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Sample records for rabbit corneal endothelial

  1. Human Bone Derived Collagen for the Development of an Artificial Corneal Endothelial Graft. In Vivo Results in a Rabbit Model.

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    Natalia Vázquez

    Full Text Available Corneal keratoplasty (penetrating or lamellar using cadaveric human tissue, is nowadays the main treatment for corneal endotelial dysfunctions. However, there is a worldwide shortage of donor corneas available for transplantation and about 53% of the world's population have no access to corneal transplantation. Generating a complete cornea by tissue engineering is still a tough goal, but an endothelial lamellar graft might be an easier task. In this study, we developed a tissue engineered corneal endothelium by culturing human corneal endothelial cells on a human purified type I collagen membrane. Human corneal endothelial cells were cultured from corneal rims after corneal penetrating keratoplasty and type I collagen was isolated from remnant cancellous bone chips. Isolated type I collagen was analyzed by western blot, liquid chromatography -mass spectrometry and quantified using the exponentially modified protein abundance index. Later on, collagen solution was casted at room temperature obtaining an optically transparent and mechanically manageable membrane that supports the growth of human and rabbit corneal endothelial cells which expressed characteristic markers of corneal endothelium: zonula ocluddens-1 and Na+/K+ ATPase. To evaluate the therapeutic efficiency of our artificial endothelial grafts, human purified type I collagen membranes cultured with rabbit corneal endothelial cells were transplanted in New Zealand white rabbits that were kept under a minimal immunosuppression regimen. Transplanted corneas maintained transparency for as long as 6 weeks without obvious edema or immune rejection and maintaining the same endothelial markers that in a healthy cornea. In conclusion, it is possible to develop an artificial human corneal endothelial graft using remnant tissues that are not employed in transplant procedures. This artificial endothelial graft can restore the integrality of corneal endothelium in an experimental model of

  2. [Transplantation of corneal endothelial cells].

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    Amano, Shiro

    2002-12-01

    endothelial cells. Cultured rabbit corneal endothelial cells that endocytosed iron were injected into the anterior chamber of rabbits whose corneal endothelium was cryo-injured, and were pulled to Descemet's membrane by putting a magnet on the eyelid. In these rabbits, corneal edema decreased more quickly than in the control group and no intraocular pressure rise was observed during 8 weeks after the operation, suggesting that the direct delivery of cultured HCECs into the anterior chamber can be an alternative method of choice. The following obstacles should be addressed to make the transplantation of cultured corneal endothelial cells clinically applicable. 1. To reconstruct a cornea that is the same as or superior to the normal cornea, more innovation is necessary in the method of culturing and seeding HCECs. We should consider utilizing HCECs obtained from fetuses after clearing ethical issues. Moreover, we need to develop a method to enhance the cell density and the cell functions. 2. Porcine corneal stroma is promising as a carrier of HCECs instead of human corneal stroma, which is in very limited supply. The usefulness of porcine corneal stroma acellularized to prevent retrovirus infection should be evaluated. 3. To make the self immature cells applicable to corneal transplantation, we should elucidate the corneal endothelial cell specific markers and the factors that are necessary to induce self immature cells to become corneal endothelial cells. 4. The direct delivery of cultured HCECs into the anterior chamber can be an alternative method of choice when its long-term safety is confirmed.

  3. Animal study on transplantation of human umbilical vein endothelial cells for corneal endothelial decompensation

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    Li Cui

    2014-06-01

    Full Text Available AIM: To explore the feasibility of culturing human umbilical vein endothelial cells(HUVECon acellular corneal stroma and performing the posterior lamellar endothelial keratoplasty(PLEKtreating corneal endothelial decompensation.METHODS: Thirty New-Zealand rabbits were divided into three groups randomly, 10 rabbits for experimental group, 10 for stroma group and 10 for control group. Corneal endothelial cells were removed to establish animal model of corneal endothelial failure. PLEK was performed on the rabbits of experimental group and stroma group, and nothing was transplantated onto the rabbits of control group with the deep layer excised only. Postoperative observation was taken for 3mo. The degree of corneal edema and central corneal thickness were recorded for statistical analysis.RESULTS: Corneas in experimental group were relieved in edema obviously compared with that in stroma group and the control group, and showed increased transparency 7d after the operation. The average density of endothelial cells was 2 026.4±129.3cells/mm2, and average central corneal thickness was 505.2±25.4μm in experimental group, while 1 535.6±114.5μm in stroma group and 1 493.5±70.2μm in control group 3mo after operation.CONCLUSION:We achieved preliminary success in our study that culturing HUVEC on acellular corneal stroma and performing PLEK for corneal endothelial decompensation. HUVEC transplanted could survive in vivo, and have normal biological function of keeping cornea transparent. This study provides a new idea and a new way clinically for the treatment of corneal endothelial diseases.

  4. Corona sign: manifestation of peripheral corneal epithelial edema as a possible marker of the progression of corneal endothelial dysfunction.

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    Inoue, Tomoyuki; Hara, Yuko; Kobayashi, Takeshi; Zheng, Xiaodong; Suzuki, Takashi; Shiraishi, Atsushi; Ohashi, Yuichi

    2016-09-01

    To describe a characteristic form of the corona sign and its clinical relevance to the degree of corneal endothelial decompensation and investigate the underlying mechanism using a rabbit model. These observational cases include 31 patients undergoing penetrating keratoplasty (PKP) and 15 patients undergoing Descemet stripping automated endothelial keratoplasty (DSAEK) with special attention to the circumferentially developed corneal epithelial edema. We also conducted a laboratory observation of horizontal water flow in the rabbit cornea. We consistently observed the corona sign at the superior periphery during the initial stage of corneal endothelial decompensation after PKP. With progressive corneal endothelial cellular loss, the epithelial edema gradually expanded circumferentially in the periphery. The endothelial cellular density associated with the corona sign significantly (P corona sign compared with a superior corona sign. After DSAEK, however, the corneal epithelial edema subsided from the center but persisted peripherally as a corona sign in all cases. By 3 months postoperatively, the epithelial edema was confined to the superior periphery along with uneventful corneal endothelial healing. Rabbit experiments showed that total corneal endothelial decompensation decreased the horizontal intracorneal water migration (Inoue-Ohashi phenomenon) in the corneal periphery and induced peripheral corneal edema. The slit-lamp microscopic findings of the corona-like epithelial edema in the peripheral cornea are associated with the stage of corneal endothelial function. To support this, the developmental mechanism of the corona sign was demonstrated experimentally.

  5. Development of a rabbit corneal equivalent using an acellular corneal matrix of a porcine substrate.

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    Xu, Yong-Gen; Xu, Yong-Sheng; Huang, Chen; Feng, Yun; Li, Ying; Wang, Wei

    2008-01-01

    cultured rabbit corneal cells were positive for K3, vimentin, and aquaporin A. CFE and BrdU (5-bromo-2'-deoxyuridine) staining showed no statistical difference. The cultured rabbit corneal limbal epithelial cells, keratocyte cells, and endothelial cells formed a confluent cell sheet on the ACMP, which consisted of one to two cell layers. Immunofluorescence and scanning electron microscopy examination showed that the cells steadily adhered to the surface of the ACMP and maintained their conformation and special molecule expression such as K3, vimentin, and aquaporin A. Rabbit corneal epithelium-ACMP, keratocytes-ACMP, and endothelium-ACMP scaffold was built in vitro. The rabbit corneal scaffold was made by the ACMP as a frame with three types of allogeneic rabbit corneal cells. This is a new concept in treating injured corneas.

  6. Gamma-irradiated sterile cornea for use in corneal transplants in a rabbit model

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    Junko Yoshida; Thomas Heflin; Andrea Zambrano; Qing Pan; Huan Meng; Jiangxia Wang; Stark, Walter J.; Daoud, Yassine J.

    2015-01-01

    Purpose: Gamma irradiated corneas in which the donor keratocytes and endothelial cells are eliminated are effective as corneal lamellar and glaucoma patch grafts. In addition, gamma irradiation causes collagen cross inking, which stiffens collagen fibrils. This study evaluated gamma irradiated corneas for use in corneal transplantations in a rabbit model comparing graft clarity, corneal neovascularization, and edema. Methods: Penetrating keratoplasty was performed on rabbits using four ty...

