Squamous Metaplasia Grade Research Articles

Increased expression of neutrophil CD15 correlates with the severity of anterior ocular surface damage in type II diabetes mellitus.

Diabetes mellitus is associated with increased risk of ocular surface diseases in elderly. We consider neutrophil CD15 as a potential marker of ocular surface damage in type II diabetes mellitus patients. We aimed to evaluate expression of neutrophil CD15 and correlate it with results of conjunctival impression cytology and routine objective anterior ocular surface tests (TMH, NIBUT, LLT, MGD) in T2DM patients. We prospectively enrolled sixty type II diabetes mellitus patients (120 eyes) into a study group. The control group included forty (80 eyes) age- and sex-matched healthy individuals. All patients underwent comprehensive ophthalmological examination, and tear meniscus height test (TMH), noninvasive tear break-up time (NIBUT), lipid layer thickness measurement (LLT), Meibomian gland dysfunction evaluation (MGD), conjunctival impression cytology (CIC) and expression of CD15. Abnormal Nelson's grades of squamous metaplasia (grades 2 and 3) were observed in 50% (60 eyes) of the study group, and 13.8 (11 eyes) of the control group. Fifteen patients with type II diabetes mellitus suffered from grade 3 squamous metaplasia. Nelson's grades of squamous metaplasia have shown a positive correlation with the level of CD15 expression either in the study and control groups (rs = 0.628, p = <0.0001; rs = 0.746, p < 0.0001; respectively). The research shows significantly reduced values of routine objective ocular tests in type II diabetes mellitus patients in comparison to healthy participants older than 60 y.o. Increased CD15 in the peripheral blood is associated with the development of squamous metaplasia and may be used to evaluate the severity of ocular surface damage in type II diabetes mellitus patients.

Prevalence of dry eye, its categorization (Dry Eye Workshop II), and pathological correlation: A tertiary care study.

To study the prevalence of dry eye disease (DED), further categorize using DEWS II protocol, grade squamous metaplasia in each group, and determine associated risk factors in a tertiary care hospital. This cross-sectional hospital-based study screened 897 patients ≥30 years via systematic random sampling. Patients with both symptoms and signs as defined by the Dry Eye Workshop II protocol were considered as DED, further categorized, and subjected to impression cytology. Categorical data were assessed using the Chi-square test. P value < 0.05 was considered statistically significant. In total, 265 (of 897) patients were defined as DED based on the presence of symptoms (DEQ-5 ≥6) and at least one positive sign (fluorescein breakup time [FBUT] <10 s or OSS ≥4). DED prevalence was thus 29.5% with aqueous deficient dry eye (ADDE), evaporative dry eye (EDE), and mixed type seen in 92 (34.71%), 105 (39.62%), and 68 (25.7%) patients, respectively. The risk of developing dry eye was higher in the age above 60 years (33.74%) and in the third decade. Females, urban dwellers, diabetics, smokers, history of previous cataract surgery, and usage of visual display terminal devices were found to be significantly associated with risk factors of DED. Squamous metaplasia and goblet cell loss were more severe in mixed compared to EDE and ADDE. Hospital-based prevalence of DED is 29.5% with a preponderance of EDE (EDE 39.62%, ADDE 34.71%, and mixed 25.71%). A higher grade of squamous metaplasia was seen in the mixed type compared to other sub-types.

Ocular surface characteristics in pediatric vernal keratoconjunctivitis: a clinico-cytological study

