Corneal reshaping: an experiment with a type I collagen-based vitrigel for remodeling porcine corneas / Remodelamento corneano: relato de um experimento com um vitrigel a base de colágeno tipo I para remodelamento de córneas porcinas

ABSTRACT Purpose: This study aimed to report an experiment designed to determine anatomical changes in porcine corneas following placement of a novel polymer implant into the cornea. Methods: An ex vivo porcine eye model was used. A novel type I collagen-based vitrigel implant (6 mm in diameter) was shaped with an excimer laser on the posterior surface to create three planoconcave shapes. Implants were inserted into a manually dissected stromal pocket at a depth of approximately 200 μm. Three treatment groups were defined: group A (n=3), maximal ablation depth 70 μm; Group B (n=3), maximal ablation depth 64 μm; and group C (n=3), maximal ablation depth 104 μm, with a central hole. A control group (D, n=3) was included, in which a stromal pocket was created but biomaterial was not inserted. Eyes were evaluated by optical coherence tomography (OCT) and corneal tomography. Results: Corneal tomography showed a trend for a decreased mean keratometry in all four groups. Optical coherence tomography showed corneas with implants placed within the anterior stroma and visible flattening, whereas the corneas in the control group did not qualitatively change shape. Conclusions: The novel planoconcave biomaterial implant described herein could reshape the cornea in an ex vivo model, resulting in the flattening of the cornea. Further studies are needed using in vivo animal models to confirm such findings.

RESUMO Objetivo: Relatar um experimento projetado para determinar alterações anatômicas em córneas porcinas após a colocação de um novo implante de polímero na córnea. Métodos: Foi utilizado olho de porco ex vivo. Um novo agente modelador biocompatível, de colágeno tipo 1, com 6mm de diâmetro foi moldado com excimer laser em sua face posterior, para criar três formatos planocôncavos. Os implantes foram inseridos dentro de um bolsão, dissecado manualmente, a 200 micrômetros (μm). Foram definidos três grupos de tratamento: grupo A (n=3), teve a profundidade máxima de ablação de 70 μm; o grupo B (n=3), profundidade máxima de ablação de 64 μm; e o grupo C (n=3), profundidade máxima de ablação de 104 μm, com buraco central. O grupo controle, D (n=3), foi incluído, com a criação do bolsão estromal, porém sem inserir o material. A avaliação desses olhos foi realizada por tomografia de coerência óptica (OCT) e por tomografia corneana. Resultados: A tomografia corneana mostrou uma tendência para diminuição da ceratometria média em todos os 4 grupos. A tomografia de coerência óptica mostrou córneas com implantes localizados no estroma anterior e aplanamento visível, enquanto as córneas não mudaram qualitativamente o formato no grupo controle. Conclusões: O novo implante de biomaterial planocôncavo descrito aqui foi capaz de remodelar a córnea em modelo de animal ex vivo, resultando no aplanamento corneano. Novos estudos são necessários usando modelos animais in vivo para confirmar tais achados.

Corneal reshaping: an experiment with a type I collagen-based vitrigel for remodeling porcine corneas.

PURPOSE: This study aimed to report an experiment designed to determine anatomical changes in porcine corneas following placement of a novel polymer implant into the cornea. METHODS: An ex vivo porcine eye model was used. A novel type I collagen-based vitrigel implant (6 mm in diameter) was shaped with an excimer laser on the posterior surface to create three planoconcave shapes. Implants were inserted into a manually dissected stromal pocket at a depth of approximately 200 µm. Three treatment groups were defined: group A (n=3), maximal ablation depth 70 µm; Group B (n=3), maximal ablation depth 64 µm; and group C (n=3), maximal ablation depth 104 µm, with a central hole. A control group (D, n=3) was included, in which a stromal pocket was created but biomaterial was not inserted. Eyes were evaluated by optical coherence tomography (OCT) and corneal tomography. RESULTS: Corneal tomography showed a trend for a decreased mean keratometry in all four groups. Optical coherence tomography showed corneas with implants placed within the anterior stroma and visible flattening, whereas the corneas in the control group did not qualitatively change shape. CONCLUSIONS: The novel planoconcave biomaterial implant described herein could reshape the cornea in an ex vivo model, resulting in the flattening of the cornea. Further studies are needed using in vivo animal models to confirm such findings.