  7. TSG-6 protects corneal endothelium from transcorneal cryoinjury in rabbits.

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    Kim, Jeong-Ah; Ko, Jung Hwa; Ko, Ah Young; Lee, Hyun Ju; Kim, Mee Kum; Wee, Won Ryang; Lee, Ryang Hwa; Fulcher, Samuel F; Oh, Joo Youn

    2014-07-17

    To investigate the effect of an anti-inflammatory protein, TNF-α stimulated gene/protein (TSG)-6 and an antiapoptotic protein, stanniocalcin (STC)-1 on corneal endothelium in rabbits with transcorneal cryoinjury. Transcorneal freezing (-80°C) was applied to rabbit corneas for 30 seconds. Immediately post injury, either TSG-6 (10 μg/100 μL), STC-1 (10 μg/100 μL), or the same volume of balanced salt solution (BSS) was injected into the anterior chamber. Each eye was examined for corneal opacity, corneal thickness, endothelial cell density, and endothelial hexagonality every 2 to 6 hours for 48 hours post injury. The concentrations of myeloperoxidase (MPO) and IL-1β were measured in the aqueous humor every 6 hours. At 48 hours post injury, each cornea was assayed for TNF-α, IL-1β, IL-6, and MPO, and histologically evaluated with alizarin red-trypan blue staining, hematoxylin-eosin staining, and immunostaining for neutrophils. Tumor necrosis factor-α stimulated gene/protein-6 significantly decreased the development of corneal opacity and edema after cryoinjury compared with STC-1 or BSS. The corneal endothelial cell density and hexagonality were markedly preserved by TSG-6. The mRNA levels of TNF-α, IL-1β, and IL-6 in the cornea and the protein levels of MPO and IL-1β in the aqueous humor and cornea were significantly lower in TSG-6-treated eyes than BSS-treated controls. Similarly, the expression of fibroblast growth factor-2 was reduced by TSG-6 treatment. Histologic evaluation demonstrated that neutrophil infiltration of the cornea was decreased in TSG-6-treated eyes. Tumor necrosis factor-α stimulated gene/protein-6 protected corneal endothelial cells from transcorneal cryoinjury through suppression of inflammation. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

  8. Molecular expression in transfected corneal endothelial cells

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    Wang, Fan; Miao, Zhuang; Lu, Chengwei; Hao, Jilong

    2017-10-01

    To investigate the capability of human corneal endothelial cells serving as immunological cells. Expression of HLA-DP, -DQ, -DR, CD40, CD80, and CD86 was determined by immunohistochemical methods. Meanwhile, purified peripheral blood mononuclear cells were cocultured with human corneal endothelial cells which were pre-treated with and without -IFN respectively, activation of lymphocytes was determined by FACS analysis. In coculture system, T lymphocyte was activated by corneal endothelial cells, HLA-DP, -DQ, -DR and CD40 expression were increased by - IFN induction. Costimulatory molecular CD80 was shown on the endothelial cells. Human corneal endothelial cells were assumed to be involved in the corneal transplantation rejection process as potential antigen presenting cells.

  9. Corneal donor tissue preparation for endothelial keratoplasty.

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    Woodward, Maria A; Titus, Michael; Mavin, Kyle; Shtein, Roni M

    2012-06-12

    Over the past ten years, corneal transplantation surgical techniques have undergone revolutionary changes. Since its inception, traditional full thickness corneal transplantation has been the treatment to restore sight in those limited by corneal disease. Some disadvantages to this approach include a high degree of post-operative astigmatism, lack of predictable refractive outcome, and disturbance to the ocular surface. The development of Descemet's stripping endothelial keratoplasty (DSEK), transplanting only the posterior corneal stroma, Descemet's membrane, and endothelium, has dramatically changed treatment of corneal endothelial disease. DSEK is performed through a smaller incision; this technique avoids 'open sky' surgery with its risk of hemorrhage or expulsion, decreases the incidence of postoperative wound dehiscence, reduces unpredictable refractive outcomes, and may decrease the rate of transplant rejection. Initially, cornea donor posterior lamellar dissection for DSEK was performed manually resulting in variable graft thickness and damage to the delicate corneal endothelial tissue during tissue processing. Automated lamellar dissection (Descemet's stripping automated endothelial keratoplasty, DSAEK) was developed to address these issues. Automated dissection utilizes the same technology as LASIK corneal flap creation with a mechanical microkeratome blade that helps to create uniform and thin tissue grafts for DSAEK surgery with minimal corneal endothelial cell loss in tissue processing. Eye banks have been providing full thickness corneas for surgical transplantation for many years. In 2006, eye banks began to develop methodologies for supplying precut corneal tissue for endothelial keratoplasty. With the input of corneal surgeons, eye banks have developed thorough protocols to safely and effectively prepare posterior lamellar tissue for DSAEK surgery. This can be performed preoperatively at the eye bank. Research shows no significant difference in

  10. Corneal Donor Tissue Preparation for Endothelial Keratoplasty

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    Woodward, Maria A.; Titus, Michael; Mavin, Kyle; Shtein, Roni M.

    2012-01-01

    Over the past ten years, corneal transplantation surgical techniques have undergone revolutionary changes1,2. Since its inception, traditional full thickness corneal transplantation has been the treatment to restore sight in those limited by corneal disease. Some disadvantages to this approach include a high degree of post-operative astigmatism, lack of predictable refractive outcome, and disturbance to the ocular surface. The development of Descemet's stripping endothelial keratoplasty (DSEK...

  11. Cataract phacoemulsification and corneal endothelial cell damage

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    Ni Zhu

    2013-07-01

    Full Text Available Phacoemulsification with small incision, reduced number of inflammation cells, and better postoperative recovery has been recognized as the world's most popular option for cataract surgery. Modern cataract surgery is developing gradually from sight rehabilitating to refractive surgery with better vision acuity. Being the most important part of the eye refractive system, maintenance of the cornea's transparency relies heavily upon the healthy endothelial cells. It is well known that there will be endothelial cell loss after phacoemulsification and the damage of the endothelial cells may lead to corneal swellings and opacity, or even the corneal descompensation, which often severely influenced the postoperative vision recovery. This is a review of phacoemulsification and the risk factors of corneal endothelial damage pre-and postoperation.

  12. Effects of argon laser iridotomy on the corneal endothelium of pigmented rabbit eyes.

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    Youm, Jie Hyun; Heo, Jeong-Hwa; Kim, Hyo Myung; Song, Jong-Suk

    2014-02-01

    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. 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. 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 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. Argon laser iridotomy induces corneal endothelial cell apoptosis in pigmented rabbit eyes, resulting in decreased endothelial cell density.

  13. Phototoxic effects of 8-methoxypsoralen on rabbit corneal endothelium

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    Menon, I.A.; Basu, P.K.; Hasany, S.M.; Persad, S.D. (Univ. of Toronto, Ontario (Canada))

    1989-01-01

    The phototoxic effects of 8-methoxypsoralen (8-MOP) were investigated using the rabbit corneal endothelium in organ culture. The corneas were divided into four groups: (a) irradiated with a mercury vapor lamp (emitting UVA and visible radiation) in the presence of 8-MOP (experimental), (b) irradiated without 8-MOP (control A), (c) incubated with 8-MOP (control B) and (d) incubated without 8-MOP (control C). Specular and light microscopic examination showed that the experimental corneas had greater cellular damage compared to the control corneas. The effects of 8-MOP were restricted to certain localized areas of the cornea. However there was no significant difference in the amounts of 51Cr released from the labelled experimental and control corneas. These results show phototoxic damage of the corneal endothelial cells.