To compare the ocular surface parameters of children with vernal keratoconjunctivitis (VKC) with those of healthy controls and to correlate cytological characteristics with clinical findings and disease severity. Newly diagnosed cases of VKC, not currently being treated, were recruited, along with age-matched controls with no ocular comorbidities. The Ocular Surface Disease Index questionnaire (OSDI) was administered to all children. Slit lamp biomicroscopy for meibomian gland dysfunction and ocular surface analysis was performed, including tear meniscus height, noninvasive tear film break-up time, lipid layer thickness, meibomian gland morphology, and meibomian gland duct distortion on meibography imaging. Conjunctival impression cytology was also performed. A total of 68 VKC patients and 33 controls were included. Statistically significant difference was seen in the mean OSDI score (30 ± 13.7 vs 16.1 ± 3 [P ≤ 0.01]), lipid layer thickness (24.2 ± 7.9 nm vs 69.9 ± 15.1 nm [P <0.001]), and noninvasive tear film break-up time (6.8 ± 1.7 vs 12.5 ± 1.8 sec [P < 0.01]) between groups. Mean tear meniscus height was 0.22 ± 0.06 mm in the VKC and 0.24 ± 0.04 mm in the control group (P = 0.096). Significant association was seen between grade of squamous metaplasia and severity of VKC (P<0.01). Severity of VKC was found to be positively correlated with OSDI score (r=0.767), grade of squamous metaplasia (r = 0.64) and negatively correlated with noninvasive tear film break-up time (r = -0.468), and lipid layer thickness (r = -0.253). This study highlights the poor ocular surface health of children with VKC, with severe disease being associated with worse forms of dry eye disease.

Evaluation and comparison of subjective and objective anterior ocular surface damage in patients with type 2 diabetes mellitus and dry eye disease.

To evaluate anterior ocular surface damage in patients with type 2 diabetes mellitus and dry eye disease in comparison to non-diabetic controls based on conjunctival impression cytology, objective scales (Efron, Oxford) and OSDI, to correlate vision-related quality of life with grades of squamous metaplasia in T2DM patients suffering from DED. All participants underwent complete ophthalmologic examination including Shirmer test, TBUT, conjunctival/corneal staining (Oxford scheme), evaluation of conjunctival redness (Efron grading scale), and conjunctival impression cytology (Nelson's scale). The OSDI questionnaire was completed by both groups of patients to assess severity of DED and QoL. Squamous metaplasia was observed in 94% of the study group and 19.3% of controls (p = 0.0000). Based on the OSDI scores, 73.5% of patients reported mild DED and 26.5% suffered from moderate DED in the study group. The mean OSDI score for the study group with Nelson's grade 2 was 18 ± 3.52 and 20.8 ± 4.68 for Nelson's grade 3, respectively (p = 0.0745). Hence, no significant difference in QoL between grade 2 and grade 3 of squamous metaplasia was observed in patients of the study group. Impression cytology is a reliable minimally invasive tool for an accurate evaluation of the ocular surface damage in patients with DED and type 2 diabetes mellitus. Severe squamous metaplasia (Nelson's grade 3) was observed in 29.4% (10/34) of T2DM patients. In contrast, it was not detected in the control group (p = 0.0032). The absence of goblet cells in T2DM patients nether significantly reduces QoL nor contributes to the subjective DED severity (OSDI) due to complex pathways leading to DED. Thus, diagnosis of DED severity should not be solely based on subjective symptoms in this population.

Conjunctival impression cytology and tear film parameters in patients with multiple sclerosis.

To evaluate conjunctival impression cytology (CIC) findings and tear film parameters in patients with multiple sclerosis (MS) compared with controls. Thirty-three patients with MS (MS group) and 33 age- and sex-matched healthy subjects (control group) were included in this cross-sectional comparative study. CIC grades, tear break-up time (TBUT), Schirmer 1 test results, and Ocular Surface Disease Index (OSDI) scores were compared between the two groups, and correlations between CIC grade, TBUT, Schirmer 1 test result, OSDI score, Expanded Disability Status Scale score, and disease duration were analyzed. Mean CIC grade was higher in the MS group than in the control group (1.48 ± 0.71 and 0.39 ± 0.56, respectively; p < 0.001). In the MS group, CIC of the 14 participants (42.4%) was grade 2-3. In the control group, CIC of the only one participant (3.3%) was grade 2, and none of them was grade 3. TBUT (8.12 ± 3.16, 13.06 ± 4.23s in MS and control groups, respectively; p < 0.001) and Schirmer 1 test results (8.45 ± 5.75, 17.36 ± 10.89mm in MS and control groups, respectively; p < 0.001) were lower, and OSDI score (36.36 ± 19.19, 13.70 ± 15.36 in MS and control groups, respectively; p < 0.001) was higher in the MS group compared to the control group. In patients with MS, objective findings of dry eye, subjective symptoms related to dry eye, and CIC abnormalities, including high grades of conjunctival squamous metaplasia and goblet cell loss, are more common. Patients with MS should be monitored for ocular surface alterations and dry eye disease.

