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To compare the cyclosporine 0.05 % exposure effect on fibroblasts from primary and recurrent pterygium. Primary culture of fibroblasts from primary and recurrent pterygium was performed until the third passage, which was exposed to cyclosporine 0.05 % in a group and the other remaining unexposed (control group), in triplicates. After 3, 6, 12, and 17 days of exposure the viable cell counting was performed by hemocytometer. The results were statistically analyzed using the technique of analysis of non-parametric variance model for repeated measures with three factors. There was a significant reduction in both fibroblast proliferation, in primary as in the recurrent pterygium cultures exposed to cyclosporine when compared not exposed cultures, with statistical significance (P < 0.05). Comparing primary and recurrent pterygium that received the drug, there was no significant difference in cell proliferation in relation to primary or recurrent pterygium. Cyclosporine 0.05 % is effective in inhibiting fibroblast proliferation in culture, both in primary and as in recurrent pterygium.
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Ciclosporina/farmacología , Fibroblastos/efectos de los fármacos , Inmunosupresores/farmacología , Pterigion/tratamiento farmacológico , Análisis de Varianza , Recuento de Células , Proliferación Celular/efectos de los fármacos , Humanos , Estudios ProspectivosRESUMEN
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.
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Técnicas de Cultivo de Célula , Diferenciación Celular , Epitelio Corneal , Citometría de Flujo , Células Madre Mesenquimatosas , Humanos , Medios de Cultivo Condicionados , Epitelio Corneal/citología , Diferenciación Celular/fisiología , Citometría de Flujo/métodos , Células Madre Mesenquimatosas/citología , Técnicas de Cultivo de Célula/métodos , Amnios/citología , Células Cultivadas , Queratina-3/metabolismo , Queratina-3/análisis , Queratina-12/metabolismo , Reproducibilidad de los ResultadosRESUMEN
BACKGROUND: The pathogenesis and treatment of lateral elbow epicondylitis (LEE) are still controversial. The purpose of the current study was to evaluate the production of inflammatory cytokines by LEE-derived cells and to compare the anti-inflammatory effect of triamcinolone acetonide with platelet-rich plasma (PRP) on cytokines production in primary culture of these cells. METHODS: Third passage cells from primary cultures of LEE were assessed for the production of the cytokines IL-1ß, IL-6, IL-8, IL-10 and TNF-α by immune-enzymatic assay (ELISA), after the treatment with 1, 10 and 100 µM triamcinolone compared to no treated controls at the time points 6, 12, 18, 24, 48, 72 and 96 h, and to PRP at 48, 72 and 96 h. RESULTS: The cytokines IL-6 and IL-8 were produced in high concentrations by LEE cells. One, 10 and 100 µM triamcinolone induced significant decrease in the production of IL-6 and IL-8 at 48, 72 and 96 h, adding the time point 12 h for IL-8. Compared to controls, PRP caused a significant increase in the production of IL-6 and IL-8 and there was a significant increase in IL-10 production with the use of 100 µM triamcinolone at 48 h. The production of IL1-ß and TNF-α was very low and did not change when the cultures were treated with triamcinolone or PRP. CONCLUSION: LEE-derived cells produce IL-6 and IL-8, confirming the inflammatory nature of this condition. While triamcinolone inhibited the production of IL-6 and IL-8 by LEE cells, PRP induced an increase in these cytokines compared with controls.
