Impact of Smoking on Visual Field Progression in a Long-term Clinical Follow-up.

PURPOSE: To investigate the effect of smoking on rates of progressive visual field (VF) damage over time in glaucoma. DESIGN: Retrospective cohort study. PARTICIPANTS: Five hundred eleven eyes of 354 patients with glaucoma followed up from multicenter glaucoma registries. METHODS: In this longitudinal study, 354 patients with primary open-angle glaucoma with a minimum of 3 years of follow-up and 5 VF tests were enrolled from the Diagnostic Innovations in Glaucoma Study and the African Descent and Glaucoma Evaluation Study. Univariate and multivariate linear mixed models were used to investigate the effects of smoking on rates of 24-2 VF mean deviation loss. Visual field progression was defined using pointwise linear and significant negative VF mean deviation loss. Logistic regression was used to identify baseline factors and whether different levels of smoking intensity were associated with VF progression. Kaplan-Meier survival analysis and the log-rank test were used to compare the cumulative risk ratio of progression between smoker and never smoker groups. MAIN OUTCOME MEASURES: Visual field progression. RESULTS: Five hundred eleven eyes of 354 patients were included over the median follow-up of 12.5 years. Median baseline age was 64.8 years. Of the 354 patients, 124 (35%) were Black, and 149 (42.1%) and 168 (59.8%) had reported a history of smoking or alcohol consumption, respectively. In a multivariate model, higher smoking intensity was associated with faster VF loss (coefficient, -0.05 decibels (dB)/year per 10 pack-years; 95% confidence interval [CI], -0.08 to -0.01 dB/year per 10 pack-years; P = 0.010). Developing VF progression in eyes of heavy smokers (≥ 20 pack-years) was 2.2 times more than in eyes of patients without smoking history (odds ratio, 2.21; 95% CI, 1.02-4.76; P = 0.044). Statistically significant differences were found between heavy smokers (≥ 20 pack-years) and never smokers by Kaplan-Meier analysis (P = 0.011, log-rank test). CONCLUSIONS: Heavy smokers are more likely to sustain VF loss in eyes with glaucoma. The prospective longitudinal design of this study supports the hypothesis that levels of smoking may be a significant predictor for glaucoma progression. Additionally, this information can be used for clinically relevant tobacco prevention and intervention messages.

Cornea-Derived Mesenchymal Stromal Cells Therapeutically Modulate Macrophage Immunophenotype and Angiogenic Function.

Macrophages are crucial drivers of inflammatory corneal neovascularization and thus are potential targets for immunomodulatory therapies. We hypothesized that therapeutic use of cornea-derived mesenchymal stromal cells (cMSCs) may alter the function of macrophages. We found that cMSCs can modulate the phenotype and angiogenic function of macrophages. In vitro, cMSCs induce apoptosis of macrophages while preferentially promoting a distinct CD14hi CD16hi CD163hi CD206hi immunophenotype that has significantly reduced angiogenic effects based on in vitro angiogenesis assays. In vivo, application of cMSCs to murine corneas after injury leads to reduced macrophage infiltration and higher expression of CD206 in macrophages. Macrophages cocultured ("educated") by cMSCs express significantly higher levels of anti-angiogenic and anti-inflammatory factors compared with control macrophages. In vivo, injured corneas treated with cMSC-educated macrophages demonstrate significantly less neovascularization compared with corneas treated with control macrophages. Knocking down the expression of pigment epithelial derived factor (PEDF) in cMSCs significantly abrogates its modulating effects on macrophages, as shown by the reduced rate of apoptosis, decreased expression of sFLT-1/PEDF, and increased expression of vascular endothelial growth factor-A in the cocultured macrophages. Similarly, cMSCs isolated from PEDF knockout mice are less effective compared with wild-type cMSCs at inhibiting macrophage infiltration when applied to wild-type corneas after injury. Overall, these results demonstrate that cMSCs therapeutically suppress the angiogenic capacity of macrophages and highlight the role of cMSC secreted PEDF in the modulation of macrophage phenotype and function. Stem Cells 2018;36:775-784.

Staphylococcus aureus alpha-hemolysin impairs corneal epithelial wound healing and promotes intracellular bacterial invasion.

