Three-dimensional structure of the mammalian limbal stem cell niche.

Although the existence of the limbal stem cell (LSC) niche is accepted, precise knowledge of its three-dimensional (3D) architecture remains incomplete. The LSC niche was explored on freshly excised and organ-cultured corneoscleral rims from human donors (n = 47), pigs (n = 15) and mice (n = 27) with full-field optical coherence microscopy (FFOCM). Limbal crypt features were detected in 90% of organ-cultured human corneoscleral rims, extending between the palisades of Vogt as radially oriented rectangular (74% of eyes) and/or rounded (23% of eyes) forms, often branching off to, or becoming interconnected by, sub-scleral radially or circumferentially oriented crypts (in 56% of eyes). Mean crypt volume represented 16% of sampled limbal volume on the vertical axis and 8% on the horizontal axis. In pigs, palisades were finer and crypts wider with relatively uniform distribution around the eye, and radial orientation, connecting to numerous narrow criss-crossing invaginations beneath the scleral surface. In mice, only a circumferential limbal trough was detected. Mean crypt volume represented 13% of sampled limbal volume in humans and 9% in pigs. FFOCM combined with fluorescence, and confocal fluorescence microscopy, showed presence of p63-α+ cells and cytokeratin-3+ cells in the limbal crypts. To assess colony forming efficiency (CFE), limbal epithelial cells were cultured at low density with mitomycin-arrested 3T3 feeders. CFE increased with limbal crypt volume and was not significantly decreased in organ-cultured cornea, despite degradation of the epithelial roof, suggesting that stem cells remain protected at the base of crypts during organ culture. CFE in human samples was significantly greater than in pig, and CFE in mouse was zero. Crypt architecture in the three species appears associated with eye exposure to light. LSC density increased with percentage limbal volume occupied by crypts.

Simultaneous penetrating keratoplasty and amniotic membrane transplantation in eyes with a history of limbal stem cell deficiency.

PURPOSE: To describe the outcomes of simultaneous penetrating keratoplasty (PK) and amniotic membrane transplantation (AMT) performed both as a ring-shaped graft and as a temporary patch in eyes with a history of limbal stem cell deficiency (LSCD). METHODS: Prospective observational case series including 48 simultaneous PK/AMT procedures (48 patients) in eyes with a history of partial or total LSCD. Patients with total LSCD were first treated with limbal stem cell transplantation. The preoperative indication was graft failure in 58.3% of cases. Most recipients (89.6%) were at high-risk for rejection. RESULTS: The mean graft reepithelialization time was 29.2±30.8 days. Graft reepithelialization was achieved in 30 days in 70.8% of cases. No AMT-related adverse events were observed. The mean time from keratoplasty-to-last visit was 84.5±54.5 months. The 3-year graft survival rate was 62.5%. Recurrence of corneal epithelial defects after graft reepithelialization (47.9%) was associated with lower graft survival (P=0.004). In eyes with successful grafts at the last visit, the mean LogMAR visual acuity was 1.90 (20/1575)±5 lines before keratoplasty and 0.89 (20/155)±10 lines at 5 years. A ring of amniotic membrane was visible between the graft stroma and the corneal epithelium on slit-lamp examination and optical coherence tomography in all successful cases. CONCLUSIONS: In this series of eyes with a history of LSCD and at high-risk of rejection, simultaneous PK and AMT were associated with satisfactory graft survival and no additional adverse events.

Microbiologic study of organ-cultured donor corneas.

BACKGROUND: Our purpose was to evaluate the sterility of organ-cultured human donor corneas at the time of surgery. METHODS: We studied 603 organ-cultured corneas. Of these 603 corneas, 409 (68%) were grafted and 69 (11%) were contaminated during storage. RESULTS: Contamination during preservation was either bacterial (65%) or fungal (35%). None of the tested antibiotics were effective against all of the 45 isolated bacteria. The risk of contamination decreased with death-to-organ culture time (P=0.008) and was higher for corneas excised in situ than for those enucleated (P=0.02). Corneoscleral rims were sterile in 99.3% of the grafted corneas. Deswelling media were sterile in 100% of cases. A 19- to 53-fold decrease in the percentage of rim contamination was assessed with organ culture as compared with hypothermic storage (previous studies, P < 0.0001). CONCLUSIONS: These results demonstrate the benefit of organ culture over hypothermic storage, because it allows contaminated tissue to be discarded.