Comparison of 20% sulfur hexafluoride with air for intraocular tamponade in Descemet membrane endothelial keratoplasty (DMEK).

PURPOSE:: To compare the effect of 20% sulfur hexafluoride (SF6) with that of air on graft detachment rates for intraocular tamponade in Descemet membrane endothelial keratoplasty (DMEK). METHODS:: Forty-two eyes of patients who underwent DMEK by a single surgeon (A.S.J.) at Wilmer Eye Institute between January 2012 and 2014 were identified; 21 received air for intraocular tamponade and the next consecutive 21 received SF6. The main outcome measure was the graft detachment rate; univariate and multivariate analyses were performed. RESULTS:: The graft detachment rate was 67% in the air group and 19% in the SF6 group (p<0.05). No complete graft detachments occurred, and all partial detachments underwent intervention with injection of intraocular air. The percentages of eyes with 20/25 or better vision were not different between the groups (67% vs. 71%). Univariate analysis showed significantly higher detachment rates with air tamponade (OR, 8.50; p<0.005) and larger donor graft size (OR, 14.96; p<0.05). Multivariate analysis with gas but not graft size included showed that gas was an independent statistically significant predictor of outcome (OR, 6.65; p<0.05). When graft size was included as a covariate, gas was no longer a statistically significant predictor of detachment but maintained OR of 7.81 (p=0.063) similar to the results of univariate and multivariate analyses without graft size. CONCLUSION:: In comparison with air, graft detachment rates for intraocular tamponade in DMEK were significantly reduced by 20% SF6.

Comparison of 20% sulfur hexafluoride with air for intraocular tamponade in Descemet membrane endothelial keratoplasty (DMEK) / Comparação de hexafluoreto de enxofre a 20% com ar para tamponamento intraocular na ceratoplastia endotelial de membrana Descemet (DMEK)

ABSTRACT Purpose: To compare the effect of 20% sulfur hexafluoride (SF6) with that of air on graft detachment rates for intraocular tamponade in Descemet membrane endothelial keratoplasty (DMEK). Methods: Forty-two eyes of patients who underwent DMEK by a single surgeon (A.S.J.) at Wilmer Eye Institute between January 2012 and 2014 were identified; 21 received air for intraocular tamponade and the next consecutive 21 received SF6. The main outcome measure was the graft detachment rate; univariate and multivariate analyses were performed. Results: The graft detachment rate was 67% in the air group and 19% in the SF6 group (p<0.05). No complete graft detachments occurred, and all partial detachments underwent intervention with injection of intraocular air. The percentages of eyes with 20/25 or better vision were not different between the groups (67% vs. 71%). Univariate analysis showed significantly higher detachment rates with air tamponade (OR, 8.50; p<0.005) and larger donor graft size (OR, 14.96; p<0.05). Multivariate analysis with gas but not graft size included showed that gas was an independent statistically significant predictor of outcome (OR, 6.65; p<0.05). When graft size was included as a covariate, gas was no longer a statistically significant predictor of detachment but maintained OR of 7.81 (p=0.063) similar to the results of univariate and multivariate analyses without graft size. Conclusion: In comparison with air, graft detachment rates for intraocular tamponade in DMEK were significantly reduced by 20% SF6.