Colonization by Staphylococcus aureus (S. aureus) has been implicated in many infectious and wound healing disorders. This study was performed to characterize the pathogenic role of S. aureus alpha-hemolysin (alpha-toxin) in corneal epithelial wound healing and infectious keratitis in the setting of a corneal wound. The effect of wild-type and isogenic Hla mutant (α-hemolysin gene deleted) S. aureus bacteria and conditioned media on corneal epithelial wound healing was tested in vitro using a scratch assay and in vivo using a murine epithelial debridement model. The invasiveness of wild-type and Hla mutant S. aureus was evaluated in vitro in human corneal epithelial cells and in vivo in a murine model of infectious keratitis following total epithelial debridement. S. aureus and its conditioned media significantly delayed epithelial wound closure both in vitro (P < 0.05) and in vivo (P < 0.05). The effect of S. aureus on wound healing was significantly diminished with the Hla mutant strain (P < 0.05). Likewise, compared to the wild-type strain, the Hla mutant strain demonstrated significantly reduced ability to invade corneal epithelial cells in vitro (P < 0.05) and infect murine corneas following total epithelial debridement in vivo (P < 0.05). In conclusion, S. aureus alpha-hemolysin plays a major role in the pathologic modulation of corneal epithelial wound healing and the intracellular invasion of the bacteria. Limiting colonization by S. aureus and/or blocking alpha-hemolysin may provide a therapeutic approach for corneal wound healing and infectious disorders.

Stability of limbal stem cell deficiency after mechanical and thermal injuries in mice.

We studied the reproducibility and stability of limbal stem cell deficiency (LSCD) in mice following controlled injuries to the corneal and limbal epithelia. In one method, corneal and limbal epithelia were entirely removed with a 0.5 mm metal burr. In the other, limbus to limbus epithelial removal with the burr was followed by thermal injury to the limbus. These two methods were compared with a previously published one. Unwounded corneas were used as control. The corneas were examined monthly for three months by slit lamp with fluorescein staining. Immunofluorescence staining for cytokeratin 12 and 8 on corneal wholemount and cross sections were performed to determine the phenotype of the epithelium. Mechanical shaving of the epithelium, with or without thermal injury, resulted in a reproducible state of LSCD marked by superficial neovascularization, reduce of keratin 12 expression and presence of goblet cells on the cornea. The phenotype was stable in 100% of the eyes up to at least three months. Thermal injury produced a more severe phenotype with more significant stromal opacification. These corneal injury models may be useful for studying the mechanisms leading to limbal stem cell deficiency.

Operating Room Waste Generated From Corneal and Conjunctival Surgeries.

PURPOSE: The purposes of this study were to bring awareness to the surgical waste generated from corneal and conjunctival surgeries and to compare those findings with the waste generated from cataract surgeries. METHODS: This was an observational prospective pilot cohort study at a tertiary corneal/anterior segment private practice. All waste related to cataract, cornea, and conjunctival surgical procedures (including anesthesia waste and corneal tissue storage) was weighed. The primary outcome was total waste generated while other outcomes included surgical setting (ambulatory surgical center, hospital, and minor operating room) and comparison of corneal/conjunctival surgeries with cataract surgery. RESULTS: Surgical waste data were collected from 119 surgeries (82 corneal/conjunctival surgeries and 37 cataract surgeries). Hospital surgeries produced more waste than ambulatory surgical center and minor operating room surgeries. Penetrating keratoplasty (2.22 kg, P = 0.483) and Descemet stripping only (2.11 kg, P = 0.326) procedures generated comparable mean waste with cataract surgery (2.07 kg) while endothelial keratoplasties produced more ( P < 0.001, 0.002). (Deep) anterior lamellar keratoplasty results depended on the surgical setting. All conjunctival surgeries produced less waste than cataract surgery. CONCLUSIONS: In comparison with cataract surgery, keratoplasties overall produced comparable or more waste while conjunctival surgeries produced less waste. The surgical setting and type of anesthesia played a substantial role in the amount of waste generated. Assessing waste production from different ophthalmic surgeries may increase awareness of the negative environmental impact of surgical waste and promote practice or legal changes to improve environmental sustainability.

Detection of glaucoma progression on longitudinal series of en-face macular optical coherence tomography angiography images with a deep learning model.

