Investigating the effect of intracameral cefuroxime on oxidative stress and apoptosis in the rat cornea / Investigando o efeito de cefuroxima intracameral sobre o estresse oxidativo e a apoptose na córnea de ratos

ABSTRACT PURPOSE: We examined the effect of intracameral administration of cefuroxime on oxidative stress and endothelial apoptosis in rat corneal tissue. METHODS: In total, 30 rats were divided into 3 groups of 10 rats each (intracameral administration of cefuroxime 0.1 mg/0.01 mL (cefuroxime group); intracameral administration of balanced salt solution 0.01 mL (control group); or absence of intracameral injection (sham group). Corneal endothelial apoptosis was assessed by immunohistochemical analysis using caspase-3 and caspase-8. Total oxidant status, total antioxidant status, oxidative stress index, and paraoxonase and arylesterase levels were examined in corneal endothelial tissue and serum. RESULTS: Paraoxonase levels in the serum were significantly different between the sham and cefuroxime groups (p=0.027). A significant difference was also observed in total oxidant status levels between the cefuroxime and balanced salt solution groups (p=0.023). In addition, there were significant differences in total antioxidant status levels in corneal tissue between the cefuroxime and sham groups (p<0.001) and between the cefuroxime and balanced salt solution groups (p<0.001). Furthermore, significant differences were also observed in oxidative stress index levels between the cefuroxime and balanced salt solution groups (p=0.001) and between the cefuroxime and sham groups (p=0.026). According to the immunohistochemical staining results, a significant association with caspase-3 activity existed between the cefuroxime and balanced salt solution groups (p=0.007), while no significant difference was found with caspase-8 activity (p=0.541). Caspase-3 activity exhibited a significant relationship between the sham and balanced salt solution groups (p=0.018), but no relationship was found with caspase-8 activity (p=0.623). CONCLUSION: Immunohistochemical examination revealed that intracameral cefuroxime increased apoptosis when compared to the sham and balanced salt solution groups. Moreover, intracameral cefuroxime increased oxidative stress in the cornea and simultaneously induced apoptosis.

RESUMO OBJETIVO: Examinamos o efeito da administração intracameral da cefuroxima sobre o estresse oxidativo e a apoptose endotelial no tecido corneano de ratos. MÉTODOS: No total, 30 ratos foram divididos em 3 grupos de 10 ratos cada (administração intracameral de cefuroxima 0,1 mg/0,01 mL (grupo cefuroxima), administração intracameral de solução salina balanceada 0,01 mL (grupo controle) ou ausência de injeção intracameral (grupo sham)). A apoptose endotelial da córnea foi avaliada por análise imuno-histoquimica usando caspase-3 e -8. O status oxidante total, o status antioxidante total, o índice de estresse oxidativo e os níveis de a paraoxonase e arilesterase foram investigados no tecido endotelial da córnea e no soro. RESULTADOS: Os níveis de paraoxonase no soro foram significativamente diferentes entre os grupos sham e cefuroxima (p=0,027). Foi também observada uma diferença significativa nos níveis de estado oxidante total entre os grupos cefuroxima e solução salina balanceada (p=0,023). Além disso, houve diferenças significativas nos níveis de status antioxidante total no tecido da córnea entre os grupos cefuroxima e sham (p<0,001) e entre os grupos cefuroxima e solução salina balanceada (p<0,001). Diferenças significativas também foram observadas nos níveis do índice de estresse oxidativo entre os grupos cefuroxima e solução salina balanceada (p=0,001) e entre os grupos cefuroxima e sham (p=0,026). De acordo com os resultados de coloração imuno-histoquimica, houve associação significativa com a atividade da caspase-3 entre os grupos cefuroxima e solução salina balanceada (p=0,007), enquanto não houve diferença significativa com a atividade da caspase-8 (p=0,541). A atividade da caspase-3 exibiu uma relação significativa entre os grupos sham e solução salina balanceada (p=0,018), mas nenhuma relação foi encontrada com a atividade da caspase-8 (p=0,623). CONCLUSÃO: O exame imuno-histoquímico revelou que a cefuroxima intracameral aumentou a apoptose quando comparada com os grupos sham e solução salina balanceada. Além disso, a cefuroxima intracameral aumentou o estresse oxidativo na córnea e induziu simultaneamente a apoptose.

Effects of atracurium besylate on corneal endothelium of chickens: in vitro study / Avaliação dos efeitos do besilato de atracúrio no endotélio corneano de galinhas: estudo in vitro

The aim of this study was to investigate the acute effects of atracurium besylate on cellular damage in corneal endothelium of chickens. Twenty healthy chicken eyes were assigned to the following groups: Group 1 (G1), experimental group (n=10); and Group 2 (G2), control (n=10). Excised corneoscleral buttons were immediately placed on glass microscopy slides with endothelial region faced up. Corneal endothelium of eyes in G1 were covered with AB (0.2mL, 10mg/mL) for 3 min and then rinsed with balanced salt solution (BSS), while the corneal endothelium of eyes in G2 were covered with BBS for 3 min. Corneas from both groups were stained with alizarin red/trypan blue and visualized by light microscopy. Ten random photographs were taken from each cornea. The area of cellular damage was measured by software in all samples and cell loss of each group was averaged and compared. Endothelial area of denudation and Descemet's membrane exposure were higher in G1 than G2. In conclusion, atracurium besylate induced an acute damage on corneal endothelium of chickens.(AU)'