The results of successful penetrating keratoplasty using donor organ-cultured corneal tissue.

BACKGROUND: The aim of this study was identification of predictive factors for postoperative visual acuity in patients with a clear organ-cultured graft and to analyze the change in visual acuity between 12 and 24 months after transplantation. METHODS: The study design was a prospective cohort study. A total of 342 consecutive penetrating keratoplasties using donor organ-cultured grafts, performed in 324 patients, were included. Visual acuity, graft thickness, and graft endothelial cell density were recorded in patients with clear transplants. RESULTS: At 24 months postoperatively, 25 (18.7%) of 134 patients had 20/200 or worse visual acuity and 66 (49.3%) had 20/40 or better visual acuity. Graft thickness took 1 month to decrease to normal values. A temporary graft thinning occurred at 6 months postoperatively, followed by recovery of normal graft thickness by 18 months. The average postoperative endothelial cell density was 1,533+/-598 cells/mm2 during the second year. The 24-month LogMAR (logarithm of minimal angle of resolution) visual acuity correlated with preoperative LogMAR visual acuity (beta=0.26, P=0.005), postoperative lens status (beta=-0.34, P=0.008), preoperative intraocular pressure (beta=0.50, P=0.020), and postoperative astigmatism (beta=0.17, P=0.040). Visual acuity (P=0.022) significantly improved between 12 and 24 months. Preoperative diagnosis (P < 0.0001) and postoperative lens status (P < 0.0001) significantly influenced the change in LogMAR visual acuity between 12 and 24 months. CONCLUSIONS: Donor variables do not influence the visual acuity results of penetrating keratoplasty using organ-cultured donor tissue, whereas they have a strong influence on graft survival and graft endothelial cell density. Visual acuity improves during the first 2 years after transplantation. After keratoplasty, organ-cultured corneal grafts undergo dramatic modifications of their thickness and probably of their transparency.

Corneal endothelial cell apoptosis in patients with Fuchs' dystrophy.

PURPOSE: To investigate whether apoptosis plays a notable role in degeneration of corneal endothelial cells in patients with Fuchs' dystrophy. METHODS: Forty-seven corneal buttons from 41 patients with Fuchs' dystrophy were studied. Nucleus labeling, transmission electron microscopy (TEM), and TdT-dUTP terminal nick-end labeling (TUNEL) were used to detect apoptosis. TEM and TUNEL were performed on sections of all 47 corneal buttons, and nucleus labeling was performed on the last 10 corneas. Seven human donor corneas, two corneal buttons from two patients with keratoconus, and one corneal button from a patient with interstitial keratitis were used as negative controls for detection of apoptotic endothelial cells. Negative controls were studied by means of nucleus labeling, TUNEL, and TEM. RESULTS: In the nucleus labeling assay, the average percentage of apoptotic endothelial cells was 2.65% in the Fuchs' dystrophy group (n = 10) and 0.23% in the control group (n = 10; P = 0.0003). In the TUNEL assay, labeling of some endothelial cells was observed on 42 of 47 corneas in the Fuchs' dystrophy group, whereas it was absent on most specimens of the control group. In TEM, most endothelial cell nuclei had a normal appearance, and apoptotic endothelial cells featuring condensed nucleus and decreased cell size could be observed exceptionally. Some apoptotic cells were found in the basal epithelial cell layer by means of nucleus labeling, TUNEL, and TEM in the Fuchs' dystrophy group but not in the control group. CONCLUSIONS: This study suggests that apoptosis plays an important role in endothelial cell degeneration in Fuchs' dystrophy. Because of a lack of conclusive evidence of increased endothelial apoptosis by TEM, further studies are needed to ascertain this finding.

Cryopreservation and culture of human corneal keratocytes.