BACKGROUND/AIMS: To design a deep learning (DL) model for the detection of glaucoma progression with a longitudinal series of macular optical coherence tomography angiography (OCTA) images. METHODS: 202 eyes of 134 patients with open-angle glaucoma with ≥4 OCTA visits were followed for an average of 3.5 years. Glaucoma progression was defined as having a statistically significant negative 24-2 visual field (VF) mean deviation (MD) rate. The baseline and final macular OCTA images were aligned according to centre of fovea avascular zone automatically, by checking the highest value of correlation between the two images. A customised convolutional neural network (CNN) was designed for classification. A comparison of the CNN to logistic regression model for whole image vessel density (wiVD) loss on detection of glaucoma progression was performed. The performance of the model was defined based on the confusion matrix of the validation dataset and the area under receiver operating characteristics (AUC). RESULTS: The average (95% CI) baseline VF MD was -3.4 (-4.1 to -2.7) dB. 28 (14%) eyes demonstrated glaucoma progression. The AUC (95% CI) of the DL model for the detection of glaucoma progression was 0.81 (0.59 to 0.93). The sensitivity, specificity and accuracy (95% CI) of DL model were 67% (34% to 78%), 83% (42% to 97%) and 80% (52% to 95%), respectively. The AUC (95% CI) for the detection of glaucoma progression based on the logistic regression model was lower than the DL model (0.69 (0.50 to 0.88)). CONCLUSION: The optimised DL model detected glaucoma progression based on longitudinal macular OCTA images showed good performance. With external validation, it could enhance detection of glaucoma progression. TRIAL REGISTRATION NUMBER: NCT00221897.

Anwar versus Melles deep anterior lamellar keratoplasty for keratoconus: a prospective randomized clinical trial.

PURPOSE: To compare the outcomes of 2 techniques (Anwar vs. Melles) of deep anterior lamellar keratoplasty (DALK) in patients with keratoconus. DESIGN: Randomized, double-blind clinical trial. PARTICIPANTS: Fifty-seven eyes of 57 patients 20 to 35 years of age were enrolled. METHODS: Patients with clinical diagnosis of keratoconus who were contact lens intolerant and whose corrected distance visual acuity (CDVA) was less than 20/80 were enrolled. Eligible eyes were allocated randomly into 2 groups: the Anwar technique (23 eyes) or the Melles technique (25 eyes). MAIN OUTCOME MEASURES: The primary outcome measure was CDVA. Secondary outcomes were spherical equivalent, contrast sensitivity, corneal aberrations, corneal biomechanical properties, endothelial cell count, and central corneal thickness. All outcomes were compared 15 months after surgery. RESULTS: The CDVA was 0.17 ± 0.09 logarithm of the minimum angle of resolution (logMAR) units and 0.18 ± 0.11 logMAR units in the Anwar and Melles groups, respectively (P = 0.803). Spherical equivalent was -1.82 ± 2.7 diopters (D) and -2.69 ± 3.94 D in the Anwar and Melles groups, respectively (P = 0.155). Overall, the difference in photopic and mesopic contrast sensitivity function between the 2 groups was statistically significant (P<0.05). There was no significant difference between 2 groups in total and higher-order aberrations up to the fifth order (P>0.05 for all parameters). Corneal hysteresis was not significantly different between the 2 groups (9.9 ± 0.8 vs. 9.9 ± 0.6; P = 0.606). The corneal resistance factor was 10.02 ± 0.8 and 10.13 ± 0.76 (P = 0.509). There was no significant difference in percentage of endothelial cell loss between the 2 groups (1 ± 2% vs. 1 ± 3% in the Anwar and Melles groups, respectively; P = 0.869). Mean central corneal thickness was 525.56 ± 47.87 µm versus 504.64 ± 54.20 µm in the Anwar and Melles groups, respectively (P = 0.155). CONCLUSIONS: The Anwar and Melles techniques of DALK have comparable visual acuity and refractive outcomes, aberrometric profiles, biomechanical properties, corneal thicknesses, and endothelial cell densities. However, patients who underwent the Anwar technique showed better contrast sensitivity.

Perimetric Comparison Between the IMOvifa and Humphrey Field Analyzer.