Objetivou-se avaliar os efeitos agudos do besilato de atracúrio sobre o endotélio corneano de galinhas. Vinte olhos saudáveis de galinhas foram aleatoriamente separados em dois grupos com 10 olhos cada, sendo G1 o grupo controle e G2 o grupo tratamento. Imediatamente após a excisão dos botões corneoesclerais estes foram colocados em lâminas de microscopia de vidro com o lado endotelial voltado para cima. No Grupo 1, o endotélio corneano foi recoberto com 0,2ml de besilato de atracúrio (10mg/ml) durante 3 minutos e depois lavado com solução salina balanceada. No Grupo 2, o endotélio corneano foi recoberto apenas com solução salina balanceada durante 3 min. As córneas de ambos os grupos foram coradas com vermelho de alizarina e azul de tripano e visualizadas com microscópio óptico. Foram obtidas dez fotografias aleatórias de cada amostra. As imagens foram analisadas e com auxílio de um software as áreas com ausência de células endoteliais calculadas. A perda celular endotelial foi significativamente maior no grupo tratamento comparativamente ao grupo controle. Com base nos resultados apresentados foi possível concluir que o besilato de atracúrio induziu dano agudo nas células do endotélio da córnea de galinhas.(AU)

Aspectos qualitativos das rupturas e afinamentos da membrana de Descemet no exame histopatológico / Qualitative aspects of ruptures and thinnings of Descemet's membrane in histopathological examination

Resumo Objetivo: Descrever os diferentes tipos de rupturas e afinamentos da membrana de Descemet encontrados no exame histopatológico, questionando uma possível relação entre eles. Métodos: Estudo observacional, transversal, retrospectivo e descritivo de botões corneanos provenientes de ceratoplastia penetrante, durante o período escolhido de forma aleatória de quatro anos: 2006, 2010, 2014 e 2015. A coloração foi realizada com Hematoxilina-eosina (HE). Após o preparo, os tecidos foram examinados com microscópio óptico pelos autores. Selecionamos apenas os casos de rupturas ou afinamento da membrana de Descemet no exame histopatológico, e classificamos os diferentes tipos desses achados. Resultados: As rupturas encontradas foram classificadas em total ou comum, parcial, fratura e bisel. Os afinamentos foram divididos em generalizado, localizado e extensivo. Conclusão: Apresentamos várias nuances das rupturas e dos afinamentos da membrana de Descemet no exame histopatológico. Os achados sugerem, considerando apenas aspectos mecânicos, uma possível relação entre afinamento e ruptura como causa e efeito.

Abstract Objective: To describe the different types of ruptures and thinning of Descemet's membrane found in the histopathological examination, questioning a possible relationship between them. Methods: Observational, transversal, retrospective and descriptive study of corneal buttons from penetrating keratoplasty during the randomly chosen period of four years: 2006, 2010, 2014 and 2015. The staining was performed with hematoxylin-eosin (HE). After preparation, the authors examined the tissues with an optical microscope. We selected only the cases of rupture or thinning of Descemet's membrane in histopathological examination and classified the different types of these findings. Results: The ruptures found were classified as total or common, partial, fracture and bevel. The thinnings were divided into generalized, localized and extensive. Conclusion: We presented several nuances of Descemet's membrane ruptures and thinning in histopathological examination. Considering only mechanical aspects, the findings suggest a possible relationship between thinning and rupture as cause and effect.

Corneal endothelial cell density and pterygium: a cross-sectional study / Contagem de células endoteliais e pterígio: um estudo transversal

ABSTRACT Purpose: To investigate the effects of pterygium on corneal endothelial cell density in patients with unilateral pterygium. Methods: We performed a cross-sectional analysis of data from patients with unilateral pterygium who were selected from September 1, 2015 to July 31, 2016 at Hospital de Base do Distrito Federal to assess the corneal endothelial cell density, coefficient of variation in the cell area, hexagonality, and corneal pa­chymetric results. In all patients, noncontact specular microscopy was performed in both eyes and a minimum endothelial cell count of 75 cells/mm2 was required for inclusion in the study. The contralateral eye served as the control. Results: Sixty-one patients were included in the study. Twenty-nine (47.5%) patients were men and 32 (52.5%) were women (mean age, 50.84 ± 13.8). The percentage of pterygium that invaded the cornea ranged from 4.87% to 24.59% (median, 9.70% ± 4.99%). The mean corneal endothelial cell density (cells/mm) was lower in the pterygium eyes than in the controls (2451.83 ± 284.96 vs. 2549.95 ± 268.94, respectively; p=0.04). No differences in the mean coefficients of variation of cell size, hexagonality, and corneal pachymetric results were observed between the patients and controls. The Pearson correlation test showed a significant negative linear relationship between pterygium invasion and endothelial cell density [p<0.001, n=61, r=-0.553 (95% CI, -0.34 to -0.73)]. Conclusion: Compared with the contralateral eyes, those of patients with unilateral pterygium were associated with a decrease in corneal endothelial cell density.

