A combined approach of amniotic membrane and oral mucosa transplantation for fornix reconstruction in severe symblepharon.

PURPOSE: To evaluate the results of a combined approach of cicatrix lysis, intraoperative mitomycin C (MMC) application, oral mucosal transplantation (OMT), and amniotic membrane transplantation (AMT) for surgery of severe symblepharon. METHODS: This prospective study included 32 eyes with severe symblepharon in which after cicatrix lysis the residual conjunctiva was not enough to cover the tarsus (grade III symblepharon) or there was no residual conjunctiva (grade IV symblepharon). After symblepharon lysis and MMC application, OMT was used to cover the tarsus throughout to fornix, and AMT with fibrin glue was performed to cover the exposed sclera. Outcome was defined as complete success (restoration of an anatomically deep fornix), partial success (focal recurrence of scar), or failure (return of symblepharon). RESULTS: Etiology of symblepharon included chemical burn (n = 16), thermal burn (n = 7), Stevens-Johnson syndrome (n = 5), mucous membrane pemphigoid (n = 2), xeroderma pigmentosum (n = 1), and graft-versus-host disease (n = 1). Motility restriction was present in 87.5% preoperatively. After a mean follow-up of 16.4 ± 7.6 months, the anatomical outcome included complete success in 84.4%, partial success in 9.4%, and failure in 6.2%. In grades III and IV symblephara, the outcomes were complete success in 89.5% and 76.9%, partial success in 10.5% and 7.7%, and failure in none and 15.4%, respectively. No motility restriction was noted in any eye postoperatively. Complications included entropion (n = 2), ocular surface keratinization (n = 1), and pyogenic granuloma (n = 4). CONCLUSIONS: In severe symblepharon, a combined approach of cicatrix lysis, MMC application, OMT, and sutureless AMT was a safe and effective technique for fornix reconstruction.

Effects of pterygium surgery on front and back corneal astigmatism.

OBJECTIVE: To evaluate changes in front and back corneal astigmatism after pterygium surgery using the Scheimpflug imaging of Pentacam. DESIGN: Prospective interventional case series. PARTICIPANTS: We studied 96 eyes with primary pterygium that underwent surgery. METHODS: Preoperatively and at 1, 3, and 6 months after surgery, Pentacam (Oculus, Wetzlar, Germany) was used to measure astigmatism at the front and back corneal surfaces. Surgically induced astigmatism (SIA) on the front corneal surface was also calculated, using vector analysis. RESULTS: Of the eyes, 73 completed 6-month postoperative follow-up without developing pterygium recurrence. Front corneal astigmatism decreased from 3.97 ± 4.49 D preoperatively to 1.23 ± 1.88 D at 1 month (p < 0.001). Back corneal astigmatism decreased, but nonsignificantly, from 0.35 ± 0.39 D preoperatively to 0.32 ± 0.2 D at 1 month (p = 0.49). However, although back astigmatism was with-the-rule in 43.8% of patients and against-the-rule in 24.6% of patients preoperatively, these changed to 87.7% and 4.1%, respectively, at 1 month (p = 0.02). Refractive cylinder (52 eyes) reduced from 2.62 ± 2.22 D preoperatively to 1.06 ± 1.57 D at 1 month (p = 0.05). There was no significant change in any parameter after 1 month throughout the 6 months after surgery. SIA was 3.51 ± 2.66 D at 1 month postoperatively, which showed significant correlation with age, preoperative astigmatism, and pterygium length and area, but there was no correlation with gender, pterygium width, preoperative spherical power, or surgical technique. Moreover, higher grades of pterygium morphology had higher SIA postoperatively (p = 0.05). CONCLUSIONS: Pterygium surgery was associated with significant changes in front and back corneal surfaces. Eyes with more advanced pterygia achieved higher SIA.

A controlled study of amniotic membrane transplantation for acute Pseudomonas keratitis.

