Changing Indications in Penetrating Keratoplasty: A Systematic Review of 34 Years of Global Reporting.

BACKGROUND: Penetrating keratoplasty (PK) ranks among the oldest and most common kinds of human tissue transplantation. Based on the hypothesis that reported indications for PK significantly vary between global regions and over time, the present systematic review aimed to provide a thorough overview of global PK indications as reported in peer-reviewed manuscripts. METHODS: A literature search of PubMed and MEDLINE was conducted to retrieve articles published from January 1980 to May 2014. Indications for PK within 7 global regions were compared using a modified classification system for PK indications and analyzed via multivariate regression. RESULTS: A total of 141 publications from 37 countries were included, recording 180 865 PK cases. Postcataract surgery edema was the predominant indication in North America (28.0%) and ranked second in Europe (20.6%), Australia (21.1%), the Middle East (13.6%), Asia (15.5%), and South America (18.6%). Keratoconus was the leading indication in Europe (24.2%), Australia (33.2%), the Middle East (32.8%), Africa (32.4%), and South America (22.8%). It ranked third in North America (14.2%). Keratitis was the primary indication in Asia (32.3%). Fuchs endothelial corneal dystrophy was the fourth most common indication in North America (12.9%) and Europe (10.2%) and fifth in South America (3.8%). Multivariate analysis supported these results and revealed individual regional changes over time. CONCLUSIONS: Systematic analysis reveals characteristic chronological and regional differences in reported global PK indications. Leading reported indications for PK between 1980 and 2014 were keratoconus (Europe, Australia, the Middle East, Africa, and South America), pseudophakic bullous keratopathy/aphakic bullous keratopathy (North America), and keratitis (Asia).

The role of corneal endothelial morphology in graft assessment and prediction of endothelial cell loss during organ culture of human donor corneas.

PURPOSE: Endothelial assessment is crucial in the release of corneas for grafting. We retrospectively analysed the role of endothelial morphology parameters in predicting endothelial cell loss during organ culture. METHODS: Human donor corneas were cultured in minimal essential medium with 2% fetal calf serum and antibiotics. Initial endothelial morphology was assessed microscopically using score parameters polymegethism (POL), pleomorphism (PLE), granulation (GRA), vacuolization (VAC), segmentation of cell membranes (SEG), Descemet's folds (DF), trypan blue-positive cells (TBPC) and endothelial cell-free areas (ECFA). Some corneas were primarily rejected based on endothelial assessment. Endothelial cell density (ECD) was assessed at the beginning (I-ECD) and end of culture. Corneas were then placed in dehydration medium (as above + 5% dextran 500). In a subgroup, ECD was reassessed after dehydration. Endothelial cell loss during culture (ECL@Culture) and culture+dehydration (ECL-Culture&Dehydration) were calculated. Data were given as mean ± SD and analysed using multiple linear and logistic regression. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. RESULT: I-ECD was 2812 ± 360/mm2 (n = 2356). The decision to reject a cornea due to endothelial assessment was associated negatively with I-ECD (OR = 0.77/100 cells, CI 0.7-0.82) and positively with ECFA (OR = 2.7, CI 1.69-4.35), SEG (OR =1.3, CI 1.01-1.68) and donor age (OR = 1.26/decade, CI 1.33-1.41). ECL@Culture was 153 ± 201/mm2 (n = 1277), ECL@Culture&Dehydration was 169 ± 183/mm2 (n = 918). ECL@Culture was associated positively with donor age, I-ECD, GRA and TBPC, and negatively with PLE, and DF. ECL@Culture&Dehydration was associated positively with age, sex, initial ECD, POL, PLE, VAC and TBPC. CONCLUSION: Morphological parameters displayed associations with the exclusion of corneas from culture and with endothelial cell loss. Appropriate parameter selection for screening purposes may help improve graft quality.

Role of the Endothelial Layer in the Deswelling Process of Organ-Cultured Human Corneas Before Transplantation.