Evaluation of corneal topography, tear film function and conjunctival impression cytology after long-term scleral contact lens wear in keratoconus patients

ABSTRACT Purpose: To investigate the impact of long-term scleral contact lens (ScCL) wear on corneal curvature, corneal thickness, tear film function, and ocular surface in patients with keratoconus.Methods: Sixteen keratoconus patients wearing ScCLs for 6 months were enrolled in the study. Corneal topography, tear osmolarity test, Schirmer 1 test, tear film break-up time (TBUT) test, and impression cytology analysis were assessed at baseline and follow-up examinations.Results: There were no significant differences in visual acuity, keratometric and pachymetric values after 6 months of ScCL wear compared to baseline (p>0.05 for all). Tear osmolarity, Schirmer 1 test, and TBUT test results showed no significant change during follow-up (p>0.05 for all). Median goblet cell density and grade of squamous metaplasia did not differ significantly at 1-month. However, there was a gradual deterioration in goblet cell density and Nelson grade until the third-month visit compared to baseline (p for goblet cell, p=0.003; p for Nelson grade, p=0.003). These impaired cytological features observed at 3-month visit persisted at 6-month visit (p for goblet cell, p=0.008; p for Nelson grade, p<0.001).Conclusion: Six months of ScCL wear did not induce any changes in corneal curvature and thickness and also did not affect tear function tests in keratoconic eyes. The only significant changes observed were a decrease in goblet cell density and metaplastic changes in conjunctival epithelium in impression cytology analysis. Further investigations may be needed to better understand the cause of impairment in cytological features of ocular surface and its clinical implications.

The Efficacy of 2% Topical Rebamipide on Conjunctival Squamous Metaplasia and Goblet Cell Density in Dry Eye Disease.

Purpose: To clarify the pharmacological effects of 2% rebamipide eye drops on mucosal membrane functions of the ocular surface epithelium, we investigated keratoconjunctival alterations at the cellular level in this study.Methods: Fifteen patients with definite dry eye disease were recruited from outpatient clinics of the Department of Ophthalmology, Ichikawa General Hospital. The patients received treatment with 2% rebamipide eye drops q.i.d for 12 weeks. Symptom score assessment, tear film breakup time, fluorescein and lissamine green ocular surface vital staining, grading of lid wiper epitheliopathy, Cochet–Bonnet corneal sensitivity, assessment of squamous metaplasia grades, and goblet cell density calculations from conjunctival impression cytology samples, as well as evaluation of nucleocytoplasmic ratios and corneal epithelial cells from in vivo confocal microscopy images before and 3 months after treatment were performed.Results: The mean symptom scores, tear film breakup time values, ocular surface fluorescein and lissamine green vital staining scores, and lid wiper scores showed a significant improvement after treatment (P < 0.01). The mean squamous metaplasia grade also showed a significant improvement (1.2 ± 0.1 → 0.3 ± 0.1) 3 months after treatment (P = 0.004). There were similar significant improvements in the mean corneal epithelial cell density (660.1 ± 62.6 → 1015.5 ± 43.5 cells/mm2) (P = 0.002) and nucleocytoplasmic ratios (0.1 ± 0.0 → 0.2 ± 0.0) (P = 0.0042) after treatment.Conclusions: Topical use of 2% rebamipide for 3 months was associated with improvements in ocular surface differentiation due to changes of mucosal functions at the cellular level. These alterations may explain objective and subjective improvements in dry eye disease.

Efficacy of polyethylene glycol-propylene glycol-based lubricant eye drops in reducing squamous metaplasia in patients with dry eye disease.

