Modulation of corneal transplant rejection using gene therapy shows promise in experimental models but the most appropriate vector for gene transfer is yet to be determined. The overarching aim of the thesis was to evaluate the potential of a lentiviral vector for use in human corneal transplantation. Specific aims were: (i) to assess the ability of an HIV-1-based lentiviral vector to mediate expression of the enhanced yellow fluorescent protein (eYFP), and a model secreted protein interleukin-10 (IL10), in ovine and human corneal endothelium; and (ii) to examine the influence of lentivirus-mediated IL10 expression on the survival of ovine corneal allografts. Four lentiviral vectors expressing eYFP under the control of different promoters, were tested: the simian virus type-40 (SV40) early promoter, the phosphoglycerate kinase (PGK) promoter, the elongation factor-1alpha (EF) promoter, and the cytomegalovirus (CMV) promoter. Two lentiviral vectors expressing IL10 were tested: one containing the SV40 promoter and another containing a steroid-inducible promoter (GRE5). Lentivirus-mediated expression in transduced ovine and human corneal endothelium was assessed by fluorescence microscopy, real-time quantitative RT-PCR and ELISA, following alterations of transduction period duration (2–24 hr) and vector dose, as well as in the presence or absence of polybrene or dexamethasone (GRE5 vector). It was also compared to expression mediated by adenoviral vectors. Orthotopic transplantation of ex vivo transduced donor corneas was performed in outbred sheep. Allografts were reviewed daily for vascularisation and signs of immunological rejection. Lentivirus-mediated eYFP expression was delayed in ovine corneal endothelium compared to human. However, in both species the final transduction rate was greater than 80% and expression was stable for at least 14 d in vitro. Lentivirus-mediated expression in ovine and human corneal endothelium was higher with the viral promoters in comparison to the mammalian promoters. A 24 h transduction of ovine corneal endothelium with the lentiviral vector encoding IL10 resulted in expression levels which were increasing after 15 d of organ culture but logarithmically lower than those achieved by adenovirus. Shortening the lentiviral transduction period to 2 h led to a reduction in expression, but the addition of polybrene (40 micrograms / ml) to the transduction mixture restored expression to levels comparable to those attained after a 24 h transduction period. Lentivirus-mediated IL10 expression was higher and more rapid in human corneal endothelium compared to ovine corneas. Dexamethasone-responsive transgene expression was observed in both ovine and human corneal endothelium using the lentiviral vector containing the GRE5 promoter. Lentivirus-mediated expression in ovine corneal endothelium was stable for 28 d in vivo. A modest prolongation of ovine corneal allograft survival (median of 7 d) was achieved by transduction of donor corneas for 2–3 h with the lentivirus expressing IL10. Attempts to increase the expression of IL10 by the addition of polybrene (40 micrograms / ml) to the transduction mixture, resulted in a toxic effect on corneal allografts which abrogated the beneficial effect of IL10. The lentiviral vector shows potential for the stable expression of therapeutic transgenes in human corneal transplantation. However, the mechanisms underlying the species-specific differences in HIV-1-mediated transgene expression will need to be elucidated and overcome if the ovine preclinical model is to provide justification for a clinical trial.