PURPOSE: Before corneal transplant surgery, a deswelling process of organ-cultured corneas is required. This study compares the deswelling kinetics of corneas with an intact endothelial cell layer and disrupted or removed endothelium by measuring central corneal thickness (CCT) over time using anterior segment spectral domain optical coherence tomography. METHODS: Ten donor pairs were cultured in organ culture. The right and left corneas were alternately assigned to one of 2 deswelling groups. Deswelling in the first group [endothelial group (EG)] was induced using a medium with dextran 5%. Corneas of the second group [nonendothelial group (NEG)] were deprived of their endothelial cell layer by trypsinization and were then placed in the same deswelling medium. CCT (mean ± SD) was measured by anterior segment spectral domain optical coherence tomography before deswelling (0 hours) and after 1, 2, 3, 6, 12, 24, 48, 72, and 144 hours. Deswelling kinetics was analyzed through the nonlinear platform in SAS/JMP11 Pro. RESULTS: Before deswelling, CCT was 1071.0 µm (±129.6 µm) and 1133.8 µm (±124.3 µm) in the EG and NEG, respectively. Minimum corneal thickness was obtained after 24 hours in the EG (531.9 ± 47.5 µm) and 6 hours in the NEG (645 ± 81.2 µm). CCT was significantly (P < 0.01) higher in the NEG than EG after more than 6 hours. CONCLUSIONS: Corneal dehydration after organ culture seems to be a multifactorial process, which not only depends on osmotic effects of the deswelling compound but also requires the presence of an intact endothelial cell layer.

Gebauer SLc Original and Moria One-Use Plus automated microkeratomes for ultrathin Descemet's stripping automated endothelial keratoplasty preparation.

PURPOSE: We compared the SLc Original (SLc) and One-Use Plus (OUP) microkeratomes for ultrathin Descemet's stripping automated endothelial keratoplasty (DSAEK) lamella preparation and storage, vis-à-vis accuracy, endothelial cell loss (ECL) and lamellar surface roughness (LSR). METHODS: Twenty-five human corneas were dissected with single-use heads of different sizes aiming for a posterior lamella (PL) thickness of 85 µm, after which they were incubated for 6 days in a 5% dextran medium. Before preparation (0 hr) and 1, 24, and 144 hr after dissection, ECL and corneal thickness (CCT) were measured by ultrasound pachymetry (USP) and optical coherence tomography (OCT). Lamellar surface roughness (LSR) was assessed by scanning electron microscopy (SEM) and evaluated by two masked observers. RESULTS: Prior to cutting, CCTs did not differ between OCT and USP measurements, with a high correlation between the two modalities (r2  = 0.8; p < 0.0001). Both systems succeeded in UT lamella preparation (CCT 40-130 µm) in 88% of cases. The OUP heads cut significantly deeper than the according SLc counterparts (p = 0.001), while the variance did not differ. The mean PL thickness increased significantly in the following incubation period (p = 0.01) with no difference between the keratome groups. Endothelial cell density (ECD) decreased significantly from before to 1 hr after preparation (-5.6%; p = 0.04), with no changes in the following 144-hr incubation period and no differences between the OUP and SLc group. Lamellar surface roughness (LSR) did not differ between both systems. CONCLUSIONS: The SLc and the OUP system are both suited for the preparation of UT-DSAEK lamellae. Neither system differed significantly in variability, LSR or ECL, which did not increase during a 6-day incubation period.

Comparison of Gebauer SLc and Moria CBm Carriazo-Barraquer ALK Microkeratomes for Descemet's Stripping Automated Endothelial Keratoplasty Preparation.

PURPOSE: We compared the hand-guided Moria Carriazo-Barraquer (CBm) microkeratome with the fully automatic SLc microkeratome for Descemet's stripping automated endothelial keratoplasty (DSAEK)-lamella preparation and storage, vis-à-vis accuracy, endothelial cell loss (ECL), and lamellar surface roughness (LSR). METHODS: A total of 18 human corneas were dissected with both the 300 µm CBm multi-use (n = 9) and the 300 µm SLc (n = 9) single-use heads, after which they were incubated for 6 d in a 5% dextran medium. Before preparation (0 h) and 1, 24, and 144 h after dissection, ECL and corneal thickness (CT) were measured by ultrasound pachymetry (USP) and optical coherence tomography (OCT). LSR was assessed by scanning electron microscopy (SEM) and evaluated by three masked observers. RESULTS: Prior to cutting, CTs did not differ significantly between OCT or USP measurements, with a high correlation between the two modalities (r(2)= 0.94, p < 0.0001). One hour after preparation the anterior lamella showed a significantly higher dissection depth with the CBm (429.4 ± 21.8 µm) than the SLc (311.7 ± 54.8 µm, p = 0.0006), with the variance of the SLc system showing a trend towards higher values (p = 0.07). Anterior and posterior lamellae swelled significantly in the subsequent culture period. Both groups showed a significant ECL 1 h after preparation (p < 0.0001) with no significant difference between the systems (1 h: p = 0.44; CBm: - 9.4%, SLc: -11.7%), which stabilized over 144 h (144 h CBm: -13.9%, 144 h SLc: -10.3%). LSR did not differ significantly between both systems (p = 0.60). CONCLUSIONS: The SLc system agrees more with the designated cutting depth than the CBm. The dissection produced a comparable LSR and a ∼10% ECL independently of the system. Further incubation of the prepared lamellae led to a swelling, but no further ECL.