PURPOSE: To assess the effects of two different concentrations of albumin in a cryoprotective solution and two freezing methods on human corneal keratocyte ctyopreservation. METHODS: Isolated keratocytes were used for cryopreservation. Solutions of 10% dimethylsulfoxide with either 2% or 10% human albumin were used as cryoprotective agents. Cells either were transferred directly into a -80 degrees C freezer (freezing rate, 2 degrees C/min) or were cooled in a programmed freezer (1 degrees C/min until -40 degrees C and then 10 degrees C/min), which resulted in four different cryopreservation protocols. Cells were stored at -80 degrees C, then were thawed at 37 degrees C, and subsequently were cultured. Keratocytes were studied by means of trypan blue staining, growth assay, apoptosis assays, transmission electron microscopy, and immunochemistry. RESULTS: The percentage of cells that were alive after thawing ranged from 80% to 99% by trypan blue staining and from 45% to 60% by flow cytometry. The ratio of the number of living cells at the end of primary culture after cryopreservation to that before cryopreservation was significantly (P=0.04) higher after direct transfer into the -80 degrees C freezer than after controlled-rate freezing, whereas the albumin concentration had no significant influence on this ratio (P=0.45). The percentage of apoptotic cells was significantly higher after cryopreservation than in the control group of noncryopreserved cells; more than 5% 24 hours after thawing. Cryopreservation did not modify the keratocyte ultrastructure. Fibroblast growth factor dramatically decreased the serum-induced cell expression of alpha smooth muscle actin, whereas cryopreservation had no influence on this cell expression. CONCLUSIONS: A freeze-thaw trauma, which was related to cryopreservation-induced cell apoptosis, was revealed during primary culture after thawing. Direct transfer into the -80 degrees C freezer resulted in better postcryopreservation growth in the culture than controlled-rate freezing. A change in albumin concentration from 2% to 10% did not affect the results.

The complication of retained intraocular recipient corneal button after penetrating keratoplasty.

PURPOSE: To describe the complication of retained intraocular recipient corneal button after penetrating keratoplasty. METHOD: Case report. A 27-year-old man was referred to us because of complications after penetrating keratoplasty on the right eye. Slit-lamp examination disclosed a clear graft with a second clear corneal button behind. RESULTS: A second penetrating keratoplasty was performed successfully on the right eye. Histologic examination showed an intact epithelium and stroma vascularization of the recipient corneal button. Immunochemistry of the recipient corneal button and the first donor corneal button was performed. CONCLUSION: Although the recipient corneal button remained inside the anterior chamber for 5 months, no epithelial ingrowth occurred.

Measurement of irregular astigmatism using semimeridian data from videokeratographs.

BACKGROUND: Irregular astigmatism is one of the most troublesome side effects of corneal surgery. We attempted to quantify irregular astigmatism in a simple way, using semimeridian data from videokeratographs of corneas with different amounts of irregular astigmatism. METHODS: We studied the corneal topography of 30 normal corneas to be used as controls, 30 corneas from patients after radial keratotomy, 30 corneas from patients after nonfreeze myopic keratomileusis, and 30 corneas with clinically apparent keratoconus. We developed two diagnostic formulas, a refractive power symmetry index and an angle symmetry index, using the absolute-scale, color-coded map, and semimeridian data from the EyeSys Corneal Analysis System, and used them to compare the four groups of corneas. RESULTS: In the corneas with irregular astigmatism, values for the refractive power symmetry index and angle symmetry index were significantly higher than in the normal corneas. The refractive power symmetry index confidence interval set at 95% was 0.58 diopters (D) (range 0.00 D to 0.58 D) in the control group. Using these limits, most values in the radial keratotomy, keratomileusis, and keratoconus groups were abnormal. Spectacle-corrected visual acuity correlated negatively with the refractive power symmetry index (rs = -0.58, p < .001). The refractive power symmetry index sensitivity and specificity were respectively 97% and 66%. Spectacle-corrected visual acuity did not correlate with the angle symmetry index. CONCLUSIONS: Semimeridian data from videokeratographs can be used to quantify irregular astigmatism.

Videokeratography, keratometry, and refraction after penetrating keratoplasty.