PurposeSquamous metaplasia in dry eye disease (DED) manifesting as the loss of conjunctival goblet cells results in reduced mucin secretion and tear film instability. The objective of this study was to evaluate the efficacy of a polyethylene glycol–propylene glycol/hydroxypropyl-guar (PEG-PG/HP-guar) artificial tear formulation in reducing the squamous metaplasia in patients with DED using conjunctival impression cytology (CIC).MethodsIn this Phase IV, single-arm, open-label study, DED patients (aged ≥18 years) with a corneal staining sum score ≥3 and tear film break-up time (TFBUT) <7 s self-administered the PEG-PG/HP-guar artificial tears, 3 times a day for a period of 90 days. The primary end point was the change from baseline in goblet cell density (Nelson’s CIC grading score) over the treatment period. Other end points were change in the corneal and conjunctival staining scores, and TFBUT. Statistical evaluation was performed using a paired t-test.ResultsIn total, 49 patients (n=98 eyes) completed the study. Compared with baseline, there was a significant reduction in the mean CIC scores (ie, improvement in goblet cell density) at Days 30, 60, and 90 (1.6±0.5 vs 1.2±0.5, 0.9±0.5, and 0.8±0.5; P<0.0001). At Day 90, 22% of eyes demonstrated squamous metaplasia Grade 0 (ie, normal epithelium). Similar improvements were observed in the corneal staining scores (5.7 vs 3.1, 1.1, and 0.5; P<0.0001), conjunctival staining scores (5.5 vs 3.6, 1.6, and 0.9; P<0.0001), and TFBUT (4.8 vs 5.8, 6.3, and 6.8 s; P<0.0001) at Days 30, 60, and 90, respectively.ConclusionIn this study, treatment with PEG-PG/HP-guar artificial tears for 90 days decreased CIC score, reduced corneal and conjunctival staining, and increased TFBUT in patients with DED. These results suggest that PEG-PG/HP-guar artificial tears can improve the ocular surface health and reverse the changes induced by squamous metaplasia in DED.

Effect of diquafosol tetrasodium 3% on the conjunctival surface and clinical findings after cataract surgery in patients with dry eye.

To investigate the effects of diquafosol tetrasodium (DT) 3% on conjunctival impression cytologic findings in addition to clinical symptoms and signs after cataract surgery in patients with preexisting dry eye disease (DED). Ninety-four eyes of 94 patients with DED who underwent uneventful cataract surgery were included. In total, 50 patients were treated with DT 3% (group A), while 44 patients were treated with sodium hyaluronate 0.1% (group B) postoperatively, along with topical antibiotics and steroids. Conjunctival impression cytology was performed at baseline and at 4 and 12weeks after surgery. Visual acuity, ocular surface disease index (OSDI), tear film breakup time (TBUT), keratoepitheliopathy score, Schirmer's test, and tear clearance rate were measured at baseline and at 1, 4, and 12weeks, and corneal aberration was analyzed at baseline and at 4 and 12weeks. The grade of conjunctival squamous metaplasia was lower at 12weeks, and goblet cell density was higher at 4 and 12weeks in group A than in group B (P<0.05). Compared with group B, group A showed significantly lower OSDI scores at 4 and 12weeks, longer TBUT at 1, 4, and 12weeks, lower keratoepitheliopathy scores at 1 and 12weeks, and lower total root-mean-square score and spherical aberrations at 4weeks after surgery (P<0.05). DT 3% eye drops application after cataract surgery was effective in improving conjunctival epithelial morphology and goblet cell density as well as clinical findings in patients with DED.