Orientador: Silvana Artioli Schellini / Resumo: O objetivo do estudo foi avaliar a influência do diabetes experimental sobre a ultra-estrutura do endotélio corneal de ratos. O estudo foi prospectivo, utilizando 20 ratos da raça Wistar, com 3 meses de idade, divididos em: grupo controle (GC), contendo 10 ratos, e grupo diabético (GD), contendo 10 ratos. A indução do diabetes foi feita com injeção de Aloxana endovenosa 42 mg/kg de peso (M0), após o que os animais foram observados por 15 dias para confirmar a presença de diabetes grave (M1). Um mês após M1 (M2) e 12 meses após M1 (M3) os animais foram sacrificados, sendo removidos e preparados os olhos para avaliação à microscopia eletrônica de transmissão. Os animais do GD mostraram importante redução de peso, aumento da injestão hídrica e aumento da diurese em relação aos ratos do GC. Na avaliação morfológica observou-se nos animais do GC corpos densos e figuras de Mielina no M3. Os ratos do GD apresentaram as mesmas alterações encontradas no GC em M3, em maior intensidade, com alterações nucleares e citoplasmáticas, como mitocôndrias bastante alteradas na forma e tamanho, rarefação do citoplasma e aumento de vesículas. Os ratos do GD em M3 apresentaram mais alterações que os do GDM2. Conclui-se que o diabetes experimental causa dano ultra-estrutural no endotélio corneal de ratos e que as alterações são evolutivas. / Abstract: The objective of study was to make na assessment of experimental diabetes influence on of rats corneal endothelium ultra-structure. The study was prospective, using 20 Wistar 3-month-old rats, divided (by draw) into groups: control group (GC), with 10 rats, and diabetic group (GD), with 10 rats. The diabetes induction was made by means of intravenous injection of Aloxan 42 mg/Kg weigth. After the diabetes induction (M1), the animals had been observed for 15 days, and then, 1 month after M1 (M2) and 12 months after M1 (M3) to confirm the diagnosis of severe diabetes. At experimental moments M2 and M3, the animals eyes enucleation and preparation were carried out for assessment trough transmission eletronic microscopy. GD animals had shown significant reduction of weigth, increasing of hydric and nourishing injection and increasing of diuresis in relation to GC rats. In the morphological assessment, dense bodies and Myelin figures were observed in M3 GC animals. GC rats had presented the same alterations found in GC animals in M3, in major intensity, beyond mitochondrias rather modified in their form and size, cytoplasm rarefaction, vesicles increasing and nuclear alterations. It is concluded that experimental diabets causes ultra-structural of rats corneal endothelium. / Mestre

Made available in DSpace on 2014-06-11T19:23:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2002Bitstream added on 2014-06-13T19:09:41Z : No. of bitstreams: 1 martins_lc_me_botfm.pdf: 1919468 bytes, checksum: d4a42d395d3517a07ebafa18c26a4e02 (MD5) / O objetivo do estudo foi avaliar a influência do diabetes experimental sobre a ultra-estrutura do endotélio corneal de ratos. O estudo foi prospectivo, utilizando 20 ratos da raça Wistar, com 3 meses de idade, divididos em: grupo controle (GC), contendo 10 ratos, e grupo diabético (GD), contendo 10 ratos. A indução do diabetes foi feita com injeção de Aloxana endovenosa 42 mg/kg de peso (M0), após o que os animais foram observados por 15 dias para confirmar a presença de diabetes grave (M1). Um mês após M1 (M2) e 12 meses após M1 (M3) os animais foram sacrificados, sendo removidos e preparados os olhos para avaliação à microscopia eletrônica de transmissão. Os animais do GD mostraram importante redução de peso, aumento da injestão hídrica e aumento da diurese em relação aos ratos do GC. Na avaliação morfológica observou-se nos animais do GC corpos densos e figuras de Mielina no M3. Os ratos do GD apresentaram as mesmas alterações encontradas no GC em M3, em maior intensidade, com alterações nucleares e citoplasmáticas, como mitocôndrias bastante alteradas na forma e tamanho, rarefação do citoplasma e aumento de vesículas. Os ratos do GD em M3 apresentaram mais alterações que os do GDM2. Conclui-se que o diabetes experimental causa dano ultra-estrutural no endotélio corneal de ratos e que as alterações são evolutivas. / The objective of study was to make na assessment of experimental diabetes influence on of rats corneal endothelium ultra-structure. The study was prospective, using 20 Wistar 3-month-old rats, divided (by draw) into groups: control group (GC), with 10 rats, and diabetic group (GD), with 10 rats. The diabetes induction was made by means of intravenous injection of Aloxan 42 mg/Kg weigth. After the diabetes induction (M1), the animals had been observed for 15 days, and then, 1 month after M1 (M2) and 12 months after M1 (M3) to confirm the diagnosis of severe diabetes. At experimental moments M2 and M3, the animals eyes enucleation and preparation were carried out for assessment trough transmission eletronic microscopy. GD animals had shown significant reduction of weigth, increasing of hydric and nourishing injection and increasing of diuresis in relation to GC rats. In the morphological assessment, dense bodies and Myelin figures were observed in M3 GC animals. GC rats had presented the same alterations found in GC animals in M3, in major intensity, beyond mitochondrias rather modified in their form and size, cytoplasm rarefaction, vesicles increasing and nuclear alterations. It is concluded that experimental diabets causes ultra-structural of rats corneal endothelium.