PRCIS: IMO visual function analyzer (IMOvifa), a binocular perimeter, has similar output to the Humphrey Field Analyzer (HFA), but reduced the measurement time. PURPOSE: The purpose of this study is to evaluate the performance of IMOvifa, a perimeter that performs binocular visual field (VF) testing, and to compare its results with standard automated perimetry. METHODS: All patients underwent HFA 24-2 SITA-Fast and IMOvifa 24-2 AIZE-Rapid on the same day. Mean deviation (MD), pattern SD (PSD), foveal threshold, and visual field index (VFI) were compared between the 2 perimeters using Wilcoxon signed-rank tests, Pearson correlation, and Bland-Altman plot. Measurement time for performing VF for both eyes was also collected for each device. RESULTS: In this cross-sectional study, 138 eyes (including 25 healthy, 48 glaucoma suspects, and 65 primary open angle glaucoma) of 69 patients were evaluated. Measurement time was significantly faster for IMOvifa compared with HFA (256 vs. 419 s, P <0.001). No significant differences were seen in MD and VFI between HFA and IMOvifa (both P >0.05). Significant differences were seen in mean PSD 3.2 (2.7, 3.6) dB for HFA versus 4.1 (3.5, 4.6) for IMOvifa ( P <0.001), and foveal threshold 33.9 (33.1, 34.6) dB for HFA versus 30.6 (29.3, 31.9) dB for IMOvifa ( P <0.001). Pearson r was strong for MD ( r =0.90, P <0.001), PSD ( r =0.78, P <0.001), and VFI ( r =0.94, P <0.001). The mean difference (95% limits of agreement) was -0.1 (-3.8, 3.5) dB for MD, -0.4 (-3.4, 2.5) dB for PSD, and 0.1 (-8.9, 9.1) dB for VFI, respectively. CONCLUSIONS: IMOvifa reduced measurement time by 39%. MD, PSD, and VFI values for IMOvifa showed good agreement with HFA SITA-Fast strategy. This perimeter reduced fatigue for both patient and examiner. Additional studies are needed to determine whether it will be useful for routine VF testing.

Comparison of Persistent Epithelial Defect Treatment With Sutureless Cryopreserved and Dehydrated Amniotic Membrane.

PURPOSE: To compare cryopreserved sutureless amniotic membrane (C-SAM) and dehydrated SAM (D-SAM) outpatient treatment outcomes for persistent epithelial defects (PEDs), analyze risk factors for treatment failure, and identify adverse events. DESIGN: Retrospective, interventional comparative clinical study. METHODS: This study was a multicenter retrospective interventional cohort from 2 tertiary corneal referral practices from 2016 to 2020. The inclusion criteria were as follows: (1) PEDs treated (2) outpatient with (3) either C-SAM or D-SAM. PEDs were defined as epithelial defects present for ≥7 days after failing prior conservative therapy. The primary outcome measure was the resolution or improvement of a PED. The secondary outcomes included analysis of treatment failures and identification of adverse events. A total of 220 PEDs from 204 eyes (197 patients) treated with either C-SAM or D-SAM met the inclusion criteria. RESULTS: A total of 100 PEDs (45.5%) resolved after single amniotic membrane administration, 46.5% (59 of 127) in the C-SAM group and 44.1% (41 of 93) in the D-SAM group (P = .727). Forty-nine PEDs neither improved nor resolved without a significant difference between the C-SAM (21.3%) and D-SAM groups (23.7%, P = .673). There was no statistically significant difference for PED resolution, PED improvement, PEDs that did not resolve/improve, or those requiring surgery between the 2 groups for initial SAM. CONCLUSIONS: C-SAM and D-SAM were both effective for treating PEDs with comparable outcomes for resolution, improvement, and need for additional surgical intervention. Specific differences in adverse events may help dictate clinical use. Inflammatory disease was a risk factor for nonresolution of all PEDs.

Smoking and progressive retinal nerve fibre layer thinning in glaucoma.

BACKGROUND/AIMS: To investigate the relationship between smoking and smoking intensity, and the rate of retinal nerve fibre layer (RNFL) thinning in patients with primary open angle glaucoma (POAG). METHODS: In this longitudinal study, patients with POAG who had at least 3 years of follow-up with a minimum of 5 visits of optical coherence tomography (OCT) were enrolled. The smoking intensity was calculated as the pack-year at the baseline OCT. Univariable and multivariable linear mixed models were used to determine the effect of each parameter on the rates of RNFL thinning over time. Non-linear least-squares estimation with piecewise regression model was used to investigate the cut-off point for the relationship between circumpapillary RNFL thinning and smoking intensity. RESULTS: A total of 466 eyes of 314 patients were included over the mean (95% CI) follow-up of 6.6 (6.4 to 6.7) years. Of the 314 patients, 121 (39%) had reported any history of smoking. Greater smoking intensity was associated with faster RNFL thinning (-0.06 (95% CI -0.11 to 0.00) µm/year per 10 pack-year higher; p=0.031) after adjusted for confounding factors. RNFL thinning became significantly faster when smoking intensity was >8 pack-year. CONCLUSIONS: Smoking intensity is associated with faster rates of RNFL thinning. Evaluation of smoking intensity might add information to the assessment of risk of glaucoma progression. Future studies are required to explore if withdrawing smoking as a modifiable risk factor can decrease progression in patients with glaucoma.