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Citocinas/sangre , Plasma Rico en Plaquetas , Codo de Tenista/terapia , Triamcinolona/uso terapéutico , Humanos , Interleucina-10 , Interleucina-6 , Interleucina-8 , Codo de Tenista/tratamiento farmacológico , Triamcinolona/farmacología , Factor de Necrosis Tumoral alfaRESUMEN
Background: Mesenchymal stem cells (MSCs) are an excellent treatment option for a wide variety of orthopaedic conditions. This study aimed to establish if bone marrow MSCs obtained from proximal humerus fractures can be an alternative source for obtaining primary cultures of human MSCs. Methods: Human bone marrow was obtained during osteosynthesis surgeries on closed proximal humerus fractures within 48 hours of injury. MSCs were harvested using the Ficoll gradient separation protocol and in vitro cultured until the third passage. Then, the cells were immunophenotyped by flow cytometry using stem cell specific surface markers. The cells were also induced to differentiate into osteoblasts and adipocytes for the characterization and confirmation of MSCs. The production of cytokines interleukin (IL)-1ß, IL-6, IL-8, IL-10, tumor necrosis factor α (TNF-α), and interferon γ (IFN-γ) was assessed using enzyme-linked immunosorbent assay (ELISA) in the supernatant of the cultures after 3, 5 or 7 days. Results: Immunophenotyping showed high expression of the stem cell surface markers CD73, CD90, and CD105 and negative or very low expression of CD34, CD45, CD11b, CD19, and human leukocyte antigen (HLA)-DR. The bone marrow derived MSCs were able to differentiate into osteoblasts and adipocytes. The quantification of secreted cytokines revealed that IL-8 was the most produced cytokine, followed by IL-6 and IL-10 at similar quantities and lower levels of IL-1ß. TNF-α and IFN-γ were not detected. Conclusions: Proximal humerus fractures can be an alternative source for the collection of bone marrow MSCs. The cytokine production of these cells is very similar to the production profile of fracture haematomas previously reported and may be used for improving bone repair.
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AIM: To investigate the anti-inflammatory effects of the sesquiterpenes α-humulene and ß-caryophyllene on pterygium fibroblasts. METHODS: Primary cultures of pterygium fibroblasts were established. Third passage pterygium fibroblasts were exposed to α-humulene and ß-caryophyllene separately and together. The cell viability was assessed by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay at 12, 24, 48, and 72h after exposure. The levels of the inflammatory cytokines interleukin (IL)-1ß, IL-6, IL-8, tumor necrosis factor (TNF)-α and IL-10 in the conditioned culture medium were assessed by enzyme-linked immunosorbent assay (ELISA) at 12, 24 and 48h after exposure. Data were statistically analyzed using Friedman repeated measures analysis of variances on ranks. RESULTS: The 25 µmol/L ß-caryophyllene induced significant decrease in the IL-6 production by pterygium fibroblasts 48h after the exposure (P=0.041). The levels of IL-1ß, IL-8, IL-10, and TNF-α were very low and had no statistically significant variations after exposure to α-humulene, ß-caryophyllene, or both compounds together. CONCLUSION: The exposure to 25 µmol/L of ß-caryophyllene significantly reduce the production of IL-6 by pterygium fibroblasts after 48h. This sesquiterpene may be a potential alternative adjuvant agent for the treatment of pterygium.
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ABSTRACT 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.
RESUMO Objetivos: Foram estudadas cinco técnicas de cultivo primário de células epiteliais de córnea humana para se determinar o melhor protocolo para a obtenção do maior rendimento de meio de cultivo condicionado para ser utilizado na diferenciação de células tronco mesenquimais para células epiteliais de córnea. Métodos: As técnicas de cultivo estudadas foram: explantes em frascos de cultivo com e sem tratamento hidrofílico de superfície, sobre membrana amniótica, com digestão enzimática e por raspado de córnea. O meio de cultivo condicionado foi coletado e as células tronco mesenquimais induzidas a se diferenciarem em células epiteliais da córnea utilizando o meio de cultivo condicionado. As células foram caracterizadas por citometria de fluxo com pan-citoqueratina e com os marcadores específicos da córnea, citoqueratina 3 e citoqueratina 12. Resultados: A técnica utilizando frascos com o tratamento de superfície apresentou o maior rendimento de meio de cultivo condicionado. Os frascos sem tratamento de superfície levaram a uma taxa de sucesso muito baixa. A digestão enzimática e a raspagem da córnea mostraram contaminação das culturas com fibroblastos de córnea. A cultura sobre membranas amnióticas só permitiu a coleta do meio de cultivo condicionado durante a 1ª confluência celular. A análise de citometria de fluxo confirmou o sucesso da diferenciação celular utilizando o meio de cultivo condicionado coletado, demonstrada pela expressão de citoqueratina 3 (95,3%), citoqueratina 12 (93,4%) e pan-citoqueratina (95,3%). Conclusão: O cultivo de explantes de células tronco mesenquimais em frascos com tratamento hidrofílico de superfície é a melhor técnica para a obtenção de um alto rendimento de meio de cultivo condicionado.
