Comparison of techniques for corneal epithelium cell culture for the collection of conditioned medium.

PURPOSES: To determine the best protocol in obtaining the higher yield of conditioned culture medium to be used for the bone marrow mesenchymal stem cell differentiation into corneal epithelial cells, five techniques for the primary culture of human corneal epithelial cells were evaluated. METHODS: The studied culture techniques of corneal epithelial cells were: explants in culture flasks with and without hydrophilic surface treatment, on amniotic membrane, with enzymatic digestion, and by corneal scraping. The conditioned culture medium collected from these cultures was used to differentiate human bone marrow mesenchymal stem cells into corneal epithelial cells, which were characterized using flow cytometry with pan-cytokeratin and the corneal-specific markers, cytokeratin 3 and cytokeratin 12. RESULTS: The culture technique using flasks with hydrophilic surface treatment resulted in the highest yield of conditioned culture medium. Flasks without surface treatment resulted to a very low success rate. Enzymatic digestion and corneal scraping showed contamination with corneal fibroblasts. The culture on amniotic membranes only allowed the collection of culture medium during the 1st cell confluence. The effectiveness of cell differentiation was confirmed by cytometry analysis using the collected conditioned culture medium, as demonstrated by the expressions of cytokeratin 3 (95.3%), cytokeratin 12 (93.4%), and pan-cytokeratin (95.3%). CONCLUSION: The culture of corneal epithelial cell explants in flasks with hydrophilic surface treatment is the best technique for collecting a higher yield of conditioned culture medium to be used to differentiate mesenchymal stem cells.

Nasolacrimal obstruction concomitant to cataract: diagnosis and management in the preoperative period of cataract surgery / Obstrução lácrimo-nasal concomitante a catarata - diagnóstico e conduta no período pré-operatório da facectomia

ABSTRACT Purpose: Concomitant nasolacrimal duct obstruction can occur in cataract carriers, which increases the risk of postoperative endophthalmitis. The primary aim of this study is to evaluate the knowledge of Brazilian cataract surgeons on the diagnosis and management of cataracts associated with nasolacrimal duct obstruction. Methods: This survey was based on a questionnaire involving Brazilian cataract surgeons that was conducted from March to April 2018. Data were collected on the participant's profile, time and experience in ophthalmic practice, previous training in diagnosis and management of nasolacrimal duct obstruction, and background with endophthalmitis after cataract surgery in patients with nasolacrimal duct obstruction. All data were entered into an Excel spreadsheet and analyzed according to the frequency of occurrence. Results: Ninety-one ophthalmologists answered the questionnaire. Most (63.7%) had been performing cataract surgery for >10 years, and most (84.6%) received training to diagnose and handle nasolacrimal duct obstruction during their medical residence training. Nasolacrimal duct obstruction was investigated in the preoperative period of the cataract by lacrimal sac expression test (53.8%) or by irrigation of the tear pathways (23.1%). Nasolacrimal duct obstruction was treated with antibiotic eye drops by 47.2% of respondents. Seventy-eight percent of surgeons indicate usually performing lacrimal surgery prior to the intraocular surgery, waiting for 4 to 6 weeks to proceed with the cataract surgery. The procedure of choice for treating nasolacrimal duct obstruction prior to cataract surgery was dacryocystorhinostomy (88.4%). Most participants recognized the need for a protocol to assist in the detection and management of nasolacrimal duct obstruction in cataract carriers. Conclusion: Improvement in the diagnosis and management of nasolacrimal duct obstruction concomitant to cataract is needed, as this is a risk factor for endophthalmitis.

RESUMO Objetivo: Portadores de catarata podem apresentar concomitantemente obstrução do ducto lacrimo-nasal (DLN), com risco de desenvolver endoftalmite no pós-operatório da facectomia. O objetivo do presente estudo é apresentar as percepções dos cirurgiões de catarata sobre a propedêutica e a conduta frente a pacientes com obstrução do ducto lacrimo-nasal concomitante com catarata. Métodos: Trata-se de uma pesquisa baseada em um questionário envolvendo cirurgiões brasileiros de catarata, realizado no período de março a abril de 2018. Foram levantados dados sobre o perfil dos participantes, o tempo e a experiencia da prática oftalmológica, o treinamento prévio para diagnóstico e tratamento da obstrução do ducto lacrimo-nasal e os conhecimentos de endoftalmite após cirurgia de catarata. Todos os dados foram inseridos em planilha Excel e analisados de acordo com a frequência de ocorrência. Resultados: Noventa e um oftalmologistas responderam ao questionário. A maioria (63,7%) deles realiza cirurgias de catarata há mais de 10 anos e a maioria (84,6%) recebeu treinamento para diagnóstico e tratamento da obstrução do ducto lacrimo-nasal durante o curso de residência médica. A pesquisa da obstrução crônica do ducto lacrimo-nasal no pré-operatório da catarata é feita pelo teste do refluxo de secreção pelos pontos lacrimais (53,8%) ou por irrigação das vias lacrimais (23,1%). A obstrução do ducto lacrimo-nasal é tratada com colírios antibióticos por 47,2% dos respondentes. Para os portadores de obstrução do ducto lacrimo-nasal , 78% indicam a desobstrução das vias lacrimais previamente à facectoma, aguardando de 4 a 6 semanas para tal. O procedimento de escolha para tratar a obstrução do ducto lacrimo-nasal antes da facectomia é a dacriocistorrinostomia (88,4%). A necessidade de um protocolo para auxiliar na detecção e tratamento da obstrução do ducto lacrimo-nasal em portadores de catarata é reconhecida pela maioria dos participantes deste estudo. Conclusão: É necessário melhorar a propedêutica e o manejo da catarata em portador de obstrução do ducto lacrimo-nasal porque esse é um fator de risco para endoftalmite.

Nasolacrimal obstruction concomitant to cataract: diagnosis and management in the preoperative period of cataract surgery.

PURPOSE: Concomitant nasolacrimal duct obstruction can occur in cataract carriers, which increases the risk of postoperative endophthalmitis. The primary aim of this study is to evaluate the knowledge of Brazilian cataract surgeons on the diagnosis and management of cataracts associated with nasolacrimal duct obstruction. METHODS: This survey was based on a questionnaire involving Brazilian cataract surgeons that was conducted from March to April 2018. Data were collected on the participant's profile, time and experience in ophthalmic practice, previous training in diagnosis and management of nasolacrimal duct obstruction, and background with endophthalmitis after cataract surgery in patients with nasolacrimal duct obstruction. All data were entered into an Excel spreadsheet and analyzed according to the frequency of occurrence. RESULTS: Ninety-one ophthalmologists answered the questionnaire. Most (63.7%) had been performing cataract surgery for >10 years, and most (84.6%) received training to diagnose and handle nasolacrimal duct obstruction during their medical residence training. Nasolacrimal duct obstruction was investigated in the preoperative period of the cataract by lacrimal sac expression test (53.8%) or by irrigation of the tear pathways (23.1%). Nasolacrimal duct obstruction was treated with antibiotic eye drops by 47.2% of respondents. Seventy-eight percent of surgeons indicate usually performing lacrimal surgery prior to the intraocular surgery, waiting for 4 to 6 weeks to proceed with the cataract surgery. The procedure of choice for treating nasolacrimal duct obstruction prior to cataract surgery was dacryocystorhinostomy (88.4%). Most participants recognized the need for a protocol to assist in the detection and management of nasolacrimal duct obstruction in cataract carriers. CONCLUSION: Improvement in the diagnosis and management of nasolacrimal duct obstruction concomitant to cataract is needed, as this is a risk factor for endophthalmitis.