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.

Anti-inflammatory effects of α-humulene and ß-caryophyllene on pterygium fibroblasts.

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.

Expression of inflammatory cytokines in mesenchymal stem cells derived from proximal humerus fractures.

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.

In vitro effect of triamcinolone and platelet-rich plasma on cytokine levels of elbow lateral epicondylitis-derived cells.

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.

Topical cyclosporine A 0. 05% before and after surgery to prevent pterygium recurrence / Ciclosporina A 0, 05% antes e após a cirurgia do pterígio para a prevenção da recorrência

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.

Topical cyclosporine A 0.05% before and after surgery to prevent pterygium recurrence.

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.

Eyelid fissure dimensions in Japanese and in Brazilians of European descent over 50 years of age / Medidas das fendas palpebrais em japoneses e brasileiros de descendência européia após os 50 anos

ABSTRACT 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 Bra­­­zilian 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.

RESUMO Objetivo: Investigar as diferenças no posicionamento da fenda palpebral em ja­po­neses e brasileiros de descendência européia maiores de 50 anos. Métodos: Estudo transversal avaliando-se 50 japoneses e 50 brasileiros de descendência européia maiores que 50 anos de idade, avaliando-se o efeito do envelhe­cimento sobre a posição palpebral. As fotografias digitais foram realizadas em posição primária do olhar e transferidas para um computador para serem processadas pelo software de imagem Scion, avaliando-se as medidas da altura e comprimento da fenda palpebral e distância margem-reflexo (MRD). Os dados foram analisados pela estatística segundo sexo, idade e raça. Resultados: O comprimento da fenda palpebral em japoneses foi maior do que o dos brasileiros, enquanto que a altura da fenda palpebral e MRD foram maiores nos brasileiros (p<0,01). O comprimento, altura e MRD foram associados à idade li­nearmente apenas em japoneses (p<0,05) e esta associação foi observada apenas quanto ao comprimento no grupo dos brasileiros (p<0,05). Não houve diferença quanto à lateralidade em ambos os grupos (p>0,05). Conclusão: Existem mudanças distintas no processo de envelhecimento palpebral de acordo com a etnia, que parecem ser mais intensas em pálpebras de japoneses.

Adhesion study of cultured human lens capsule cells on hydrophilic intraocular lenses coated with polyethylene glycol.

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.

Expression of vascular endothelial growth factor (VEGF) in macrophages, fibroblasts, and endothelial cells in pterygium treated with 5-Fluorouracil.

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.

Exposição de fibroblastos de pterígios recidivados e da cápsula de Tenon normal à triancinolona / Exposure of recurrent pterygium and normal Tenon's capsule fibroblasts to triamcinolone

OBJETIVO: Avaliar a taxa de proliferação de fibroblastos provenientes de pterígios recidivados e da cápsula de Tenon normal, quando expostos in vitro à triancinolona. MÉTODOS: Foram realizadas culturas primárias de explantes de pterígios recidivados e da cápsula de Tenon normal do mesmo portador do pterígio. Os fibroblastos foram cultivados e subcultivados até a terceira passagem e posteriormente expostos à triancinolona, com avaliação da taxa de proliferação celular após três, seis, 12 e 18 dias. RESULTADOS: Os fibroblastos expostos à triancinolona apresentaram taxa de proliferação significativamente menor (P<0,05) se comparados aos não expostos quando a avaliação foi feita após três, seis e 12 dias. Na avaliação do 18º dia houve retomada da taxa de proliferação, com significância estatística, somente nos fibroblastos provenientes de pterígios. CONCLUSÃO: Tanto os fibroblastos provenientes da cápsula de Tenon normal, quanto os de pterígios recidivados, apresentaram taxa de proliferação significativamente menor após a exposição à triancinolona. Os fibroblastos de pterígio retomaram a proliferação após 18 dias da exposição. Os resultados apontaram que a triancinolona pode ser útil como adjuvante no tratamento do pterígio.

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 pterigyum 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.

[Safety and efficacy of intraoperative 5-fluorouracil infiltration in pterygium treatment]. / Segurança e efetividade no tratamento do pterígio usando infiltração de 5-fluoruracila no intraoperatório.