Conjunctival and intraocular swabs for the microbiological assessment of donor corneas.

PURPOSE: In this study, we investigated the associations between conjunctival (co) and intraocular (io) swabs and their implications for the contamination rates of organ-cultured corneas. METHODS: A total of 4177 swabs from 1054 corneas of 527 donors were acquired from the conjunctiva, after disinfection with 5% polyvinylpyrrolidone-iodine solution, and also from the anterior chamber after corneoscleral trepanation (io). Samples were incubated at 22.5 ± 2.5°C and 32.5 ± 2.5°C in thioglycollate broth for 14 days. Donor corneas were cultured in a closed system at 31°C. Microbial differentiation was performed for positive cultures. RESULTS: A higher temperature (32.5°C) and the intraocular swab retrieving localization led to significantly higher swab positive rates (32.5°C versus 22.5°C, odds 1.65, p < 0.0001; io versus co, odds 1,53, p < 0.0001). Death-to-collection time and laterality (left or right eye) had no significant influence on swab positivity. The cause of death significantly influenced the positive rates (p < 0.0001). Detection at 32.5°C occurred significantly earlier than at 22.5°C (p < 0.0001). The overall comparison of detected species showed no significant differences in the variety between intraocular and conjunctival swabs. During the study period, six contaminations of organ-cultured corneas occurred: four times Pseudomonas aeruginosa and once each Candida albicans and Staphylococcus hominis were found. Swap results and cornea contaminations were not significantly correlated. CONCLUSIONS: Co and io swabs show high microbial colonization rates, even after standard disinfection. Io swabs generally reproduce the co microbial range, most likely due to a mobilization and diversion of microorganisms during the trepanation procedure. Swab results do not yield a valuable tool to predict contaminations of organ-cultured corneas.

The Role of Specifically Tailored Communication Training Among Factors Influencing Consent for Cornea Donation Requested Via Telephone.

BACKGROUND: Consent for cornea donation from non-heart-beating donors is often requested by means of telephone interviews with relatives of the deceased. The purpose of this study was to identify the effect of specifically tailored interviewer communications training among other factors related to obtaining consent. METHODS: A retrospective analysis of consent requests made by the Aachen Cornea Bank was performed. Interviews were conducted via telephone by 26 ophthalmic residents or fellows in accordance with German and European laws and guidelines. Multiple logistic regression was used to identify factors related to the consent. Results were expressed as odds ratios (OR), 95% confidence interval (95% CI), and Wald P value. RESULTS: In 1694 interviews, the overall consent rate was 50.12%. Multivariate analysis identified 4 significant factors associated with the donation decision. Completion of a specific communication training seminar increased odds of consent by 1.533 (95% CI, 1.250-1.880; P < 0.0001). The individual interviewer had an impact on the OR of up to 1.255 (95% CI, 1.113-1.417; P = 0.0002). The odds for consent were inversely correlated with the clinical experience of the interviewer (OR, 0.884; 95% CI, 0.831-0.938; P < 0.0001). The consulted family member was 0.894 times less likely to grant consent for donation with each increase in degree of blood relation (95% CI, 0.849-0.940; P < 0.0001). CONCLUSIONS: A telephone interview is effective for obtaining consent for cornea donation. The consent decision may be associated with factors relating to the interviewer as well as the consulted family member. Specific training for cornea bank staff may increase the odds of obtaining consent.

A disposable system for rapid purification of autologous plasmin as an adjunct to vitrectomy - performance and safety profile.

