Impression cytology of ocular surface in xeroderma pigmentosum.

PURPOSE: To describe cellular alterations detected by impression cytology of the ocular surface in patients with xeroderma pigmentosum. The secondary objective was to assess the reliability of impression cytology in diagnosing ocular surface squamous neoplasia. METHODS: Patients with xeroderma pigmentosum underwent a single-day complete ophthalmological examination and impression cytology for ocular surface evaluation using 13 mm diameter mixed cellulose esters membrane filters and combined staining with Periodic Acid Schiff, Hematoxylin and Eosin, and Papanicolaou stains followed by microscopic analysis. The cytological findings were correlated with the clinical diagnosis. The impression cytology findings at baseline and one-year follow-up were correlated with the clinical course (no tumor, treated tumor, residual tumor recurrent tumor, new tumor). RESULTS: Of the 42 patients examined, impression cytology was performed in 62 eyes of 34 participants (65% females). The mean age of patients was 29.6 ± 17 years (range 7-62). Fifteen eyes had a clinical diagnosis of ocular surface squamous neoplasia. Impression cytology showed goblet cells (47, 75%), inflammatory cells (12, 19%), keratinization (5, 8%), and squamous metaplasia (30, 48%). Impression cytology was positive for atypical cells in 18 patients (12 with and 6 without ocular surface squamous neoplasia). The sensitivity, specificity, positive predictive value, and negative predictive value of impression cytology (at baseline) for diagnosis of ocular surface squamous neoplasia were 80%, 87%, 67%, and 93%, respectively, using clinical diagnosis of ocular surface squamous neoplasia as the reference standard. CONCLUSION: Impression cytology has a moderate positive predictive value for the diagnosis of ocular surface squamous neoplasia in patients with xeroderma pigmentosum. However, the lack of detection of atypical cells on impression cytology has a high negative predictive value for ocular surface squamous neoplasia. Integration of impression cytology in the long-term management of high-risk patients, such as patients with xeroderma pigmentosum, can avoid unnecessary diagnostic biopsies.

Impression cytology of ocular surface in xeroderma pigmentosum

ABSTRACT Purpose: To describe cellular alterations detected by impression cytology of the ocular surface in patients with xeroderma pigmentosum. The secondary objective was to assess the reliability of impression cytology in diagnosing ocular surface squamous neoplasia. Methods: Patients with xeroderma pigmentosum underwent a single-day complete ophthalmological examination and impression cytology for ocular surface evaluation using 13 mm diameter mixed cellulose esters membrane filters and combined staining with Periodic Acid Schiff, Hematoxylin and Eosin, and Papanicolaou stains followed by microscopic analysis. The cytological findings were correlated with the clinical diagnosis. The impression cytology findings at baseline and one-year follow-up were correlated with the clinical course (no tumor, treated tumor, residual tumor recurrent tumor, new tumor). Results: Of the 42 patients examined, impression cytology was performed in 62 eyes of 34 participants (65% females). The mean age of patients was 29.6 ± 17 years (range 7-62). Fifteen eyes had a clinical diagnosis of ocular surface squamous neoplasia. Impression cytology showed goblet cells (47, 75%), inflammatory cells (12, 19%), keratinization (5, 8%), and squamous metaplasia (30, 48%). Impression cytology was positive for atypical cells in 18 patients (12 with and 6 without ocular surface squamous neoplasia). The sensitivity, specificity, positive predictive value, and negative predictive value of impression cytology (at baseline) for diagnosis of ocular surface squamous neoplasia were 80%, 87%, 67%, and 93%, respectively, using clinical diagnosis of ocular surface squamous neoplasia as the reference standard. Conclusion: Impression cytology has a moderate positive predictive value for the diagnosis of ocular surface squamous neoplasia in patients with xeroderma pigmentosum. However, the lack of detection of atypical cells on impression cytology has a high negative predictive value for ocular surface squamous neoplasia. Integration of impression cytology in the long-term management of high-risk patients, such as patients with xeroderma pigmentosum, can avoid unnecessary diagnostic biopsies.

Ocular surface findings in impression cytology after interferon a2b or mitomycin C in rabbits / Achados em citologia de impressão da superfície ocular após uso de interferon alfa-2b ou mitomicina C: estudo em coelho

ABSTRACT Objective: To describe ocular surface findings in impression cytology obtained from healthy rabbit conjunctiva treated with interferon alpha-2b eyedrop, and compare them to findings after use of mitomycin C 0.02%. Methods: An experimental study using a rabbit model was performed between September 2013 and October 2014 at the Faculdade de Medicina de Marília, Universidade Federal de São Paulo, Clínica de Olhos Moacir Cunha. Thirty New Zealand white rabbits were divided into 6 groups and received interferon alpha-2b or mitomycin C 0.02%. Impression cytology (IC) was performed prior to topical applications and at15, 30 and 60 days of use. The following variables were analyzed in impression cytology: goblet cells, cellularity, cell-to-cell adhesion, nucleus/cytoplasm ratio, nuclear chromatin, inflammatory cells keratinization, and cytomegaly. Results: The major findings in impression cytology after us of interferon alpha-2b included loss of goblet cells (50.8%), reduced cell-to-cell adhesion (26.2%), abnormal nucleus/cytoplasm ratio (20%) and reduced cellularity (15.4%). After use of mitomycin C 0.02%, the most common changes included loss of goblet cells (46.2%), abnormal nucleus/cytoplasm ratio (25.6%), less cell-to-cell adhesion (23.1%), and reduced cellularity (20.5%). There were no significant differences in any variable when comparing impression cytology after interferon alpha-2b and after mitomycin C 0.02%. Goblet cell loss was more pronounced at days 30 and 60, as compared to impression cytology at day 15 for both drugs. Conclusion: The loss of goblet cells, reduced cell-to-cell adhesion and cellularity, along with abnormal nucleus/cytoplasm ratio were the most common findings in impression cytology after use of interferon alpha-2b. These findings are similar to those described for use of mitomycin C 0.02%. ..