RESUMO Objetivo: Comparar as taxas de descolamento do botão endotelial com o uso de gás hexafluoreto de enxofre a 20% (SF6) em relação ao ar para o tamponamento intraocular na ceratoplastia endotelial da membrana de Descemet (DMEK). Métodos: Quarenta e dois olhos foram operados com a técnica de DMEK por um único cirurgião (A.S.J.) no Wilmer Eye Institute entre janeiro de 2012 a 2014. Os primeiros 21 olhos receberam ar para o tamponamento intraocular após o enxerto do botão endotelial e os 21 olhos seguintes receberam SF6. O desfecho primário medido foi a taxa de descolamento do botão endotelial por análise univariada e multivariada. Resultados: A taxa de descolamento do botão endotelial foi de 67% no grupo que recebeu ar vs 19% no grupo que recebeu SF6 (p<0,05). Não houve nenhum descolamento total de botão e todos os parciais foram tratados com injeção de ar intraocular. Não houve diferença estatística significativa entre os grupos em relação a AV de 20/25 ou melhor (67% vs 71%). A análise univariada demonstrou maior taxa de descolamento com o tamponamento por ar intraocular (OR 8,50, p<0,005) e com botões doadores maiores (OR 14,96, p<0,05). Na análise multivariada, incluindo gás, mas não o tamanho do botão doador, o tipo de gás usado permaneceu sendo um fator preditivo independente e estatisticamente significativo para o desfecho primário, com OR de 6,65 (p<0,05). Porém, quando o tamanho do botão doador foi incluso como covariável, o gás perdeu a sua significância como preditor de descolamento, mantendo o OR de 7,81 (p=0,063), semelhante as análises univariada e multivariada excluindo o tamanho do botão doador. Conclusão: O uso de gás hexafluoreto de enxofre a 20% (SF6) para o tamponamento intraocular reduz a taxa de descolamento do botão endotelial quando comparado ao uso de ar no DMEK.

Characterization of cryopreserved primary human corneal endothelial cells cultured in human serum-supplemented media.

PURPOSE: To compare cryopreserved human corneal endothelial cells (HCECs) grown in human serum-supplemented media (HS-SM) with cryopreserved HCECs grown in fetal bovine serum-supplemented media (FBS-SM). METHODS: Three pairs of human corneas from donors aged 8, 28, and 31 years were obtained from the eye bank. From each pair, one cornea was used to start a HCEC culture using HS-SM; the other cornea was grown in FBS-SM. On reaching confluence, the six cell populations were frozen using 10% dimethyl sulfoxidecontaining medium. Thawed cells grown in HS-SM were compared with those grown in FBS-SM with respect to morphology, growth curves, immunohistochemistry, real time-reverse transcriptase polymerase chain reaction (RT-PCR) for endothelial cell markers, and detachment time. RESULTS: No difference in morphology was observed for cells grown in the two media before or after cryopreservation. By growth curves, cell counts after thawing were similar in both media, with a slight trend toward higher cell counts in FBS-SM. Cells grown in both the media demonstrated a similar expression of endothelial cell markers when assessed by immunohistochemistry, although HCEC marker gene expression was higher in cells grown in HS-SM than in those grown in FBS-SM as assessed by RT-PCR. With FBS-SM, there was a tendency of longer detachment time and lower cell passages. CONCLUSIONS: HS-SM was similar to FBS-SM for cryopreservation of cultured HCECs as assessed by analysis of cell morphology, proliferation, and protein expression, although marker gene expression was higher in cells grown in HS-SM than in those grown in FBS-SM. Detachment time was longer with FBS-SM and in lower passages.

Quantitative analysis of iris parameters in keratoconus patients using optical coherence tomography.

PURPOSE: To investigate the relationship between quantitative iris parameters and the presence of keratoconus. METHODS: Cross-sectional observational study that included 15 affected eyes of 15 patients with keratoconus and 26 eyes of 26 normal age- and sex-matched controls. Iris parameters (area, thickness, and pupil diameter) of affected and unaffected eyes were measured under standardized light and dark conditions using anterior segment optical coherence tomography (AS-OCT). To identify optimal iris thickness cutoff points to maximize the sensitivity and specificity when discriminating keratoconus eyes from normal eyes, the analysis included the use of receiver operating characteristic (ROC) curves. RESULTS: Iris thickness and area were lower in keratoconus eyes than in normal eyes. The mean thickness at the pupillary margin under both light and dark conditions was found to be the best parameter for discriminating normal patients from keratoconus patients. Diagnostic performance was assessed by the area under the ROC curve (AROC), which had a value of 0.8256 with 80.0% sensitivity and 84.6% specificity, using a cutoff of 0.4125 mm. The sensitivity increased to 86.7% when a cutoff of 0.4700 mm was used. CONCLUSIONS: In our sample, iris thickness was lower in keratoconus eyes than in normal eyes. These results suggest that tomographic parameters may provide novel adjunct approaches for keratoconus screening.

