Confocal microscopy study of donor-recipient interface after Descemet's stripping with endothelial keratoplasty.

AIM: To evaluate the in vivo confocal microscopic corneal features seen in patients who underwent Descemet's stripping with endothelial keratoplasty (DSEK) and to correlate these findings with the postoperative best spectacle-corrected visual acuity (BSCVA) obtained at the time of confocal microscopy. METHODS: Observational case series. A review of charts and confocal microscopy images and videos (Confoscan 4; Nidek Technologies America, Greensboro, North Carolina, USA) of 25 consecutive eyes from 21 patients who underwent DSEK for Fuchs' dystrophy or pseudophakic bullous keratopathy and had a follow-up of at least 6 months post-DSEK. Time from DSEK to confocal microscopy ranged from 6 to 22 months. Subepithelial haze, donor-recipient interface haze, interface birefringent particles and presence of stromal folds were evaluated by a blinded observer and correlated to the postoperative BSCVA. RESULTS: 25 eyes (21 patients) with a mean age of 71.3 (11.8) years were included. A significant negative correlation was found between postoperative BSCVA and subepithelial corneal haze (Spearman correlation coefficient) (r(s)=-0.651; p=0.0004). Donor-recipient interface haze (r(s)=-0.0908; p=0.6658), interface particles (r(s)=-0.2403; p=0.2472) or the presence of stromal folds (r(s)=0.111; p=0.6055) did not correlate with BSCVA. CONCLUSION: Subepithelial haze, donor-recipient stromal interface haze, birefringent particles and stromal folds were present at different degrees in all patients who underwent DSEK. However, subepithelial haze was the only factor that correlated to postoperative BSCVA.

Urrets-Zavalia syndrome as a complication of argon laser peripheral iridoplasty.

OBJECTIVE: To report on 8 patients who developed prolonged mydriasis after argon laser peripheral iridoplasty (ALPI). DESIGN: Retrospective case series. METHODS: A review of the charts of 8 patients with persistent occludable angles after laser iridotomy who developed mydriasis after undergoing ALPI. RESULTS: 12 eyes of 8 patients (mean age 50.2, standard deviation 6.9, range 40-61 years) developed persistent dilatation. 7 of 8 patients had plateau iris syndrome. Only one patient had a decrease in visual acuity. Intraocular pressure increased only in 1 eye and remained stable or decreased in the others. Of the 8 patients, 7 had blurred vision, 2 had photophobia, 4 had glare and 1 had discomfort. The pupillary response to pilocarpine instillation was minimal or absent. Mydriasis eventually resolved in all eyes without treatment. CONCLUSION: ALPI can be complicated by mydriasis unresponsive to pilocarpine. Mydriasis and accompanying symptoms resolved spontaneously within 1 year in most patients.

Keratolimbal allograft in corneal reconstruction.

The replenishment of corneal epithelial SC is a crucial step for reconstructing the ocular surface in patients suffering from devastating ocular surface diseases manifesting with total LSCD. KLAL is one of such procedures and has a long track record and a long follow-up for patients with bilateral total LSCD. This review summarizes the literature experiences and outline new strategies that are important to enhance the success of this procedure. Further research is needed to fully understand the biological processes involved in allogeneic tissue transplantation for preserving epithelial SC adhesion, migration, and survival.

Rat enterocyte cultures on human amniotic membranes.

BACKGROUND: Rat enterocytes were cultured on human amniotic membranes. METHODS: Intestine of neonatal DA rats was digested using collagenase and dispase according to the technique developed by Evans. The harvested enterocytes were cultured on human amniotic membranes using standard cell culture techniques. RESULTS: After the second day of culture, some intestinal epithelial units started to gradually detach from the membrane, dispersing as single cells and disappearing within a few days. On the contrary, other units showed signs of cell proliferation. The cultured cells underwent morphologic changes, survived, and remained attached to the amniotic membrane for 3 weeks. Paraffin sections of the membrane showed cultured cytokeratin-positive cells attached to the membrane as a monolayer. CONCLUSIONS: Human amniotic cell membranes help to maintain rat enterocytes in culture for a long time period (3 weeks), possibly via secretion of trophic factors. This technique may provide a valuable tool to study the development and the properties of these epithelial cells in a culture environment.

Clinical characteristics of conjunctivochalasis with or without aqueous tear deficiency.