RESUMO Objetivo: Descrever os achados em citologia de impressão de conjuntiva sadia de coelho submetida ao uso de colírio de interferon alfa-2b e compará-los ao que foi encontrado após uso da mitomicina C 0,02%. Métodos: Estudo experimental realizado em modelo animal no período entre setembro de 2013 e outubro de 2014 nas dependências da Faculdade de Medicina de Marília, da Universidade Federal de São Paulo e da Clínica de Olhos Moacir Cunha. Trinta coelhos albinos da raça Nova Zelândia foram divididos em seis grupos e receberam interferon alfa-2b ou mitomicina C. A citologia de impressão foi realizada antes do início dos colírios e após 15, 30, 60 dias de seu uso. As seguintes variáveis foram analisadas na citologia de impressão: células caliciformes, celularidade, adesão intercelular, razão núcleo/citoplasma, cromatina, células inflamatórias, queratinização e citomegalia. Resultados: Os principais achados na citologia de impressão após o uso do interferon alfa-2b foram a redução de células caliciformes (50,8%), a diminuição da adesão intercelular (26,2%), a alteração da razão N/C (20%) e a redução da celularidade (15,4%). Após o uso da mitomicina C 0,02%, foram mais frequentes a redução das células caliciformes (46,2%), a alteração da razão N/C (25,6%), a adesão intercelular (23,1%) e a redução da celularidade (20,5%). Não houve diferença estatisticamente significante para nenhuma das variáveis estudas quando se compararam as citologias de impressão após interferon alfa-2b com as citologias de impressão após mitomicina C 0,02%. Independentemente da substância utilizada, as citologias colhidas 30 e 60 dias após início das drogas apresentaram maior redução de células caliciformes quando comparadas com as citologias de impressão colhidas após 15 dias. Conclusão: A redução das células caliciformes, a diminuição da adesão intercelular, a alteração da razão N/C e a diminuição da celularidade foram as alterações mais frequentes na citologia de impressão colhida após o uso de interferon alfa-2b. Os achados em citologias de impressão após o uso de interferon alfa-2b são semelhantes àqueles encontrados após o uso da mitomicina C 0,02%.

Novel low-cost approach to the treatment of ocular surface squamous neoplasia using pattern scanning laser photocoagulation / Nova abordagem de baixo custo para neoplasia escamosa da superficie ocular usando fotocoagulação tipo pattern scanning

ABSTRACT Purpose: To evaluate the safety and 12-month effect of treatment with pattern scanning laser photocoagulation for ocular surface squamous neoplasia in a low-resource setting with extremely limited access to an operating room. Methods: Adult patients with a clinical diagnosis of ocular surface squamous neoplasia underwent a complete ophthalmologic examination. After topical anesthesia and instillation of toluidine blue 1%, the lesion was treated using pattern scanning photocoagulation for a duration time that varied from 20 to 100 ms and power from 600 to 1,800 mW. Patients were examined on a weekly basis for the first month and underwent weekly retreatment of the remaining lesions, as necessary. Patients had a minimum follow-up of 12 months. Results: Thirty-eight patients (38 eyes) were included. All patients had clinical ocular surface squamous neoplasia that was confirmed by impression cytology. The age of patients ranged from 40 to 83 years (average: 65.5 years) and 28 of them were males (74%). The patients were divided into two groups: group I (immunocompetent) and group II (immuno­suppressed). In group I, 23 patients (74%) presented complete response with lesion control after laser treatment alone. In group II, two of seven patients (28%) showed treatment response during the follow-up. The average number of treatments was 2.5 (one to six laser treatments). Procedures were well tolerated. Conclusion: Short-term results of the laser photocoagulation approach for the treatment of ocular surface squamous neoplasia conjunctival lesions were favorable, with a 74% success rate observed in immunocompetent patients. This novel strategy is a less resource-intensive alternative that could demonstrate its usefulness in settings with shortages in operating rooms and in recurrent cases. Studies with longer follow-ups and larger sample sizes are warranted to confirm our findings and evaluate the effectiveness of laser treatment in association with topical chemotherapy.