PURPOSE: To investigate the safety and efficacy of intraoperative infiltration of 5-fluorouracil (5-FU) as an adjuvant drug in pterygium treatment. METHODS: Of 125 consecutive patients, 125 eyes with primary and recurrent pterygium underwent pterygium excision with intraoperative 5-FU (25 mg/mL) infiltration. The superior and inferior conjunctiva was approximated to cover the scleral bed and 0.2 mL 5-FU was injected at the end of the surgical procedure. The gender, occupation, pterygium characteristics and the follow-up at 7, 21, 60 e 180 days after surgery were evaluated and the data were statistically analyzed. RESULTS: With follow-up of 180 days the patients had no serious complications observed during or after surgery. The relapse rate was 35.8% and occurred in primary (35.7%) and recurrent (36.4%) lesions with no statistical difference. CONCLUSIONS: This study suggests that intraoperative infiltration of 5-FU is safe. However the high recurrence rate indicated that other studies would be necessary to show the concentration/dose to better prevent it.

Segurança e efetividade no tratamento do pterígio usando infiltração de 5-fluoruracila no intraoperatório / Safety and efficacy of intraoperative 5-fluorouracil infiltration in pterygium treatment

Objetivo: Avaliar a segurança e a efetividade do uso do 5-fluoruracila(5-FU) como tratamento adjuvante do pterígio, aplicado sob a forma de infiltração subconjuntival, no período intraoperatório. Métodos: Foram avaliados prospectivamente 125 indivíduos (125 olhos) portadores depterígio. Os indivíduos foram operados segundo a técnica de retalho de deslizamento e receberam, ao final do procedimento, injeção subconjuntivalde 0,2 mL de 5-FU (25 mg/mL). Foram anotados os dados do paciente como idade, sexo, profissão, características da lesão (primário ou recidivado, tamanho, carnoso ou involutivo) e feito seguimento pósoperatório, aos 7, 21, 60 e 180 dias. Os dados foram submetidos àavaliação estatística. Resultados: Não foram observados casos de complicaçãodecorrente do uso do 5-FU em injeção no intraoperatório do pterígio. A taxa de recidiva geral observada aos 180 dias de pós-operatório foi de 35,8%, sendo de 35,7% para os pterígios primários e de 36,4% para os recidivados. Conclusão: A aplicação do 5-FU no período intraoperatório sob a forma de infiltração subconjuntival é segura. Entretanto, ainda resulta em altas taxas de recidiva e novos estudos devem serrealizados a fim de conhecer a concentração/dose ideal que permitirámenores chances de recidiva da lesão.

Purpose: To investigate the safety and efficacy of intraoperative infiltration of 5-fluorouracil (5-FU) as an adjuvant drug in pterygium treatment. Methods: Of 125 consecutive patients, 125 eyes with primary and recurrent pterygium underwent pterygium excision with intraoperative 5-FU (25 mg/mL) infiltration. The superior and inferior conjunctiva was approximated to cover the scleral bed and 0.2 mL 5-FU wasinjected at the end of the surgical procedure. The gender, occupation, pterygium characteristics and the follow-up at 7, 21, 60 e 180 days after surgery were evaluated and the data were statistically analyzed. Results: With follow-up of 180 days the patients had no serious complications observed duringor after surgery. The relapse rate was 35.8% and occurred in primary (35.7%) and recurrent (36.4%) lesions with no statistical difference. Conclusions: This study suggests that intraoperative infiltration of 5-FU is safe. However the high recurrence rate indicated that other studies would be necessary to show the concentration/dose to better prevent it.

[Eyeblink rate before and after pterygium exeresis]. / Ritmo de piscar em portadores de pterígio antes e após a exérese.

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.