Smoking Cessation May Reduce Risk of Visual Field Progression in Heavy Smokers.

PRCIS: The earlier a person quits smoking the more likely is the optic nerve be spared from damage. PURPOSE: To investigate the effect of smoking cessation on visual field (VF) progression in glaucoma. METHODS: Primary open angle glaucoma patients with a minimum of 3 years follow-up and 5 VFs were included. Linear mixed models were used to investigate the effects of smoking on the rates of 24-2 VF mean deviation loss after adjusting for confounding factors. Cox proportional hazard regression was used to identify whether different levels of smoking intensity were associated with VF progression with respect to different duration of quitting. RESULTS: Five hundred eleven eyes of 354 patients were included over the mean follow-up of 12.4 years. Mean baseline age (95% confidence interval) was 62.3 (61.2, 63.4) years. One hundred forty nine (42.1%) patients were smokers. In a multivariable model, smoking intensity was associated with faster VF loss (-0.06, 95% confidence interval (-0.10, -0.01) dB/year per 10 pack-years, P =0.01) among smokers. Heavy smokers (≥20 pack-years) who had quit ˂25 years prior had significantly greater odds of VF progression compared with never smokers (odds ratio=2.49 (1.01, 6.08); P =0.046). There was no significant difference in odds of VF progression in heavy smokers who had quit smoking more than 25 years compared with never smokers ( P =0.43). A significantly higher proportion of VF progression was found in heavy smokers who quit < 25 years compared with heavy smokers who quit ≥25 years by Kaplan-Meier analysis ( P =<0.001). CONCLUSIONS: After ≥25 years of smoking cessation, the risk of VF progression in former heavy smokers becomes similar to never smokers. Long-term smoking cessation may be associated with lower VF progression in glaucoma patients.

Effects of Smoking on Optic Nerve Head Microvasculature Density in Glaucoma.

PRCIS: Decreased superficial whole image capillary density was observed in open angle glaucoma (OAG) patients with high smoking intensity. PURPOSE: To investigate the effects of smoking on optic nerve head capillary density measured by optical coherence tomography angiography in patients with OAG. METHODS: In this retrospective cross-sectional study, perimetric and preperimetric glaucoma patients enrolled in the Diagnostic Innovations in Glaucoma Study (DIGS) with optical coherence tomography angiography follow-up were included. Univariable and multivariable linear mixed analysis were performed to determine the effects of different variables on the superficial whole image capillary density. RESULTS: A total of 432 eyes of 271 glaucoma patients comprising 63 preperimetric (106 eyes) and 208 perimetric OAG (326 eyes) were included. A history of tobacco consumption was reported in 105 (38.8%). Among smokers, mean (95% confidence interval) smoking intensity was 12.8 (10.2, 15.5) pack-years. After adjusting for age, glaucoma severity and other confounders, each 10 pack-year increase in smoking intensity (95% confidence interval) was associated with -0.54 (-1.06, -0.02) % lower whole image capillary density ( P =0.041). CONCLUSIONS: Smoking intensity is associated with reduced optic nerve vessel density in glaucoma.

A Prospective Longitudinal Study to Investigate Corneal Hysteresis as a Risk Factor of Central Visual Field Progression in Glaucoma.