RESUMO Objetivo: Avaliar a segurança e o efeito de 12 meses de tratamento com fotocoagulação pelo pattern scanning laser para neoplasia escamosa da superfície ocular em um ambiente com poucos recursos e acesso extremamente limitado a um tratamento cirúrgico. Métodos: Pacientes adultos com diagnóstico de neoplasia escamosa de superfície ocular foram submetidos a exame oftalmológico completo. Após anestesia tópica e instilação de azul de toluidina 1%, a lesão foi tratada com laser por um tempo de duração que variou de 20 a 100 ms e potência de 600 a 1800 mW. Os pacientes foram examinados semanalmente durante o primeiro mês e fo­ram retratados semanalmente das lesões restantes, conforme necessário. Os pacientes tiveram um seguimento mínimo de 12 meses. Resultados: Trinta e oito pacientes (38 olhos) foram incluídos no estudo. Todos os pacientes apresentaram neoplasia escamosa da superfície ocular clínica, confirmada por citologia de impressão. A idade dos pacientes variou entre 40 e 83 anos (média de 65.5 anos) e 28 deles eram do sexo masculino (74%). Os pacientes foram divididos em dois grupos: Grupo I (imunocompetente) e grupo II (imunossuprimido). No grupo I, 23 pacientes (74%) apresentaram resposta completa com o controle da lesão após o tratamento com laser. No grupo II, dois dos sete pacientes (28%) apresentaram resposta ao tratamento durante o acompanhamento. A média de aplicações de laser foi de 2,5 (1 a 6 aplicações). Os procedimentos foram bem tolerados. Conclusões: Os resultados a curto prazo da abordagem de fotocoagulação a laser para o tratamento das lesões conjuntivais de neoplasia escamosa de superfície ocular foram favoráveis, com uma taxa de sucesso de 74% observada em pacientes imunocompetentes. Essa nova estratégia é uma alternativa menos intensiva em recursos que pode demonstrar sua utilidade em ambientes com escassez de salas cirúrgicas e em casos recorrentes. Estudos com acompanhamentos mais longos e amostras maiores são necessários para confirmar nossos achados e avaliar a eficácia do tratamento a laser asso­ciado à quimioterapia tópica.

Novel low-cost approach to the treatment of ocular surface squamous neoplasia using pattern scanning laser photocoagulation.

PURPOSE: To evaluate the safety and 12-month effect of treatment with pattern scanning laser photocoagulation for ocular surface squamous neoplasia in a low-resource setting with extremely limited access to an operating room. METHODS: Adult patients with a clinical diagnosis of ocular surface squamous neoplasia underwent a complete ophthalmologic examination. After topical anesthesia and instillation of toluidine blue 1%, the lesion was treated using pattern scanning photocoagulation for a duration time that varied from 20 to 100 ms and power from 600 to 1,800 mW. Patients were examined on a weekly basis for the first month and underwent weekly retreatment of the remaining lesions, as necessary. Patients had a minimum follow-up of 12 months. RESULTS: Thirty-eight patients (38 eyes) were included. All patients had clinical ocular surface squamous neoplasia that was confirmed by impression cytology. The age of patients ranged from 40 to 83 years (average: 65.5 years) and 28 of them were males (74%). The patients were divided into two groups: group I (immunocompetent) and group II (immuno-suppressed). In group I, 23 patients (74%) presented complete response with lesion control after laser treatment alone. In group II, two of seven patients (28%) showed treatment response during the follow-up. The average number of treatments was 2.5 (one to six laser treatments). Procedures were well tolerated. CONCLUSION: Short-term results of the laser photocoagulation approach for the treatment of ocular surface squamous neoplasia conjunctival lesions were favorable, with a 74% success rate observed in immunocompetent patients. This novel strategy is a less resource-intensive alternative that could demonstrate its usefulness in settings with shortages in operating rooms and in recurrent cases. Studies with longer follow-ups and larger sample sizes are warranted to confirm our findings and evaluate the effectiveness of laser treatment in association with topical chemotherapy.

Impression cytology in the evaluation of ocular surface tumors: review article.

Impression cytology (IC) has been widely used as a method for evaluating the ocular surface and superficial cells layers in the diagnosis and follow-up after treatment of several ocular surface tumors of both epithelial and melanocytic origin. Information regarding this can be found in the English-language literature since 1992. Using either cellulose acetate or Biopore membranes for specimen collection, a high correlation has been found between IC and tissue histology. Compared with exfoliative cytology with spatula, IC is less traumatic to the patient's eye, provides a precise location of the area being studied, and allows accurate observation of the cells the way they exist in vivo. The additional advantage of IC is the preservation of limbal stem cells responsible for continuous corneal epithelium renewal; these can be affected after incisional or excisional biopsy at the corneoscleral limbus, which is the most frequent site of appearance of tumors in the stratified epithelium. Treatment for ocular surface squamous neoplasia has historically included surgery, but nonsurgical interventions have also been adopted. Hence, in certain cases, ophthalmologists may prefer interventions less invasive than surgical biopsy such as of impression cytology for both initial diagnosis and therapeutic monitoring of treatment for ocular surface lesions. Nevertheless, it should be considered that IC may be less helpful if the results conflict with the clinical picture or if the clinical diagnosis is uncertain and results are negative. In such cases, surgical biopsy is required for accurate diagnosis. The purpose of this review is to examine the published literature on the utilization of IC for the diagnosis and management of ocular surface tumors and to discuss the requirement for further investigation on the subject.