No description available.

In a 1982 paper, Gould and Vrba argue that a conflation of the two components of adaptation of a feature, historical development of the feature and present utility, has caused evolutionists to overlook a missing term in the science of form, which they call 'exaptation'. In the present project, I show that evolutionary biology still contains a confusion in the use of 'adaptation' due to an inappropriate perception of the interaction between the two components of adaptation. Because of the confusion, evolutionists have missed another term in the science of form. Evolutionary theory, specifically the treatment of adaptation, would profit from the introduction of a term referring to features that have a selective history which causes them to appear overly well adapted to their present function. I suggest we refer to these features as hyperadaptations, since they appear to be hyperbolized adaptations. By introducing hyperadaptation into the conceptual framework of adaptation, we can sharpen our understanding of related concepts (adaptation to function, exaptation, maladaptation, etc.) and remove or reduce some confusion regarding the interplay between analysis of historical pathways and ascriptions of (current) function in the diagnosis of adaptation. Furthermore, the improved framework should allow evolutionists to more adequately explain biological phenomena. / Master of Arts

A manutenção da morfologia normal do endotélio da córnea é um importante indicador da integridade funcional. A reparação endotelial frente a um trauma, em coelhos, ocorre por migração, hipertrofia e mitose celular. Objetivou-se comparar a morfologia das células do endotélio, da região periférica perincisional à região central, da córnea de coelhos (Oryctolagus cuniculus) em diferentes períodos pós-operatórios. Foram designados três grupos, com 5 animais cada, para avaliação pós-operatória, sendo G1 (7 dias); G2 (15 dias) e G3 (45 dias). Trinta bulbos dos olhos de coelhos, da raça Nova Zelândia, foram submetidos à incisão de córnea clara uniplanar com 3,2 mm. Ao fim dos períodos determinados, procedeu-se a avaliação da morfologia endotelial valendose da microscopia eletrônica de varredura. Realizaram-se seis eletromicrografias de varredura, de cada região da córnea, com aumento de 1000 vezes. Para análise do percentual do número de lados celular, foram analisadas 100 células endoteliais. Na região periférica perincisional, avaliada ao 7° dia de pós-operatório, foram encontradas células com 6 lados (47,8%), 5 lados (31,3%), 7 lados (13,9%), 3 lados (0,1%), 4 lados (4,9%), 8 lados (1,8%) e 9 lados (0,2%). Na avaliação ao 15° dia de pós-operatório, observaram-se células com 6 lados (45,6%), 5 lados (32,6%), 7 lados (17,4%), 4 lados (1,7%) e 8 lados (2,7%). No 45° dia de pós-operatório, verificou-se a presença de células com 6 lados (57%), 5 lados (24%), 7 lados (17,2%), 4 lados (0,1%), 8 lados (1,6%) e 9 lados (0,1%). Na área central, ao 7° dia de pós-operatório, detectaram-se células com 6 lados (75,6%), 5 lados (13,3%), 7 lados (10,8%) e 8 lados (0,3%). Na avaliação, ao 15° dia de pós-operatório, foi possível observar células com 6 lados (78,9%), 5 lados (11,5%) e 7 lados (9,6%). No 45° dia de pós-operatório identificaram-se células com 6 lados (74,8%), 5 lados (13,6%) e 7 lados (11,6%). Os resultados demonstraram que na região periférica perincisional ocorreu diminuição das células com seis lados e aumento do número de células com cinco e sete lados. Na região central manteve-se o padrão regular de hexagonalidade das células endoteliais nos diferentes períodos pós-operatórios. Conclui-se que houve alteração na morfologia das células endoteliais, da região periférica perincisional comparada à região central, da córnea de coelhos nos diferentes períodos pós-operatórios. / The maintenance of the normal corneal endothelium morphology is an important indicator of its functional