Ritmo de piscar em portadores de pterígio antes e após a exérese / Eyeblink rate before and after pterygium exeresis

OBJETIVO: Avaliar o ritmo de piscar de portadores de pterígio antes e depois da exérese. MÉTODOS: Foram avaliados os movimentos palpebrais de 41 pacientes antes e 60 dias depois da remoção cirúrgica da lesão. Os movimentos palpebrais foram capturados durante 1 minuto, usando filmadora Sony Digital 8 DCR - TRV110, sob iluminação artificial, com o indivíduo em posição primária do olhar, tendo como ponto de fixação a própria filmadora. As imagens obtidas foram processadas por computador, quantificando-se o total de movimentos de piscar, o número de piscar completo e incompleto, e as respectivas durações. Os dados foram submetidos à análise estatística. RESULTADOS: Antes da cirurgia 36,36 por cento dos pacientes queixavam-se de sensação de corpo estranho e após a cirurgia, 61,02 por cento estavam assintomáticos. A avaliação do ritmo de piscar revelou que a freqüência do piscar incompleto aumentou no pós-operatório. A duração do piscar não se alterou antes e após a cirurgia. CONCLUSÃO: A exérese do pterígio leva à diminuição dos sintomas irritativos. Após a remoção da lesão, houve discreto aumento da freqüência de piscar incompleto. Há a possibilidade da presença do pterígio não estar relacionada com alterações do filme lacrimal, considerando que a alteração do ritmo de piscar foi discreta. No entanto, outros estudos deverão ser realizados para afirmar ou contestar esta hipótese.

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 ilumination. 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 percent of the patients with pterygium complained of foreign body and after that, 61.02 percent 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.

[Exposure of normal Tenon's capsule fibroblasts from pterygium to 5-fluorouracil and mitomycin C]. / Exposição de fibroblastos da cápsula de Tenon normal de portadores de pterígio ao 5-fluorouracil e à mitomicina C.

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.

Exposição de fibroblastos provenientes de pterígios recidivados e da cápsula de Tenon normal ao tacrolimus (FK-506) / Fibroblasts from recurrent pterygium and normal Tenon's capsule exposed to tacrolimus (FK-506)

OBJETIVO: Avaliar a taxa de proliferação de fibroblastos provenientes de pterígios recidivados e da cápsula de Tenon normal, quando expostos in vitro ao tacrolimus (FK 506). MÉTODOS: Foi realizado estudo prospectivo, controlado, avaliando-se 8 amostras de explantes de cápsula de Tenon de pterígios recidivados e 6 de cápsula de Tenon normal, obtidas na Faculdade de Medicina de Botucatu - UNESP. A cápsula de Tenon normal foi colhida da região temporal inferior, do mesmo portador de pterígio. As amostras foram cultivadas em meio específico e posteriormente expostas ao tacrolimus 1M (FK 506), em única exposição, com avaliação da taxa de proliferação celular 1, 5, 12 e 19 dias após a exposição. RESULTADOS: Avaliando-se a proliferação dos fibroblastos provenientes de cápsula de Tenon de pterígios recidivados e da cápsula de Tenon normal, os fibroblastos expostos ao tacrolimus tiveram taxa de proliferação significativamente menor (p<0,05) do que quando não houve exposição a droga, quando a avaliação foi feita 1 dia após a exposição. Quando a avaliação foi feita 19 dias após a exposição, a taxa de proliferação foi maior nos grupos expostos a droga. CONCLUSÃO: Os fibroblastos provenientes da cápsula de Tenon de pterígios recidivados apresentaram taxa de proliferação significativamente menor um dia após a exposição ao tacrolimus. Novos estudos devem ser realizados para definir dose e tempo de exposição dos fibroblastos a droga, com o intuito de definir se o tacrolimus pode ser útil como tratamento coadjuvante do pterígio.

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.