PURPOSE: To identify the correlation between videokeratography, autorefractometry, autokeratometry, and keratometry measurements and the subjective manifest refraction and spectacle-corrected visual acuity after penetrating keratoplasty. METHODS: We studied 100 eyes from 100 patients that had undergone penetrating keratoplasty. All eyes were examined by videokeratography (EyeSys 2000) (axial, tangential, and refractive power maps), autorefractometry, autokeratometry, and keratometry. Measurements were made at an an average of 19 +/- 9 months after surgery. Postoperative refractive astigmatism, spherical equivalent refraction, and spectacle-corrected visual acuity were studied by regression analysis. RESULTS: Both the total topographic cylinder measured by the refractive power map and topographic cylinder measured by the axial power map showed the strongest correlation with the manifest refractive cylinder (rs = +0.89, P < .001). The axis of astigmatism determined by keratometry and autokeratometry showed the strongest correlation with the subjective manifest refraction axis (rs = +0.87, P < .001). The total topographic cylinder showed the strongest correlation with the spectacle-corrected visual acuity (rs = +0.38, P = .001); however the topographic indices of predicted corneal acuity, corneal acuity, corneal uniformity index, asphericity, and refractive power symmetry did not correlate with spectacle-corrected visual acuity. CONCLUSION: Measurement of astigmatism after penetrating keratoplasty can be made more accurate by using videokeratographic measurements to supplement retinoscopic and manifest refraction. Other useful methods for predicting the axis of refractive astigmatism include keratometry, autokeratometry, and autorefractometry. In this study, indices designed to measure corneal surface irregularity failed to predict visual acuity after penetrating keratoplasty.

Prediction of spectacle-corrected visual acuity using videokeratography.

PURPOSE: Our aim was to improve prediction of spectacle-corrected visual acuity (SCVA) using indices derived from the EyeSys System 2000 data (version 3.1). METHODS: We studied corneal topography in 182 eyes from 8 groups of patients. Holladay Diagnostic Summary indices were recorded. Nine statistical indices calculated with the first 8-ring data and refractive power symmetry index were also studied. Correlation with SCVA (LogMAR units) was studied by means of Pearson's regression. Multiple linear regression was used to obtain linear equations combining several indices. RESULTS: At a univariate level, total astigmatism cylinder showed the strongest correlation with SCVA (r = .63, P = .0001). At a multivariate level, the predicted visual acuity obtained by linear equation combining the asphericity coefficient, the predicted corneal acuity, the mean of the means, and the total astigmatism cylinder was closely associated with SCVA (r = .72, P = .0001). It was identical to SCVA in 58.2% of the cases, within one line in 75.8%, and within two lines in 91.2%. CONCLUSION: Multiple linear regression resulted in the best prediction of spectacle-corrected visual acuity, giving notable improvement in prediction of spectacle-corrected visual acuity as compared to the predicted corneal acuity available in the EyeSys System 2000.

Tissue crossmatch before corneal transplantation.

AIM: To investigate the influence of anticorneal antibodies on the outcome of transplantation in recipients before penetrating keratoplasty. METHODS: 100 consecutive penetrating keratoplasties performed between July 1995 and June 1996 were included in this study. Donor corneoscleral rim cryosections were incubated with recipient serum. Fixation of recipient IgM on donor corneas was revealed with a polyclonal anti-human mu-chain antibody. The mean follow up time was 61 months. Five years after transplantation, 63% of patients were available for follow up. During the follow up period, 15% of keratoplasties failed because of irreversible rejection. RESULTS: Anti-donor stromal lamellae IgM were found in 14 recipients. Anti-donor keratocyte IgM were present in 28 recipients. Dilution of recipient serum (up to 1/800) did not modify the results of positive crossmatch assays. Crossmatch assays performed with positive recipient sera and negative donor corneoscleral rims were negative. No significant influence of IgM crossmatch results on graft survival and rejection-free graft survival was found. CONCLUSION: Donor specific anticorneal stroma IgM were found in 28% of recipients before transplantation. This pre-immunisation was not associated with a higher risk of transplantation rejection.

The triple procedure: in the bag placement versus ciliary sulcus placement of the intraocular lens.