PURPOSE: To evaluate the role of corneal hysteresis (CH) as a risk factor of central visual field (VF) progression in a cohort of glaucoma suspect and glaucoma patients. DESIGN: Prospective cohort study. METHODS: Two hundred forty-eight eyes of 143 subjects who were followed for an average of 4.8 years with a minimum of 5 visits with 10-2 and 24-2 VF tests were included. Univariable and multivariable linear mixed-effects models were used to identify characteristics associated with the rate of change over time in 10-2 and 24-2 mean deviation (MD). Mixed-effects logistic regression was used to evaluate characteristics associated with an increased likelihood of event-based 10-2 VF progression based on the clustered pointwise linear regression criterion. RESULTS: CH was significantly associated with 10-2 and 24-2 VF progression in the univariable trend-based analysis. In multivariable trend-based analyses, lower CH was associated with a faster rate of decline in 10-2 MD (0.07 dB/y per 1 mm Hg, P < .001) but not with 24-2 MD (P = .490). In multivariable event-based analysis, lower CH was associated with an increased likelihood of 10-2 VF progression (odds ratio = 1.35 per 1 mm Hg lower, P = .025). Similar results were found in eyes with early glaucomatous damage at the baseline (baseline: 24-2 MD ≥ -6 dB). CONCLUSIONS: Lower CH was associated with a statistically significant, but relatively small, increased risk of central VF progression on the 10-2 test grid. Given the substantial influence of central VF impairment on the quality of life, clinicians should consider using CH to assess the risk of progression in patients with primary open-angle glaucoma including those with early disease.

Longitudinal Structure-Function Relationship between Macular Vessel Density and Thickness and Central Visual Field in Early Glaucoma.

PURPOSE: To investigate the relationship of longitudinal changes in macular vessel density (VD) from OCT angiography and in ganglion cell complex (GCC) from OCT with central visual field (VF) in eyes with early glaucoma. DESIGN: Observational cohort. PARTICIPANTS: A total of 95 eyes, 37 preperimetric and 58 with early glaucoma (24-2 VF mean deviation [MD] ≥ -6 decibels), with an average follow-up of 3.8 years and 5.3 visits, were included. METHODS: Whole-image VD (wiVD) and whole-image GCC (wiGCC) and parafoveal scans, as well as localized regions of interest (LROIs), hemiretinae of whole images, and superior, inferior, temporal, and nasal sectors of parafoveal maps, were matched with central VF locations. Age-adjusted rates of change of VD, GCC, mean sensitivity of VF locations, and 10-2 VF MD were calculated using linear mixed-effect models. Normalized rates of change were calculated for comparison of change rates in wiVD and wiGCC. MAIN OUTCOME MEASURES: Structure-function (SF) correlations of VD and GCC with central VF measurement change rates and comparison of different correlations of SF relationships after bootstrapping the difference of the correlation coefficients. RESULTS: Vessel density loss and GCC thinning demonstrated significant correlations with central VF damage, globally and with most LROIs. The SF correlation (r, 95% confidence interval [CI]) between wiVD and 10-2 VF MD change rates was 0.42 [0.24, 0.58], whereas it was 0.27 [0.08, 0.45] between wiGCC and 10-2 VF MD changes rates (all P < 0.05). In contrast to GCC thinning, VD loss in the parafoveal sectors demonstrated significant correlations with central VF damage in inferior and temporal sectors. Differences in the relationship of SF with central VF damage were not significant between VD loss and GCC thinning. The mean (95% CI) normalized change rates of wiVD (-7.40 [-7.71 to 7.09] %/year) was faster than that of wiGCC (-2.39 [-2.94 to 1.84] %/year) (P < 0.05). CONCLUSIONS: Rates of VD loss and GCC thinning are associated with central VF loss over time. Assessment of both macular VD and GCC thickness should be considered for evaluation of glaucoma progression.

Mechanical versus alcohol-assisted epithelial debridement during photorefractive keratectomy: a confocal microscopic clinical trial.

PURPOSE: To compare the effects of mechanical versus alcohol-assisted epithelial debridement on corneal cellular elements after photorefractive keratectomy (PRK) using confocal microscopy. METHODS: This randomized, clinical trial included 66 eyes from 33 patients with spherical equivalent refraction <-4.00 diopters (D). Mechanical versus alcohol-assisted epithelial debridement was performed during PRK. The right eye of each patient was randomly assigned to one group (mechanical group or alcohol-assisted group) and the fellow eye to the alternate group. Confocal examination was performed preoperatively and at 3 and 6 months postoperatively. Main outcome measures were keratocyte density and maximum anterior stromal light reflectivity. RESULTS: Mean epithelial healing time was 3.2 ± 0.4 and 3.0 ± 0.3 days in the mechanical and alcohol-assisted groups, respectively (P=.001). Anterior retroablation stromal keratocyte density was 704.3 ± 119.9 cells/mm² and 734.3 ± 103.7 cells/mm² at 3 months (P=.05) and 643.8 ± 134.4 cells/mm² and 696.7 ± 129.6 cells/mm² at 6 months (P=.02) in the mechanical and alcohol-assisted groups, respectively. No significant difference was noted in midstromal and posterior keratocyte density between the two groups. Maximum reflectivity was 61.56 ± 12.64 international units (IU) and 56.93 ± 7.86 IU in the mechanical and alcohol-assisted groups, respectively, 3 months after surgery (P=.018). Corresponding values were 49.46 ± 4.97 IU and 48.98 ± 4.60 IU, respectively, 6 months after surgery (P=.628). CONCLUSIONS: Due to more adverse effects of mechanical epithelial debridement on anterior keratocyte density and anterior stromal reflectivity, alcohol-assisted epithelial debridement is recommended as the procedure of choice for epithelial removal during PRK in patients with mild myopia.