BACKGROUND: The generation of an atraumatic posterior vitreous detachment (PVD), a common goal in vitreoretinal surgery, is a challenge particularly in children and young trauma patients. Plasmin has been proposed as a surgical adjunct to enzymatically generate a PVD. This study assesses the performance and safety of a new system for rapid purification of plasmin as an adjunct to vitrectomy. METHODS: Plasminogen was isolated from human plasma by affinity chromatography using a disposable rapid purification kit, and activated to plasmin with streptokinase. Activities were assessed spectrophotometrically. For safety studies, 38 rabbits received intravitreal injections of one of the following compounds in 0.1 ml respectively: 4.7, 12.7 and 24 IU plasmin, 15 mg dextran, 4,100 U streptokinase, 500 µg ε-aminocaproic acid, 0.1 M potassium phosphate or balanced salt solution (BSS). Thirty min after injection, a two-port vitrectomy was performed. Rabbits were followed clinically and with bright flash electroretinography (ERG) for up to 9 months. The eyes were investigated by light and transmission electron microscopy. RESULTS: The specific plasmin activity obtained from blood of healthy volunteers averaged 42.3 ± 6.6 IU/ml (range 21.6 IU/ml to 54.5 IU/ml). The identity and purity of the enzyme was confirmed by several methods. Clinically, a mild to moderate inflammatory response was seen in most eyes on day 1, but had disappeared by day 7. ERG showed moderate depressions of a- and b-wave amplitudes on day 2, particularly in the potassium phosphate (a: -29.16 ± 4.56, b: -21.23 ± 6.31), 4.7 (a: -34.38 ± 6.64, b: -26.66 ± 6.06) and 24 IU (a: -38.25 ± 4.05, b: -23.38 ± 4.29) plasmin groups, but also in the BSS- (a: -11.19 ± 21.78, b: -11.41 ± 15.47) and dextran- (a: -17.86 ± 14.18, b: -6.67 ± 18.14) treated eyes. ERG changes recovered during follow-up. One rabbit each from the 12.7 and the 24 IU plasmin groups showed a minimal discoloration of one medullary ray after 9 months. Histology did not reveal morphologic signs of toxicity. CONCLUSION: The isolation system generated plasmin with a high degree of purity. A failure-mode analysis did not reveal significant risks of toxicity. A single preparation can provide a maximum dose of 10.9 IU/200 µl, the likely target clinical dose being 1.88 IU. Plasmin doses of at least 12.7 IU appear be safe when injected into rabbit eyes, followed by vitrectomy.

New treatment option for adult-onset limbal xanthogranuloma.

PURPOSE: The purposes of this study are to report a case of adult-onset juvenile xanthogranuloma and suggest a new treatment option. METHODS: A 35-year-old man presented with an isolated mass of the limbus. Biopsy revealed a histiocytic tumor with Touton's giant cells staining positive for CD68 and S-100 but negative for CD1a, consistent with JXG. Two intratumoral injections of triamcinolone acetonide 6 weeks apart were performed. The main outcome measure was 6-months follow up clinically and by ultrasound biomicroscopy. RESULTS: The lesion regressed, and corneal thickness returned to normal 3 months after the first injection. An increase in intraocular pressure required topical treatment. No recurrence was seen throughout follow up. CONCLUSION: Limbal juvenile xanthogranuloma can be successfully treated with intralesional triamcinolone acetonide, thus avoiding a lamellar keratectomy approach.

Detection of contamination during organ culture of the human cornea.

PURPOSE: Corneas harvested post-mortem are at risk of contamination, therefore antibiotic additives are used in cold storage and organ culture systems. In the latter, sterility testing of the medium is part of the standard protocol. Intuitively, testing after longer organ culture periods should be more likely to detect contaminations than early testing, but may delay allocation. This study evaluates whether an optimal time for detection of donor cornea contamination can be identified. METHODS: The study complies with the Declaration of Helsinki. All procedures were supervised using a certified quality management system (ISO 9001:2000). Donor corneas harvested by enucleation or corneoscleral excision over 5 consecutive years were processed according to German and EC laws and guidelines. The corneas were stored in a closed organ culture system in 100 ml MEM containing penicillin, streptomycin and amphotericin B at 31 degrees C for up to 28 days without media exchange. In 762 corneas, 10 ml samples of medium, obtained between days 3 and 8 of culture, were tested for sterility in an automated detection system (BacT/ALERT, bioMérieux). In 424 corneas, a second sterility test was performed from the same medium before release. Contamination detection probabilities were related to the culture duration before the primary test (Cochran-Armitage). RESULTS: Overall, 19 contaminations were found. Contaminations were bilateral in four donors. One contamination was apparent by macroscopic inspection of the medium prior to the primary sterility test; 12 were detected upon primary sterility testing. Furthermore, six primarily undetected contaminations were observed: five in the secondary sterility test and one suspected microscopically. In most cases, contamination could also be seen by medium turbidity and acidification, but in three cases macroscopic medium changes were significantly delayed or absent. No trends were found between the times of primary sterility sampling and both positive and false negative test outcome probabilities. CONCLUSION: Detection probability of contaminations in organ culture media does not increase between days 3 and 8; therefore, sterility can be tested on day 3. With the recommended follow-up after sterility testing being 7 days, microbiologic release can take place after 10 days of culture. Nevertheless, the testing is not failsafe, and should always be combined with macroscopic inspection of the media.