OBJECTIVE: To evaluate the efficacy of amniotic membrane transplantation (AMT) to improve the outcomes of acute Pseudomonas keratitis as compared with a control group. DESIGN: Prospective interventional case series with retrospective controls. PARTICIPANTS: We studied 14 eyes with Pseudomonas keratitis as the AMT group and 11 eyes with Pseudomonas keratitis as the control group. METHODS: Eyes in the AMT group were treated with antibiotic therapy followed by single-layer AMT at 2 to 3 days. Eyes in the control group received only antibiotic therapy. Patients were followed for 11.1 ± 2.4 months. RESULTS: In the AMT group, pain significantly decreased from a mean score of 2.4 ± 0.5 preoperatively to 1.1 ± 0.9 at day 2 postoperatively (p < 0.001). Corneal epithelial defects healed completely within 13.2 ± 2.6 days in the AMT group compared with 15.5 ± 3.4 days in the control group (p = 0.07). At final follow-up visits, the sizes of corneal opacity and deep neovascularization were not different between the 2 groups. However, the mean score for density of the corneal opacity was significantly less in the AMT group compared with the control group (2.1 ± 0.4 vs 2.5 ± 0.7, respectively, p = 0.04). Although the best corrected visual acuity using hard contact lenses was not different between the 2 groups, uncorrected visual acuity was better in the AMT group (0.45 ± 0.22 logMAR) than in the control group (0.71 ± 0.32 logMAR, p = 0.03). No patient in either group developed significant corneal thinning or perforation. CONCLUSIONS: AMT in acute Pseudomonas keratitis was associated with immediate pain relief, less density of the final corneal opacity, and better uncorrected visual acuity at the final follow-up visit.

Correlation between ocular Demodex infestation and serum immunoreactivity to Bacillus proteins in patients with Facial rosacea.

PURPOSE: To investigate correlation between ocular Demodex infestation and serum. DESIGN: A prospective study to correlate clinical findings with laboratory data. PARTICIPANTS: We consecutively enrolled 59 patients: 34 men and 25 women with a mean age of 60.4+/-17.6 years (range, 17-93). METHODS: Demodex counting was performed based on lash sampling. Serum immunoreactivity to two 62-kDa and 83-kDa proteins derived from B oleronius was determined by Western blot analysis. Facial rosacea, lid margin, and ocular surface inflammation were documented by photography and graded in a masked fashion. MAIN OUTCOME MEASURES: Statistical significance based on correlative analyses of clinical and laboratory data. RESULTS: These 59 patients were age matched, but not gender matched, regarding serum immunoreactivity, ocular Demodex infestation, or facial rosacea. There was a significant correlation between serum immunoreactivity and facial rosacea (P = 0.009), lid margin inflammation (P = 0.040), and ocular Demodex infestation (P = 0.048), but not inferior bulbar conjunctival inflammation (P = 0.573). The Demodex count was significantly higher in patients with positive facial rosacea (6.6+/-9.0 vs. 1.9+/-2.2; P = 0.014). There was a significant correlation of facial rosacea with lid margin inflammation (P = 0.016), but not with inferior bulbar conjunctival inflammation (P = 0.728). Ocular Demodex infestation was less prevalent in patients with aqueous tear-deficiency dry eye than those without (7/38 vs. 12/21; P = 0.002). CONCLUSIONS: The strong correlation provides a better understanding of comorbidity between Demodex mites and their symbiotic B oleronius in facial rosacea and blepharitis. Treatments directed to both warrant future investigation.

Surgical strategies for fornix reconstruction based on symblepharon severity.