integrity. In rabbits, endothelial repair in the event of traumas is made through cell migration, hypertrophy and mitosis. The purpose of this study was to compare the morphology of endothelial cells of the perincisional area with the central area of the cornea of rabbits (Oryctolagus cuniculus), in different post-operative periods. Three groups containing 5 animals each were designed for post-operative evaluation: G1 (7 days); G2 (15 days) and G3 (45 days). The clear cornea of thirty New Zealand rabbits was subjected to a single-planed incision of 3.2 mm. At the end of the established periods, a morphological evaluation of the endothelium was carried out using scanning electron microscopy. Six scanning electron micrographs of each corneal area were performed using a magnification of 1000 x. One hundred endothelial cells were analyzed to obtain the cell side count percentage. In the perincisional peripheral area, which was evaluated at the 7 post-operative day, 6-sided (47.8%), 5-sided (31.3%) and 7-sided (13.9%) cells were found, in addition to, 3-sided cells (0.1%), 4-sided cells (4.9%), 8-sided cells (1.8%) and 9-sided cells (0.2%). In the evaluation made on the 15th post-operative day, 6-sided (45.6%), 5-sided (32.6%) and 7-sided (17.4%) cells were observed, as well as 4-sided (1.7%) and 8-sided cells (2.7%). On the 45th postoperative day, the presence of 6-sided (57%), 5-sided (24%), 7-sided (17.2%), 4-sided (0,1%), 8-sided (1.6%) and 9-sided cells (0.1%) was verified. On the 7th post-operative day, 6-sided (75.6%), 5-sided (13.3%), 7-sided (10.8%) and 8-sided cells (0.3%) were observed in the central area .Upon evaluation made on the 15th post-operative day, it was possible to observe 6-sided (78.9%), 5-sided (11.5%) and 7-sided (9.6%) cells. Results have shown that there was a reduction of six-sided cells and an increase in the number of five and seven-sided cells in the perincisional peripheral area. The regular hexagonal standard of the endothelial cells was maintained in the central area in different post-operative periods. In comparison to the central area, there was a morphological alteration of the endothelial cells of the peripheral perincisional area in different post-operative periods of the cornea of rabbits.

A manutenção da morfologia normal do endotélio da córnea é um importante indicador da integridade funcional. A reparação endotelial frente a um trauma, em coelhos, ocorre por migração, hipertrofia e mitose celular. Objetivou-se comparar a morfologia das células do endotélio, da região periférica perincisional à região central, da córnea de coelhos (Oryctolagus cuniculus) em diferentes períodos pós-operatórios. Foram designados três grupos, com 5 animais cada, para avaliação pós-operatória, sendo G1 (7 dias); G2 (15 dias) e G3 (45 dias). Trinta bulbos dos olhos de coelhos, da raça Nova Zelândia, foram submetidos à incisão de córnea clara uniplanar com 3,2 mm. Ao fim dos períodos determinados, procedeu-se a avaliação da morfologia endotelial valendose da microscopia eletrônica de varredura. Realizaram-se seis eletromicrografias de varredura, de cada região da córnea, com aumento de 1000 vezes. Para análise do percentual do número de lados celular, foram analisadas 100 células endoteliais. Na região periférica perincisional, avaliada ao 7° dia de pós-operatório, foram encontradas células com 6 lados (47,8%), 5 lados (31,3%), 7 lados (13,9%), 3 lados (0,1%), 4 lados (4,9%), 8 lados (1,8%) e 9 lados (0,2%). Na avaliação ao 15° dia de pós-operatório, observaram-se células com 6 lados (45,6%), 5 lados (32,6%), 7 lados (17,4%), 4 lados (1,7%) e 8 lados (2,7%). No 45° dia de pós-operatório, verificou-se a presença de células com 6 lados (57%), 5 lados (24%), 7 lados (17,2%), 4 lados (0,1%), 8 lados (1,6%) e 9 lados (0,1%). Na área central, ao 7° dia de pós-operatório, detectaram-se células com 6 lados (75,6%), 5 lados (13,3%), 7 lados (10,8%) e 8 lados (0,3%). Na