  14. Corneal endothelial cytotoxicity of the Calotropis procera (ushaar) plant.

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    Al-Mezaine, Hani S; Al-Amry, Mohammed A; Al-Assiri, Abdullah; Fadel, Talal S; Tabbara, Khalid F; Al-Rajhi, Ali A

    2008-05-01

    To report 6 eyes of 5 patients with transient corneal edema after exposure to the milky latex of Calotropis procera (ushaar). Interventional case series. Intracorneal penetration of ushaar latex can lead to permanent endothelial cell loss with morphologic alteration. Corneal edema resolved completely in approximately 2 weeks in all cases, despite reduced endothelial cell count and abnormal morphology. Corneal endothelial toxicity of ushaar latex is caused by its ability to penetrate the corneal stroma and induce permanent loss of endothelial cells. Corneal edema resolves if sufficient endothelial cell viability is still present after resolution of ushaar keratitis.

  15. Investigation of Overrun-Processed Porous Hyaluronic Acid Carriers in Corneal Endothelial Tissue Engineering.

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    Jui-Yang Lai

    Full Text Available Hyaluronic acid (HA is a linear polysaccharide naturally found in the eye and therefore is one of the most promising biomaterials for corneal endothelial regenerative medicine. This study reports, for the first time, the development of overrun-processed porous HA hydrogels for corneal endothelial cell (CEC sheet transplantation and tissue engineering applications. The hydrogel carriers were characterized to examine their structures and functions. Evaluations of carbodiimide cross-linked air-dried and freeze-dried HA samples were conducted simultaneously for comparison. The results indicated that during the fabrication of freeze-dried HA discs, a technique of introducing gas bubbles in the aqueous biopolymer solutions can be used to enlarge pore structure and prevent dense surface skin formation. Among all the groups studied, the overrun-processed porous HA carriers show the greatest biological stability, the highest freezable water content and glucose permeability, and the minimized adverse effects on ionic pump function of rabbit CECs. After transfer and attachment of bioengineered CEC sheets to the overrun-processed HA hydrogel carriers, the therapeutic efficacy of cell/biopolymer constructs was tested using a rabbit model with corneal endothelial dysfunction. Clinical observations including slit-lamp biomicroscopy, specular microscopy, and corneal thickness measurements showed that the construct implants can regenerate corneal endothelium and restore corneal transparency at 4 weeks postoperatively. Our findings suggest that cell sheet transplantation using overrun-processed porous HA hydrogels offers a new way to reconstruct the posterior corneal surface and improve endothelial tissue function.

  16. Substrates for Expansion of Corneal Endothelial Cells towards Bioengineering of Human Corneal Endothelium

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    Jesintha Navaratnam

    2015-09-01

    Full Text Available Corneal endothelium is a single layer of specialized cells that lines the posterior surface of cornea and maintains corneal hydration and corneal transparency essential for vision. Currently, transplantation is the only therapeutic option for diseases affecting the corneal endothelium. Transplantation of corneal endothelium, called endothelial keratoplasty, is widely used for corneal endothelial diseases. However, corneal transplantation is limited by global donor shortage. Therefore, there is a need to overcome the deficiency of sufficient donor corneal tissue. New approaches are being explored to engineer corneal tissues such that sufficient amount of corneal endothelium becomes available to offset the present shortage of functional cornea. Although human corneal endothelial cells have limited proliferative capacity in vivo, several laboratories have been successful in in vitro expansion of human corneal endothelial cells. Here we provide a comprehensive analysis of different substrates employed for in vitro cultivation of human corneal endothelial cells. Advances and emerging challenges with ex vivo cultured corneal endothelial layer for the ultimate goal of therapeutic replacement of dysfunctional corneal endothelium in humans with functional corneal endothelium are also presented.

  17. Toxicity of intrastromal voriconazole injection on corneal endothelium in rabbits.

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    Park, Chang Hyun; Lee, Hyun Soo; Chung, Sung Kun

    2014-09-01

    The aim of this study was to evaluate the toxicity of intrastromally injected voriconazole on corneal endothelial cells in rabbits. In total, 32 eyes of 16 rabbits (8 eyes for each group) were divided into 4 groups according to the concentration of voriconazole [group A: 50 μg/0.1 mL (0.05%), group B: 100 μg/0.1 mL (0.1%), group C: 250 μg/0.1 mL (0.25%), or group D: 500 μg/0.1 mL (0.5%)]. Right eyes were injected intrastromally with voriconazole at concentrations of 50 μg/0.1 mL, 100 μg/0.1 mL, 250 μg/0.1 mL, or 500 μg/0.1 mL. Left eyes were injected intrastromally with isotonic saline as controls. Central corneal thickness and endothelial cell counts were measured before and at 6 hours, 1 day, and 1 week after the injection was given. Corneas were then harvested for transmission electron microscopy. Only the 0.5% group did not significantly recover from corneal edema 1 week after the injection (P = 0.167, P = 0.051, P = 0.086, P = 0.001 in groups A-D, respectively). There were significant differences in endothelial cell counts for the 0.1% and 0.25% groups (P = 0.077, P = 0.019, P = 0.008 in groups A-C, respectively). Transmission electron microscopy evaluation revealed definite necrotic changes in endothelial cells at concentrations of 0.1%, 0.25%, and 0.5%, but only microstructural changes at a concentration of 0.05%. Our results suggest that voriconazole could be injected safely into the intrastromal layer at a concentration of 0.05% with low endothelial cell toxicity. However, injections should be administered with caution because of the risk of microstructural damage.

  18. Heterologous corneal endothelial cell transplantation--human corneal endothelial cell transplantation in Lewis rats.

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    Tchah, H.

    1992-01-01

    A heterologous corneal endothelial transplantation was attempted using human endothelial cells and a Lewis rat penetrating keratoplasty model. Cultured human endothelial cells were seeded to a Lewis rat cornea, which was denuded of its endothelium. When grafted into the syngeneic Lewis rat, the graft remained clear for at least five days, and then became opaque and edematous because of immune rejection reaction. In contrast, corneas denuded of their endothelium became opaque and edematous imm...

  19. Volumetric and ionic regulation during the in vitro development of a corneal endothelial barrier.

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    Alaminos, M; González-Andrades, M; Muñoz-Avila, J I; Garzón, I; Sánchez-Quevedo, M C; Campos, A

    2008-05-01

    Corneal endothelium is responsible for generating an ion flux between the corneal stroma and the anterior chamber of the eye that is necessary for the cornea to remain transparent. However, the ion transport regulatory mechanisms that develop during the formation of the endothelial barrier are not known. In this study, we determined the influence of cell confluence on cell volume and intracellular ionic content on the corneal endothelial cells of rabbits. Our results demonstrate that non-confluent endothelial cells display a hypertrophic volume increase, with higher intracellular contents of potassium and chlorine than those of confluent cells. In contrast, when cells reach confluence and the endothelial barrier forms, cell volume decreases and the intracellular contents of potassium and chlorine decrease. Our genetic analysis showed a higher expression of CFTR and CA2 genes in non-confluent cells, and of the gene KCNC3 in confluent cells. These results suggest that the normal ionic current that keeps the corneal stroma dehydrated and transparent is regulated by cell-cell contacts and endothelial cell confluence, and could explain why the loss of corneal endothelial cells is often associated with corneal edema and even blindness.

  20. [Mitomycin C toxicity in rabbit corneal endothelium].

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    Silva, Maria Rosa Bet de Moraes; Gregório, Elisa Aparecida

    2009-01-01

    To evaluate corneal endothelium alterations after applying mitomycin C to the sclera using transmission and scanning electron microscopy, correlating alterations with time, concentration, and evaluation methods. The corneal endothelium of both eyes of 32 albino rabbits was evaluated and distributed into four groups of 8. Mitomycin C was applied under a scleral flap in the right eye for 5 minutes. Mitomycin C concentrations were 0.5 mg/ml for G1 and G2 and 0.2 mg/ml for G3 and G4. Examinations were performed 15 days after application to G1 and G3, and 30 days after application to G2 and G4. Four cornea in each group were prepared for transmission electron microscopy and four for scanning electron microscopy. Left eyes of all animals were used as controls. Transmission electron microscopy showed corneal endothelium alterations in all groups: rarefied cytoplasm, dilation and fragmentation of rough endoplasmic reticulum cisternae, Golgi apparatus with cisternal dilation, reduced vacuoles, and irregularities of internal membrane more noticeable in G1 and G2. Scanning electron microscopy revealed alterations in all groups except G1: changes in the shape and size of cells and longer filopodial projections. 1 - Corneal endothelium alterations were seen at both 0.5 and 0.2 mg/ml concentrations and at 15 and 30 days after mytomicin C application; 2 - Alterations were more intense with higher mytomicin C concentration by transmission electron but not by scanning electron microscopy; 3 - The alterations correlated with time by scanning electron microscopy but not by transmission electron microscopy.