RESUMO Objetivo: Investigar os efeitos do pterígio na densidade de células endoteliais cor­neanas em pacientes com pterígio unilateral. Métodos: Foi realizado um estudo do tipo transversal envolvendo pacientes com pterígio unilateral selecionados entre 1 de setembro de 2015 a 31 de julho de 2016 no Hospital de Base do Distrito Federal para avaliar a densidade de células endoteliais corneanas, coeficiente de variação da área celular, hexagonalidade, e paquimetria corneana. Em todos os pacientes foram realizadas microscopias especulares de não-contato em ambos os olhos, sendo necessário obter uma contagem endotelial mínima de 75 células/mm2 para que o paciente fosse incluído no estudo. O olho contralateral funcionou como grupo controle. Resultados: Um total de 61 pacientes foram incluídos no estudo. Vinte e nove (47,5%) eram homens e 32 (52,5%) mulheres. A média de idade era de 50,84 ±13,8. O percentual de invasão do pterígio na córnea variou entre 4,87% a 24,59%, com uma mediana de 9,70% ± 4,99%. A media de densidade de células endoteliais corneanas foi menor nos olhos com pterígio quando comparados ao grupo controle (2451,83 ± 284,96 vs 2549,95 ± 268,94; p=0,04). Não foram encontradas diferenças entre os casos e controles em relação à média do coeficiente de variação da área celular, hexagonalidade, e paquimetria. Teste de correlação de Pearson mostrou uma relação linear negativa entre a invasão do pterígio e a densidade de células endoteliais corneanas [p<0,001, n=61, r=-0,553 (95% CI -0,34 a -0,73)]. Conclusão: Em pacientes com pterígio unilateral, o olho com pterígio está asso­ciado a uma menor densidade de células endoteliais corneanas quando comparado ao olho contralateral.

Changes in Corneal Endothelial Cell after Ahmed Glaucoma Valve Implantation and Trabeculectomy: 1-Year Follow-up

PURPOSE: To compare changes in corneal endothelial cell density (CECD) after Ahmed glaucoma valve (AGV) implantation and trabeculectomy. METHODS: Changes in corneal endothelium in patients that underwent AGV implantation or trabeculectomy were prospectively evaluated. Corneal specular microscopy was performed at the central cornea using a non-contact specular microscope before surgery and 6 months and 12 months after surgery. The CECD, hexagonality of the endothelial cells, and the coefficient of variation of the cell areas were compared between the two groups. RESULTS: Forty eyes of 40 patients with AGV implantation and 28 eyes of 28 patients with trabeculectomy were studied. Intraocular pressure in the AGV implantation group was significantly higher than that in the trabeculectomy group (p < 0.001), but there was no significant difference in other clinical variables between the two groups. In the AGV implantation group, the mean CECD significantly decreased by 9.4% at 6 months and 12.3% at 12 months compared with baseline values (both, p < 0.001), while it decreased by 1.9% at 6 months and 3.2% at 12 months in the trabeculectomy group (p = 0.027 and p = 0.015, respectively). The changes at 6 months and 12 months in the AGV implantation group were significantly higher than those in the trabeculectomy group (p = 0.030 and p = 0.027, respectively). In the AGV implantation group, there was a significant decrease in the CECD between baseline and 6 months and between 6 months and 12 months (p < 0.001 and p = 0.005, respectively). However, in the trabeculectomy group, a significant decrease was observed only between baseline and 6 months (p = 0.027). CONCLUSIONS: Both the AGV implantation group and the trabeculectomy group showed statistically significant decreases in the CECD 1 year after surgery. The decrease in CECD in the AVG implantation group was greater and persisted longer than that in the trabeculectomy group.

A Comparative Study of Topical Mitomycin C, Cyclosporine, and Bevacizumab after Primary Pterygium Surgery

PURPOSE: To compare the recurrence rates and complications associated with instillation of topical mitomycin C, cyclosporine, and bevacizumab after primary pterygium surgery. METHODS: Between July 2013 and June 2014, we performed surgery using the bare sclera method on 132 eyes (132 patients) with primary pterygium. We randomly selected 33 eyes (33 patients) and treated them with artificial tears four times a day for three months, 29 eyes (29 patients) were treated with topical 0.02% mitomycin C four times a day for five days, 34 eyes (34 patients) were treated with topical 0.05% cyclosporine four times a day for three months, and 36 eyes (36 patients) were treated with topical 2.5% bevacizumab four times a day for three months after surgery. We prospectively determined the recurrence rates of pterygium and complications at the six-month follow-up examination. RESULTS: At six months after surgery, the recurrence rates in each group were as follows: 45.5% (15 eyes) in the control group, 10.3% (three eyes) in the mitomycin C group, 20.6% (seven eyes) in the cyclosporine group, and 41.7% (15 eyes) in the bevacizumab group (p = 0.004). No serious complications, except subconjunctival hemorrhages, were observed in any group. CONCLUSIONS: Groups receiving topical 0.02% mitomycin C and 0.05% cyclosporine after surgery showed lower recurrence rates than the control group; however, no difference in recurrence rate was observed between the control group and the group receiving topical 2.5% bevacizumab after surgery.