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PURPOSE: We evaluated the role of the conjunctival flap rotation technique using 5-fluorouracil and adjuvant therapy with topical cyclosporine A at 0.05% during short pre- and postoperative periods for the prevention of primary pterygium recurrence. METHODS: In this prospective study, 76 patients with primary pterygium (76 eyes) were categorized into two groups: the control group with 31 patients who did not receive cyclosporine treatment, and the cyclosporine group with 45 patients who received topical cyclosporine A (0.05%) twice a day, for 10 days before and 10 days after the pterygium excision operations. Patients were examined for disease recurrence, side effects, and complications at 10 and 21 days, and at 2 and 6 months after the operation. Data on demography, systemic diseases, and ophthalmologic histories were obtained from all patients, and these data were analyzed using descriptive statistics involving the absolute and relative percentages of frequency distribution. Goodman test was used for contrasts among multinomial populations to study the association between cyclosporine A and recurrence. RESULTS: Most patients were between 30 and 60 years of age, and 67.1% were women. We confirmed a higher recurrence in patients with occupational sunlight exposure. The cyclosporine A used topically 10 days before and 10 days after the pterygium removal did not significantly reduce the recurrence of the pterygium. CONCLUSION: Topical 0.05% cyclosporine A when used for 10 days before and 10 days after the pterygium removal does not prevent or reduce the recurrence of primary pterygium.
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Conjuntiva/anomalías , Ciclosporina/administración & dosificación , Inmunosupresores/administración & dosificación , Pterigion/prevención & control , Adulto , Terapia Combinada , Conjuntiva/cirugía , Enfermedades de la Córnea/tratamiento farmacológico , Femenino , Fluorouracilo/uso terapéutico , Humanos , Masculino , Persona de Mediana Edad , Soluciones Oftálmicas/administración & dosificación , Cuidados Posoperatorios , Cuidados Preoperatorios , Estudios Prospectivos , Pterigion/tratamiento farmacológico , Pterigion/cirugía , Recurrencia , Colgajos Quirúrgicos , Adulto JovenRESUMEN
PURPOSE: To evaluate the eyeblink rate in patients with pterygium before and after the lesion exeresis. METHODS: Forty-one patients with pterygium were evaluated before and 60 days after the lesion exeresis. They were digitally videotaped during one minute in a standard setting using a Sony Digital 8 DCR - TRV110 and the images were transferred to a personal computer (McIntosh 400) and processed with the iMovie software. The patients stayed in primary gaze position, under artificial illumination. Total blink, complete and incomplete blink and the opening and closure time were measured using frames scale. The data were submitted to statistical analysis. RESULTS: Before surgery, 36.36% of the patients with pterygium complained of foreign body and after that, 61.02% had no symptoms. The incomplete blink increased after surgery. The opening and closure time was similar before and after surgery. CONCLUSION: With pterygium exeresis the symptoms decreased and the incomplete blink increased a little. Nevertheless the blink rate barely changed. Further research needs to be provided to confirm this hypothesis.
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Parpadeo/fisiología , Pterigion/cirugía , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Periodo Posoperatorio , Estudios Prospectivos , Pterigion/fisiopatología , Estadísticas no Paramétricas , Factores de Tiempo , Grabación de Cinta de Video , Adulto JovenRESUMEN
PURPOSE: To evaluate the sequelae due to epidemic viral conjunctivitis (VC) for a possible association with inflammatory trachoma in schoolchildren. METHODS: A cross-sectional survey was performed of 2,093 schoolchildren in public schools in Sao Paulo State, Brazil. The exams were conducted according to the WHO definitions to detect trachoma. We evaluated the prevalence and the association between inflammatory trachoma and sequelae due to VC. RESULTS: The prevalence of inflammatory trachoma in schoolchildren in Botucatu City was 2.9% (CI: 1.2-17.3%). The prevalence of sequelae from VC was 0.007% (CI: 0 to 0.033%). Acute bacterial conjunctivitis was detected in 0.001%. There was no association between inflammatory trachoma and VC-related sequelae or acute bacterial conjunctivitis. CONCLUSION: An outbreak of epidemic VC can lead to definitive sequelae in the tarsal conjunctiva. The sequelae from VC were detected in 0.007% of the schoolchildren and were not associated with inflammatory trachoma.