The analysis of human conjunctival epithelium proteome in ocular surface diseases using impression cytology and 2D-DIGE

Conjunctival impression cytology samples from patients with meibomian gland dysfunction (MGD), dry eye (DE), and healthy subjects (CT) were collected for determination of the degree of squamous metaplasia (SM) by PAS-hematoxylin staining and for comparative proteomic analyses by 2D-DIGE. The protein spots with discriminant expression were identified by MALDI-TOF/TOF mass spectrometry. Three independent statistical studies were conducted: i). Analysis of differential protein expression between study groups: We observed increased expression of proteins S100A4, S100A8, retinal dehydrogenase-1, peroxiredoxin-1, annexin-A1, annexin-A2, α-enolase, and glutathione S-transferase-P in DE, whereas the highest expression of peroxiredoxin-6, actin cytoplasmic-1, peroxiredoxin-2, and heat shock protein HSP-90-α was observed in MGD; ii). Correlation between changes in the proteome profile and the grade of SM: The expression of 5 different cytokeratins (KRT1, KRT4, KRT8, KRT10, and KRT13) correlated with the degree of SM; iii). Proteome profile differences between pathological and CT groups: An overall proteome analysis revealed upregulation of 9 proteins in the pathological groups (Annexin-A1, α-enolase, Annexin-A2, S100A8, cytokeratin-1, Peroxiredoxin-2 and Leukocyte elastase inhibitor) and downregulation of 2 proteins (Galectin-3 and Lipocalin-1). In conclusion, a sensitive proteomic approach to study conjunctival tissue collected from minimally invasive impression cytology was implemented. Differential proteomics analyses showed that in comparison with the MGD, the DE patients presented higher overexpression of proteins related to antimicrobial defense, tissue-damage response, and regulation of body fluid secretions. Changes in MGD proteome were associated with oxidative stress and anti-apoptotic processes. We found a correlation between the grade of SM and expression of proteins associated with cytoskeleton and keratinization. The studied pathological groups shared elements related to the defense and inflammatory responses. Dot blot assays of proteins ANXA1, S100A8, and S100A4 validated the proteomic results obtained from 2D-DIGE experiments and confirmed the correlation between the expression of these proteins and the clinical parameters.

Conjunctival cytologic features in patients with unilateral primary acquired nasolacrimal duct obstruction.

To evaluate the cytologic features of conjunctival epithelium in patients with unilateral primary acquired nasolacrimal duct obstruction (PANDO) using impression cytology. Twenty-four patients with unilateral PANDO for at least 1year were enrolled in this study. The healthy eyes of the patients are assessed as control group. All patients were subjected to ophthalmic surface examination with conjunctival impression cytology analysis. Squamous metaplasia and goblet cell density were graded according to Nelson grading system. The mean age was 52.7±16.3 (range 21-70) years. The mean period for complaints of epiphora was 3.2±2.4 (range 1-8) years. The eye with PANDO had a mean squamous metaplasia grade of 2.38±0.59 versus 1.91±0.82 for control eyes (p=0.011). The mean grade of goblet cell density was 2.0±0.51 for eyes with PANDO and 2.38±0.65 for control eyes (p=0.013). There was no statistically significant correlation between conjunctival cytological features in terms of squamous metaplasia and goblet cell density grades and duration of epiphora complaint (r=-0.04, p=0.82; r=0.09, p=0.66, respectively). PANDO may cause alterations in conjunctival impression cytology. Successful dacryocystorhinostomy surgeries will help to protect ocular surface health.

Cytomorphology of Conjunctival Epithelium in Ocular Surface Disorders

Introduction: To study cytomorphological changes in various ocular surface disorders and find out the sensitivity, specificity and the diagnostic accuracy of impression cytology. Materials and Methods: One hundred and ten patients (84 cases and 26 controls) were included in this study. Out of these 84 cases, 25 cases having vitamin A deficiency (VAD), 19 cases are having vernal keratoconjunctivitis (VKC), 30 of dry eye and 10 of chemical burn. Impression cytology was performed with each patient and the results were noted. Biopsy was done in 2 patients from each group. Patients were examined for goblet cell density in the form of clumping of goblet cells and degree of squamous metaplasia in terms of loss of cell cohesion, altered cell morphology and keratinisation. Results: On impression cytology, all cases of vitamin A deficiency showed loss of goblet cells and 92% showed squamous metaplasia. In case of vernal conjunctivitis showed clumping (37%), loss of goblet cells (63%) and squamous metaplasia (64%). In dry eye group, severe reduction of goblet cells (73%) with moderate to severe grade of squamous metaplasia (88%) was seen. All cases of chemical burn showed marked reduction of goblet cells and squamous metaplasia. Control group, showed mild reduction in goblet cells (23%) and mild grade of squamous metaplasia (15%). Sensitivity, specificity and diagnostic accuracy of impression cytology was 89.28%, 76.92% and 86.36% respectively. Cytological findings are well correlated with histological findings. Conclusion: Impression cytology was found to be good technique for diagnosing metaplastic changes in various ocular surface disorders.