Ultrasound biomicroscopy after palliative surgical procedures for bullous keratopathy: a descriptive comparative study / Biomicroscopia ultrassônica após procedimentos cirúrgicos paliativos na ceratopatia bolhosa: estudo comparativo descritivo

Purpose: To describe quantitative and qualitative features of eyes with advanced bullous keratopathy assessed using ultrasound biomicroscopy, before and after anterior stromal puncture (ASP) or amniotic membrane transplantation (AMT) procedures to relieve chronic pain. Methods: The present descriptive comparative study included 40 eyes of 40 patients with chronic intermittent pain due to bullous keratopathy who were randomly assigned to one of the two treatments (AMT or ASP). Ultrasound biomicroscopy (Humphrey, UBM 840, 50 MHz transducer, immersion technique) was used, and a questionnaire about pain intensity was completed preoperatively and postoperatively at days 90 and 180, respectively. Exclusion criteria were age<18 years, presence of concurrent infection, ocular hypertension, and absence of pain. Results: In a 180-day follow-up, the AMT group exhibited mean central corneal thickness (CCT), 899.4 µm preoperatively and 1122.5 µm postoperatively (p<0.001); mean epithelial thickness (ET), 156.4 µm preoperatively and 247.8 µm postoperatively (p<0.001); and mean stromal thickness (ST), 742.9 µm preoperatively and 826.3 µm postoperatively (p=0.005). The ASP group exhibited mean CCT, 756.7 µm preoperatively and 914.8 µm postoperatively (p<0.001); mean ET, 102.1 µm preoperatively and 245.2 µm postoperatively (p<0.001); and mean ST, 654.6 µm preoperatively and 681.5 µm postoperatively (p<0.999). Correlations between CCT and pain intensity in the AMT group (p=0.209 pre- and postoperatively) and the ASP group (p=0.157 preoperatively and p=0.426 at the 180-day follow-up) were not statistically significant. Epithelial and stromal edema, Descemet’s membrane folds, epithelial bullae, and the presence of interface fluid were frequently observed qualitative features. Conclusion: CCT increased over time in both groups. The magnitude of CCT did not correlate with pain intensity in the sample studied. The presence of interface ...

Objetivo: Descrever as características quantitativas e qualitativas da biomicroscopia ultrassônica (UBM) em olhos com ceratopatia bolhosa avançada, antes e após os procedimentos de punção estromal anterior (ASP) ou transplante de membrana amniótica (AMT) para alívio de dor crônica. Métodos: Estudo comparativo descritivo incluindo 40 olhos de 40 pacientes com dor crônica intermitente devido a ceratopatia bolhosa, randomizados em duas modalidades de tratamento (AMT e ASP). Biomicroscopia ultrassônica (Humphrey, UBM 840, transdutor de 50 MHz, técnica de imersão) foi utilizada, e um questionário de avaliação da intensidade da dor foi aplicado no pré-operatório, e após 90 e 180 dias de pós-operatório. Critérios de exclusão foram: idade abaixo de 18 anos, presença de infecção, hipertensão ocular, e ausência de dor. Resultados: No seguimento de 180 dias, o grupo Transplante de membrana amniótica apresentou: média da espessura corneana central (CCT): 899,4 µm (pré), 1.122,5 µm (pós-operatório) (p<0,001); média da espessura epitelial (ET): 156,4 µm (pré), 247,8 µm (pós-operatório) (p<0,001); média da espessura estromal (ST): 742,9 µm (pré), 826,3 µm (pós-operatório) (p=0,005), e, grupo ASP apresentou: CCT média: 756.7 µm (pré), 914,8µm (pós-operatório) (p<0,001); ET média: 102,1 µm (pré), 245,2 µm (pós-operatório) (p<0,001); ST média: 654,6 µm (pré), 681.5 µm (pós-operatório) (p<0,999). A correlação entre intensidade da dor e espessura corneana central no grupo AMT (p=0,209 pré e pós-operatórios) e no grupo ASP (p=0,157 pré-operatório e p=0,426 aos 180 dias de seguimento) não foi significativa. Edema epitelial e estromal, dobras na membrana de Descemet, bolhas epiteliais, e presença de fluido na interface foram características qualitativas frequentemente observadas. Conclusão: A espessura corneana central aumentou ao longo do tempo em ambos os grupos. A magnitude da espessura corneana central não interfere na intensidade da dor ...

O papel da membrana de Descemet na patogenia do edema corneano após cirurgia de segmento anterior / The role of Descemet's membrane in the pathogeny of corneal edema following anterior segment surgery

Objetivo: Encontrar fatores importantes na patogenia do edema corneano pós-cirúrgico, em casos de pós-facectomia e pósceratoplastia, por meio do estudo dos achados histopatológicos, a fim de ver o que pode ser feito para evitar sucessivas ceratoplastias. Métodos: Estudo retrospectivo descritivo das alterações histopatológicas de casos de edema corneano pós-cirúrgicos. Os tecidos foram provenientes de ceratoplastia penetrante no período compreendido entre setembro de 2009 e agosto de 2013. Foi realizada revisão de prontuários em busca principalmente de informações sobre cirurgias prévias. Resultados: Foram incluídos 70 botões corneanos, sendo 34 de pacientes do sexo masculino e 36 do sexo feminino. A média das idades foi de 63,1±17,20 (média ± DP) anos. A maioria dos casos era de falência após transplante (71,43%). A rarefação celular foi a principal alteração encontrada no endotélio (58 casos), sendo também a alteração histopatológica mais frequente. Na membrana de Descemet, predominaram as alterações de integridade (53 casos), seja na forma de ruptura, de descolamento isolado ou de descolamento associado à ruptura. Foi frequente a associação de alterações endoteliais à ausência da integridade da membrana de Descemet. Conclusão: Descolamento da membrana de Descemet é um achado histopatológico frequente nos casos de edema corneano pós-cirúrgicos estudados, devendo ser considerado um fator importante na patogenia dos mesmos. Essa alteração deve ser procurada com atenção nos pós-operatórios, a fim de ser diagnosticada e tratada precocemente, vindo possivelmente a evitar muitas indicações de ceratoplastia. .