AIMS: To evaluate the influence of intraocular lens (IOL) placement on triple procedure clinical results and to investigate whether it is appropriate to use phacoemulsification in patients with large lens nucleus. METHODS: 40 consecutive penetrating keratoplasties combined with cataract extraction performed in a single institution were studied. Whenever possible a capsulorhexis was performed and the IOL was placed into the capsular bag. Phacoemulsification was used when the nucleus was too large to pass through the capsulorhexis. RESULTS: Out of 25 patients with an intact capsulorhexis phacoemulsification was used in 13 (52.0%) whereas the entire nucleus passed through the capsulorhexis in the remaining 12 patients (48%). The average 12 month visual acuity was 0.46 (SD 0.21) in patients with in the bag IOL (n = 23) and 0.29 (0.08) in patients with ciliary sulcus IOL (n = 13) (p = 0.04). Elevated intraocular pressure occurred in 26.1% (6/23) of patients with in the bag IOL and 61.5% (8/13) of patients with ciliary sulcus IOL (p = 0.08). The average postoperative graft thickness at 18 months was 552 (27) microns in the former group and 650 (29) microns in the latter group (p = 0.04). No significant difference in graft survival, postoperative endothelial cell density, astigmatism, and videokeratoscopic measurements was found between both groups. CONCLUSION: In the bag placement of the intraocular lens during the triple procedure results in better outcome of transplantation than ciliary sulcus placement of the IOL. Phacoemulsification allows removal of large nuclei through a 5 mm capsulorhexis without performing relaxing incisions out towards the periphery of the capsule.

Donor organ cultured corneal tissue selection before penetrating keratoplasty.

AIMS: Donor organ cultured corneal tissue selection before penetrating keratoplasty is carried out by taking into account different variables. The objective was to identify preoperative variables which are significantly and independently associated with transplant outcome and should effectively be taken into account before transplantation. METHODS: 231 consecutive penetrating keratoplasties were prospectively studied using organ cultured tissue. Morphometric analysis of the donor corneal endothelium was performed before transplantation. Graft survival and endothelial cell density, during the second year following transplantation, were studied both at a univariate and multivariate level. RESULTS: Recipient age, recipient rejection status, and preoperative diagnosis significantly influenced graft survival. Graft survival was higher when using corneal tissue from donors older than 80 years. Postoperative endothelial density decreased with preservation time and coefficient of variation after preservation. It increased with endothelial cell density after preservation and deswelling time, and correlated with preoperative diagnosis. CONCLUSION: Organ cultured corneas with endothelial cell density after preservation < 2000 cells/mm2, and high coefficient of variation, may be discarded before transplantation. Corneas should be preserved for less than 3 weeks, and allowed to deswell before transplantation for 2 or 3 days rather than 1 day.

Screening human donor corneas during organ culture for the presence of guttae.

AIMS: To detect the presence of guttae by means of light microscopy during organ culture and to evaluate the influence of the presence of guttae in the donor tissue on transplantation outcome. METHODS: Donor corneas were investigated for the presence of guttae by means of light microscopy at the end of organ culture. Recipient corneal buttons from patients with severe Fuchs' dystrophy and donor corneas with advanced guttae were first studied by light microscopy and subsequently by transmission electron microscopy. Lastly, 168 consecutive donor corneas were evaluated for the presence of guttae and issued for transplantation. RESULTS: Corneal specimens with Fuchs' dystrophy displayed numerous round highly reflecting guttae at the level of the corneal endothelium. Donor corneas with advanced guttae showed less numerous guttae. Among 168 organ cultured donor corneas issued for transplantation, low density guttae were found in 43 (25.6%) corneas. The endothelial cell density and figure coefficient were significantly lower and organ culture time was significantly higher in the cornea guttata group than in the control group. The presence of grouped guttae significantly decreased the adjusted graft survival. The incidence of postoperative stage 3 cornea guttata was significantly higher when grouped guttae were found (5/6) than when no guttae or scattered guttae were found (8/101). CONCLUSION: Cornea guttata can be detected during organ culture by means of light microscopy. It is associated with a decrease in endothelial cell figure coefficient and cell density. The presence of grouped guttae is associated with poorer graft survival and more frequent stage 3 cornea guttata in the graft after transplantation.

Surgical correction of postkeratoplasty astigmatism with the Hanna arcitome.