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Conjuntivitis Viral/epidemiología , Epidemias/estadística & datos numéricos , Infecciones Virales del Ojo/epidemiología , Tracoma/epidemiología , Población Urbana , Brasil/epidemiología , Niño , Conjuntivitis Viral/diagnóstico , Estudios Transversales , Progresión de la Enfermedad , Infecciones Virales del Ojo/diagnóstico , Femenino , Humanos , Incidencia , Masculino , Prevalencia , Tracoma/diagnósticoRESUMEN
PURPOSE: To evaluate the fibroblast proliferation activity of normal Tenon's capsule from primary and recurrent patients with pterygium. METHODS: A randomized prospective study was performed with 41 normal Tenon's capsule fragments from 21 primary and 20 recurrent patients with pterygium. The sample was collected from the inferior cul-de-sac. Proliferation rate from fibroblasts were evaluated after mitomycin C and 5-fluorouracil exposition. Data were submitted to statistical analysis. RESULTS: Of the 41 cultivated normal Tenon's capsules, only 1 from primary and 2 from recurrent pterygium patients proliferated. After antimitotic exposition, the proliferation rate was similar with both drugs. CONCLUSION: Mitomycin and 5-fluorouracil promote similar inhibition regarding proliferation of normal Tenon's fibroblast cultures.
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Antimetabolitos Antineoplásicos/farmacología , Proliferación Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Fluorouracilo/farmacología , Mitomicina/farmacología , Pterigion/patología , Recuento de Células , Técnicas de Cultivo de Célula , Fibroblastos/citología , Humanos , Estudios Prospectivos , Pterigion/cirugía , RecurrenciaRESUMEN
PURPOSE: To evaluate fibroblast proliferation activity of normal Tenon's capsule and of recurrent pterygia exposed in vitro to tacrolimus (FK506). METHODS: A randomized prospective study was performed with 8 samples of recurrent pterygia and 6 samples of normal Tenon's capsule from the same patient with pterygium at the Faculdade de Medicina de Botucatu - UNESP. The samples were cultivated and exposed once to 1M tacrolimus (FK 506) and the proliferation rate was evaluated 1, 5, 12 and 19 days after the exposure. The data were statistically analyzed. RESULTS: The fibroblasts from pterygia exposed in vitro to tacrolimus had a significantly lower proliferation rate than controls after one day of exposure (p<0.05). Nineteen days after exposure to tacrolimus, exposed cells proliferated more than the non-exposed. CONCLUSION: Tacrolimus is effective to inhibit proliferation of Tenon's capsule fibroblasts from pterygia one day after exposure. Further studies are needed to define the role of tacrolimus in pterygium treatment.
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Fibroblastos/efectos de los fármacos , Inmunosupresores/farmacología , Pterigion/patología , Tacrolimus/farmacología , Técnicas de Cultivo de Célula , Proliferación Celular/efectos de los fármacos , Fibroblastos/patología , Humanos , Estudios Prospectivos , Recurrencia , Factores de TiempoRESUMEN
PURPOSE: To investigate differences in the palpebral fissure eyelid position in Japanese subjects and in Brazilian subjects of European descent over 50 years of age. METHODS: A cross-sectional study evaluating 50 Japanese subjects and 50 Bra---zilian subjects of European descent over 50 years of age was performed to evaluate the effect of aging on eyelid position. Digital photographs were taken with the eyes in the primary position and transferred to a computer to be processed by Scion imaging software for evaluation of palpebral fissure width, height, and margin reflex distance (MRD). The data were statistically analyzed according to sex, age, and race. RESULTS: The width of the eyelid fissure in Japanese subjects was higher than that in Brazilian subjects, whereas the height of the eyelid fissure and the MRD1 was higher in Brazilians than in Japanese (p<0.01). Width, height, and MRD1 were linearly associated with age only in the Japanese group (p<0.05); this association was observed only with width in the Brazilian group (p<0.05). There were no differences according to laterality in either group (p>0.05). CONCLUSION: There are racial differences in age-related changes in the eyelids, which are more marked in Japanese subjects.