PURPOSE: To identify surgical strategies of fornix reconstruction for symblepharon graded according to the length from the limbus to the lid margin, to the width, and to associated inflammation. DESIGN: Retrospective, comparative, interventional case series. METHODS: In 61 eyes with symblepharon, cicatrix lysis and amniotic membrane transplantation (AMT) were performed with sutures (n = 34) or fibrin glue (n = 27) together with (n = 47) or without (n = 14) intraoperative mitomycin C (MMC), plus fornix reconstruction using anchoring sutures without (n = 30) or with (n = 7) oral mucosal graft or with conjunctival autograft (n = 4). Overall, success was defined as an outcome of complete success (restoration of an anatomically deep fornix) or partial success (focal recurrence of scar), and failure was defined as the return of symblepharon. RESULTS: For a follow-up of 25 +/- 10.8 months, the overall success was achieved by the first attempt in 52 eyes (85.2%) and failure resulted in nine eyes (14.8%); however, the success rate improved to 59 eyes (96.7%) with additional attempts. At the first attempt, AMT alone achieved overall successes in 92.8% of grade I eyes and in 100% of grade II eyes. Additional anchoring sutures achieved successes in 100% of grade I eyes, 70% of grade II eyes, and 71.4% of grade III/IV eyes. Additional oral mucosa or conjunctival autograft achieved successes in 100% of grade III/IV eyes. The complete success was correlated positively with lower grades of symblepharon or intraoperative use of MMC, but negatively correlated with younger ages, canthal involvement, or use of anchoring sutures. Anatomic improvement was accompanied by reduction of preoperative conjunctival inflammation (n = 40), improved visual acuity (n = 14), improved ocular motility (n = 18), improved eyelid closure (n = 3), and feasibility of contact lens wear (n = 10). CONCLUSIONS: Successful outcome can be achieved by selectively deploying cicatrix lysis and AMT, intraoperative MMC, anchoring sutures, and oral mucosal or conjunctival autograft based on the severity of pathogenic symblepharon.

Minimal conjunctival limbal autograft for total limbal stem cell deficiency.

PURPOSE: To report the results of one 60 degrees conjunctival limbal autograft (CLAU) combined with amniotic membrane (AM) transplantation for an eye with total limbal stem cell deficiency (LSCD). METHODS: One eye of a patient with chronic total LSCD and symblepharon caused by chemical burn was subjected to symblepharon lysis, removal of pannus from corneal surface, AM transplantation to cover the conjunctival and corneal surfaces as a permanent graft, one 60 degrees CLAU to the superior limbal area, and insertion of ProKera as a temporary AM patch to cover the CLAU. RESULTS: After surgery, corneal epithelialization over the AM was evident adjacent to the CLAU on day 6, progressed to pass the horizontal midline by day 11, and was completed by day 18. During a follow-up of 1 year, the corneal surface remained stable and smooth, and the stroma considerably regained clarity with regression of midstromal vascularization. The best-corrected visual acuity improved from 20/400 to 20/50. The conjunctival inflammation completely resolved, and the fornices were deep. CONCLUSIONS: One 60 degrees CLAU combined with AM transplantation as both a permanent graft and a temporary patch can restore the entire corneal surface in an eye with total LSCD caused by chemical burn.

Sutureless amniotic membrane transplantation for partial limbal stem cell deficiency.

PURPOSE: To evaluate the results of sutureless amniotic membrane (AM) transplantation using fibrin glue for reconstructing corneal surfaces with partial limbal stem cell deficiency (LSCD). DESIGN: Retrospective noncomparative interventional case series. METHODS: Eleven eyes of nine patients that had LSCD with 120 degrees to almost 360 degrees of limbal involvement underwent superficial keratectomy to remove the conjunctivalized pannus followed by AM transplantation using fibrin glue. Additional sutureless AM patch (ProKera; Bio-Tissue, Inc, Miami, Florida, USA) was used in seven patients, and mitomycin C was applied on the cornea in four eyes and during fornix reconstruction in seven eyes. The surgery was repeated in three eyes for residual pannus. RESULTS: During a mean follow-up of 14.2 +/- 7.7 months (range, six to 26 months), all eyes maintained a smooth and stable corneal epithelial surface without recurrent erosion or persistent epithelial defect, and showed less stromal cloudiness and vascularization. Best-corrected visual acuity improved in nine eyes (81.8%). Corneal epithelialization proceeded by epithelial growth over AM (n = 4), accompanied by dissolution of AM (n = 4) or a combination of both (n = 3). No complication was noted regarding initial or repeated uses of fibrin glue. CONCLUSION: AM transplantation using fibrin glue appears to be a safe and effective method of restoring a stable corneal epithelium for cases with partial LSCD. This approach avoids the need of transplanting limbal epithelial stem cells.

Role of conjunctival inflammation in surgical outcome after amniotic membrane transplantation with or without fibrin glue for pterygium.