  1. Long-Term Topical Ganciclovir and Corticosteroids Preserve Corneal Endothelial Function in Cytomegalovirus Corneal Endotheliitis.

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    Fan, Nai-Wen; Chung, Yu-Chien; Liu, Yao-Chung; Liu, Catherine Jui-Ling; Kuo, Yih-Shiuan; Lin, Pei-Yu

    2016-05-01

    To report the long-term outcomes of topical ganciclovir (GCV) and corticosteroids as a maintenance therapy for cytomegalovirus (CMV) corneal endotheliitis. This retrospective study included 10 eyes of 9 patients diagnosed with CMV corneal endotheliitis with a minimum 1-year follow-up at a tertiary referral hospital between 2008 and 2014. CMV corneal endotheliitis was defined by corneal edema associated with typical keratic precipitates (KPs) and a positive CMV polymerase chain reaction from aqueous humor taps. Patients receiving long-term topical 0.5% GCV and topical corticosteroids without discontinuation were included. The final corneal condition and endothelial cell density (ECD) were reported. The mean age was 45.6 ± 11.7 years. The mean follow-up duration was 48 ± 25 months. All patients exhibited typical coin-shaped and/or linear KPs. A significant resolution of corneal edema and decreased KPs were achieved within 1 month in all patients after initiating topical 0.5% GCV every 2 hours and topical corticosteroids twice a day. The dose frequency was gradually tapered to GCV 4 times and corticosteroids once or twice a day as a maintenance therapy. All 10 eyes had a clear graft or corneas at the end of this study. The mean ECD was 1630 ± 699 cells per millimeter square before treatment and 1776 ± 834 cells per millimeter square at the end of the study period. Topical 0.5% GCV and corticosteroids as a maintenance regimen without interruption effectively preserved long-term corneal endothelial function.

  2. Diurnal rhythm of mitosis in rabbit corneal epithelium.

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    Fogle, J A; Yoza, B K; Neufeld, A H

    1980-01-01

    Incorporation of 3H-thymidine by rabbit corneal epithelium during the course of a one-hour incubation in vitro varies according to the time of day, suggesting a diurnal rhythm of mitotic activity. Adrenergic decentralization of the cornea does not affect this rhythm. Furthermore, there is no diurnal variation in the basal or sympathomimetically-stimulated cyclic AMP production by freshly excised rabbit corneas, incubated in vitro. Therefore, the diurnal rhythm of corneal epithelial mitotsis in the rabbit is not paced by catecholamines.

  3. Role of Corneal Epithelium in Riboflavin/Ultraviolet-A Mediated Corneal Cross-Linking Treatment in Rabbit Eyes

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    Xiangchen Tao; Haiqun Yu; Yong Zhang; Zhiwei Li; Vishal Jhanji; Shouxiang Ni; Ya Wang; Guoying Mu

    2013-01-01

    Purpose. To evaluate the role of corneal epithelium in riboflavin/ultraviolet-A (UVA) mediated corneal collagen cross-linking treatment. Methods. Fifty New Zealand rabbits were divided into 5 groups: UVA treatment with or without corneal epithelium, UVA+riboflavin treatment with or without corneal epithelium, and control without any treatment. All rabbits were sacrificed after irradiation and subsequently 4?mm???10?mm corneal strips were harvested for biomechanical evaluation. Results. UVA ir...

  4. Preliminary studies of rabbit corneal structure using laser beams

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    Carstocea, Benone D.; Gafencu, Otilia L.; Apostol, Silvia; Ionita, Marcel A.; Pascu, Mihail-Lucian; Moise, N.; Popescu, Gheorghe; Roman, Miruna; Hogiu, S.

    1996-12-01

    A new method for corneal structure investigation is reported. The light reflected form rabbit cornea is recorded by a video-camera so that the images of epithelium and endothelium are obtained. Because of the great magnification factor due to the corneal curvature, the characteristics of the cells can be directly visualized. The theoretical evaluation of the optical magnification fits reasonably well the experimental results for collimated laser beams; therefore in the future it will be possible to determine with a good accuracy the corneal cell dimensions. Corneal diagnosis based on this method is discussed.

  5. Does corneal hysteresis correlate with endothelial cell density?

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    Akova-Budak, Berna; Kıvanç, Sertaç Argun

    2015-05-21

    Our aim was to determine if there is a correlation between corneal biomechanical properties, endothelial cell count, and corneal pachymetry in healthy corneas. Ninety-two eyes of all subjects underwent complete ocular examination, including intraocular pressure measurement by Goldmann applanation tonometer, objective refraction, and slit-lamp biomicroscopy. Topographic measurements and corneal pachymetry were performed using a Scheimpflug-based (Pentacam, Oculus, Germany) corneal topographer. Corneal hysteresis (CH) and corneal resistance factor (CRF) were measured with an Ocular Response Analyzer (ORA, Reichert Ophthalmic Instruments, Buffalo, NY). Endothelial cell count measurement was done using a specular microscope (CellChek, Konan, USA). Right eye values of the subjects were taken for the study. The mean CH was 11.5±1.7 mmHg and the mean CRF was 11.2±1.4 mmHg. Mean intraocular pressure was 15.3±2.3 mmHg. The mean endothelial cell count was 2754±205 cells/mm2. No correlation was found between biomechanical properties of cornea and endothelial cell count. There was a significant positive correlation between CH, CRF, and corneal thickness (pcorneal biomechanical properties significantly correlated with corneal thickness. We found no correlation between CH and CRF with the endothelial cell density in normal subjects.

  6. Inhibitory effect of polysulfated heparin endostatin onalkali burn induced corneal neovascularization in rabbits

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    Zhao-Na Li

    2015-04-01

    Full Text Available AIM: To investigate anti-angiogenic effects of polysulfated heparin endostatin (PSH-ES on alkali burn induced corneal neovascularization (NV in rabbits. METHODS: An alkali burn was made on rabbit corneas to induce corneal NV in the right eye of 24 rabbits. One day after burn creation, a 0.2 mL subconjunctival injection of 50 μg/mL PSH-ES, 50 μg/mL recombinant endostatin (ES, or normal saline was administered every other day for a total of 14d (7 injections. Histology and immunohistochemisty were used to examine corneas. Corneal NV growth was evaluated as microvessel quantity and corneal vascular endothelial growth factor (VEGF expression was measured by immunohistochemical assay. RESULTS: Subconjunctival injection of ES and PSH-ES resulted in significant corneal NV suppression, but PSH-ES had a more powerful anti-angiogenic effect than ES. Mean VEGF concentration in PSH-ES treated corneas was significantly lower than in ES treated and saline treated corneas. Histological examination showed that corneas treated with either PSH-ES or ES had significantly fewer microvessels than eyes treated with saline. Additionally corneas treated with PSH-ES had significantly fewer microvessels than corneas treated with ES. CONCLUSION: Both PSH-ES and recombinant ES effectively inhibit corneal NV induced by alkali burn. However, PSH-ES is a more powerful anti-angiogenic agent than ES. This research has the potential to provide a new treatment option for preventing and treating corneal NV.

  7. Localization of angiotensin converting enzyme in rabbit cornea and its role in controlling corneal angiogenesis in vivo.