Exposição de fibroblastos da cápsula de Tenon normal de portadores de pterígio ao 5-fluorouracil e à mitomicina C / Exposure of normal Tenon's capsule fibroblasts from pterygium to 5-fluorouracil and mitomycin C

OBJETIVO: Avaliar a atividade proliferativa dos fibroblastos da cápsula de Tenon normal, proveniente de portadores de pterígios primários e recidivados. MÉTODOS: Foi realizado estudo prospectivo, aleatório, avaliando-se fragmentos da cápsula de Tenon normal, removidos de 20 portadores de pterígios primários e 21, recidivados. A taxa de proliferação foi avaliada em fibroblastos de terceira passagem, quando as culturas foram expostas a agentes antimitóticos: mitomicina C e 5-fluorouracil. Os dados obtidos foram submetidos à análise estatística. RESULTADOS: Dentre os 41 espécimes cultivados, apenas 1 de pterígio primário e 2 de recidivado proliferaram. Quando expostos a mitomicina C e ao 5-fluorouracil não houve diferença estatisticamente significativa quanto a inibição de proliferação celular. CONCLUSÃO: Desta forma, in vitro, ambos os antimitóticos estudados têm a mesma eficácia na inibição da proliferação sobre os fibroblastos de cápsulas de Tenon normal.

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.

Exposição de fibroblastos da cápsula de Tenon de pterígios à ciclosporina 0.05 por cento / Exposure of Tenon's capsule fibroblasts of pterygium to cyclosporin 0.05 percent

OBJETIVO: Avaliar o comportamento dos fibroblastos da cápsula de Tenon de pterígios e normais em cultura, quando expostos a ciclosporina 0,05 por cento. MÉTODOS: Estudo prospectivo, controlado, do qual participaram 20 portadores de pterígio primário. Foram colhidas amostras da cápsula de Tenon normal e proveniente do pterígio, ambas do mesmo indivíduo, colocadas para crescimento em cultura e expostas a ciclosporina 0,05 por cento. As avaliações foram feitas aos 3, 6, 12 e 17 dias após a exposição. RESULTADOS: Das amostras colhidas, 7 foram utilizadas para exposição a ciclosporina - 6 provenientes de pterígios e 1 da Tenon normal. Houve redução significativa na proliferação celular das culturas de fibroblastos expostos a ciclosporina (p<0,05). CONCLUSÃO: A ciclosporina 0,05 por cento é capaz de inibir a proliferação de fibroblastos provenientes da cápsula de Tenon de pterígio e também da Tenon normal. Novos estudos devem ser providenciados para definir o papel da ciclosporina no tratamento do pterígio.

PURPOSE: To evaluate normal Tenon capsule fibroblasts and from pterygia in culture when exposed to cyclosporin 0.05 percent. 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 percent. 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 percent 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.

Análise do cultivo de fibroblastos de pterígios primários e recidivados e da cápsula de Tenon normal / Fibroblasts from primary and recurrent pterygia and Tenon's capsule in cell culture

OBJETIVO: Avaliar a atividade proliferativa dos fibroblastos da cápsula de Tenon, provenientes de explantes de pterígios primários e recidivados e da conjuntiva normal. MÉTODOS: Foi realizado estudo prospectivo, randomizado, avaliando-se 43 pecas cirúrgicas, produto da exérese de 30 pterígios primários e 13 recidivados, além de fragmentos da cápsula de Tenon normal, obtida dos próprios portadores de pterígio. Foram avaliadas a taxa de proliferacão, migracão e confluência, analisadas segundo dados dos portadores como: idade, localizacão da lesão, tipo de lesão (tamanho, involutivo ou carnoso), primário ou recidivado. Os dados obtidos foram submetidos à análise estatística. RESULTADOS: Dentre os 30 pterígios primários cultivados, 70 por cento migraram e proliferaram e 60 por cento chegaram à confluência. A migracão, proliferacão e confluência iniciaram-se mais precocemente nas culturas de fibroblastos provenientes de pterígios em comparacão àquelas provenientes da Tenon normal. Os pterígios recidivados apresentaram migracão mais precoce que os primários. Não houve diferenca estatisticamente significativa quanto ao início da migracão, proliferacão e confluência entre os pterígios carnosos ou involutivos, assim como entre os pterígios de graus I-II e os de graus III-IV. CONCLUSAO: O cultivo celular de fibroblastos de pterígios é mais viável que o de Tenon normal. A migracão, proliferacão e confluência diferem em pterígios primários e recidivados. Pterígios carnosos e involutivos são semelhantes em cultura, assim como não existe diferenca entre os pterígios segundo o tamanho da lesão.