Impression cytology in the evaluation of ocular surface tumors: review article / Citologia de impressão na avaliação de tumores da superfície ocular: artigo de revisão

Impression cytology (IC) has been widely used as a method for evaluating the ocular surface and superficial cells layers in the diagnosis and follow-up after treatment of several ocular surface tumors of both epithelial and melanocytic origin. Information regarding this can be found in the English-language literature since 1992. Using either cellulose acetate or Biopore membranes for specimen collection, a high correlation has been found between IC and tissue histology. Compared with exfoliative cytology with spatula, IC is less traumatic to the patient’s eye, provides a precise location of the area being studied, and allows accurate observation of the cells the way they exist in vivo. The additional advantage of IC is the preservation of limbal stem cells responsible for continuous corneal epithelium renewal; these can be affected after incisional or excisional biopsy at the corneoscleral limbus, which is the most frequent site of appearance of tumors in the stratified epithelium. Treatment for ocular surface squamous neoplasia has historically included surgery, but nonsurgical interventions have also been adopted. Hence, in certain cases, ophthalmologists may prefer interventions less invasive than surgical biopsy such as of impression cytology for both initial diagnosis and therapeutic monitoring of treatment for ocular surface lesions. Nevertheless, it should be considered that IC may be less helpful if the results conflict with the clinical picture or if the clinical diagnosis is uncertain and results are negative. In such cases, surgical biopsy is required for accurate diagnosis. The purpose of this review is to examine the published literature on the utilization of IC for the diagnosis and management of ocular surface tumors and to discuss the requirement for further investigation on the subject.

A citologia de impressão (CI) tem sido amplamente utilizada como um método de avaliação da superfície ocular e das camadas de células superficiais no diagnóstico e no seguimento após tratamento de vários tumores da superfície ocular de origem epitelial ou melanocítica. As informações podem sem encontradas na literatura em língua inglesa desde 1992. Utilizando-se de membranas de acetato de celulose ou Biopore na coleta dos espécimes, uma alta correlação tem sido encontrada entre a CI e a histologia do tecido. Comparando-se com a citologia esfoliativa, a citologia de impressão é menos traumática para o olho do paciente, fornece uma localização precisa da área estudada e permite ver as células da forma como elas organizam-se in vivo. A vantagem adicional da citologia de impressão é a preservação das células- tronco germinativas responsáveis pela renovação contínua do epitélio da córnea. Elas podem ser afetadas após biópsia cirúrgica na região do limbo que é o sítio mais frequentemente acometido pelos tumores do epitélio estratificado. O tratamento para a neoplasia escamosa da superfície ocular tem sido historicamente a cirurgia, mas intervenções não cirúrgicas também foram adotadas. Por esta razão, em certos casos, oftalmologistas podem recorrer a formas menos invasivas que a biópsia cirúrgica (como a citologia de impressão) tanto para o diagnóstico inicial quanto para o monitoramento terapêutico das lesões da superfície ocular. No entanto, deve-se ter em mente que a citologia de impressão deixa de ser útil quando seu resultado não coincide com o quadro clínico ou quando o diagnóstico clínico é incerto e o resultado da citologia de impressão negativo. Nesses casos, a biópsia cirúrgica deve ser realizada para o diagnóstico. O objetivo desta revisão é examinar a literatura sobre a utilização da citologia de impressão no diagnóstico e tratamento dos tumores da superfície ocular bem como discutir a necessidade de uma investigação mais aprofundada sobre o assunto.

Use of impression cytology for the detection of unsuspected ocular surface squamous neoplasia cells in pterygia.

Purpose: To evaluate the agreement between the methodologies of impression cytology (IC) and histopathology regarding epithelial lesions clinically diagnosed as pterygium and also regarding the detection of unsuspected and associated ocular surface squamous neoplasia (OSSN). Methods: Thirty-two Brazilian patients were included and IC was performed on all pterygia before excision. Histopathogical examination was considered the gold standard and was performed by two experienced ocular pathologists in which consensus existed regarding pterygia diagnosis. IC accuracy was assessed by sensitivity and specificity with a 95% confidence interval. Results: From the 32 primary lesions studied, histopathological examination confirmed the diagnosis of pterygium without atypical cells in 19 cases (60%) and showed unsuspected and associated OSSN cells in 13 cases (40%). IC demonstrated one false-negative and one false-positive result for atypia. Statistical analysis showed an estimated sensitivity of 92%, specificity of 94%, positive predictive value of 92%, and negative predictive value of 94%. Conclusion: IC demonstrated high agreement with histopathological analysis in the detection of atypical epithelial cells in unsuspected OSSN in Brazilian pterygia patients.

Use of impression cytology for the detection of unsuspected ocular surface squamous neoplasia cells in pterygia / Uso da citologia de impressão para a detecção de células de neoplasia escamosa da superfície ocular insuspeita em pterígios

Purpose: To evaluate the agreement between the methodologies of impression cytology (IC) and histopathology regarding epithelial lesions clinically diagnosed as pterygium and also regarding the detection of unsuspected and associated ocular surface squamous neoplasia (OSSN). Methods: Thirty-two Brazilian patients were included and IC was performed on all pterygia before excision. Histopathogical examination was considered the gold standard and was performed by two experienced ocular pathologists in which consensus existed regarding pterygia diagnosis. IC accuracy was assessed by sensitivity and specificity with a 95% confidence interval. Results: From the 32 primary lesions studied, histopathological examination confirmed the diagnosis of pterygium without atypical cells in 19 cases (60%) and showed unsuspected and associated OSSN cells in 13 cases (40%). IC demonstrated one false-negative and one false-positive result for atypia. Statistical analysis showed an estimated sensitivity of 92%, specificity of 94%, positive predictive value of 92%, and negative predictive value of 94%. Conclusion: IC demonstrated high agreement with histopathological analysis in the detection of atypical epithelial cells in unsuspected OSSN in Brazilian pterygia patients. .