avaliação, ao 15° dia de pós-operatório, foi possível observar células com 6 lados (78,9%), 5 lados (11,5%) e 7 lados (9,6%). No 45° dia de pós-operatório identificaram-se células com 6 lados (74,8%), 5 lados (13,6%) e 7 lados (11,6%). Os resultados demonstraram que na região periférica perincisional ocorreu diminuição das células com seis lados e aumento do número de células com cinco e sete lados. Na região central manteve-se o padrão regular de hexagonalidade das células endoteliais nos diferentes períodos pós-operatórios. Conclui-se que houve alteração na morfologia das células endoteliais, da região periférica perincisional comparada à região central, da córnea de coelhos nos diferentes períodos pós-operatórios. / The maintenance of the normal corneal endothelium morphology is an important indicator of its functional integrity. In rabbits, endothelial repair in the event of traumas is made through cell migration, hypertrophy and mitosis. The purpose of this study was to compare the morphology of endothelial cells of the perincisional area with the central area of the cornea of rabbits (Oryctolagus cuniculus), in different post-operative periods. Three groups containing 5 animals each were designed for post-operative evaluation: G1 (7 days); G2 (15 days) and G3 (45 days). The clear cornea of thirty New Zealand rabbits was subjected to a single-planed incision of 3.2 mm. At the end of the established periods, a morphological evaluation of the endothelium was carried out using scanning electron microscopy. Six scanning electron micrographs of each corneal area were performed using a magnification of 1000 x. One hundred endothelial cells were analyzed to obtain the cell side count percentage. In the perincisional peripheral area, which was evaluated at the 7 post-operative day, 6-sided (47.8%), 5-sided (31.3%) and 7-sided (13.9%) cells were found, in addition to, 3-sided cells (0.1%), 4-sided cells (4.9%), 8-sided cells (1.8%) and 9-sided cells (0.2%). In the evaluation made on the 15th post-operative day, 6-sided (45.6%), 5-sided (32.6%) and 7-sided (17.4%) cells were observed, as well as 4-sided (1.7%) and 8-sided cells (2.7%). On the 45th postoperative day, the presence of 6-sided (57%), 5-sided (24%), 7-sided (17.2%), 4-sided (0,1%), 8-sided (1.6%) and 9-sided cells (0.1%) was verified. On the 7th post-operative day, 6-sided (75.6%), 5-sided (13.3%), 7-sided (10.8%) and 8-sided cells (0.3%) were observed in the central area .Upon evaluation made on the 15th post-operative day, it was possible to observe 6-sided (78.9%), 5-sided (11.5%) and 7-sided (9.6%) cells. Results have shown that there was a reduction of six-sided cells and an increase in the number of five and seven-sided cells in the perincisional peripheral area. The regular hexagonal standard of the endothelial cells was maintained in the central area in different post-operative periods. In comparison to the central area, there was a morphological alteration of the endothelial cells of the peripheral perincisional area in different post-operative periods of the cornea of rabbits.

A manutenção da morfologia normal do endotélio da córnea é um importante indicador da integridade funcional. A reparação endotelial frente a um trauma, em coelhos, ocorre por migração, hipertrofia e mitose celular. Objetivou-se comparar a morfologia das células do endotélio, da região periférica perincisional à região central, da córnea de coelhos (Oryctolagus cuniculus) em diferentes períodos pós-operatórios. Foram designados três grupos, com 5 animais cada, para avaliação pós-operatória, sendo G1 (7 dias); G2 (15 dias) e G3 (45 dias). Trinta bulbos dos olhos de coelhos, da raça Nova Zelândia, foram submetidos à incisão de córnea clara uniplanar com 3,2 mm. Ao fim dos períodos determinados, procedeu-se a avaliação da morfologia endotelial valendose da microscopia eletrônica de varredura. Realizaram-se seis eletromicrografias