Objective: To find relevant factors in the pathogeny of postoperative corneal edema in post-cataract surgery and post-keratoplasty cases, through the study of histopathological findings in order to see what can be done to avoid successive keratoplasties. Methods: Retrospective descriptive study of histopathological findings in postoperative corneal edema cases. Tissues were obtained from penetrating keratoplasty in the period between september 2009 and august 2013. A medical record review was conducted primarily looking for information about previous surgeries. Results: Seventy corneal buttons were included, out of which 34 were from male patients and 36 were from female patients. The mean age was 63.1±17.20 (mean ± SD) years. Most of cases were corneal failure after keratoplasty (71.43%). The main change found in endothelium was cellular rarefaction (58 cases), and it was also the most common histopathological change. Changes in integrity predominated in Descemet's membrane (53 cases), whether in the form of rupture, isolated detachment or detachment associated with rupture. Endothelial changes associated with the absence in Descemet's membrane integrity were frequent. Conclusion: Descemet's membrane detachment is a frequent histopathological finding in postoperative corneal edema cases studied, thus it should be considered an important factor in the pathogeny of such cases. This change should be carefully researched in the postoperative period in order to be diagnosed and treated early, possibly avoiding many keratoplasty indications. .

Effects of Argon Laser Iridotomy on the Corneal Endothelium of Pigmented Rabbit Eyes

PURPOSE: In Asian countries, laser iridotomy for the treatment of angle-closure glaucoma is a common cause of bullous keratopathy, which may be associated with a shallow anterior chamber and dark iris pigmentation in Asians. Several cases of corneal decompensation after argon laser iridotomy have been reported. In the present study, we evaluated the harmful effects of argon laser iridotomy on the corneal endothelium. METHODS: Argon laser iridotomy was performed on the right eyes of pigmented rabbits. Changes in corneal thickness and endothelial cell density after laser iridotomy were evaluated. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was performed for assessment of corneal endothelial cell apoptosis. Combined staining with alizarin red and trypan blue, as well as a live/dead cell assay, were performed for evaluation of damage to the corneal endothelium induced by laser iridotomy. RESULTS: Corneal thickness did not change immediately after laser iridotomy; however, a significant increase was observed 24 hours after iridotomy (p = 0.001). The endothelial cell density of laser-treated eyes four days after laser iridotomy was significantly decreased compared with control eyes (p < 0.001). TUNEL staining showed many TUNEL-positive cells in the corneal endothelium and corneal stroma. No endothelial trypan blue-stained cell nuclei were observed after laser iridotomy; however, several large endothelial cells with damaged membrane integrity were observed. The live/dead cell assay clearly showed a large number of dead cells stained red in several areas throughout the entire corneal button 24 hours after iridotomy. CONCLUSIONS: Argon laser iridotomy induces corneal endothelial cell apoptosis in pigmented rabbit eyes, resulting in decreased endothelial cell density.

Effects of Argon Laser Iridotomy on the Corneal Endothelium of Pigmented Rabbit Eyes

PURPOSE: In Asian countries, laser iridotomy for the treatment of angle-closure glaucoma is a common cause of bullous keratopathy, which may be associated with a shallow anterior chamber and dark iris pigmentation in Asians. Several cases of corneal decompensation after argon laser iridotomy have been reported. In the present study, we evaluated the harmful effects of argon laser iridotomy on the corneal endothelium. METHODS: Argon laser iridotomy was performed on the right eyes of pigmented rabbits. Changes in corneal thickness and endothelial cell density after laser iridotomy were evaluated. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was performed for assessment of corneal endothelial cell apoptosis. Combined staining with alizarin red and trypan blue, as well as a live/dead cell assay, were performed for evaluation of damage to the corneal endothelium induced by laser iridotomy. RESULTS: Corneal thickness did not change immediately after laser iridotomy; however, a significant increase was observed 24 hours after iridotomy (p = 0.001). The endothelial cell density of laser-treated eyes four days after laser iridotomy was significantly decreased compared with control eyes (p < 0.001). TUNEL staining showed many TUNEL-positive cells in the corneal endothelium and corneal stroma. No endothelial trypan blue-stained cell nuclei were observed after laser iridotomy; however, several large endothelial cells with damaged membrane integrity were observed. The live/dead cell assay clearly showed a large number of dead cells stained red in several areas throughout the entire corneal button 24 hours after iridotomy. CONCLUSIONS: Argon laser iridotomy induces corneal endothelial cell apoptosis in pigmented rabbit eyes, resulting in decreased endothelial cell density.