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    Sharma, Ajay; Bettis, Daniel I; Cowden, John W; Mohan, Rajiv R

    2010-04-23

    The renin angiotensin system (RAS) has been shown to modulate vascular endothelial growth factor and angiogenesis. In this study we investigated (i) the existence of the RAS components angiotensin converting enzyme (ACE) and angiotensin II receptors (AT(1) and AT(2)) in the rabbit cornea using in vitro and ex vivo models and (ii) the effect of enalapril, an ACE inhibitor, to inhibit angiogenesis in rabbit cornea in vivo. New Zealand White rabbits were used. Cultured corneal fibroblasts and corneal epithelial cells were used for RNA isolation and cDNA preparation using standard molecular biology techniques. PCR was performed to detect the presence of ACE, AT(1), and AT(2) gene expression. A corneal micropocket assay to implant a vascular endothelial growth factor (VEGF) pellet in the rabbit cornea was used to induce corneal angiogenesis. Rabbits of the control group received sterile water, and the treated group received 3 mg/kg enalapril intramuscularly once daily for 14 days starting from day 1 of pellet implantation. The clinical eye examination was performed by slit-lamp biomicroscopy. We monitored the level of corneal angiogenesis in live animals by stereomicroscopy at days 4, 9, and 14 after VEGF pellet implantation. Collagen type IV and lectin immunohistochemistry and fluorescent microscopy were used to measure corneal angiogenesis in tissue sections of control and enalapril-treated corneas of the rabbits. Image J software was used to quantify corneal angiogenesis in the rabbit eye in situ. Our data demonstrated the presence of ACE, AT(1), and AT(2) expression in corneal fibroblasts. Cells of corneal epithelium expressed AT(1) and AT(2) but did not show ACE expression. Slit-lamp examination did not show any significant difference between the degree of edema or cellular infiltration between the corneas of control and enalapril-treated rabbits. VEGF pellet implantation caused corneal angiogenesis in the eyes of vehicle-treated control rabbits, and the mean area

  8. Gamma-Irradiated Sterile Cornea for Use in Corneal Transplants in a Rabbit Model.

    Science.gov (United States)

    Yoshida, Junko; Heflin, Thomas; Zambrano, Andrea; Pan, Qing; Meng, Huan; Wang, Jiangxia; Stark, Walter J; Daoud, Yassine J

    2015-01-01

    Gamma irradiated corneas in which the donor keratocytes and endothelial cells are eliminated are effective as corneal lamellar and glaucoma patch grafts. In addition, gamma irradiation causes collagen cross inking, which stiffens collagen fibrils. This study evaluated gamma irradiated corneas for use in corneal transplantations in a rabbit model comparing graft clarity, corneal neovascularization, and edema. Penetrating keratoplasty was performed on rabbits using four types of corneal grafts: Fresh cornea with endothelium, gamma irradiated cornea, cryopreserved cornea, and fresh cornea without endothelium. Slit lamp examination was performed at postoperative week (POW) one, two, and four. Corneal clarity, edema, and vascularization were graded. Confocal microscopy and histopathological evaluation were performed. A P cornea with endothelium compared to the other three groups (P cornea scored better than the cryopreserved and fresh cornea without endothelium groups in clarity (0.9 vs. 1.5 and 2.6, respectively), and edema (0.6 vs. 0.8 and 2.0, respectively). The gamma irradiated corneas, cryopreserved corneas and the fresh corneas without endothelium, developed haze and edema after POW 2. Gamma irradiated cornea remained statistically significantly clearer than cryopreserved and fresh cornea without endothelium during the observation period (P cornea. Gamma irradiated corneas remained clearer and thinner than the cryopreserved cornea and fresh cornea without endothelium. However, this outcome is transient. Gamma irradiated corneas are useful for lamellar and patch grafts, but cannot be used for penetrating keratoplasty.

  9. Subacute effects of rose Bengal/Green light cross linking on rabbit thin corneal stability and safety.

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    Wang, Ti; Zhu, Lu; Zhu, Jingyin; Peng, Yinbo; Shen, Nianci; Yu, Yan; Yao, Min

    2017-11-02

    The purpose of this study was to investigate the subacute effects of Rose Bengal (RB) and 532 nm green light-induced photochemical crosslinking (RB-PCL) on rabbit thin corneal stability and safety in vivo. Rabbit thin corneal models with 250 μm thickness were created by photorefractive keratectomy surgery. Photochemical crosslinking with green light (wavelength 532 nm) at an illumination intensity of 0.4 W/cm2 for 250 s (100 J/cm2 ) was performed, followed by antibiotic treatment and slit lamp monitoring for four weeks. At the end of week four, corneal biomechanical stiffness, biochemical resistance to collagenase digestion, and corneal cellular morphology were assessed. The penetration depth of RB into the corneal stromal was measured by confocal microscopy. At the end of week 4, RB-PCL had increased corneal tensile strength by an average 2.5-fold and had extended the corneal collagenase digestion time from 10.17 ± 2.93 to 15.83 ± 2.64 days. RB penetrated approximately 90 µm into the corneal stroma. RB-PCL did not alter the corneal endothelial and stromal morphology at the cellular or subcellular levels, according to electron microscopic examination. RB and 532 nm green light irradiation effectively induced crosslinking in rabbit thin cornea, by increasing both the biomechanical stiffness and the biochemical resistance without evidence of morphological damage to the corneal endothelium or stroma. This study demonstrated the efficacy of RB-PCL in strengthening thin cornea at four weeks after the treatment, providing a potential and possibly better option for treating corneal ectasia disorders in cases where corneal thickness is less than 400 µm. Lasers Surg. Med. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  10. Variety of corneal endothelial cell in glaucoma by confocal microscope

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    Hong-Liang Gao

    2014-10-01

    Full Text Available ATM: To define the causes of corneal endothelial cell damage, to investigate the preventive methods, and to observe the variety of corneal endothelial cell in glaucoma using confocal microscope.METHODS: Totally, 143 eyes of 97 patients with different types of glaucoma, and matched normal people were 20 cases, all 40 eyes. The cell density, cell area and cell variable coefficient were measured used confocal microscope. These indicatives of every kind of glaucoma were compared.RESULTS: The corneal endothelial cell density of normal group was 2 893.88±255.026/mm2, the group of acute angle-closure glaucoma(AACGwas 1 674.11±683.95/mm2, and the group of open angle glaucoma(OAGwas 2 687.22±391.87/mm2, the group of chronic angle-closure glaucoma(CACGwas 2 706.97±351.27/mm2. In all index the average cell density of corneal endothelial and the average area have statistical significance(F=62.950, 8.795; P=0.000, especially the group of AACG. CONCLUSION: The index of corneal endothelial cell in AACG is lower than that of normal. All index in OAG and CACG is difference with that of normal, but the difference has no statistical significance. And the dominant factor of damaged corneal endothelial is the time of intraocular hypertension.

  11. Generation and Feasibility Assessment of a New Vehicle for Cell-Based Therapy for Treating Corneal Endothelial Dysfunction.

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    Naoki Okumura

    Full Text Available The corneal endothelium maintains corneal transparency by its pump and barrier functions; consequently, its decompensation due to any pathological reason causes severe vision loss due to corneal haziness. Corneal transplantation is the only therapeutic choice for treating corneal endothelial dysfunction, but associated problems, such as a shortages of donor corneas, the difficulty of the surgical procedure, and graft failure, still need to be resolved. Regenerative medicine is attractive to researchers as a means of providing innovative therapies for corneal endothelial dysfunction, as it now does for other diseases. We previously demonstrated the successful regeneration of corneal endothelium in animal models by injecting cultured corneal endothelial cells (CECs in combination with a Rho kinase (ROCK inhibitor. The purpose of the present study was to optimize the vehicle for clinical use in cell-based therapy. Our screening of cell culture media revealed that RELAR medium promoted CEC adhesion. We then modified RELAR medium by removing hormones, growth factors, and potentially toxic materials to generate a cell therapy vehicle (CTV composed of amino acid, salts, glucose, and vitamins. Injection of CECs in CTV enabled efficient engraftment and regeneration of the corneal endothelium in the rabbit corneal endothelial dysfunction model, with restoration of a transparent cornea. The CECs retained >85% viability after a 24 hour preservation as a cell suspension in CTV at 4°C and maintained their potency to regenerate the corneal endothelium in vivo. The vehicle developed here is clinically applicable for cell-based therapy aimed at treating the corneal endothelium. Our strategy involves the generation of vehicle from a culture medium appropriate for a given cell type by removing materials that are not favorable for clinical use.

