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

Slit lamp microscopy and anterior segment optical coherence tomography (AS-OCT) images of pre- and postoperative corneas.

<p>(A) Slit lamp (top panel) and retro illumination photographs (bottom panel) of the cornea before ReLEx and on week 8 and 16 after ReLEx and refractive lenticule re-implantation. (B) Temporal AS-OCT images of postoperative corneas shows thinning of the cornea after ReLEx and restoration of corneal thickness after lenticule re-implantation. (C) Mean corneal thickness before ReLEx and 8 and 16 weeks after ReLEx and lenticule re-implantation. (D) Post-reimplantation corneal haze graded based on observation of the slit lamp photographs found in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0067058#pone.0067058.s003" target="_blank">Figure S3</a>. Corneal clarity or haze is graded on a scale of 0–4 (from 0 being completely clear to 4 being completely obscured). Statistical significance was obtained by comparing post-operative to pre-operative corneal clarity. Error bars in the bar graphs represent standard deviation. Asterisk (*) and double asterisk (**) denote p<0.001 and p<0.05, respectively. PR: post-ReLEx, PLR: post-lenticule re-implantation.</p

RNA transcripts that were down-regulated in the SPB placentas, relative to the STB placentas, according to the microarray experiment (Mann-Whitney test, adjusted by the Benjamini and Hochberg method, adjusted p<0.05).

a<p>If a probeset interrogates a RNA transcript with no gene symbol (“—”) approved by the HUGO Gene Nomenclature Committee at the European Bioinformatics Institute (<a href="http://www.genenames.org" target="_blank">http://www.genenames.org</a>), it is shown as a “transcribed locus”. Other details on the probesets are freely accessible at <a href="http://www.netaffx.com" target="_blank">http://www.netaffx.com</a>.</p>b<p>Only 15 transcripts with the greatest fold-change values are shown. The remaining 171 transcripts are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034328#pone.0034328.s007" target="_blank">Table S6</a>.</p><p>SPB, spontaneous preterm birth; STB, spontaneous term birth.</p

Gene ontology (GO) terms over-represented in the list of genes that were down-regulated in the SPB placentas, compared with the STB placentas.

<p>Terms with the highest fold of over-representation from each category are shown. The remaining terms are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034328#pone.0034328.s009" target="_blank">Table S8</a>.</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

Gene ontology (GO) terms over-represented in the list of genes that were up-regulated in the SPB placentas, compared with the STB placentas.

<p>Terms with the highest fold of over-representation from each category are shown. The remaining terms are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034328#pone.0034328.s008" target="_blank">Table S7</a>.</p

In vivo confocal micrographs of pre- and post-operative corneas.

<p><b>(</b>A) Horizontal surgical plane between the flap and stromal bed on week 8 and 16 after ReLEx. This region is normally indicated by a relatively higher light reflective layer. Keratocyte repopulation could be observed on week 8 and 16. (B) The top panel shows the anterior interface of the re-implanted lenticule. The middle panel shows the presence of keratocytes within the lamellae of the lenticule and the bottom panel shows the posterior interface of the lenticule. Keratocyte repopulation of anterior and posterior borders of the lenticule occurred by week 8 after lenticule re-implantation. (C) Mean reflectivity level of the laser incision site on day 3, and 2, 4, 8 and 16 weeks after ReLEx. The representative in vivo confocal images of the cornea at the stated time point can be found in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0067058#pone.0067058.s004" target="_blank">Figure S4A</a>. (D) Mean reflectivity level of the re-implanted lenticule’s anterior interface on day 3, and 2, 4, 8 and 16 weeks. (E) Mean reflectivity level of the lenticule’s posterior interface post-reimplantation. The representative in vivo confocal images of the cornea at the stated time point in panes D and E can be found in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0067058#pone.0067058.s004" target="_blank">Figure S4B</a>. Error bars in the bar graphs represent standard deviation. Asterisk (*) and double asterisk (**) indicate p<0.05 and p<0.001, respectively. PR: post-ReLEx, PLR: post-lenticule re-implantation.</p

RNA transcripts that were up-regulated in the SPB placentas, relative to the STB placentas, according to the microarray experiment (Mann-Whitney test, adjusted by the Benjamini and Hochberg method, adjusted p<0.05).

a<p>The RNA transcript interrogated by each probeset is shown as the gene symbol and name approved by the HUGO Gene Nomenclature Committee at the European Bioinformatics Institute (<a href="http://www.genenames.org" target="_blank">http://www.genenames.org</a>). Certain probesets interrogating the same gene may be listed as different GenBank accession numbers. This may reflect the different isoforms being interrogated. Other details on the probesets are freely accessible at <a href="http://www.netaffx.com" target="_blank">http://www.netaffx.com</a>.</p>b<p>Only 15 transcripts with the greatest fold-change values are shown. The remaining 225 transcripts are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034328#pone.0034328.s006" target="_blank">Table S5</a>.</p>c<p>Up-regulated transcripts with higher expression levels than the <i>PLAC4</i> mRNA in the placenta, according to the microarray data.</p><p>SPB, spontaneous preterm birth; STB, spontaneous term birth.</p

Expression of fibronectin, tenascin, collagen type I and CD18 in post-operative central corneas.

<p>(A–D) Fibronectin predominantly appeared along the laser incision site or lenticular interface. The expression was reduced over time after either ReLEx or refractive lenticule re-implantation. (E–H) Tenascin was absent along the flap interface on week 8 and 16 following ReLEx, but was present along the borders of the stromal lenticule after re-implantation. The intensity of the staining was attenuated over time. (I–L) Collagen type I was expressed uniformly in the full thickness of corneal stroma. No significant anomaly in collagen arrangement was observed in the corneas post-ReLEx and post-reimplantation. (M–P) CD18-positive cells were not seen in all post-operative corneas. 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

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