de varredura, de cada região da córnea, com aumento de 1000 vezes. Para análise do percentual do número de lados celular, foram analisadas 100 células endoteliais. Na região periférica perincisional, avaliada ao 7° dia de pós-operatório, foram encontradas células com 6 lados (47,8%), 5 lados (31,3%), 7 lados (13,9%), 3 lados (0,1%), 4 lados (4,9%), 8 lados (1,8%) e 9 lados (0,2%). Na avaliação ao 15° dia de pós-operatório, observaram-se células com 6 lados (45,6%), 5 lados (32,6%), 7 lados (17,4%), 4 lados (1,7%) e 8 lados (2,7%). No 45° dia de pós-operatório, verificou-se a presença de células com 6 lados (57%), 5 lados (24%), 7 lados (17,2%), 4 lados (0,1%), 8 lados (1,6%) e 9 lados (0,1%). Na área central, ao 7° dia de pós-operatório, detectaram-se células com 6 lados (75,6%), 5 lados (13,3%), 7 lados (10,8%) e 8 lados (0,3%). Na avaliação, ao 15° dia de pós-operatório, foi possível observar células com 6 lados (78,9%), 5 lados (11,5%) e 7 lados (9,6%). No 45° dia de pós-operatório identificaram-se células com 6 lados (74,8%), 5 lados (13,6%) e 7 lados (11,6%). Os resultados demonstraram que na região periférica perincisional ocorreu diminuição das células com seis lados e aumento do número de células com cinco e sete lados. Na região central manteve-se o padrão regular de hexagonalidade das células endoteliais nos diferentes períodos pós-operatórios. Conclui-se que houve alteração na morfologia das células endoteliais, da região periférica perincisional comparada à região central, da córnea de coelhos nos diferentes períodos pós-operatórios. / The maintenance of the normal corneal endothelium morphology is an important indicator of its functional integrity. In rabbits, endothelial repair in the event of traumas is made through cell migration, hypertrophy and mitosis. The purpose of this study was to compare the morphology of endothelial cells of the perincisional area with the central area of the cornea of rabbits (Oryctolagus cuniculus), in different post-operative periods. Three groups containing 5 animals each were designed for post-operative evaluation: G1 (7 days); G2 (15 days) and G3 (45 days). The clear cornea of thirty New Zealand rabbits was subjected to a single-planed incision of 3.2 mm. At the end of the established periods, a morphological evaluation of the endothelium was carried out using scanning electron microscopy. Six scanning electron micrographs of each corneal area were performed using a magnification of 1000 x. One hundred endothelial cells were analyzed to obtain the cell side count percentage. In the perincisional peripheral area, which was evaluated at the 7 post-operative day, 6-sided (47.8%), 5-sided (31.3%) and 7-sided (13.9%) cells were found, in addition to, 3-sided cells (0.1%), 4-sided cells (4.9%), 8-sided cells (1.8%) and 9-sided cells (0.2%). In the evaluation made on the 15th post-operative day, 6-sided (45.6%), 5-sided (32.6%) and 7-sided (17.4%) cells were observed, as well as 4-sided (1.7%) and 8-sided cells (2.7%). On the 45th postoperative day, the presence of 6-sided (57%), 5-sided (24%), 7-sided (17.2%), 4-sided (0,1%), 8-sided (1.6%) and 9-sided cells (0.1%) was verified. On the 7th post-operative day, 6-sided (75.6%), 5-sided (13.3%), 7-sided (10.8%) and 8-sided cells (0.3%) were observed in the central area .Upon evaluation made on the 15th post-operative day, it was possible to observe 6-sided (78.9%), 5-sided (11.5%) and 7-sided (9.6%) cells. Results have shown that there was a reduction of six-sided cells and an increase in the number of five and seven-sided cells in the perincisional peripheral area. The regular hexagonal standard of the endothelial cells was maintained in the central area in different post-operative periods. In comparison to the central area, there was a morphological alteration of the endothelial cells of the peripheral perincisional area in different post-operative periods of the cornea of rabbits.