Effect of Omega-3 Fatty Acids Dietary Supplementation on Ocular Surface and Tear Film in Diabetic Patients with Dry Eye

Objective: The objective of this study was to investigate the effect of dietary supplementation with omega-3 fatty acids on ocular surface and tear film in patients with type 2 diabetes suffering from dry eye.Methods: Thirty-six patients suffering from type 2 diabetes and moderate to severe dry eye syndrome were included in the study. Patients were assigned to receive omega-3 long-chain polyunsaturated fatty acids for 3 months. Tear film break-up time test, Schirmer-I test, and conjunctival impression cytology analysis were performed on all patients at baseline and after 1 and 3 months. The subjective symptoms of dry eye were evaluated with the Ocular Surface Disease Index (OSDI) questionnaire at the same time points.Results: Patients’ average age was 65.57 ± 4.27 years and the mean duration of diabetes was 14.85 ± 5.4 years. There was a statistically significant increase in Schirmer-I test results and tear break-up time score after 3 months of supplementary intake of omega-3 fatty acids compared to baseline (p < 0.05 and p < 0.001, respectively). Impression cytology demonstrated a significantly lower grade of conjunctival squamous cell metaplasia after 1 and 3 months of omega-3 fatty acids intake compared to baseline (p < 0.05 and p < 0.01, respectively). The OSDI score was statistically significant lower both at 1 and 3 months after omega-3 fatty acids supplementation compared to baseline (p < 0.001).Conclusions: Omega-3 fatty acids may effectively improve tear film characteristics, reverse ocular surface features, and alleviate the subjective symptoms associated with dry eye syndrome in patients with type 2 diabetes.

A comparative study of tear film function and tear secretion among diabetics

The present work aims to study the changes in tear film function by studying tear film stability, amount of tear secretion and conjunctival impression cytology among diabetics and to compare with that in non-diabetics. In this prospective case control study, 100 eyes of 50 patients with type II Diabetes Mellitus and 100 eyes of 50 normal patients coming to OPD of a tertiary hospital were chosen. The tests done included Tear film Break Up Time, Rose Bengal staining, Schirmer’s test and Conjunctival Impression Cytology (CIP). For CIP, samples were taken from nasal and temporal conjunctiva and smeared on slides and stained with Papanicolau stain and observed under light microscope of 400x magnification. The mean age among diabetics was 53.4±11.2 years and non-diabetics was 48.8±15.3years. The TBUT was significantly reduced in diabetics with a mean of 9.8±7.01 seconds and 12.8±5.71 seconds in non-diabetics (p<0.005). 30% patients had values less than 10 seconds. The Rose Bengal test did not show a statistically significant difference. Schirmer’s test was positive in 26% in diabetics and 10% in non- diabetics. The mean was 7.7±3.9 in diabetics and 13.4±5.7 in non-diabetics (p=0.003). The mean grade of squamous metaplasia in diabetics was 1.58±0.62 and 0.52±0.57 in non-diabetics (p=0.001). Corneal sensitivity was found to be significantly lower in diabetics (29.17mm) when compared to controls (54.58mm). Dry eye is a significant ocular surface disorder in diabetes. The tear film parameters were significantly reduced in diabetics but Rose Bengal test did not show much difference.

The effects of topical antiglaucoma drugs as monotherapy on the ocular surface: a prospective study.