Comparison of morphological and functional endothelial cell changes after cataract surgery: phacoemulsification versus manual small-incision cataract surgery

To compare the morphological [cell density, coefficient of variation and standard deviation] and functional [central corneal thickness] endothelial changes after phacoemulsification versus manual small-incision cataract surgery [MSICS]. Prospective randomized control study. In this prospective randomized control study, patients were randomly allocated to undergo phacoemulsification [Group 1, n = 100] or MSICS [Group 2, n = 100] using a random number Table. The patients underwent complete ophthalmic evaluation and specular microscopy preoperatively and at land 6 weeks postoperatively. Functional and morphological endothelial evaluation was Noncon ROBO PACHY SP-9000 specular microscope. Phacoemulsification was performed, the chop technique and MSICS, by the viscoexpression technique. The mean difference in central corneal thickness at baseline and 1 week between Group 1 and Group 2 was statistically significant [P- 0.027]. However, this difference at baseline when compared to 6 week and 1 week, 6 weeks was not statistically significant [P> 0.05]. The difference in mean endothelial cell density between groups at 1 week and 6 weeks was statistically significant [P = 0.016]. The mean coefficient of variation and mean standard deviation between groups were not statistically significant [P> 0.05, both comparisons]. The central corneal thickness, coefficient of variation, and standard deviation were maintained in both groups indicating that the function and morphology of endothelial cells was not affected despite an initial reduction in endothelial cell number in MSICS. Thus, MSICS remains a safe option in the developing world

Variación del endotelio corneal en pacientes operados de alta miopía con implante de lentes fáquicas / Variation of the corneal endothelium in patients operated on from high myopia, with phakic lens implantation

Objetivo: determinar la variación que evidencia el endotelio corneal, en pacientes operados de alta miopía con la utilización de lentes intraoculares fáquicas de cámara anterior con soporte iridiano. Método: se realizó un estudio longitudinal, prospectivo y descriptivo en una muestra conformada por 26 ojos de 15 pacientes, operados de alta miopía con la utilización de lentes intraoculares fáquicas Artisan y Artiflex de cámara anterior con soporte iridiano, que se atendieron en el Servicio de Córnea y Cirugía Refractiva del Instituto Cubano de Oftalmología Ramón Pando Ferrer de septiembre de 2009 a mayo de 2010. Resultados: se evidenció una pérdida celular endotelial posterior a la cirugía seis meses de 6,9 por ciento. En el posoperatorio la hexagonalidad y el coeficiente de variación disminuyeron, mientras que la paquimetría no mostró diferencias significativas. Conclusiones: en el presente estudio se observó la presencia de un daño mínimo en el endotelio corneal con el uso de lentes fáquicas de cámara anterior, en la corrección quirúrgica de las altas miópias

Objective: to determine the variation of the corneal endothelium in those patients operated on from high myopia, with the use of iris claw anterior chamber phakic intraocular lenses. Methods: a prospective, longitudinal and descriptive study was carried out in a sample of 26 eyes from 15 patients. They had been operated on from high myopia with the use of iris claw anterior chamber phakic intraocular lenses Artisan and Artiflex at the Cornea and Refractive Surgery Service of Ramon Pando Ferrer Cuban Institute of Ophthalmology from September 2009 through May 2010. Results: it was evinced that there was endothelial cell loss after surgery six months of 6.9 percent. In the postoperative phase, hexagonality and variation coefficient diminished whereas pakimetry did not show significant differences. Conclusions: the present study indicated the existence of minimal damage in the corneal endothelium with the use of the anterior chamber phakic lenses for the surgical correction of high myopias

Caracterización del endotelio corneal en pacientes con indicación de cirugía de catarata / Characterization of the corneal endothelium in patients requiring cataract surgery

Objetivo: describir la morfología del endotelio corneal en pacientes con indicación de cirugía de catarata. Métodos: se estudiaron 200 ojos de 127 pacientes, 86 mujeres y 41 hombres, con indicación de cirugía de catarata en una investigación de tipo descriptiva y transversal realizada entre octubre y diciembre de 2011. Se examinaron los pacientes en lámpara de hendidura buscando alteraciones de la morfología del endotelio y se les realizó microscopia especular con el Topcon SP3000P. Se analizaron las imágenes y se evaluaron la densidad celular, el coeficiente de variabilidad, la hexagonalidad celular y la paquimetría. Resultados: predominó el sexo femenino con 86 pacientes y en los dos grupos la mayoría tenían entre 60 y 79 años. La densidad celular media fue de 2 238,19 cél/mm2, disminuyó significativamente con la edad y no tuvo diferencias con el sexo. El coeficiente de variabilidad, la hexagonalidad y la paquimetría no tuvieron diferencias significativas con la edad, ni con el sexo. Las guttas corneales predominaron en las féminas (103; 76,9 por ciento) con diferencia significativa entre los sexos. Conclusiones: a mayor edad hay disminución de la densidad celular, así como un aumento del coeficiente de variabilidad sin diferencias entre sexos. Se comprueba la estrecha relación entre la disminución de la densidad celular y el aumento de la paquimetría. Además, se muestran los parámetros de normalidad del endotelio corneal en pacientes con indicación de cirugía de catarata en nuestro medio