PURPOSE: To determine the clinical significance of postoperative conjunctival inflammation noted at the third or fourth week after intraoperative application of mitomycin C and amniotic membrane transplantation for pterygium. METHODS: This retrospective study included 27 eyes of 23 patients with primary (n = 12) or recurrent (n = 15) pterygia. All cases were operated by extensive removal of subconjunctival fibrovascular tissue and intraoperative application of 0.04% mitomycin C in the fornix, followed by amniotic membrane transplantation by using either fibrin glue (14 eyes) or sutures (13 eyes). Main outcome measures included development of conjunctival inflammation, pyogenic granuloma, and pterygium recurrence after surgery. RESULTS: For a follow-up of 29.6 +/- 17.2 months (range, 6-56 months), 16 (59.3%) eyes without postoperative conjunctival inflammation resulted in favorable outcomes. Conjunctival inflammation around the surgical site was noted in the remaining 11 (40.7%) eyes and was significantly more common in eyes with sutures than those with fibrin glue (61.5% vs. 21.4%, respectively; P = 0.05). Among those with this inflammation, 7 eyes receiving subconjunctival injection of triamcinolone resulted in complete resolution and a good aesthetic outcome. Four eyes without this injection gradually developed conjunctival (n = 1) or corneal (n = 1) recurrence and/or pyogenic granuloma (n = 3). CONCLUSIONS: Host conjunctival inflammation is still common after intraoperative application of mitomycin C and amniotic membrane transplantation, especially when sutures are used in pterygium surgery. If left untreated, persistent inflammation may lead to a poor surgical outcome.

Corneal manifestations of ocular demodex infestation.

PURPOSE: To report the corneal manifestations in eyes with Demodex infestation of the eyelids. DESIGN: Noncomparative, interventional case series. METHODS: This retrospective review included six patients with Demodex blepharitis who also exhibited corneal abnormalities, which led to suspicion of limbal stem cell deficiency in three cases. All patients received weekly lid scrubs with 50% tea tree oil and a daily lid scrubs with tea tree shampoo for a minimum of six weeks. Improvement of symptoms and corneal and conjunctival signs were evaluated. RESULTS: All six patients exhibited ocular irritation and conjunctival inflammation, while meibomian gland dysfunction (n = 5), rosacea (n = 4), and decreased vision (n = 3) also were noted despite prior treatments with oral tetracycline, topical steroids with antibiotics, and lid scrub with baby shampoo. These patients were proven to have Demodex folliculorum (n = 6) and Demodex brevis (n = 3) by microscopic examination of epilated lashes. Their corneal manifestation included superficial corneal vascularization (six eyes of five cases), marginal corneal infiltration (two eyes of two cases), phlyctenule-like lesion (one eye of one case), superficial corneal opacity (two eyes of two cases), and nodular corneal scar (two eyes of two cases). After treatment, the Demodex count was reduced from 6.8 +/- 2.8 to 1 +/- 0.9 (standard deviation; P = .001). All patients showed dramatic resolution of ocular irritation, conjunctival inflammation, and all inflammatory, but not scarred, corneal signs; three patients showed improved vision. CONCLUSIONS: A variety of corneal pathologic features together with conjunctival inflammation, commonly noted in rosacea, can be found in patients with Demodex infestation of the eyelids. When conventional treatments for rosacea fail, one may consider lid scrub with tea tree oil to eradicate mites as a new treatment.

High prevalence of Demodex in eyelashes with cylindrical dandruff.

PURPOSE: To determine the prevalence of Demodex in eyelashes with cylindrical dandruff (CD). METHODS: A modified sampling and counting method was applied to 55 clinical cases. Patients were divided in to group A (n = 20) with diffuse CD, group B (n = 12) with sporadic CD, and group C (n = 23) with clean lashes or greasy scales, of which the latter was divided into subgroup C1 (n = 15) without lid hygiene and subgroup C2 (n = 8) using daily lid hygiene for the past year. Each patient underwent a routine complete eye examination and modified counts of Demodex. RESULTS: Demodex was found in all group A and B patients (n = 32) with CD, which was significantly higher than the 22% of group C patients (n = 23) without CD (P < 0.001). The Demodex counts were 4.1 +/- 1.0 and 2.0 +/- 1.2 per epilated lash with retained CD, significantly higher than the 0.2 +/- 0.5 and 0.2 +/- 0.4 per lash without retained CD in groups A and B, respectively (each P < 0.001) and than the 0.01 +/- 0.09 and 0.12 +/- 0.41 per lash in subgroups C1 and C2, respectively (each P < 0.001). Demodex was still found in CD fragments left on the lid skin after epilation. Five Demodex brevis mites were found among the 422 Demodex specimens. CONCLUSIONS: The modified sampling and counting method showed that the prior controversy regarding Demodex has resulted from miscounting and confirmed that lashes with CD are pathognomonic for ocular Demodex infestation. Lid hygiene with shampoo reduces Demodex counts but does not eradicate the mites.