  12. Expansion and cryopreservation of porcine and human corneal endothelial cells.

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    Marquez-Curtis, Leah A; McGann, Locksley E; Elliott, Janet A W

    2017-08-01

    Impairment of the corneal endothelium causes blindness that afflicts millions worldwide and constitutes the most often cited indication for corneal transplants. The scarcity of donor corneas has prompted the alternative use of tissue-engineered grafts which requires the ex vivo expansion and cryopreservation of corneal endothelial cells. The aims of this study are to culture and identify the conditions that will yield viable and functional corneal endothelial cells after cryopreservation. Previously, using human umbilical vein endothelial cells (HUVECs), we employed a systematic approach to optimize the post-thaw recovery of cells with high membrane integrity and functionality. Here, we investigated whether improved protocols for HUVECs translate to the cryopreservation of corneal endothelial cells, despite the differences in function and embryonic origin of these cell types. First, we isolated endothelial cells from pig corneas and then applied an interrupted slow cooling protocol in the presence of dimethyl sulfoxide (Me 2 SO), with or without hydroxyethyl starch (HES). Next, we isolated and expanded endothelial cells from human corneas and applied the best protocol verified using porcine cells. We found that slow cooling at 1 °C/min in the presence of 5% Me 2 SO and 6% HES, followed by rapid thawing after liquid nitrogen storage, yields membrane-intact cells that could form monolayers expressing the tight junction marker ZO-1 and cytoskeleton F-actin, and could form tubes in reconstituted basement membrane matrix. Thus, we show that a cryopreservation protocol optimized for HUVECs can be applied successfully to corneal endothelial cells, and this could provide a means to address the need for off-the-shelf cryopreserved cells for corneal tissue engineering and regenerative medicine. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  13. Biosynthetic corneal implants for replacement of pathologic corneal tissue : performance in a controlled rabbit alkali burn model

    OpenAIRE

    Joanne M. Hackett; Lagali, Neil; Merrett, Kimberley; Edelhauser, Henry; Sun, Yifei; Gan, Lisha; Griffith, May; Fagerholm, Per

    2011-01-01

    Purpose: To evaluate the performance of structurally reinforced, stabilized recombinant human collagen-phosphorylcholine (RHCIII-MPC) hydrogels as corneal substitutes in a rabbit model of severe corneal damage. Methods: One eye each of 12 rabbits received a deep corneal alkali wound. Four corneas were implanted with RHCIII-MPC hydrogels. The other eight control corneas were implanted with either allografts or a simple crosslinked RHCIII hydrogel. In all cases, 6.25 mm diameter, 350 µm thick b...

  14. CMV endotheliitis: a cause for recurrent failed corneal transplant

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    Shahrudin, Nurul Ain

    2017-12-01

    Full Text Available Objective: To highlight the clinical presentation of CMV endotheliitis and the challenge in diagnosing this condition in recurrent failed penetrating keratoplasty (PK. Methods: Case seriesResults: There are 3 cases of recurrent failure in PK secondary to CMV endotheliitis presented. Case 1 and 2 were pseudophakic patients, while in case 3, the patient had a previous history of recurrent anterior uveitis. Case 1 and 3 had four and one previous failed PK respectively, while case 2 had endothelial keratoplasty twice before the diagnosis of CMV endotheliitis was made, following positive culture of aqueous humour. The visual acuity ranged from 1/60 to hand movement. All patients had pigmented KP, and two of them had typical coin-shaped KP. Oral valganciclovir was instituted for all patients consisting of bidaily for two weeks, followed by 900 mg once daily for six months. Additionally, topical ganciclovir eyedrop 0.5% was given every four hours with topical dexaminim four times a day. Repeated anterior chamber (AC tap after six months of treatment was negative for CMV in case 3 while cases 1 and 2 are still on treatment. CMV endotheliitis is an increasingly important cause of failed corneal transplant. We recommend anterior chamber tap in suspicious cases of repeatedly failed corneal transplant, regardless of the presence of coin-shaped KP or not. Minim treatment with oral valganciclovir is important to eradicate the problem, before proceeding with another corneal transplant.Conclusion: It is important to make an accurate early diagnosis by good clinical judgement in preventing loss of corneal endothelial cells. High index of suspicion for CMV endotheliitis as a cause of graft failure must be made especially when the patient presents with coin-shaped KP. Therefore unnecessary treatment resulting from misdiagnosis in these patients can be prevented. Early recognition and treatment of this condition is important to prevent permanent endothelial cell

  15. 3D map of the human corneal endothelial cell

    OpenAIRE

    Zhiguo He; Fabien Forest; Philippe Gain; Damien Rageade; Aurélien Bernard; Sophie Acquart; Michel Peoc’h; Dennis M. Defoe; Gilles Thuret

    2016-01-01

    Corneal endothelial cells (CECs) are terminally differentiated cells, specialized in regulating corneal hydration and transparency. They are highly polarized flat cells that separate the cornea from the aqueous humor. Their apical surface, in contact with aqueous humor is hexagonal, whereas their basal surface is irregular. We characterized the structure of human CECs in 3D using confocal microscopy of immunostained whole corneas in which cells and their interrelationships remain intact. Hexa...

  16. [Corneal endothelial decompensation of iridocorneal endothelial syndrome treated by penetrating keratoplasty].

    Science.gov (United States)

    Chen, J; Liu, Z; Yu, L

    1996-07-01

    To evaluate the effect of penetrating keratoplasty in treatment of iridocorneal endothelial syndrome. The clinical and pathologic evaluation records were retrospectively analyzed for 8 cases with iridocorneal endothelial syndrome treated by penetrating keratoplasty. Postoperatively, they were followed for 3 months to 6 years. Seven cases had visual improvement, five buttons kept being transparent and three buttons became opaque. The penetrating keratoplasty is an effective measure for treatment of corneal endothelial decompensation of iridocorneal endothelial syndrome. However, the successful operative rate of this syndrome is lower than that in cases with corneal leucoma without vascularization.

  17. Endothelial replacement without surface corneal incisions or sutures: topography of the deep lamellar endothelial keratoplasty procedure.

    Science.gov (United States)

    Terry, M A; Ousley, P J

    2001-01-01

    To evaluate the immediate postoperative corneal topography after the deep lamellar endothelial keratoplasty procedure. Eight eye bank eyes underwent deep lamellar endothelial keratoplasty through a 9.0-mm limbal incision replacing the central 7.0 mm of posterior stroma and endothelium through the lamellar pocket wound. Orbscan topography was performed before and after surgery, and simulated keratometry readings and central corneal diopter power were recorded. The change in astigmatism and corneal power from preoperative to postoperative readings was then determined. The net change in corneal astigmatism averaged 0.4+/-0.5 diopters (range, -0.1 to 1.1 diopters). The net change in corneal power averaged -0.2+/-0.4 diopters of flattening (range, -0.9 to +0.2 diopters). Neither the astigmatism nor the corneal power levels after this surgery were significantly different from the preoperative topography (p = 0.22 and 0.27, respectively). The deep lamellar endothelial keratoplasty procedure, with its absence of corneal surface incisions or sutures, has no significant effect on immediate postoperative corneal topography. The potential advantages of this procedure over penetrating keratoplasty in the treatment of endothelial dysfunction are considerable.

  18. Corneal Stroma Regeneration with Acellular Corneal Stroma Sheets and Keratocytes in a Rabbit Model.

    Science.gov (United States)

    Ma, Xiao Yun; Zhang, Yun; Zhu, Dan; Lu, Yang; Zhou, Guangdong; Liu, Wei; Cao, Yilin; Zhang, Wen Jie

    2015-01-01

    Acellular corneal stroma matrix has been used for corneal stroma engineering. However, because of its compact tissue structure, regrowth of keratocytes into the scaffold is difficult. Previously, we developed a sandwich model for cartilage engineering using acellular cartilage sheets. In the present study, we tested this model for corneal stroma regeneration using acellular porcine corneal stroma (APCS) sheets and keratocytes. Porcine corneas were decellularized by NaCl treatment, and the APCS was cut into 20-μm-thick sheets. A rabbit corneal stroma defect model was created by lamellar keratoplasty and repaired by transplantation of five pieces of APCS sheets with keratocytes. Six months after transplantation, transparent corneas were present in the experimental group, which were confirmed by anterior segment optical coherence tomography examination and transmittance examination. The biomechanical properties in the experimental group were similar to those of normal cornea. Histological analyses showed an even distribution of keratocytes and well-oriented matrix in the stroma layer in the experimental group. Together, these results demonstrated that the sandwich model using acellular corneal stroma sheets and keratocytes could be potentially useful for corneal stroma regeneration.