Objetivo: Avaliar a concordância entre a citologia de impressão (CI) e os achados histopatológicos de lesões epiteliais clinicamente diagnosticadas como pterígio, no que tange a detecção de células de neoplasia escamosa da superfície ocular (NESO) insuspeita associada. Métodos: Trinta e dois pacientes brasileiros foram incluídos e a CI foi obtida de todos os pterígios antes da excisão. O exame histopatológico foi realizado por dois patologistas oculares experientes em consenso de opinião e considerado o padrão-ouro para o diagnóstico. A acurácia da CI foi avaliada pela sensibilidade e especificidade com intervalo de confiança de 95% Resultados: Das 32 lesões estudadas, o exame histopatológico confirmou o diagnóstico de pterígio sem atipia em 19 casos (60%) e mostrou células de NESO insuspeita associada em 13 espécimes (40%). A CI demonstrou um resultado falso-positivo e um falso-negativo para atipia. A análise estatística mostrou uma sensibilidade estimada de 92%, especificidade de 94%, valor preditivo positivo de 92% e valor preditivo negativo de 94%. Conclusão: A CI apresentou alta concordância com o estudo histopatológico na detecção de células epiteliais atípicas de NESO insuspeita em pterígios do Brasil. .

Amelanotic corneally displaced malignant conjunctival melanoma: a case report evaluated with impression cytology.

Here we describe the case of a 65-year-old Caucasian female who presented with an amelanotic malignant conjunctival melanoma and highlight the clinical and pathological features of this rare entity that displayed exclusive corneal invasive growth without evidence of conjunctival tumors other than primary acquired melanosis. Impression cytology aided in the initial diagnosis. The patient underwent surgical treatment. Histopathology and immunohistochemistry revealed an invasive amelanotic melanoma limited to the cornea and exhibiting S-100, Melan A, and HMB-45 positivity. The absence of pigmentation delayed early clinical detection and treatment. Awareness of this nonpigmented melanoma is important for early recognition and appropriate management.

Amelanotic corneally displaced malignant conjunctival melanoma: a case report evaluated with impression cytology / Melanoma amelanótico conjuntival maligno deslocado sobre a córnea com citologia de impressão: relato de caso

Here we describe the case of a 65-year-old Caucasian female who presented with an amelanotic malignant conjunctival melanoma and highlight the clinical and pathological features of this rare entity that displayed exclusive corneal invasive growth without evidence of conjunctival tumors other than primary acquired melanosis. Impression cytology aided in the initial diagnosis. The patient underwent surgical treatment. Histopathology and immunohistochemistry revealed an invasive amelanotic melanoma limited to the cornea and exhibiting S-100, Melan A, and HMB-45 positivity. The absence of pigmentation delayed early clinical detection and treatment. Awareness of this nonpigmented melanoma is important for early recognition and appropriate management.

Os autores descrevem o caso de uma mulher branca de 65 anos que apresentava um melanoma amelanótico maligno conjuntival e destacam as características clínicas e patológicas desta entidade rara com crescimento invasivo exclusivo na córnea sem evidência de tumores na conjuntiva além de melanose adquirida primária sem pigmento. A citologia de impressão auxiliou no diagnóstico inicial. A paciente foi submetida a tratamento cirúrgico. A histopatologia e a imuno-histoquímica revelaram um melanoma amelanótico invasivo limitado sobre a córnea exibindo positividade para proteína S-100, Melan A e HMB-45. A ausência de pigmentação retardou sua identificação clínica e seu tratamento precoce. O conhecimento deste melanoma não pigmentado é importante para o reconhecimento precoce e a conduta apropriada.

Ocular surface evaluation in patients treated with a fixed combination of prostaglandin analogues with 0.5% timolol maleate topical monotherapy: a randomized clinical trial.

OBJECTIVES: To compare ocular surface changes induced via glaucoma treatment in patients using fixed combinations of prostaglandin analogues (travoprost, latanoprost and bimatoprost) with 0.5% timolol maleate METHODS: A prospective, multicenter, randomized, parallel group, single-blind clinical trial was performed in 33 patients with ocular hypertension or open angle glaucoma who had not been previously treated. The ocular surface was evaluated prior to and three months after treatment, with a daily drop instillation of one of the three medications. The main outcome measurements included the tear film break-up time, Schirmer's test, Lissamine green staining, the Ocular Surface Disease Index questionnaire, impression cytology using HE and PAS and immunocytochemistry for interleukin-6 and HLA-DR. Ensaiosclinicos.gov.br: UTN - U1111-1129-2872 RESULTS: All of the drugs induced a significant reduction in intraocular pressure. Decreases in the Schirmer's test results were observed with all of the drugs. Decreases in tear-film break-up time were noted with travoprost/timolol and latanoprost/timolol. An increase in the Lissamine green score was noted with travoprost/timolol and bimatoprost/timolol. The Ocular Surface Disease Index score increased after treatment in the travoprost/timolol group. Impression cytology revealed a significant difference in cell-to-cell contact in the same group, an increase in cellularity in all of the groups and an increase in the number of goblet cells in all of the groups. The fixed combinations induced an increase in IL-6 expression in the travoprost/timolol group, in which there was also an increase in HLA-DR expression. CONCLUSIONS: All of the fixed combinations induced a significant reduction in intraocular pressure, and the travoprost/timolol group showed increased expression of the inflammatory markers HLA-DR and interleukin-6. All three tested medications resulted in some degree of deterioration in the ocular surface after three months of glaucoma treatment.