PURPOSE: To report the results of arcuate keratotomy performed with the Hanna arcitome in patients with postkeratoplasty astigmatism. SETTING: Department of Ophthalmology, Saint-Antoine Hospital, Paris VI University, Paris, France. METHODS: This retrospective study comprised 22 eyes (22 patients) with postkeratoplasty astigmatism. Paired symmetrical arcuate keratotomy was performed with the Hanna arcitome. Outcome measures included refraction, videokeratography, and keratometry. RESULTS: At 6.6 months +/- 8.9 (SD) after surgery, the mean increase in best spectacle-corrected visual acuity (BSCVA) was 2.1 +/- 2.4 lines. Thirteen eyes gained 2 lines or more of BSCVA, and 15 gained 3 lines or more of uncorrected visual acuity. Two patients had a decrease in BSCVA: 1 had lens opacification unrelated to arcuate keratotomy and 1, increased corneal irregularity. Mean refractive astigmatism was 6.94 +/- 2.11 diopters (D) preoperatively and 3.85 +/- 1.95 D postoperatively (P < .01). Mean change in keratometric astigmatism was -51 +/- 36%. Astigmatism decreased in 21 eyes as measured by manifest refraction, keratometry, and videokeratography; it increased in 1 cornea with a microperforation. CONCLUSIONS: The results of arcuate keratotomy performed with the Hanna arcitome were comparable to those with freehand relaxing incisions. The instrument made safer and more uniform arcuate incisions than a freehand technique.

Automated keratoconus detection using the EyeSys videokeratoscope.

PURPOSE: To evaluate the effectiveness of indices derived from the EyeSys System 2000 in detecting keratoconic corneas. SETTING: Department of Ophthalmology, Hôpital Saint Antoine, Paris VI University, Paris, France. METHODS: Topographies of 208 corneas were evaluated. The corneas were from 8 groups of patients classified by the following diagnoses: normal, regular astigmatism, cataract, radial keratotomy, photorefractive keratectomy, myopic keratomileusis, penetrating keratoplasty (PKP), and keratoconus. Nine statistical indices derived from EyeSys data, 2 Holladay Diagnosis Summary indices (coefficient of uniformity and coefficient of asphericity [Asph]), and our refractive power symmetry index were studied. A training set of 104 corneas was used to determine the most efficient threshold value of each index based on sensitivity and specificity curves. Decision trees combining 2 indices were generated. Sensitivity and specificity were calculated in a validation set composed of the remaining 104 corneas. RESULTS: Based on the results of the training set, the optimum indices were SDSD (standard deviation of the standard deviations of the radii of curvature of each ring) and Asph. In the validation set, the decision tree using these indices featured 88.5% sensitivity and 94.9% specificity; the 4 false-positive cases were in corneas in the PKP group of patients. CONCLUSIONS: Clinically apparent keratoconus can be detected among normal corneas and irregular corneal shape patterns using the EyeSys System 2000 data and a decision tree combining 2 indices.

Ultrastructure of cultured and cryopreserved human corneal keratocytes.

PURPOSE: To describe the ultrastructural features of cultured and cryopreserved keratocytes. METHODS: Isolated human keratocytes were cultured with 10% fetal calf serum and 10 ng/ml acidic fibroblast growth factor. The 10% Me2SO and 10% human albumin were used as cryoprotective agents. Cells were cooled at 2 degrees C/min, then thawed at 37 degrees C, and subsequently recultured. They were studied by means of transmission electron microscopy (TEM). RESULTS: TEM of cultured keratocytes before cryopreservation showed a network of intact connecting cells. The average cell thickness was 2.4 microm in cross sections and 5.8 microm in frontal sections. The average nuclear thickness was 1.6 microm in cross sections and 3.7 microm in frontal sections. Nuclei appeared regular and oval in cross sections and indented in frontal sections. Organelles were found in greater amounts in frontal sections than in cross sections. Gap junctions, fenestrations along the cell surface, omega-shaped structures, fibrils, and filamentous networks also were found. Most of the just-thawed, suspended cells were elongated and condensed but had intact plasma membranes. These cells were surrounded by a granular material, corresponding to the albumin-containing thawing medium. Scattered isolated round cells displayed nuclear damage, cell edema, loss of organelles, and cell-membrane disruption. By the end of reculture after cryopreservation, cultured keratocytes displayed the same ultrastructural features as before cryopreservation. CONCLUSION: Cultured human keratocytes display many ultrastructural features of in situ keratocytes. These features are still present after reculture after cryopreservation. Cryopreservation induces necrosis in a small percentage of cells, which seems to be related to a relative lack of cell-membrane protection by the cryoprotectants used.