Objective: to describe the morphology of corneal endothelium in patients requiring cataract surgery.Methods: adescriptive, cross-sectional study was conducted in 200 eyes from 127 patients distributed into 86 females and 41 males, who required cataract surgery. All the patients were examined with slit-lamp method to look for corneal guttas and were performed Topcon specular microscopy SP3000P. Images were analyzed and the cell density, the variability coefficient, the cell hexagonality and pachymetry were all evaluated. Results: females predominated with 86 patients in both groups and most aged 60-79 years. The average cell density was 2238.19 cell/mm2 that significantly decreased with the age increase, and it did not show any differences between the sexes. The average variability coefficient, hexagonality and pachymetry did not show significant differences in terms of age and sex. The presence of corneal guttas predominated in females (103; 76.9 percent.) but the difference between the sexes was significant.Conclusions: as the age increases, the cell density diminishes but the variability coefficient increases and there are no differences between the sexes. The close relationship between the decreased cell density and the increased pachymetry is proved. Additionally, the study presents the normal parameters of the corneal endothelium in patients requiring cataract surgery in our setting

Bilateral lineal endotheliitis: case report / Endotelite linear bilateral: relato de caso

To report the case of a patient with bilateral herpetic lineal endotheliitis successfully treated with topic steroids and systemic antiviral. 17 year old female with blurred vision, at evaluation localized edema was observed on both corneas associated to Descemet folds and a line of pigmented precipitates. Topic prednisolone and oral acyclovir are initiated with complete resolution of signs and symptoms. Lineal endotheliitis is produced as an answer of endotelial cells to viral infection; maybe due to an immune reaction against some antigens from herpes virus family. It has the potential of relapses even in the absence of viral replication, with secondary untreatable stromal edema. It responds well to antiviral and steroids treatment, although, on those patients who don't improve, is necesary to make additional tests.

Relatar o caso de uma paciente com endotelite linear herpética bilateral tratado com sucesso por meio de corticoides tópicos e antivirais sistêmicos. Paciente do sexo feminino, 17 anos de idade, com a visão turva, na avaliação foi observado edema localizado em ambas as córneas associadas a dobras de Descemet e uma linha de precipitados ceráticos pigmentados. Prednisolona tópica e aciclovir oral foram utilizados com resolução completa dos sinais e sintomas. A endotelite linear é uma resposta das células endoteliais à infecção viral, talvez devido a uma reação imunológica contra alguns antígenos do vírus da família do herpes. Tem o potencial de recidiva, mesmo na ausência de replicação viral, com edema estromal secundário intratável. Ela responde bem ao tratamento antiviral e esteroides, embora, em pacientes que não melhoram, é necessária a realização de testes adicionais.

Long-term Evaluation of Endothelial Cell Changes in Fuchs Corneal Dystrophy: The Influence of Phacoemulsification and Penetrating Keratoplasty

PURPOSE: To evaluate the natural course of the long-term endothelial cell changes in Fuchs corneal dystrophy (FCD) patients and investigate the effects of phacoemulsification on the annual rate of change in endothelial indices in FCD patients. METHODS: Thirty-four patients diagnosed with FCD at Seoul National University Hospital from 1994 to 2010 were retrospectively reviewed. Sixteen patients who had been followed up for more than 1 year were selected and classified into 3 groups: group A, patients with no ocular surgery; group B, patients who had undergone phacoemulsification only; and group C, patients who had undergone penetrating keratoplasty with cataract surgery. Endothelial cell density, polymegethism, pleomorphism, and pachymetry were measured and the exponential rates of endothelial cell and pachymetry change were analyzed. RESULTS: A non-linear mixed model of non-operated FCD patients showed that only pachymetric data tended to increase with statistical significance (p = 0.001) with a mean follow-up period of 4.15 years. Using an exponential regression analysis fitting curve, the mean rates of annual endothelial cell loss were 0.82%/yr, 20.39%/yr, and 29.27%/yr in groups A, B, and C respectively, and statistical significance was seen only in group C (p < 0.05). CONCLUSIONS: Retrospective long-term follow-up data showed that changes in endothelial density did not significantly decrease over at least 4 years in middle-aged FCD patients. The changes in pachymetric corneal thickness appeared to increase over the same period. Considering that no exponential changes were aggravated after performing cataract surgery alone, cataract surgery would be a preferable option in FCD patients compared to an approach of "wait-and-do" penetrating keratoplasty combined with cataract surgery.

Corneal endothelium after deep anterior lamellar keratoplasty and penetrating keratoplasty for keratoconus: A four-year comparative study.

Purpose: To compare the status of corneal endothelium and central corneal thickness within the first four postoperative years after deep anterior lamellar keratoplasty (DALK) and penetrating keratoplasty (PK) in patients with keratoconus. Materials and Methods: Thirty-nine eyes (Group A) which had PK and 44 eyes (Group B) which had DALK for the treatment of keratoconus were included in this retrospective study. The endothelial cell density (ECD), the mean endothelial cell area and the coefficient of variation of cell area were assessed with a non-contact specular microscope, and the central corneal thickness (CCT) was measured with an ultrasound pachymeter. Results: Mean ECD loss rate at two years was 36.24% in Group A and 18.12% in Group B (P<0.001). Mean ECD loss rate at four years was 47.82% in Group A and 21.62% in Group B (P<0.001). Mean annual ECD loss rate was calculated 14.12% per year in Group A and 5.78% per year in Group B. In the PK group, increase in mean CCT was 15.60% in two years and 15.03% in four years, while in the DALK group, mean CCT increased by 8.05% in two years and 9.31% in four years. Conclusions: As the majority of ectatic disorders such as keratoconus occur in young people, long-term endothelial cell survival following treatment with keratoplasty is essential for the long-term visual ability. Our finding that corneal endothelial cell loss in the DALK group occurs at a slower rate than in the PK group suggests DALK as a safer alternative to PK in these selected patients.