Effect of intravitreal plasmin on vitreous removal through a 25-gauge cutting system in the rabbit in vivo.

PURPOSE: Intravitreal plasmin creates a posterior vitreous detachment, but may also liquefy the vitreous. This study measures the rate of vitreous removal from rabbit eyes after plasmin injection in vivo. METHODS: Intravitreal injections of 150 IU hyaluronidase (n = 5), 0.5 activity units (AU, n = 6) or 0.9 AU of streptokinase-activated human plasmin (n = four groups of 6) in 0.1 ml were performed in rabbits, the fellow eyes received 0.1 ml BSS. After 30 min (hyaluronidase), 30 min, 4 h, 12 h or 24 h (0.9 AU plasmin) or 24 h (0.5 AU plasmin), 1 ml of vitreous was removed from each eye without infusion, using a 25-gauge cutter and a standardized protocol. Animals were sacrificed after surgery. RESULTS: Compared to fellow eyes, the average rate of vitreous removal was increased by hyaluronidase by 68.9 +/- 6.3% (p < 0.05) and by 0.5 AU plasmin (24 h) by 26.8 +/- 3.3% (p < 0.05). 0.9 AU of plasmin increased removal rates by 0.8 +/- 10% (n.s.), 15.4 +/- 6.3% (p < 0.05), 40.3 +/- 3.1% (p < 0.05), and 71.9 +/- 32.4% (p < 0.05) after 30 min, 4 h, 12 h and 24 h incubation respectively. The ratios of removal rates of treated/control eyes in the 0.9 AU groups showed a linear correlation with incubation time (r = 0.783, p < 0.0001). CONCLUSION: Intravitreal plasmin increases the rate of vitreous removal in rabbits.

Recurrent suprachoroidal dislocation of an anterior chamber intraocular lens.

We present a patient with a history of posterior chamber intraocular lens (PC IOL) implantation and trabeculotomy 9 years previously who recently had IOL exchange for an angle-based anterior chamber IOL. Twenty-four hours after the exchange, he was referred to us with hypotony and "loss" of the IOL. Ultrasound revealed a rigid choroidal detachment and echoes suggestive of a suprachoroidally dislocated IOL. Anterior vitrectomy and resuturing of the incision were performed. Subsequently, the IOL repeatedly migrated in and out of the suprachoroidal space, allowing atraumatic removal. Angle-based IOL implementation should be avoided in eyes with a history of trabeculotomy or cyclodialysis.

Sterilization and preservation influence the biophysical properties of human amnion grafts.

The introduction of amniotic membrane (AM) transplantation in ophthalmic surgery holds great promise and in many clinical situations it offers an alternative to existing management options. The purpose of this study was to examine the influence of established sterilization and preservation procedures on biophysical and histological properties of AM grafts. Amnion was sterilized by peracetic acid/ethanol sterilization [PES] and preserved by air-drying (sterile laminar flow) [AD] or in glycerol [GLYC]. Unsterilized AM were preserved at -80 degrees C [-80 degrees C] and served as an experimental control. Amnion allografts were characterized by the determination of their thickness, moisture vapour permeability (MVP), oxygen permeability (OPERM), tensile strength and sulphur content. Immunostaining for tissue-specific and basement membrane-related proteins was performed. Differences in biophysical parameters were found between the unsterilized allografts and the sterilized, air-dried or glycerol-preserved allografts. [PES/AD] showed the highest MVP and OPERM, the highest tensile strength and the lowest sulphur content and thickness. [PES/GLYC] exhibited the lowest OPERM and the highest thickness compared to [-80 degrees C] and [PES/AD]. Collagen types V and VII were preserved the best in the control group. Sterilization and preservation affect biophysical properties important for the use of AM as allogenic grafts. It has to be determined if any change, as noted, has a clinical impact.

Efficacy of plasmin enzymes and chondroitinase ABC in creating posterior vitreous separation in the pig: a masked, placebo-controlled in vivo study.