ABO antigen blood-group compatibility in corneal transplantation.

Our objective was to evaluate the effect of ABO antigen blood-group compatibility on corneal allograft rejection. The 199 penetrating keratoplasties performed between 1985 and 1994 were analyzed retrospectively for ABO compatibility and the occurrence of irreversible allograft rejection. Of these, 72 were considered high-risk transplants, as there was significant vascularization of the recipient cornea or a history of irreversible corneal allograft rejection or both. One hundred twenty-seven were low-risk transplants. The data were analyzed by using the Kaplan-Meier method and compared with the log-rank test. Overall, the estimated 1-year graft survival was 83.9% in the low-risk group and 61.4% in the high-risk group (p = 0.001). The estimated 1-year rejection-free graft survival was 89.8% in the low-risk group and 67.1% in the high-risk group (p = 0.0002). In the high-risk group, graft survival (p = 0.008) and rejection-free graft survival (p = 0.0002) were higher in the ABO-compatible subgroup than in the ABO-incompatible subgroup. In the low-risk group, graft survival and rejection-free graft survival of the ABO-compatible and ABO-incompatible subgroups were similar. ABO compatibility may be effective in preventing irreversible allograft rejection in high-risk recipients.

Effects of penetrating keratoplasty suture removal on corneal topography and refraction.

PURPOSE: To study the refractive and topographic modifications induced by penetrating keratoplasty suture removal. METHODS: We retrospectively studied 50 corneas from 50 patients who had undergone penetrating keratoplasty. All of the eyes were examined before and after all sutures were removed at 10.8+/-4.7 months and 20.3+/-7.9 months after keratoplasty. Subjective refraction, best spectacle-corrected visual acuity (LogMAR units), and corneal topography (EyeSys 2000R device) were recorded. RESULTS: Suture removal decreased the subjective cylinder by an average of 0.91+/-2.32 D (p = 0.004) and modified the corneal topographic pattern (p = 0.03) and shape (p < 0.001) distribution. The change in subjective cylinder correlated with the variation of the steepest meridian power (r(s) = 0.46; p < 0.001). It correlated with the change in topographic pattern (r(s) = 0.59; p < 0.001). The subjective spherical equivalent increased (hyperopization) by an average of 0.61+/-2.24 D (p = 0.01). After suture removal, the prolate shape was less frequent, and the oblate shape was more frequent than before suture removal. Best spectacle-corrected visual acuity increased by an average of 0.8+/-2.0 lines (p = 0.004). The change in subjective cylinder correlated with the change in visual acuity (r(s) = 0.36; p = 0.006). CONCLUSION: Suture removal modifies the corneal topographic pattern and shape. It decreases the subjective cylinder and induces an hyperopization. Visual-acuity improvement after suture removal mainly is explained by the decrease in astigmatism. Suture removal seems particularly helpful in corneas with a bow-tie pattern.

Evaluation of the deswelling period in dextran-containing medium after corneal organ culture.

PURPOSE: To evaluate corneal structural modifications induced by the deswelling period in dextran-containing medium following organ culture. METHODS: Twenty human corneas were organ-cultured for 2 weeks and subsequently incubated in Exosol deswelling medium (Opsia, Toulouse, France) for 1-4 days. Corneas were studied by means of light microscopy, morphometry, and transmission electron microscopy. RESULTS: The deswelling period induced a statistically significant 8.4% endothelial cell loss and a 27.4% increase in the coefficient of variation. The endothelial layer remained intact. The basal epithelial cells displayed a flat appearance. Thin endothelial cells were observed in addition to dark vacuoles (the number of which increased with incubation time) with dense material and no mitochondrial swelling. Some basal epithelial cells and keratocytes were damaged on day 3 or 4. CONCLUSIONS: Preservation injuries induced by 1 or 2 days of incubation in deswelling medium are moderate. Three or 4 days of incubation result in more severe dextran-induced injuries. Consequently, the deswelling period should not exceed 2 days.