Ocular surface evaluation in patients treated with a fixed combination of prostaglandin analogues with 0.5% timolol maleate topical monotherapy: a randomized clinical trial

OBJECTIVES: To compare ocular surface changes induced via glaucoma treatment in patients using fixed combinations of prostaglandin analogues (travoprost, latanoprost and bimatoprost) with 0.5% timolol maleate METHODS: A prospective, multicenter, randomized, parallel group, single-blind clinical trial was performed in 33 patients with ocular hypertension or open angle glaucoma who had not been previously treated. The ocular surface was evaluated prior to and three months after treatment, with a daily drop instillation of one of the three medications. The main outcome measurements included the tear film break-up time, Schirmer's test, Lissamine green staining, the Ocular Surface Disease Index questionnaire, impression cytology using HE and PAS and immunocytochemistry for interleukin-6 and HLA-DR. Ensaiosclinicos.gov.br: UTN - U1111-1129-2872 RESULTS: All of the drugs induced a significant reduction in intraocular pressure. Decreases in the Schirmer's test results were observed with all of the drugs. Decreases in tear-film break-up time were noted with travoprost/timolol and latanoprost/timolol. An increase in the Lissamine green score was noted with travoprost/timolol and bimatoprost/timolol. The Ocular Surface Disease Index score increased after treatment in the travoprost/timolol group. Impression cytology revealed a significant difference in cell-to-cell contact in the same group, an increase in cellularity in all of the groups and an increase in the number of goblet cells in all of the groups. The fixed combinations induced an increase in IL-6 expression in the travoprost/timolol group, in which there was also an increase in HLA-DR expression. CONCLUSIONS: All of the fixed combinations induced a significant reduction in intraocular pressure, and the travoprost/timolol group showed increased expression of the inflammatory markers HLA-DR and interleukin-6. All three tested medications resulted in some degree of deterioration in ...

The use of 1% toluidine blue eye drops in the diagnosis of ocular surface squamous neoplasia.

PURPOSE: To evaluate the use of in vivo toluidine blue (TB) dye in the diagnosis of ocular surface squamous neoplasia and to correlate staining intensity with the histopathological diagnosis. METHODS: A prospective study was performed at the Federal University of Sao Paulo, Brazil. Patients with conjunctival epithelial lesions were examined by slit-lamp biomicroscopy, with and without 1% TB eye drops, and the results were photo documented. Before the instillation of the dye, 1% proxymetacaine HCl eye drops were used. All patients were submitted to surgery and histopathological analysis to confirm the diagnosis. The patients were grouped according to the histopathological aspects of the lesions into 3 groups: group 1-patients with ocular surface squamous neoplasia; group 2-patients with premalignant lesions; and group 3-patients with pterygium. The digital images were analyzed by 2 masked examiners who had no previous access to the histopathological results. The photographs were classified according to the positivity and intensity of the staining. The statistical analysis method chosen depended on the type of data, with the level of significance set as P < 0.05. RESULTS: Forty-seven patients were included in the study: 10 had benign lesions (pterygium), 10 had premalignant lesions (actinic keratosis), and 27 had malignant lesions (conjunctival intraepithelial neoplasia and conjunctival squamous cell carcinoma). Agreement between observers regarding the analysis of the digital photographs was 100% for positivity and 82.9% for intensity of staining (κ = 0.938). Ninety percent of patients with premalignant lesions and all patients with malignant lesions showed positive staining with 1% TB. One patient had positive staining, but histopathological examination revealed a benign lesion (false positive). CONCLUSIONS: The use of 1% TB eye drops is an efficient method for the clinical diagnosis of ocular surface squamous neoplasia and premalignant lesions. Nevertheless, the intensity of the staining does not correlate with the degree of malignancy of these tumors.

Impression cytologic analysis after corneal cross-linking and insertion of corneal ring segments for keratoconus: two-year results.

PURPOSE: To report the impression cytologic results after corneal cross-linking and insertion of intrastromal corneal ring segments for keratoconus. METHODS: Thirty-nine eyes were distributed into two groups: 1) cross-linking group (patients underwent corneal cross-linking procedure), and 2) riboflavin eyedrops group (patients received 0.1% riboflavin (w/v) eyedrops in 20% dextran solution for 1 month). After 3 months, all patients underwent insertion of intrastromal corneal ring segments. Impression cytologic specimens were obtained from all eyes at baseline, at 1 month and 3 months after cross-linking or riboflavin eyedrops, and again at 6 months, 1 year, and 2 years after intrastromal corneal ring segment insertion. RESULTS: Patients in the cross-linking group demonstrated improvement in the cell-to-cell contact of epithelial cells and the nucleus-to-cytoplasm ratio on the temporal conjunctiva after treatment (P=0.008 and P=0.047), respectively. On the superior conjunctiva, increases in goblet cell density (P=0.037) and level of organization of nuclear chromatin (P=0.010) after treatment were noted. Patients in the riboflavin eyedrops group demonstrated improvement in the cell-to-cell contact of epithelial cells on the superior conjunctiva after treatment (P=0.021). On the temporal conjunctiva, an improvement in the cell-to-cell contact of epithelial cells (P<0.001) and increases in the nucleus-to-cytoplasm ratio (P<0.001), goblet cell density (P=0.001), and less keratinization (P=0.011) were noted. No changes were identified on the cornea for either group. Fisher's exact test comparison of the impression cytologic total scores after treatment revealed no difference between groups. CONCLUSION: Despite changes in some conjunctival parameters (e.g., cell-to-cell contact of epithelial cells, nucleus-to-cytoplasm ratio, level of organization of nuclear chromatin, goblet cell density, and keratinization), comparison of the total impression cytologic scores revealed no difference between groups.