Quantitative analysis of iris parameters in keratoconus patients using optical coherence tomography / Uso de tomografia de coerência óptica na análise quantitativa dos parâmetros irianos em pacientes com ceratocone

ABSTRACTPurpose:To investigate the relationship between quantitative iris parameters and the presence of keratoconus.Methods:Cross-sectional observational study that included 15 affected eyes of 15 patients with keratoconus and 26 eyes of 26 normal age- and sex-matched controls. Iris parameters (area, thickness, and pupil diameter) of affected and unaffected eyes were measured under standardized light and dark conditions using anterior segment optical coherence tomography (AS-OCT). To identify optimal iris thickness cutoff points to maximize the sensitivity and specificity when discriminating keratoconus eyes from normal eyes, the analysis included the use of receiver operating characteristic (ROC) curves.Results:Iris thickness and area were lower in keratoconus eyes than in normal eyes. The mean thickness at the pupillary margin under both light and dark conditions was found to be the best parameter for discriminating normal patients from keratoconus patients. Diagnostic performance was assessed by the area under the ROC curve (AROC), which had a value of 0.8256 with 80.0% sensitivity and 84.6% specificity, using a cutoff of 0.4125 mm. The sensitivity increased to 86.7% when a cutoff of 0.4700 mm was used.Conclusions:In our sample, iris thickness was lower in keratoconus eyes than in normal eyes. These results suggest that tomographic parameters may provide novel adjunct approaches for keratoconus screening.

RESUMOObjetivo:Investigar a relação entre os parâmetros quantitativos irianos e a presença de ceratocone.Métodos:Estudo observacional transversal com quinze olhos de 15 pacientes com ceratocone e 26 olhos de 26 indivíduos normais, pareados por idade e gênero. Parâmetros da íris (área, espessura e diâmetro da pupila) de olhos com ceratocone e olhos sem ceratocone foram medidos usando tomografia de coerência óptica do segmento anterior (AS-OCT), em condições padronizadas de alta luminosidade e ambiente escuro. Com o objetivo de maximizar a sensibilidade, especificidade e identificar o melhor ponto de corte na diferenciação entre ceratocone e indivíduos normais, foi realizada a análise quantitativa da curva característica operacional do receptor (ROC) dos parâmetros de espessura da íris.Resultados:A área e espessura da íris estavam reduzidas nos olhos de pacientes com ceratocone. Observamos que o melhor parâmetro para discriminar indivíduos normais de pacientes com ceratocone foi a espessura média na margem pupilar, tanto em condições de alta luminosidade quanto em ambiente escuro. O desempenho diagnóstico deste parâmetro foi avaliado pela análise quantitativa da área sob a curva ROC (AROC), mostrando AROC de 0,8256, com sensibilidade de 80,0% e especificidade de 84,6% usando um ponto de corte de 0,4125 milímetros de espessura da íris. A sensibilidade aumentou para 86,7%, com um corte de 0,4700 milímetros de espessura da íris.Conclusões:Na amostra deste estudo, a espessura da íris demostrou-se reduzida nos olhos com ceratocone. Estes resultados sugerem que os parâmetros tomográficos podem proporcionar novas informações no auxilio da triagem de pacientes com ceratocone.

Novel spatula and dissector for safer deep anterior lamellar keratoplasty / Uso de espátula e dissector para otimização da ceratoplastia lamelar anterior profunda (DALK)