Sulfur mustard-induced ocular surface disorders.

Sulfur mustard is a vesicant agent with severe irritating effects on living tissues, including skin, mucous membranes, eyes, and respiratory tract. The eyes are the most susceptible tissue to mustard gas effects, and varying degrees of ocular involvement are seen in 75% to 90% of exposed individuals. Most cases resolve uneventfully; however, a minority of exposed patients will have a continuous process, which manifests clinically either as a persistent smoldering inflammation (chronic form) or late-onset lesions appearing many years after a variable "silent" period (delayed form). Distinctive features common to most cases with chronic involvement include chronic blepharitis, meibomian gland dysfunction, dry eye, limbal ischemia, limbal stem cell deficiency, aberrant conjunctival vessels, corneal neovascularization, and secondary degenerative changes, including lipid and amyloid deposition and corneal irregularity, thinning and scarring. Most cases can be managed with conservative measures, eg, preservative-free artificial tears, lubricants, and topical steroids. Punctal plugs or punctal cauterization is helpful in moderate and severe forms of injury. Surgical modalities, including lateral or medial tarsorrhaphies, amniotic membrane transplantation, lamellar or penetrating keratoplasty, and stem cell transplantation have been used.

Clinical Outcomes of Allogeneic Ocular Surface Stem Cell Transplantation in Pediatric Patients.

PURPOSE: To describe the outcomes of allograft ocular surface stem cell transplantation (OSST) and the complication profile of systemic immunosuppression (SI) in pediatric patients with limbal stem cell deficiency. METHODS: This was a retrospective interventional case series from a single tertiary referral institution of 20 eyes from 13 patients who 1) underwent allograft OSST surgery, 2) were 18 years or less at time of OSST, and 3) received SI with 4) a minimum of 12-months follow-up. The main outcome measures were ocular surface stability, visual acuity, and SI adverse events. RESULTS: The mean age of patients was 15.1 ± 3.2 years (range 9-18 years). The mean follow-up was 5.6 ± 5.0 years after OSST. At the last follow-up, 15 eyes (75%) had a stable ocular surface, 1 eye (5%) developed partial failure, and 4 eyes (20%) developed total surface failure. Preoperative mean logarithm of the minimum angle of resolution visual acuity 1.5 improved to 1.1 at the last follow-up (P = 0.1); when 4 eyes of 3 nonadherent patients were excluded, the results were more pronounced and statistically significant (1.5 improved to 1.0, P = 0.002). SI was tolerated well by all patients with minimal adverse events. CONCLUSIONS: OSST provides a stable ocular surface and is a successful treatment option for pediatric patients with limbal stem cell deficiency. SI is well-tolerated with a minimal complication profile.

Region of Interest Densitometry Analysis of Descemet Membrane Endothelial Keratoplasty Dehiscence on Anterior Segment Optical Coherence Tomography.