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Párpados/anatomía & histología , Factores de Edad , Anciano , Anciano de 80 o más Años , Envejecimiento/fisiología , Antropometría , Pueblo Asiatico , Brasil , Estudios Transversales , Párpados/fisiología , Femenino , Humanos , Interpretación de Imagen Asistida por Computador , Japón/etnología , Masculino , Persona de Mediana Edad , Tamaño de los Órganos , Fotograbar , Valores de Referencia , Población BlancaRESUMEN
PURPOSE: To evaluate fibroblast proliferation activity of the Tenon capsule from primary and recurrent pterygia. METHODS: A randomized prospective study was performed with 43 surgical samples from 30 primary and 13 recurrent pterygia, and samples of normal Tenon capsule from the same pterygium patients. Proliferation, migration, and confluence rate, and patient age, lesion location and type (fleshy or involutive; primary or recurrent) were evaluated. Data were submitted to statistical analysis. RESULTS: Of the 30 cultivated primary pterygia, 21 migrated and proliferated, and 18 (60%) went to confluence. Migration, proliferation and confluence started much later in fibroblasts from normal Tenon than from pterygia lesion. The recurrent pterygia showed migration, proliferation, and confluence starting much earlier than the primary ones. There was no statistically significant difference in the start of migration, proliferation and confluence between fleshy and involuted pterygia, or between pterygias of Grades I-II and III-IV. CONCLUSION: Pterygium fibroblasts are more viable than normal Tenon in cell culture. Migration, proliferation, and confluence differ in primary and recurrent pterygia. Fleshy and involutive pterygia are similar in culture; there is also no difference between pterygia according to lesion size in cell culture.
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Movimiento Celular , Proliferación Celular , Conjuntiva/citología , Fibroblastos/patología , Pterigion/patología , Adulto , Técnicas de Cultivo de Célula , Método Doble Ciego , Humanos , Persona de Mediana Edad , Estudios Prospectivos , Pterigion/cirugía , Recurrencia , Índice de Severidad de la EnfermedadRESUMEN
PURPOSE: To evaluate normal Tenon capsule fibroblasts and from pterygia in culture when exposed to cyclosporin 0.05%. METHODS: A prospective, randomized study was done, involving 20 patients with primary pterygium. Samples of normal and pterygium Tenon's capsule from the same individual were collected, cultured and exposed to cyclosporin 0.05%. RESULTS: Only 7 samples could be exposed to cyclosporin--6 from pterygial Tenon's capsule and 1 from normal Tenon's capsule. There was a significant reduction of cellular proliferation in the cyclosporin exposed cultures (p<0.05). CONCLUSION: Cyclosporin 0.05% is effective to inhibit the proliferation of pterygia and normal Tenon's capsule fibroblasts. Further studies should be performed to define the role of cyclosporin in pterygium treatment.
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Ciclosporina/farmacología , Fibroblastos/efectos de los fármacos , Inmunosupresores/farmacología , Pterigion/patología , Análisis de Varianza , Recuento de Células/métodos , Células Cultivadas , Fibroblastos/citología , Humanos , Estudios Prospectivos , Pterigion/cirugía , Estadísticas no Paramétricas , Factores de TiempoRESUMEN
PURPOSE: To evaluate the possibility of epiphora relief in congenital nasolacrimal obstruction according to the age of the beginning of epiphora complaint and treatment effectiveness. METHODS: Forty children with congenital nasolacrimal obstruction underwent treatment between 1997 and 1999 and were evaluated according to: age of start of epiphora, the results of the treatment and the possibility of successful outcome. The data were submitted to statistical analysis by the binomial proportional test between and within populations. RESULTS: The success rate was worst when the epiphora started after the age of 4 months and results were similar on massage and probing, cure occurring even beyond 3 years of age. CONCLUSION: According to our results, the relief of epiphora was worst when starting at age over 4 months. Massage and probing may have good results even in children older than 3 years.