Purpose. The aim was to compare the effects of antiglaucoma eye drops on the tear functions and ocular surface. Method. Eighty-five eyes of 43 patients with glaucoma were included into this randomized prospective study. Timolol without preservative (1), timolol with benzododecinium bromide (2), latanoprost (3), bimatoprost (4), travoprost with benzalkonium chloride (5), and brimonidine with purite (6) were given to 6 groups. Schirmer I, tear film breakup time (TBUT), staining scores, and impression cytology samples were evaluated before and during 12-month-follow-up period. Results. At the end of 12 months, there was no detected change in Schirmer I and TBUT tests indicating dry eye. Corneal staining scores were higher in groups 1 and 2, while conjunctival staining scores were higher in group 6. Goblet cell count decreased in groups 1 and 5 in superior and inferior, group 2 in superior, and groups 3 and 6 in inferior conjunctiva. Squamous metaplasia grades showed a significant increase in groups 1 and 2 at 3rd, 6th, and 12th month controls (P < 0.05). Conclusion. We observed nonserious impact on tear functions and ocular surface with antiglaucoma monotherapy. Beta blockers induced more damage on the ocular surface suggesting the role of the dosing and active substances beside preservatives.

Changes in Corneal Sensation and Ocular Surface in Patients With Asymmetrical Keratoconus

To describe tear function, ocular surface changes, and corneal sensitivity in patients with asymmetrical keratoconus (KC). Thirty-one patients with asymmetrical KC (31 KC and 31 subclinical KC eyes) and 30 control subjects (1 eye in each subject) were enrolled in this prospective, case-control study. The patients and control subjects underwent ocular surface examinations including corneal sensitivity measurements, the Schirmer test using topical anesthesia, tear osmolarity test, and conjunctival impression cytology. Mean corneal sensitivity and Schirmer test values were significantly lower in the KC and subclinical KC eyes compared with the control eyes. The conjunctiva of KC and subclinical KC eyes showed significantly higher grades of squamous metaplasia and goblet cell loss compared with the control group. However, no significant difference in tear osmolarity was found among the groups. The corneal sensitivity and ocular surface changes were significant in the subclinical KC and KC eyes compared with the control subjects. Ocular surface disease in KC was characterized by tear deficiency disorder and abnormal impression cytology results. However, no significant difference in tear osmolarity was found among the groups. The decrease in corneal sensitivity and ocular surface change may be associated with the pathogenesis of ocular surface changes in KC and the progression of the disease.

Reliability of Nucleus-To-Cell and Nucleus-To-Cytoplasm Calculations for Conjunctival Impression Cytology Specimens

Purpose: To assess the outcome of two different nucleo-cytoplasmic ratio calculations in relation to the number of cells measured from conjunctival impression cytology (CIC) images.Methods: CIC from the exposed nasal bulbar conjunctiva surface was undertaken on 40 young adults using a Millcell®-CM filter; the filters were stained with Giemsa, colour images taken of the cells in a monolayer at 200 X magnification, graded by the Nelson scale and then a projection overlay method used to outline the cell and nucleus borders. Nucleo-cytoplasmic ratios were calculated based on literal interpretation of the term (i.e. N/CYT = nucleus area/[cell area − nucleus area]) as well as based on cell area (i.e. N/CELL = nucleus area/cell area).Results: A range of N/CYT or N/CELL values are expected for all cells, regardless of squamous metaplasia grade, with slightly greater heterogeneity expected from N/CYT calculations. The difference between the two calculations gets progressively less as the squamous metaplasia grade increases. For all grades of cells, these ratios could be determined to within approximately ±1% if adequate numbers of cells were measured (i.e. 75, 65, 50 and 35 for grade 0 to grade 3 cells respectively). If fewer cells are measured, then the variability in N/CELL or N/CYT calculations can be expected to increase.Conclusions: In assessments of nucleo-cytoplasmic ratios, it is important that the calculation used be specified and that the numbers of cells measured be provided.

