Table 1.
Table 2.
Primers used for gene expression analysis by real-time PCR.
Fig 1.
Components of AmnioClip and mounting device.
A) AmnioClip, disassembled, left: outer silicone ring, right: inner steel ring; B) AmnioClip, assembled without AM, view from the inner (ocular) side; C) AmnioClip, assembled with AM, view from the inner (ocular) side, D) mounting device, open position allowing placement of the rings and AM; E) mounting device, closed “mounting” position.
Table 3.
Treatment tolerance; n = 14.
Fig 2.
Sutureless application of AM for treating refractory epithelial erosion (case 1, patient #6).
A) Slit lamp examination and B) fluorescein demonstration of the corneal erosion before sutureless AM application, arrow indicates the lesioned site; C,D) slit lamp control of AM fit at days 3 and 5 of wearing, arrow in D points on AmnioClip; E) slit lamp control and F) fluorescein demonstration of the corneal surface after removal of the AM at day 7, the arrow indicates the reduced lesion.
Fig 3.
Sutureless application of AM for treating corneal ulceration and a deep stromal defect (case 2, patient #7).
A) Slit lamp examination and B,C) fluorescein demonstration of the deep corneal ulceration before sutureless AM application, the arrow in B indicates the corneal ulceration site; D) slit lamp control of AM fit before removal, the arrow points on AmnioClip which was used for applying AM in sandwich technique, E) slit lamp examination and F) fluorescein demonstration of the stable AM inlay after removal of AmnioClip at day 14, the arrow indicates the AM inlay after removal of AmnioClip.
Fig 4.
Morphology of AMs after therapeutic application.
A) AM appears condensed at the site of the ring; B) AM without epithelium (patient #3 second application); C) some AMs retained their own monolayered epithelium (patient #5 first application L); D) same AM as in C, showing a double-layered epithelium with larger cells at the stromal side which faced the patient eye during application; E) some AMs had a multi-layered stratified epithelium with big, cuboidal cells with large round nuclei on the epithelial side which faced the air during application (patient #3 first application); F) same AM as in E, here the stratified epithelium at the epithelial side appeared bulky and the cells were of irregular size. HE staining; es: epithelial side; ss: stromal side; scale bar in A 100 μm, scale bar in B-F 50 μm.
Fig 5.
Growth factor expression in AMs at different time points during processing and use.
A) Concentration of RNA from amniotic membranes directly after cryopreservation (AM), AMs after mechanical handling before transplantation (AM-transplant), or AMs after being worn on the ocular surface for one week (AM-explant) (n≥3). B) mRNA of vascular endothelial growth factor-A (isoform VEGF-A165a), hepatocyte growth factor (HGF), fibroblast growth factor 2 (FGF-2) and pigment epithelium-derived factor (PEDF) in amniotic membranes immediately after thawing (AM), or AMs after mechanical handling before transplantation (AM-transplant). Bars represent mean ± SEM of three or more measurements. *p<0.05 vs. AM.