  19. Corneal Stroma Regeneration with Acellular Corneal Stroma Sheets and Keratocytes in a Rabbit Model.

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    Xiao Yun Ma

    Full Text Available Acellular corneal stroma matrix has been used for corneal stroma engineering. However, because of its compact tissue structure, regrowth of keratocytes into the scaffold is difficult. Previously, we developed a sandwich model for cartilage engineering using acellular cartilage sheets. In the present study, we tested this model for corneal stroma regeneration using acellular porcine corneal stroma (APCS sheets and keratocytes. Porcine corneas were decellularized by NaCl treatment, and the APCS was cut into 20-μm-thick sheets. A rabbit corneal stroma defect model was created by lamellar keratoplasty and repaired by transplantation of five pieces of APCS sheets with keratocytes. Six months after transplantation, transparent corneas were present in the experimental group, which were confirmed by anterior segment optical coherence tomography examination and transmittance examination. The biomechanical properties in the experimental group were similar to those of normal cornea. Histological analyses showed an even distribution of keratocytes and well-oriented matrix in the stroma layer in the experimental group. Together, these results demonstrated that the sandwich model using acellular corneal stroma sheets and keratocytes could be potentially useful for corneal stroma regeneration.

  20. Effect of topical dorzolamide on rabbit central corneal thickness

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    Almeida Jr. G.C.

    2006-01-01

    Full Text Available Our objective was to study the effect of dorzolamide on corneal hydration in an 18-week controlled experiment using ultrasonic pachymetry. Twenty-eight male rabbits were divided randomly into four groups. The 7 rabbits in each group received eye drops containing either 2% (w/v dorzolamide or placebo in their right eye, or in their left eye. The 2% dorzolamide rabbits were treated every 8 h. Fellow eyes are defined as eyes which did not receive either dorzolamide or placebo. The study was blind for both the person who applied the drug and the one who performed the pachymetry. The effect of treatments is reported on the basis of the percentage of pachymetric variation compared to the measurement made before drug application. There was no significant difference (P = 0.061 in pachymetric variation between dorzolamide (-4.42 ± 11.71% and placebo (2.48 ± 9.63%. However, there was a significant difference (P = 0.0034 in pachymetric variation between the dorzolamide fellow eyes (-7.56 ± 10.50% and the placebo (-4.42 ± 11.71%. In conclusion, dorzolamide did not increase the corneal thickness in rabbits.

  1. Endothelin-1 enhances corneal fibronectin deposition and promotes corneal epithelial wound healing after photorefractive keratectomy in rabbits.

    Science.gov (United States)

    Lai, Yu-Hung; Wang, Hwei-Zu; Lin, Chang-Ping; Hong, Show-Jen; Chang, Shun-Jen

    2008-05-01

    The objective was to study the effects of endothelin-1 (ET1) on corneal wound healing after photorefractive keratectomy (PRK) in rabbit corneas. Following PRK, 18 New Zealand white rabbits were treated with ET1 in the right eyes and with phosphate-buffered salt solution (PBS) in the left eyes. Corneal epithelial wound size, corneal haze and corneal thickness were recorded. Corneal extracellular matrixes, including collagen types 3, 4 and 7, chondroitin sulfate and fibronectin, were investigated using immunohistochemistry study. ET1 increased the rate of healing of corneal epithelial wounds in rabbits. Anti-fibronectin fluorescence was present at week 12 and week 24 in ET1-treated eyes but not in the control eyes. There were no significant differences in corneal haze, corneal thickness and changes in other extracellular matrixes between ET1- and PBS-treated eyes. ET1 can enhance the deposition of fibronectin in corneal stroma and promote corneal epithelial wound healing after PRK. The increase in fibronectin probably explains the increased healing rate of corneal epithelial wounds.

  2. Endothelin-1 Enhances Corneal Fibronectin Deposition and Promotes Corneal Epithelial Wound Healing after Photorefractive Keratectomy in Rabbits

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    Yu-Hung Lai

    2008-05-01

    Full Text Available The objective was to study the effects of endothelin-1 (ET1 on corneal wound healing after photorefractive keratectomy (PRK in rabbit corneas. Following PRK, 18 New Zealand white rabbits were treated with ET1 in the right eyes and with phosphate-buffered salt solution (PBS in the left eyes. Corneal epithelial wound size, corneal haze and corneal thickness were recorded. Corneal extracellular matrixes, including collagen types 3, 4 and 7, chondroitin sulfate and fibronectin, were investigated using immunohistochemistry study. ET1 increased the rate of healing of corneal epithelial wounds in rabbits. Anti-fibronectin fluorescence was present at week 12 and week 24 in ET1-treated eyes but not in the control eyes. There were no significant differences in corneal haze, corneal thickness and changes in other extracellular matrixes between ET1- and PBS-treated eyes. ET1 can enhance the deposition of fibronectin in corneal stroma and promote corneal epithelial wound healing after PRK. The increase in fibronectin probably explains the increased healing rate of corneal epithelial wounds.

  3. Effects of irrigation solutions on corneal endothelial function.

    Science.gov (United States)

    Yagoubi, M I; Armitage, W J; Diamond, J; Easty, D L

    1994-04-01

    Rabbit corneas were perfused in vitro with an irrigation solution for 90 minutes. This was followed by 6 hours of perfusion with tissue culture medium TC199 during which endothelial function was assessed by monitoring rates of swelling during a period of perfusion in the absence of bicarbonate ions, and subsequent rates of thinning when bicarbonate ions were restored to the perfusate. Corneal thickness (measured with an ultrasonic pachymeter) immediately following excision was 401 microns (SD 19, n = 23). During the 90 minute perfusion at 35 degrees C, corneas exposed to balanced salt solution (BSS), Hartmann's solution or 0.9% NaCl (all initially at room temperature) swelled, respectively, at 14 (SD 2.3, n = 4), 11 (SD 2.6, n = 4), and 70 (SD 4.3, n = 4) microns/h. Cold Hartmann's solution (initially at 4 degrees C) caused corneas to swell at 9 (SD 2.3, n = 4) microns/h. On the other hand, corneas perfused with BSS Plus thinned at 9 (SD 3.4, n = 4) microns/h and TC199 with Earle's salts had little effect on thickness. Rates of swelling and thinning during the following assessment perfusion showed no apparent effects of prior exposure to any of the irrigation solutions on the barrier properties or pump function of the endothelium. Despite this, the increased thickness of corneas exposed initially to BSS, cold Hartmann's solution, or 0.9% NaCl was not fully reversed, even by the end of the 6 hour assessment perfusion. In contrast, the swelling observed in corneas exposed to Hartmann's solution at room temperature was reversed and these corneas had returned to their normal thickness by the end of the assessment period. All corneas, even those exposed to 0.9% NaCl, had an intact endothelial mosaic with no evidence of damage or cell loss, although morphological differences in cell shape and the appearance of cell borders were evident compared with freshly isolated cornea.