Comparison of Ocular Findings in Patients with H1N1 Influenza Infection versus Patients Receiving Influenza Vaccine during a Pandemic

Aim: To evaluate ocular findings during the pandemic influenza A (H1N1) and after vaccination for the same strain. Patients and Methods: This study was conducted on 89 patients with H1N1 influenza infection (group 1) and 28 subjects who received vaccination for H1N1 (group 2). All patients were subjected to history taking, ophthalmological examination, fundus examination, conjunctival impression cytology and conjunctival swabs. Results: The patients’ age ranged between 5 and 60 years (19.25 ± 11.70 years). Group 1 included 43 (48.1%) males and 46 (51.9%) females, while group 2 included 13 (46.43%) males and 15 (53.57%) females. The most common ocular finding of patients in group 1 was bilateral acute conjunctivitis in 58 cases (65.17%), while in group 2, we found 3 (10.71%) cases of mild conjunctivitis, and 2 (7.14%) cases of moderate conjunctivitis. Retinopathy, uveal affection, and optic neuritis were not statistically different between the 2 groups. Impression cytology of the conjunctiva for group 1 showed squamous metaplasia grade 3 with enlargement of epithelial cells, and fragmentation of the nucleus which is similar to virus-infected structural changes. Conclusion: Pandemic influenza H1N1 was able to induce different ocular manifestations including acute conjunctivitis, retinopathy, uveal effusion syndrome and optic neuritis.

Ocular surface changes and corneal sensitivity in Keratoconus

AbstractPurpose This study aims to describe ocular surface, tear function, corneal nerve and sensitivity in patients with keratoconus.Methods Thirty‐one patients diagnosed as having asymmetrical KC (31 keratoconus eyes, and 31 subclinical keratoconus eyes) and 31 normal control subjects (one eye) were studied in a prospective, cross‐sectional study. The subjects underwent routine ophthalmic examinations, corneal topography, corneal sensitivity measurements, Schirmer test, conjunctival impression cytology, and tear osmolarity. Western blot analysis and Immunofluorescent staining with monoclonal antibodies for Class III β‐tubulin in corneas of keratoconus and normal were done.Results The mean corneal sensitivity and Schirmer test values were significantly lower in eyes of keratoconus and subclinical keratoconus compared with control. Conjunctiva of keratoconus and subclinical keratoconus had significantly higher grades of squamous metaplasia and goblet cell loss compared with normal. Corneal nerves were significantly decreased in eyes of keratoconus and subclinical keratoconus compared with control.Conclusion Whatever the keratoconus‐related factors might be, we found marked changes on the ocular surface that affected not only the corneal, but also the conjunctival epithelium. The loss or decreae of trophic effects of corneal nerves due to primary or secondary events with the progression of keratoconus may plays a role in the pathogenesis of the ocular surface change in keratoconus.

Contact Lens Wear and the Development of Squamous Metaplasia of the Surface Cells of the Conjunctiva

To review the reported effects of contact lens wear on the surface epithelial cells of the human conjunctiva as assessed by conjunctival impression cytology (CIC). A literature search was undertaken to identify reports on the conjunctival health after contact lens wear, principally as assessed using CIC. Of 26 reports identified, 22 examined the bulbar conjunctiva, and 2 examined the tarsal conjunctiva. Just 16 reports provided data from which mean squamous metaplasia grades could be calculated, with the overall grade being just 0.7 on a 0 to 3 scale. Only 13 of these studies provided unambiguous data on the duration of contact lens wear, and only an apparent trend was evident in that grades of squamous metaplasia increased over early years of lens wear. Such a trend was not statistically significant either up to 6 years of average lens wear (P>0.05) or over all studies (P>0.5). The estimated variability in squamous metaplasia was substantially greater when low grades were reported, an observation that either reflects the heterogeneity in the cell response or highlights the difficulty in assigning low grades to cell samples. Based on subjective grading, CIC studies reveal no clearly definable relationship between the duration of contact lens wear and the extent of development of conjunctival squamous metaplasia. Logically, therefore, objective methods to assess squamous metaplasia are needed. Various options for quantitative CIC are discussed, including the use of in vivo confocal microscopy.