Impression cytologic analysis after corneal cross-linking and insertion of corneal ring segments for keratoconus: two-year results / Citologia de impressão após "cross-linking" corneano e inserção dos segmentos de anéis corneanos para ceratocone: dois anos de seguimento

PURPOSE: To report the impression cytologic results after corneal cross-linking and insertion of intrastromal corneal ring segments for keratoconus. METHODS: Thirty-nine eyes were distributed into two groups: 1) cross-linking group (patients underwent corneal cross-linking procedure), and 2) riboflavin eyedrops group (patients received 0.1% riboflavin (w/v) eyedrops in 20% dextran solution for 1 month). After 3 months, all patients underwent insertion of intrastromal corneal ring segments. Impression cytologic specimens were obtained from all eyes at baseline, at 1 month and 3 months after cross-linking or riboflavin eyedrops, and again at 6 months, 1 year, and 2 years after intrastromal corneal ring segment insertion. RESULTS: Patients in the cross-linking group demonstrated improvement in the cell-to-cell contact of epithelial cells and the nucleus-to-cytoplasm ratio on the temporal conjunctiva after treatment (P=0.008 and P=0.047), respectively. On the superior conjunctiva, increases in goblet cell density (P=0.037) and level of organization of nuclear chromatin (P=0.010) after treatment were noted. Patients in the riboflavin eyedrops group demonstrated improvement in the cell-to-cell contact of epithelial cells on the superior conjunctiva after treatment (P=0.021). On the temporal conjunctiva, an improvement in the cell-to-cell contact of epithelial cells (P<0.001) and increases in the nucleus-to-cytoplasm ratio (P<0.001), goblet cell density (P=0.001), and less keratinization (P=0.011) were noted. No changes were identified on the cornea for either group. Fisher's exact test comparison of the impression cytologic total scores after treatment revealed no difference between groups. CONCLUSION: Despite changes in some conjunctival parameters (e.g., cell-to-cell contact of epithelial cells, nucleus-to-cytoplasm ratio, level of organization of nuclear chromatin, goblet cell density, and keratinization), comparison of the total impression cytologic scores revealed no difference between groups.

OBJETIVO: Relatar os resultados de citologia de impressão após o "cross-linking" da córnea e inserção dos segmentos de anéis corneanos intraestromais para ceratocone. MÉTODOS: Trinta e nove olhos foram distribuídos em dois grupos: pacientes no grupo cross-linking foram submetidos ao procedimento do "cross-linking" corneano e pacientes no grupo colírio de riboflavina receberam o colírio de riboflavina 0,1% (w/v) -20% dextran para uso tópico por um mês. Após três meses, todos os pacientes foram submetidos à inserção dos segmentos de anéis corneanos intraestromais. Citologia de impressão foi realizada em todos os olhos no pré-operatório, um mês e três meses após o "cross-linking" ou colírio de riboflavina, e novamente com seis meses, um ano e dois anos após a inserção dos segmentos de anéis. RESULTADOS: Pacientes do grupo "cross-linking" apresentaram melhora na adesividade das células epiteliais e proporção núcleo: citoplasma na conjuntiva temporal após o tratamento (P=0,008 e P=0,047), respectivamente. Na conjuntiva superior, houve um aumento na densidade das células caliciformes (P=0,037) e nível de organização da cromatina nuclear (P=0,010) após o tratamento. Pacientes do grupo colírio de riboflavina apresentaram melhora na adesividade das células epiteliais na conjuntiva superior após o tratamento (P=0,021). Na conjuntiva temporal, houve melhora na adesividade das células epiteliais (P<0,001), aumento na proporção núcleo: citoplasma (P<0,001), células caliciformes (P=0,001) e menor queratinização (P=0,011). Ambos os grupos não apresentaram alterações corneanas. O Teste exato de Fisher para comparação de citologia de impressão no escore total após o tratamento não revelou diferença entre os grupos. CONCLUSÃO: Apesar de mudanças em alguns parâmetros conjuntivais (adesividade das células epiteliais, proporção núcleo:citoplasma, nível de organização da cromatina nuclear, células caliciformes e queratinização), a comparação do escore total na citologia de impressão não apresentou diferença entre os grupos.

Cytological features of live limbal tissue donor eyes for autograft or allograft limbal stem cell transplantation.

PURPOSE: To evaluate by impression cytology (IC) the corneal surface of live limbal tissue donor eyes for autograft or allograft limbal stem cell transplantation (LSCT). METHODS: Twenty limbal donors were enrolled (17 for autograft LSCT and 3 for allograft). Impression cytology was performed before transplantation of superior and inferior limbal grafts and after the third postoperative month. RESULTS: Impression cytology analysis showed sheets of corneal epithelial cells and goblet cell absence beyond the edge of the keratectomy sites in all patients, suggesting that conjunctival invasion towards the center did not occur in any eye. Partial conjunctivalization within 2 to 3 clock hours, confirmed by the presence of goblet cells, was limited to the keratectomy site in 10% of the cases. CONCLUSION: A clear central corneal surface was demonstrated in all eyes following surgery leading to the conclusion that limbal donation was a safe procedure in this group of patients. A small percentage of eyes can have donor sites re-epithelized with conjunctival cells at the periphery of the cornea.