Objective: We describe a novel spatula and dissector to facilitate the big-bubble technique in deep anterior lamellar keratoplasty (DALK). Methods: A 29-year-old man who was diagnosed with bilateral keratoconus underwent deep anterior lamellar keratoplasty (DALK). After 350μm partial thickness incision of the recipient cornea, the Bonfadini dissector was inserted at the deepest point in the peripheral incision and could be advanced to the center of the cornea safely because of its "semi-sharp" tip. After achieving the big-bubble (BB) separation of Descemet membrane (DM) from the overlying stroma, the anterior stromal disc was removed. Viscoelastic material was placed on the stromal bed to prevent uncontrolled collapse and perforation of DM during the paracentesis blade incision into the BB. We could detect the safe opening of the BB using the Bonfadini dissector by the leakage of air bubbles into the viscoelastic material. After injecting viscoelastic material into the BB space, we inserted the Bonfadini spatula into the bigbubble safely because of its curved profile and blunt edges. The groove along the length of the Bonfadini spatula enables safe and efficient incision or the residual stromal tissue using the pointed end of a sharp blade while protecting the underlying DM. After removal of posterior stroma, the donor button was sutured with 16 interrupted 10-0 nylon sutures. Results: This technique and the use of the Bonfadini spatula and dissector facilitate exposure of Descemet membrane. Conclusion: The smooth Bonfadini DALK spatula and dissector facilitate safe and efficient completion of DALK surgery. .

Objetivo: Descrevemos o uso de novos instrumentais cirúrgicos para facilitar a técnica de "big-bubble" na ceratoplastia lamelar anterior profunda (DALK). Métodos: Paciente masculino, 29 anos, foi diagnosticado com ceratocone bilateral e submetido à ceratoplastia lamelar anterior profunda (DALK). Após incisão da córnea receptora numa profundidade de 350μm de espessura parcial, o dissector Bonfadini foi inserido no ponto mais profundo da incisão periférica e pode avançar para o centro da córnea com segurança devido à sua ponta semiafiada. Depois de realizar a "big-bubble" (BB) e atingir a separação da Membrana de Descemet (MD) do estroma sobrejacente, o disco corneano de estroma anterior foi removido. Um viscoelástico foi colocado sobre o leito do estroma remanescente para impedir o colapso não-controlado e perfuração da MD durante a incisão na BB com lâmina de paracentese. Verificamos segurança no rompimento do estroma remanescente com o auxílio do dissector Bonfadini, para liberação da bolha de ar da BB através do viscoelástico. Depois de injetar o viscoelástico no espaço da BB, inserimos a espátula Bonfadini neste espaço, o que demonstrou-se seguro devido ao formato curvo e das bordas arredondadas do instrumental. A chanfradura ao longo do comprimento da espátula Bonfadini permite a incisão pela ponta de uma lâmina afiada, protegendo assim a MD subjacente. Após a remoção do estroma posterior, o botão doador foi suturado com 16 pontos interrompidos de fio nylon 10.0. Resultados: Esta técnica e o uso da espátula Bonfadini e dissector facilitam a exposição de membrana de Descemet. Conclusão: A superfície lisa da espátula Bonfadini e dissector, facilitam a realização segura e eficiente da ceratoplastia lamelar anterior profunda (DALK). .

Retained host Descemet membrane (Auto-DMET) during conversion of deep anterior lamellar keratoplasty to penetrating keratoplasty: a case report.

PURPOSE: The aim of this study was to relate a case of deep anterior lamellar keratoplasty (DALK) converted to penetrating keratoplasty in which the host central Descemet membrane (DM) with a large perforation was left attached to the host cornea. METHODS: This is a case report of a 34-year-old man who underwent an attempted DALK for keratoconus in his left eye, which became complicated with a large rupture in DM during dissection. The host DM was left in place, the donor cornea with DM intact was sutured onto the host bed, and air was injected into the anterior chamber. The patient was monitored by biomicroscopy, pachymetry, topography, anterior segment optical coherence tomography, and specular microscopy. RESULTS: The postoperative course resulted in improved uncorrected visual acuity, best spectacle-corrected visual acuity, and topography. Corneal edema was observed in the host cornea peripheral to the graft. Three months after the surgery, the endothelial cell density was reduced by 63% compared with the preoperative donor cell density. CONCLUSIONS: Leaving the host DM during conversion of DALK to penetrating keratoplasty was uneventful in this case, although corneal edema was observed in the area overlying the host cornea. It is possible that the retained DM could provide additional autologous endothelial cells to prolong graft survival.