BACKGROUND: Induction of posterior vitreous detachment (PVD) during vitrectomy helps to prevent proliferative complications, but can be traumatic to the retina, particularly in young patients. Adjunct enzymes have been proposed to facilitate PVD. We investigated the efficacy of enzymes in creating PVD as an adjunct to vitrectomy in the pig. METHODS: Five groups of 8 pigs received a masked intravitreal injection of chondroitinase (1 IU), human (0.4 or 1.3 activity units [AU]) or porcine plasmin (0.18 AU or 0.47 AU) into one eye, and osmolarity adjusted control into the other. After incubation, a core vitrectomy was performed on each eye at low suction, without vitreous peeling. The occurrence of spontaneous PVD and its extent were graded. Eyes were investigated using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Vitreous remnants on the retina were quantified in SEM. Data were analyzed using McNemar's test for paired observations and Wilcoxon paired signed rank test. RESULTS: Spontaneous PVD occurred more frequently in human plasmin-treated eyes (p<0.025) and all plasmin eyes (p<0.025) than in placebo controls. The extent of PVD appeared larger in human plasmin (p<0.025) and all plasmin-treated eyes (p<0.025). In plasmin-treated eyes, SEM morphometry showed a significant reduction in the vitreous-covered retina areas. Chondroitinase failed to produce an effect. CONCLUSIONS: Plasmin may prove a useful adjunct to conventional vitrectomy.

Longer-term outcomes of transconjunctival sutureless 25-gauge vitrectomy.

PURPOSE: To report longer-term outcomes in eyes undergoing 25-gauge transconjunctival sutureless vitrectomy. DESIGN: Retrospective, noncomparative, case series. METHODS: Chart review of the initial 45 consecutive patients (45 eyes) that underwent TSV by one surgeon (T.S.H.) for idiopathic epiretinal membrane (n = 15), refractory diabetic macular edema (n = 11), idiopathic macular hole (n = 10), and nonclearing vitreous hemorrhage (n = 9). All patients had at least 6-month follow-up. Main outcome measures included visual acuity (VA), intraocular pressure, intraoperative complications, and postoperative complications. RESULTS: Mean follow-up was 13 months (range 6 to 25 months). Mean overall preoperative VA vs last postoperative VA was 20/229 and 20/65, respectively (P < .0001). Statistically significant VA improvement was seen for each patient subgroup. Mean preoperative intraocular pressure was 16.9 mm Hg (range 10-26 mm Hg). On postoperative day 1, week 1, and week 4, median intraocular pressure was 14.6 mm Hg (range 8-17 mm Hg), 17.6 mm Hg (range 8-38 mm Hg), and 17.7 mm Hg (range 9-33 mm Hg), respectively. No intraoperative complications occurred. Postoperative complications were 1 inferior retinal detachment (2.2%) 4 weeks after macular hole repair, 1 macular hole (2.2%) 6 months after epiretinal membrane peel, and 23 worsening cataracts in 29 phakic eyes (79.3%). CONCLUSIONS: Less surgically complex vitreoretinal pathology may be successfully repaired with TSV. After a mean follow-up of more than 1 year, minimal complications were seen, and none was specifically related to the sutureless nature of the procedure.

Indocyanine-green-assisted internal limiting membrane peeling in macular hole surgery--a follow-up study.

BACKGROUND: Macular hole surgery including vitrectomy and peeling of epiretinal membranes and the internal limiting membrane (ILM) has become a standard procedure in retinal surgery. Poor visualization of the ILM is an obstacle for successful surgery. Recently, indocyanine green (ICG) has been reported to be a helpful intraocular substance in identifying these membranes. PATIENTS AND METHODS: Eighteen eyes with macular holes stages 2-4 were included. Intraoperatively, the ILM was stained with three drops of 1:9-diluted ICG. After 1 min incubation, the vitreous cavity was rinsed with Ringer's lactate solution, and the ILM was peeled. Autologous thrombocytes were applied to the macular hole and the eye was endotamponaded with 20% SF-6 gas. Preoperatively, 6 weeks postoperatively, and in 3-month intervals thereafter, visual acuity, fundus photographs, scanning laser ophthalmoscope imaging, and Humphrey 24-2 static perimetry was performed. RESULTS: Intraoperatively, the ILM could be nicely visualized by ICG, which allowed easier and less traumatic peeling. At 6 weeks follow-up, visual acuity had improved in 14 of 18 patients, and the macular hole was closed 6 weeks after surgery. Scanning laser imaging revealed a strong signal. During prolonged follow-up, visual acuity declined due to cataract formation. CONCLUSION: ICG as an intraocular tool for staining of the ILM is helpful in macular hole surgery. We observed no negative effects on retinal function, but patients should be followed.