Cytological features of live limbal tissue donor eyes for autograft or allograft limbal stem cell transplantation / Características citológicas do tecido límbico de doador vivo para transplante autólogo ou alógeno de células-tronco epiteliais corneais

PURPOSE: To evaluate by impression cytology (IC) the corneal surface of live limbal tissue donor eyes for autograft or allograft limbal stem cell transplantation (LSCT). METHODS: Twenty limbal donors were enrolled (17 for autograft LSCT and 3 for allograft). Impression cytology was performed before transplantation of superior and inferior limbal grafts and after the third postoperative month. RESULTS: Impression cytology analysis showed sheets of corneal epithelial cells and goblet cell absence beyond the edge of the keratectomy sites in all patients, suggesting that conjunctival invasion towards the center did not occur in any eye. Partial conjunctivalization within 2 to 3 clock hours, confirmed by the presence of goblet cells, was limited to the keratectomy site in 10 percent of the cases. CONCLUSION: A clear central corneal surface was demonstrated in all eyes following surgery leading to the conclusion that limbal donation was a safe procedure in this group of patients. A small percentage of eyes can have donor sites re-epithelized with conjunctival cells at the periphery of the cornea.

OBJETIVO: Avaliar pela citologia de impressão a superfície da córnea de doador vivo para transplante autólogo ou alógeno de células-tronco epiteliais. MÉTODOS: Vinte pacientes doadores de tecido límbico foram avaliados (17 para transplante autólogo e 3 para alógeno). Os exames citológicos foram realizados em dois momentos: antes da ceratectomia, que removeu tecido límbico dos quadrantes superior e inferior, e após o terceiro mês pós-operatório. RESULTADOS: Invasão de células da conjuntiva em direção ao centro além da margem da ceratectomia não ocorreu em nenhum olho estudado. Uma pequena área de conjuntivalização parcial, confirmada pela presença de células caliciformes, foi detectada dentro do limite da ceratectomia em 10 por cento dos casos. CONCLUSÃO: A superfície central da córnea manteve-se transparente demonstrando que a manipulação de tecido límbico em doador vivo foi um procedimento seguro neste grupo de pacientes. Uma pequena porcentagem dos olhos pode ter o local do sítio da ceratectomia re-epitelizado com células da conjuntiva sobre a periferia da córnea.

Impression cytologic analysis after corneal collagen cross-linking using riboflavin and ultraviolet-A light in the treatment of keratoconus.

PURPOSE: To report impression cytologic (IC) results after corneal cross-linking (CXL) using riboflavin and ultraviolet-A light in the treatment of keratoconus and compare the data with those from a group of subjects with the same disease. METHODS: Forty eyes were distributed into 2 groups: patients in group 1 underwent CXL, whereas patients in group 2 received riboflavin 0.1% eyedrops for 1 month of topical use. IC specimens were obtained from all eyes before treatment and 1 and 3 months after treatment. RESULTS: Patients in group 1 showed a decrease in goblet cell density on the superior conjunctiva after CXL (P = 0.008) but no difference on the temporal conjunctiva or in the cornea. Patients in group 2 demonstrated improvement in cell-to-cell contact of epithelial cells and reduced keratinization on the temporal conjunctiva after treatment (P = 0.003 and P = 0.034, respectively) but no changes on the superior conjunctiva or in the cornea. Fisher exact test comparison of IC total scores after treatment revealed no difference between groups. CONCLUSIONS: Despite changes in goblet cell density after corneal CXL in the superior conjunctiva and an improvement in the morphology of epithelial cells after the use of riboflavin eyedrops, comparison of total IC scores showed no difference between groups.

Impression cytology findings in bullous keratopathy.

AIM: To evaluate, by impression cytology (IC), the cytological features of the central area of corneal epithelial surface of patients with symptomatic bullous keratopathy (BK). Design Cross-sectional observational case series. METHODS: IC of the central cornea was performed in 72 eyes of 72 patients with symptomatic BK between June 2005 and December 2006. Patients were divided into two groups according to the presence (group GC) or not of goblet cells (group NGC). The diagnosis of limbal stem cell deficiency (LSCD) was defined by the presence of one or more intact goblet cells on the corneal surface. Squamous metaplasia was classified according to Murube and Rivas. RESULTS: Epithelial squamous metaplasia was found in 72 (100%) eyes, along with conjunctival goblet cells in 25 (34.72%) eyes. Squamous metaplasia was grade 1 in group GC and more advanced grades in group NGC. Corneal neovascularisation was present in 62 (82.11%) eyes. Vessel measure was more than 4 mm in 11 eyes (23.4%) in group NGC and 10 eyes (40%) in group GC (p=0.559), and they were mostly subepithelial in 23 eyes (48.9%) in group NGC and 12 (48%) eyes in group GC (p=0.822). CONCLUSION: Squamous metaplasia was the most frequent finding associated with advanced BK. The presence of goblet cells on the surface of corneas with squamous metaplasia grade 1, in a significant number of patients, suggests that limbal stem cell deficiency is a common condition associated with advanced cases of BK.