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Obstrucción del Conducto Lagrimal/congénito , Obstrucción del Conducto Lagrimal/terapia , Edad de Inicio , Preescolar , Humanos , Lactante , Obstrucción del Conducto Lagrimal/epidemiología , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
PURPOSE: To evaluate the adhesion of human lens capsule cells on hydrophilic acrylic intraocular lenses (IOLs) coated with polyethylene glycol (PEG). SETTING: Department of Ophthalmology, Faculty of Medicine, Universidade Estadual Paulista-Botucatu, São Paulo, Brazil. DESIGN: Experimental study. METHODS: Human anterior lens capsules obtained during cataract surgery were cultured and seeded (200 cells/IOLs) in triplicates on the surface of a copolymer comprising hydroxyethyl methacrylate, ethyl methacrylate, and methyl methacrylate IOLs (Loflex) treated or not treated with PEG. After 26 hours, the number of viable adherent cells was estimated by counting in a hemocytometer. RESULTS: The coating of hydrophilic acrylic IOLs with PEG was effective in inhibiting cell adhesion (P < .05). Cells showing 2 distinct morphologic patterns-epithelial and dendritic-like-were observed during the in vitro establishment of the cultures. A tendency toward greater adhesion of dendritic-like cells was observed in untreated IOLs compared with treated IOLs (P = .095). CONCLUSION: Coating hydrophilic acrylic IOLs with PEG was effective in inhibiting cell adhesion. This treatment might play a role in posterior capsule opacification prevention. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.
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Materiales Biocompatibles Revestidos/química , Células Epiteliales/citología , Cápsula del Cristalino/citología , Lentes Intraoculares , Polietilenglicoles/química , Adhesión Celular/fisiología , Células Cultivadas , Humanos , Microscopía de Contraste de FaseRESUMEN
PURPOSE: To evaluate the VEGF expression in macrophages, fibroblasts, and endothelial cells from pterygium before and after 5-fluorouracil (5-FU) exposure. METHODS: 68 excised pterygia (42 primary and 26 recurrent) were studied by immunohistochemistry, to analyze its expression in cells from pterygium and normal conjunctiva. VEGF expression was evaluated before and after a 5-FU subconjunctival injection 15 days prior to surgery. RESULTS: Expression in macrophages was moderate in normal conjunctiva and low to moderate in pterygium tissues. In fibroblasts, it was negative or low in both tissues. Pterygium had a higher proportion of high VEGF expression in endothelial cells compared to normal tissue (p < 0.05). 5-FU did not have any influence on expression. CONCLUSION: VEGF expression in pterygium macrophages, fibroblasts, and endothelial cells was low and similar to that normal in conjunctival tissue. High VEGF expression was found in pterygium endothelial cells compared to normal conjunctiva. 5-FU does not impact VEGF expression.
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Antimetabolitos/uso terapéutico , Células Endoteliales/metabolismo , Fibroblastos/metabolismo , Fluorouracilo/uso terapéutico , Macrófagos/metabolismo , Pterigion/tratamiento farmacológico , Factor A de Crecimiento Endotelial Vascular/metabolismo , Adulto , Anciano , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Pterigion/metabolismoRESUMEN
ABSTRACT Purpose: We evaluated the role of the conjunctival flap rotation technique using 5-fluorouracil and adjuvant therapy with topical cyclosporine A at 0.05% during short pre- and postoperative periods for the prevention of primary pterygium recurrence. Methods: In this prospective study, 76 patients with primary pterygium (76 eyes) were categorized into two groups: the control group with 31 patients who did not receive cyclosporine treatment, and the cyclosporine group with 45 patients who received topical cyclosporine A (0.05%) twice a day, for 10 days before and 10 days after the pterygium excision operations. Patients were examined for disease recurrence, side effects, and complications at 10 and 21 days, and at 2 and 6 months after the operation. Data on demography, systemic diseases, and ophthalmologic histories were obtained from all patients, and these data were analyzed using descriptive statistics involving the absolute and relative percentages of frequency distribution. Goodman test was used for contrasts among multinomial populations to study the association between cyclosporine A and recurrence. Results: Most patients were between 30 and 60 years of age, and 67.1% were women. We confirmed a higher recurrence in patients with occupational sunlight exposure. The cyclosporine A used topically 10 days before and 10 days after the pterygium removal did not significantly reduce the recurrence of the pterygium. Conclusion: Topical 0.05% cyclosporine A when used for 10 days before and 10 days after the pterygium removal does not prevent or reduce the recurrence of primary pterygium.