Seleção de córneas para transplantes / Corneal selection for transplants

O transplante de córnea é singular, pelo fato de ser habitualmente realizado em pessoas com deficiência visual, mas com expectativa de vida, mobilidade e convivência social preservadas. Um enxerto mal-sucedido pode levar à cegueira real ou ao sofrimento permanente, motivado por dor ou desconforto ocular. Um dos fatores fundamentais do sucesso do mesmo é o uso de córneas de boa qualidade. A seleção dessas córneas assenta-se em três princípios: inocuidade, transparência e vitalidade. Cada um deles é analisado detalhadamente neste trabalho.

Corneal transplantation is singular because it is habitually performed on persons with visual deficiency but with preserved life expectancy, mobility and social life. A graft not well succeeded may cause real blindness and permanent misery due to pain and discomfort. One of the fundamental determinants of the success of corneal transplantation is the use of corneas of good quality. The selection of these corneas is based on three rules: innocuousness, transparency and vitality. Each one of these concepts is considered in detail in this work.

Central Corneal Thickness and Corneal Endothelial Cell Changes Caused by Contact Lens Use in Diabetic Patients

PURPOSE: To analyze the effects of soft contact lenses on central corneal thickness and morphologic characteristics of the corneal endothelium in diabetic patients. MATERIALS AND METHODS: Ultrasound pachymetry and noncontact specular microscopy were performed on 26 diabetic patients who regularly use soft contact lenses (group 1), 27 diabetic patients who do not use soft contact lenses (group 2) and 30 normal subjects (group 3). We compared the values in each group using the Mann-Whitney test. RESULTS: The central cornea was found to be thicker in diabetic patients, both those who use and do not use contact lenses, than in the normal control group. The central corneal thickness was significantly higher in group 1 (564.73 +/- 35.41 microm) and group 2 (555.76 +/- 45.96 microm) than in the control group (534.05 +/- 27.02 microm), but there was no statistically significant difference between groups 1 and 2. Endothelial cell density was significantly different between the groups, and was smallest in the group of diabetic patients using contact lenses. The coefficient of variation of cell size was significantly higher and the percentage of hexagonal cells was significantly lower in contact lens using diabetic patients than in non-contact lens using diabetic patients and in the control group. CONCLUSION: Central corneal thickness and endothelial cell density is more affected by diabetes mellitus, and corneal endothelial cell morphology is more affected by contact lens use, when compared with normal subjects.

Confocal scan features of keratic precipitates in granulomatous versus nongranulomatous uveitis

To compare the morphologic features of keratic precipitates [KPs] by confocal microscopy in granulomatous versus nongranulomatous noninfectious uveitis. KP morphology was determined by confocal scan in patients with noninfectious granulomatous and noninfectious nongranulomatous uveitic cases. One hundred and twenty-seven eyes of 90 subjects with noninfectious uveitis were studied. Thirty-nine eyes had granulomatous and 88 had nongranulomatous uveitis. Smooth-rounded KPs were significantly more common in the granulomatous subgroup [P<0.001] while cruciform and dendritiform KPs were more frequent in nongranulomatous uveitis [P<0.001 and P<0.005 respectively]. Confocal scan may be used as an adjunctive tool for differentiating granulomatous from nongranulomatous uveitis. Smooth-rounded KPs are strongly suggestive of granulomatous inflammation

A Comparison of Posterior Lamellar Keratoplasty Modalities: DLEK vs. DSEK

PURPOSE: To compare clinical outcomes after deep lamellar endothelial keratoplasty (DLEK) with Descemet stripping endothelial keratoplasty (DSEK) performed as initial cases by a single surgeon. METHODS: Sixteen patients with corneal endothelial were enrolled. Eight patients (8 eyes) underwent DLEK and 8 patients (8 eyes) DSEK. We measured uncorrected visual acuity, best corrected visual acuity (BCVA), manifest refraction, corneal endothelial count, interface opacity via Schiempflug imaging, and contrast sensitivity, as well as tracked postoperative complications over the first postoperative year. RESULTS: Primary graft failure occurred in two DLEK cases and one DSEK case, all of which were excluded for further analysis. The average 12-month postoperative BCVA was 20/70 in the DLEK group and 20/50 in the DSEK group, with the difference not statistically significant. No significant differences were identified between the 2 groups in terms of mean spherical equivalent and refractive astigmatism, although individuals in the DSEK group tended toward hyperopia. The average endothelial cell count at postoperative month 12 was 1849+/-494 in the DLEK group and 1643+/-417 cells/mm2 in the DSEK group, representing cell losses of 26.2% and 31.9%, respectively. No significant differences in endothelial cell count or endothelial cell loss were observed between groups. Early postoperative donor disc dislocation occurred in two eyes after DLEK and one eye after DSEK. Interface opacities and contrast sensitivities were similarly not significantly different between groups. CONCLUSIONS: No significant differences in any assessed clinical outcome were observed between individuals undergoing DLEK and DSEK, when performed as initial cases by a single surgeon.