Purpose: To evaluate a region of interest (ROI) method of analyzing anterior segment optical coherence tomography (AS-OCT) corneal densitometry (CD) in the setting of Descemet membrane endothelial keratoplasty (DMEK) dehiscence. Methods: Retrospective chart review of eyes that underwent (1) DMEK for Fuchs dystrophy (2) between 2018 to 2020 with (3) a partial DMEK dehiscence on AS-OCT, (4) involvement of only one side of the graft, (5) high-quality corneal AS-OCT scan, and (6) location of dehiscence within the central 5.5 mm of the cornea. Image analysis of the ROIs with ImageJ compared the total edematous area, mean stromal CD, and ratio of anterior-to-posterior (A/P) stromal CD for regions of DMEK dehiscence compared to the contralateral side with an attached DMEK graft. Control regions (with no dehiscence) and postdehiscence resolution images were also analyzed. Results: Seventy sectors of the 21 images from 21 eyes with DMEK dehiscence were included. Compared to the contralateral side, regions of DMEK dehiscence had larger total areas (P < 0.0001), lower mean stromal CD (P = 0.0003), and higher A/P stromal CD (P < 0.0001). All control regions and postdehiscence resolution images did not show any significant differences compared to the contralateral sides. Conclusions: This technique to analyze multiple ROIs on AS-OCT can be useful to evaluate CD of specific regions of corneal pathology. Lower mean stromal CD and higher A/P stromal CD may specify corneal edema. Translational Relevance: Analyzing CD via multiple specific ROIs may be more suitable than measuring the CD of the full cornea and has broader applications extending to other corneal pathologies.

Preclinical Evaluation of the Safety and Efficacy of Cryopreserved Bone Marrow Mesenchymal Stromal Cells for Corneal Repair.

Purpose: Mesenchymal stromal cells (MSCs) have been shown to enhance tissue repair as a cell-based therapy. In preparation for a phase I clinical study, we evaluated the safety, dosing, and efficacy of bone marrow-derived MSCs after subconjunctival injection in preclinical animal models of mice, rats, and rabbits. Methods: Human bone marrow-derived MSCs were expanded to passage 4 and cryopreserved. Viability of MSCs after thawing and injection through small-gauge needles was evaluated by vital dye staining. The in vivo safety of human and rabbit MSCs was studied by subconjunctivally injecting MSCs in rabbits with follow-up to 90 days. The potency of MSCs on accelerating wound healing was evaluated in vitro using a scratch assay and in vivo using 2-mm corneal epithelial debridement wounds in mice. Human MSCs were tracked after subconjunctival injection in rat and rabbit eyes. Results: The viability of MSCs after thawing and immediate injection through 27- and 30-gauge needles was 93.1% ± 2.1% and 94.9% ± 1.3%, respectively. Rabbit eyes demonstrated mild self-limiting conjunctival inflammation at the site of injection with human but not rabbit MSCs. In scratch assay, the mean wound healing area was 93.5% ± 12.1% in epithelial cells co-cultured with MSCs compared with 40.8% ± 23.1% in controls. At 24 hours after wounding, all MSC-injected murine eyes had 100% corneal wound closure compared with 79.9% ± 5.5% in controls. Human MSCs were detectable in the subconjunctival area and peripheral cornea at 14 days after injection. Conclusions: Subconjunctival administration of MSCs is safe and effective in promoting corneal epithelial wound healing in animal models. Translational Relevance: These results provide preclinical data to support a phase I clinical study.

Limbal stem cell deficiency in chronic and delayed-onset mustard gas keratopathy.

PURPOSE: To evaluate limbal stem cell deficiency (LSCD) using impression cytology in patients with chronic and delayed-onset mustard gas keratopathy (MGK). DESIGN: Prospective observational case series. PARTICIPANTS: Thirty-five eyes of 18 patients (all male) with MGK were included. METHODS: A consecutive series of patients with MGK underwent impression cytology. Finding of goblet cells on the corneal side of specimens was considered as LSCD. Severity of corneal clinical manifestation was graded as mild, moderate, and severe in each quadrant. Relation between impression cytology findings and clinical grading was evaluated. MAIN OUTCOME MEASURES: Impression cytology findings and clinical grading. RESULTS: There was LSCD in at least 1 quadrant of cornea in all 35 eyes (100% of cases). No differences were found between impression cytology findings (positive vs. negative for corneal goblet cells) among different quadrants (P = 0.378). Clinical grading was the same between nasal and temporal quadrants (P = 0.266) and between superior and inferior quadrants (P = 0.263). By combining superior and inferior quadrants (vertical zone) and nasal and temporal quadrants (horizontal zone), corneal clinical grading was more severe in horizontal versus vertical zones (P<0.001). There was no relation between LSCD and corneal clinical severity (P = 0.893). CONCLUSIONS: A varying degree of LSCD was demonstrated in all patients with chronic or delayed-onset MGK using impression cytology. Corneal clinical manifestations are more severe in nasal and temporal quadrants. There was no relation between impression cytology findings (positive vs. negative for goblet cells) and corneal clinical grading. Other factors, such as perilimbal conjunctival ischemia, may play a role.