RESUMO Objetivo: Avaliamos os resultados da técnica de rotação de retalho conjuntival com uso de 5-fluorouracil e terapia adjuvante com ciclosporina A tópica a 0,05%, usada no pré e pós-operatório por curto período, quanto à prevenção da recidiva do pterígio primário Métodos: Estudo prospectivo, com 76 pacientes portadores de pterígio primário (76 olhos), divididos em dois grupos: controle com 31 pacientes que não receberam tratamento com ciclosporina e grupo ciclosporina com 45 pacientes que receberam ciclosporina tópica A (0,05%) duas vezes ao dia, por 10 dias antes e 10 dias após a cirurgia de excisão do pterígio. Os pacientes foram avaliados quanto à recorrência, efeitos colaterais e complicações com 10, 21 dias, 2 e 6 meses de pós-operatório. Dados demográficos, doenças sistêmicas e histórico oftalmológico foram coletados de todos os pacientes e esses dados foram analisados por meio de estatística descritiva envolvendo o percentual absoluto e relativo de distribuição de frequência. O teste de Goodman para contrastes entre populações multinomiais foi utilizado para o estudo da associação entre a ciclosporina A e a recorrência Resultados: A maioria dos pacientes tinha entre 30 e 60 anos e 67,1% eram mulheres. Confirmamos uma maior recorrência em pacientes com exposição ocupacional ao sol. A ciclosporina A tópica utilizada 10 dias antes e 10 dias após a remoção do pterígio não reduziu significativamente a sua recorrência Conclusão: A ciclosporina A tópica a 0,05% quando utilizada por 10 dias no pré e 10 dias no pós-operatório, não previne ou reduz a recidiva do pterígio primário significativamente.
Asunto(s)
Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Adulto Joven , Pterigion/prevención & control , Ciclosporina/administración & dosificación , Conjuntiva/anomalías , Inmunosupresores/administración & dosificación , Soluciones Oftálmicas/administración & dosificación , Cuidados Posoperatorios , Recurrencia , Colgajos Quirúrgicos , Cuidados Preoperatorios , Pterigion/cirugía , Pterigion/tratamiento farmacológico , Estudios Prospectivos , Terapia Combinada , Conjuntiva/cirugía , Enfermedades de la Córnea/tratamiento farmacológico , Fluorouracilo/uso terapéuticoRESUMEN
PURPOSE: To evaluate the proliferation rate of recurrent pterygium and normal Tenon's capsule fibroblasts after exposure to triamcinolone. METHODS: Explants of recurrent pterygia and normal Tenon's capsule of the same carrier were cultured. The fibroblasts were cultured and subcultured until the third passage and subsequently exposed to triamcinolone. The cell proliferation rate was evaluated 3, 6, 12 and 18 days after the exposure. RESULTS: Fibroblasts exposed to triamcinolone had significantly lower proliferation rate (P <0.05) compared to those not exposed, when the evaluation was performed 3, 6 and 12 days later. In the 18th day after exposure, there was a return in the proliferation rate, with statistical significance, only in the pterygium fibroblasts. CONCLUSION: Both the fibroblasts from normal Tenon's capsule as from recurrent pterygia showed significantly lower proliferation rate after exposure to triamcinolone. After 18 days from the exposition, pterygium fibroblasts recovered the proliferation. The results suggested the triamcinolone might be useful as adjuvant in pterygium treatment.