AIM: To show characteristic ocular surface findings caused by conjunctivochalasis (CCh) in dry eye patients with or without aqueous tear deficiency (ATD). DESIGN: Comparative non-interventional cases. PATIENTS AND METHODS: Clinical data of five ATD patients without CCh (group A), eight CCh patients with ATD (group B), and eight CCh patients without ATD (group C) were retrospectively reviewed. Presence or absence of CCh was determined by fluorescein staining to outline tear meniscus and conjunctival folds with an enhancing filter. Dry eye symptoms, history of subconjunctival haemorrhage, meibum expression, tear break up time, fluorescein and rose bengal staining, and fluorescein clearance test, and other abnormal ocular surface findings were measured. RESULTS: CCh patients were significantly older (p = 0.001). In pure ATD, the principal symptom of dryness became worse as the day progressed. In contrast, blurry vision, burning sensation, and dryness became worse during reading in all CCh patients (p = 0.0008) or worse in the morning upon awakening in the majority patients with CCh only (p = 0.02). Besides the interpalpebral exposure, which was noted in ATD, positive fluorescein or rose bengal staining was noted in the redundant conjunctival folds and the non-exposure zone in CCh (p = 0.0008). Redundant conjunctival folds were present in both lower and upper bulbar conjunctiva, obliterating both lower and upper tear meniscuses, and spatially correlated with anterior migration of the mucocutaneous junction in CCh. Delayed tear clearance was significantly more prevalent in CCh than ATD (p = 0.0008). Vigorous blinking worsened in CCh but not in ATD (p = 0.0008). Lacrimal puncta were swollen in groups B and C, but not in group A (p = 0.04). CONCLUSIONS: CCh is not restricted to the lower bulbar conjunctiva, and contributes to pathogenesis of dry eye by obliterating both lower and upper tear meniscus, causing unstable tear film and by creating delayed tear clearance. Dry eye symptoms were worsened by downgaze during reading and by vigorous blinking. Other characteristic signs including subconjunctival haemorrhage, swollen puncta, anterior migration of the mucocutaneous junction, and patterns of dye staining also help distinguish dry eye associated with CCh from that caused by ATD alone.

Correlation of long term phenotypic and clinical outcomes following limbal epithelial transplantation cultivated on amniotic membrane in rabbits.

AIM: To determine the epithelial phenotype in rabbits with total limbal stem cell deficiency (LSCD) after reconstruction with autologous limbal epithelial stem cells ex vivo expanded on rabbit amniotic membrane (AM). METHODS: Left eyes of 52 rabbits were rendered total LSCD, verified by impression cytology. The fibrovascular pannus of each cornea was removed. Group I (n = 10) received rabbit AM transplantation alone, while groups II-IV (n = 42) underwent transplantation of LSC cultured on rabbit AM (LSC-AM) from a small limbal biopsy taken from the right eye. Clinical outcome was graded as "success," "partial success," or "failure" depending on the corneal smoothness and avascularity. Epithelial phenotype was determined by immunostaining and graded as "corneal (K)," "conjunctival (J)," or "mixed (M)" depending on expression of K3 and Muc5AC. RESULTS: After 1 year follow up, group I showed 100% failure and groups II-IV showed 26% success (p<0.001). Clinical failure correlated with J phenotype p = 0.001), while clinical success correlated with K phenotype p = 0.01). When the phenotypic outcome was used for comparison, J phenotype was significantly high in group I (p = 0.003), while K phenotype was significantly high in groups II-IV (p<0.05). CONCLUSION: There is a strong correlation between clinical success and resultant corneal epithelial phenotype. Ex vivo expanded LSC can successfully reconstruct corneal surfaces with unilateral total LSCD.

Corneal stromal changes following reconstruction by ex vivo expanded limbal epithelial cells in rabbits with total limbal stem cell deficiency.

AIM: To study corneal stromal changes and the presence of myofibroblasts after transplantation of ex vivo expanded limbal epithelium. METHODS: A state of limbal deficiency was induced in 16 rabbits. After transplantation with autologous ex vivo expanded limbal epithelium on amniotic membrane (AM), their clinical outcomes were classified as success, partial success or failure according to surface smoothness, stromal clarity, and vascularisation. Clinical outcomes were correlated with phenotypic outcomes of corneal, conjunctival, or mixed epithelium, defined by expression of K3 keratin or MUC5AC. Immunostaining was performed with antibodies against collagen IV, fibronectin, and alpha-smooth muscle actin (alpha-SMA) to assess stromal wound remodelling. RESULTS: Rabbits were sacrificed after a mean follow up of 10 (SD 3.3) months. Collagen IV, expressed in the basement membrane of all three groups, was found in the stroma of the partial success, but not in that of the success or the failure. Fibronectin was absent in the success and the failure, but expressed in the stroma of the partial success. Alpha-SMA was expressed in superficial stroma of the partial success, but suppressed in areas with AM remnants. CONCLUSION: Restoration of a clear and transparent cornea is associated with a normal corneal epithelium and complete wound remodelling. In contrast, wound healing remains active and incomplete in conjunctivalised corneas, which remain opaque with myofibroblasts.

Amniotic membrane transplantation for reconstruction after excision of large ocular surface neoplasias.

AIM: To evaluate the clinical outcome of patients in whom ocular surface reconstruction was performed using amniotic membrane transplantation (AMT) after the excision of large (>20 mm square) ocular surface neoplasias (OSN). METHODS: A non-comparative interventional case series. In 16 eyes of 16 patients, excision of large OSN including conjunctival intraepithelial neoplasia (CIN), primary acquired melanosis, and malignant melanoma was followed by adjunctive cryotherapy and suturing of a single layer of amniotic membrane (AM) with the basement membrane side facing up to the healthy bordering tissue. Epithelial healing, complications, and tumour recurrences were analysed. RESULTS: During a mean follow up of 23.7 (SD 11, range 11-43) months, ocular surface healing was rapid and complete in all cases. One complication of pyogenic granuloma was noted. Tumour recurrence occurred in one out of 10 CIN cases (10%), no recurrences were observed in the patients with melanotic lesions. CONCLUSIONS: AMT in lieu of conjunctival or mucosal autograft is an effective substrate for reconstructing the ocular surface following excision of large OSN. AMT is effective in managing large OSN by avoiding the complications that may be associated with conventional removal, specifically in cases where the limbal architecture is destroyed by surgical resection or adjuvant therapies.