Characterization of corneal pannus removed from patients with total limbal stem cell deficiency.

PURPOSE: To determine the epithelial lineage of origin in corneal pannus tissue surgically removed from patients with total limbal stem cell (SC) deficiency. METHODS: The lineage of origin of the entire conjunctivalized pannus removed from eight corneas with a diagnosis of total limbal SC deficiency was characterized by anti-keratin (K)-3 and anti-K19 monoclonal antibodies. The protein and mRNA of epithelial outgrowth from segments of five such pannus specimens were analyzed by Western blot and reverse transcription-polymerase chain reaction, respectively. RESULTS: Cross sections of all eight specimens showed a stratified epithelium with goblet cells expressing mucin (MUC)-5AC, and a stroma showing blood vessels and inflammatory cell infiltrates. Immunostaining showed full-thickness expression of K19 in the entire pannus of all eight specimens. Expression of K3 was negative in seven patients, but was sporadically positive in a patient with Stevens-Johnson syndrome. In culture, all five pannus specimens generated a compact, small epithelial cell outgrowth, and except for one, reached confluence in 2 to 3 weeks. The K3/K12 pair was expressed by extracts of cell outgrowth from the control limbal epithelial explant, but not in all five pannus specimens. A 60-kDa band of DeltaNp63 was expressed in the control specimen and in all five pannus specimens. Cell outgrowth expressed K3 transcript in three, but none showed K12 transcript. CONCLUSIONS: The resultant epithelial phenotype of the pannus tissue was not corneal, as evidenced by the negative staining to cornea-specific K12 mRNA and protein, but was conjunctival, as evidenced by the presence of goblet cells, the weak expression of K3, and the strong expression of K19. The abundant expression of DeltaNp63 in such conjunctiva-derived epithelium in eyes with total limbal SC deficiency raises doubts as to its validity as a limbal SC marker.

Human keratocytes cultured on amniotic membrane stroma preserve morphology and express keratocan.

PURPOSE: To develop a new method of expanding human corneal keratocytes in serum while maintaining their characteristic morphology and keratocan expression. METHODS: Human keratocytes were isolated from central corneal buttons by digestion in 1 mg/mL of collagenase A in DMEM and seeded on plastic or the stromal matrix of human amniotic membrane (AM) in DMEM with different concentrations of FBS. On confluence, cells on AM were continuously subcultured for six passages on AM or plastic. In parallel, cells cultured on plastic at passages 3 and 11 were reseeded on AM. Cellular morphology and cell-cell networks were assessed by phase-contrast microscopy and a cell viability assay, respectively. Expression of keratocan was determined by RT-PCR and Western blot analysis. RESULTS: Trephined stroma yielded 91,600 +/- 26,300 cells (ranging from 67,000 to 128,000 cells per corneal button). Twenty-four hours after seeding, cells appeared dendritic on AM, even in 10% FBS but fibroblastic on plastic. Such a difference in morphology correlated with expression of keratocan assessed by RT-PCR and Western blot, which was high and continued at least to passage 6 on AM, even in 10% FBS, but was rapidly lost each time when cells on AM were passaged on plastic. Fibroblasts continuously cultured on plastic to passages 3 and 11 did not reverse their morphology or synthesize keratocan when reseeded on plastic in 1% FBS or on AM. CONCLUSIONS: Human keratocytes maintain their characteristic morphology and keratocan expression when subcultured on AM stromal matrix even in the presence of high serum concentrations. This method can be used to engineer a new corneal stroma.