Os corantes vitais têm sido utilizados em diversas áreas da oftalmologia para melhorar a visibilização de diferentes tecidos. Com o objetivo de avaliar os efeitos imediatos do corante Azul brilhante a 0,05% no endotélio da córnea de equinos, foram utilizadas 38 córneas de 19 cavalos, machos ou fêmeas, de diferentes idades e provenientes de abatedouro comercial licenciado. As córneas foram aleatoriamente divididas em dois grupos. Grupo1: o endotélio da córnea foi exposto a solução salina balanceada por 60 segundos. Grupo 2: o endotélio da córnea foi exposto ao corante Azul brilhante 0,05% (Opht-Blue, Ophthalmos, São Paulo, Brazil) por 60 segundos seguido por lavagem com solução salina balanceada (BSS, Ophthalmos, São Paulo, Brazil). As córneas então foram excisadas com trépano de 8mm e preparadas para avaliação com microscopia óptica (24 córneas) e microscopia eletrônica de varredura (14 córneas). As áreas de dano endotelial foram mensuradas. Devido ao aparecimento de resíduos não normais na utilização de comparação de médias via ANOVA, optou-se pelo modelo linear generalizado e, com 5% de significância, o teste Qui-quadrado demonstrou que o fator tratamento não difere do controle. Conclui-se que o corante Azul brilhante em concentração de 0,05% não provocou danos ao endotélio da córnea de equinos. / Vital dyes have been used in many areas of ophthalmology for improving visualization of different tissues. With the objective of evaluating the immediate effects of 0,05% brilliant blue on corneal endothelium of horses, 38 corneas of 19 horses, both male and female of different ages obtained from a licensed Brazilian commercial slaughterhouse were studied. Corneas were randomly divided into two groups. In group 1, corneal endothelium was perfused with BSS for 60 seconds. In group 2, corneal endothelium was perfused with 0.3 mL of brilliant blue 0.05% (Opht-Blue, Ophthalmos, São Paulo, Brazil) for 60 seconds followed by rinsing with a balanced salt solution (BSS, Ophthalmos, São Paulo, Brazil). The corneas were, subsequently, excised with an 8mm trephine and prepared to analyze posterior endothelial surface using a light microscope (24 corneas) and a scanning electron microscope (14 corneas). Due to non-normal residuals at ANOVA mean comparison, a generalized linear model was utilized at 5% level of significance. Qui-square test stated that treatment and control group were not different statistically. The 0.05 % brilliant blue did not damage equine corneal endothelium.

Os corantes vitais têm sido utilizados em diversas áreas da oftalmologia para melhorar a visibilização de diferentes tecidos. Com o objetivo de avaliar os efeitos imediatos do corante Azul brilhante a 0,05% no endotélio da córnea de equinos, foram utilizadas 38 córneas de 19 cavalos, machos ou fêmeas, de diferentes idades e provenientes de abatedouro comercial licenciado. As córneas foram aleatoriamente divididas em dois grupos. Grupo1: o endotélio da córnea foi exposto a solução salina balanceada por 60 segundos. Grupo 2: o endotélio da córnea foi exposto ao corante Azul brilhante 0,05% (Opht-Blue, Ophthalmos, São Paulo, Brazil) por 60 segundos seguido por lavagem com solução salina balanceada (BSS, Ophthalmos, São Paulo, Brazil). As córneas então foram excisadas com trépano de 8mm e preparadas para avaliação com microscopia óptica (24 córneas) e microscopia eletrônica de varredura (14 córneas). As áreas de dano endotelial foram mensuradas. Devido ao aparecimento de resíduos não normais na utilização de comparação de médias via ANOVA, optou-se pelo modelo linear generalizado e, com 5% de significância, o teste Qui-quadrado demonstrou que o fator tratamento não difere do controle. Conclui-se que o corante Azul brilhante em concentração de 0,05% não provocou danos ao endotélio da córnea de equinos. / Vital dyes have been used in many areas of ophthalmology for improving visualization of different tissues. With the objective of evaluating the immediate effects of 0,05% brilliant blue on corneal endothelium of horses, 38 corneas of 19 horses, both male and female of different ages obtained from a licensed Brazilian commercial slaughterhouse were studied. Corneas were randomly divided into two groups. In group 1, corneal endothelium was perfused with BSS for 60 seconds. In group 2, corneal endothelium was perfused with 0.3 mL of brilliant blue 0.05% (Opht-Blue, Ophthalmos, São Paulo, Brazil) for 60 seconds followed by rinsing with a balanced salt solution (BSS, Ophthalmos, São Paulo, Brazil). The corneas were, subsequently, excised with an 8mm trephine and prepared to analyze posterior endothelial surface using a light microscope (24 corneas) and a scanning electron microscope (14 corneas). Due to non-normal residuals at ANOVA mean comparison, a generalized linear model was utilized at 5% level of significance. Qui-square test stated that treatment and control group were not different statistically. The 0.05 % brilliant blue did not damage equine corneal endothelium.