onwards; Experiment C: Cell adherence analysis – xCelligence; Experiment D: Cell proliferation – Click-iT EdU; Experiment E: Immunofluorescence staining.</p

Expression ofα-smooth muscle actin (α-SMA) in the post-operative central corneas and peripheral flaps.

<p>(A–D) α-SMA (green), a marker of myofibroblasts, was not present in the central corneas on week 8 and 16 after both ReLEx and refractive lenticule re-implantation. (E–H) α-SMA (green) was expressed at the flap periphery and co-localized with F-actin (red) subepithelially on week 8 post-ReLEx and lenticule re-implantation, but was absent 16 weeks after both surgical procedures. In pane A–H, α-SMA (green) was double immunostained with F-actin marker (red), phalloidin. Nuclei were counterstained using DAPI (blue). Arrowheads indicate the location of the laser incision site or lenticular interface. PR: post-ReLEx, PLR: post-lenticule re-implantation. Scale bar: 50 µm.</p

Mean corneal curvature measured by keratometer and mean spherical error measured by refractometer (n = 7).

a<p>PR = post-ReLEx.</p>b<p>PLR = post-lenticule re-implantation.</p>c<p>D = diopter.</p>d<p><i>p</i> values relative to the keratometry before ReLEx.</p>e<p><i>p</i> values relative to the spherical error before ReLEx.</p

Atomic force microscope (AFM) images showing surface topography of native DM (DM; A–D) and DM with fibrin glue (FG) facing up (E–H).

<p>A and E: Height data; B and F: Amplitude data; C and G: Phase data. D and H represent a 3D presentation of topographical map of native DM and DM+FG, respectively. Image scale = 10 µm×10 µm.</p

Immunofluorescent staining of Ki-67, TUNEL and phalloidin in post-operative central corneas.

<p>(A–D) Ki-67-positive cells (green) were not found in the corneal stroma on week 8 and 16 after ReLEx and refractive lenticule re-implantation. (E–H) Similarly, presence of TUNEL-positive cells (green) was also not detected in the corneal stroma. In pane A–H, F-actin marker (red), phalloidin, was observed in the laser incision site or lenticular interface. Its presence was attenuated over time. Nuclei were counterstained using DAPI (blue). Unoperated corneas were used as control. Arrowheads indicate the location of the laser incision site or lenticular interface. PR: post-ReLEx, PLR: post-lenticule re-implantation. Scale bar: 50 µm.</p