  4. Relationship between Corneal Guttae and Quality of Vision in Patients with Mild Fuchs' Endothelial Corneal Dystrophy.

    Science.gov (United States)

    Watanabe, Shinya; Oie, Yoshinori; Fujimoto, Hisataka; Soma, Takeshi; Koh, Shizuka; Tsujikawa, Motokazu; Maeda, Naoyuki; Nishida, Kohji

    2015-10-01

    To investigate the effect of the severity of corneal guttae on quality of vision (QOV) in patients with mild Fuchs' endothelial corneal dystrophy (FECD). Cross-sectional study. Twenty-three eyes of 14 patients with mild FECD without corneal edema on slit-lamp examination (5 pseudophakic eyes and 18 phakic eyes with mild lens opacity; grade 1.0-2.0 nuclear opalescence, grade 1.0-2.0 nuclear color, grade 1.0 cortical cataract, and grade 1.0 posterior subcapsular cataract on the Lens Opacities Classification System, version III). The area ratio of the corneal guttae (ARCG) in the endothelial cells was measured by multifocal specular microscopy. The QOV parameters, that is, corrected distance visual acuity (CDVA), letter contrast sensitivity (LCS), and intraocular straylight, also were measured. The correlations were assessed between the ARCG and QOV parameters and between the straylight and CDVA and LCS. The ARCG, logarithm of the minimum angle of resolution CDVA, LCS, and straylight. Univariate analysis showed that the ARCG was correlated significantly with the CDVA, LCS, and straylight (R(2) = 0.41, P = 0.001; R(2) = 0.55, P = 0.001; and R(2) = 0.39, P = 0.002, respectively). Univariate analysis also showed that straylight was correlated significantly with the CDVA and LCS (R(2) = 0.47, P = 0.001 and R(2) = 0.41, P = 0.001, respectively). Corneal guttae without edema caused the QOV to deteriorate in eyes with FECD. Patients with higher straylight had worse CDVA or LCS. Intraocular forward light scatter caused by corneal guttae may result in visual disturbances. Quantification of corneal guttae can be useful to evaluate the effect of guttae on the QOV and determine the surgical indications of endothelial keratoplasty for eyes with mild FECD. Copyright © 2015 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.

  5. Specular Microscopic Features of Corneal Endothelial Vacuolation

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    Mozhgan Rezaei Kanavi

    2011-01-01

    Full Text Available Purpose: To introduce a specular microscopic reference image for endothelial vacuolation in donated corneas. Methods: Two corneas from a donor with diffuse, round to oval dark areas at the endothelial level on slit lamp biomicroscopy and one normal-appearing donor cornea underwent specular microscopy, histopathologic evaluation and transmission electron microscopy. Results: Specular microscopy of the two corneas with abnormal-looking endothelium revealed large numbers of dark, round to oval structures within the endothelium in favor of endothelial vacuolation. Light microscopy disclosed variable sized cyst-like structures within the cytoplasm. Transmission electron microscopy showed electronlucent and relatively large-sized intracytoplasmic vacuoles. These features were not observed in the endothelium of the normal cornea. Conclusion: The specular microscopic features of endothelial vacuolation in donated corneas were confirmed by light microscopy and transmission electron microscopy, therefore the specular image may be proposed as a reference to eye banks.

  6. Successful transplantation of in vitro expanded human corneal endothelial precursors to corneal endothelial surface using a nanocomposite sheet

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    Parikumar P

    2011-01-01

    Full Text Available Background: Though the transplantation of in vitro expanded human corneal endothelial precursors in animal models of endothelial damage by injecting into the anterior chamber has been reported, the practical difficulties of accomplishing such procedure in human patients have been a hurdle to clinical translation. Here we report the successful transplantation of in vitro expanded human corneal precursor cells to an animal eye using a transparent Nano-composite sheet and their engraftment.Materials and Methods: Human Corneal endothelial cells (HCEC were isolated from human cadaver eyes with informed consent and expanded in the lab using a sphere forming assay in a novel Thermoreversible Gelation Polymer (TGP for 26 days. HCEC obtained by sphere forming assay were seeded in a novel Nano-composite sheet, which was made of PNIPA-NC gels by in-situ, free-radical polymerization of NIPA monomer in the presence of exfoliated clay (synthetic hectorite “Laponite XLG” uniformly dispersed in aqueous media. After a further seven days in vitro culture of HCEC in the Nano-composite sheet, cells were harvested and transplanted on cadaver-bovine eyes (n=3. The cells were injected between the corneal endothelial layer and the Nano-composite sheet that had been placed prior to the injection in close proximity to the endothelial layer. After three hours, the transplanted Nano-composite sheets were removed from the bovine eyes and subjected to microscopic examination. The corneas were subjected to Histo-pathological studies along with controls. Results: HCEC formed sphere like colonies in TGP which expressed relevant markers as confirmed by RT-PCR. Microscopic studies of the Nanosheets and histopathological studies of the cornea of the Bull’s eye revealed that the HCEC got engrafted to the corneal endothelial layer of the bovine eyes with no remnant cells in the Nanosheet. Conclusion: Transplantation of in vitro expanded donor human corneal endothelial cells

  7. Efficacy and safety of femtosecond laser-assisted corneal endothelial keratoplasty: a randomized multicenter clinical trial.

    NARCIS (Netherlands)

    Cheng, Y.Y.; Schouten, J.S.A.G.; Tahzib, N.G.; Wijdh, R.J.; Pels, E.; Cleynenbreugel, H. van; Eggink, C.A.; Rijneveld, W.J.; Nuijts, R.M.

    2009-01-01

    BACKGROUND: To evaluate the efficacy and safety of femtosecond laser-assisted endothelial keratoplasty (FLEK) versus penetrating keratoplasty (PK) in patients with corneal endothelial disease. METHODS: A randomized multicenter clinical trial of 80 eyes of 80 patients with corneal endothelial disease

  8. Efficacy and Safety of Femtosecond Laser-Assisted Corneal Endothelial Keratoplasty : A Randomized Multicenter Clinical Trial

    NARCIS (Netherlands)

    Cheng, Yanny Y. Y.; Schouten, Jan S. A. G.; Tahzib, Nayyirih G.; Wijdh, Robert-Jan; Pels, Elisabeth; van Cleynenbreugel, Hugo; Eggink, Catharina A.; Rijneveld, Wilhelmina J.; Nuijts, Rudy M. M. A.

    2009-01-01

    Background. To evaluate the efficacy and safety of femtosecond laser-assisted endothelial keratoplasty (FLEK) versus penetrating keratoplasty (PK) in patients with corneal endothelial disease. Methods. A randomized multicenter clinical trial of 80 eyes of 80 patients with corneal endothelial disease

  9. Influence of Pre-Freezing Temperature on the Corneal Endothelial Cytocompatibility and Cell Delivery Performance of Porous Hyaluronic Acid Hydrogel Carriers.

    Science.gov (United States)

    Lai, Jui-Yang

    2015-08-11

    The development of porous hyaluronic acid (HA) hydrogels for corneal endothelial tissue engineering is attractive because they can be used as functional cell delivery carriers to help in the reconstruction of damaged areas. The purpose of this study was to investigate the corneal endothelial cytocompatibility and cell delivery performance of porous HA hydrogel biomaterials fabricated at different pre-freezing temperatures. As compared to their counterparts prepared at -80 °C, the HA samples fabricated at higher pre-freezing temperature (i.e., 0 °C) exhibited a larger pore size and higher porosity, thereby leading to lower resistance to glucose permeation. Live/dead assays and gene expression analyses showed that the restricted porous structure of HA carriers decreases the viability and ionic pump function of cultured corneal endothelial cells (CECs). The results also indicated that the porous hydrogel biomaterials fabricated at high pre-freezing temperature seem to be more compatible with rabbit CECs. In an animal model of corneal endothelial dysfunction, the wounded rabbit corneas receiving bioengineered CEC sheets and restricted porous-structured HA carriers demonstrated poor tissue reconstruction. The therapeutic efficacy of cell sheet transplants can be improved by using carrier materials prepared at high pre-freezing temperature. Our findings suggest that the cryogenic operation temperature-mediated pore microstructure of HA carriers plays an important role in corneal endothelial cytocompatibility and cell delivery performance.

  10. Applications of Biomaterials in Corneal Endothelial Tissue Engineering.

    Science.gov (United States)

    Wang, Tsung-Jen; Wang, I-Jong; Hu, Fung-Rong; Young, Tai-Horng

    2016-11-01

    When corneal endothelial cells (CECs) are diseased or injured, corneal endothelium can be surgically removed and tissue from a deceased donor can replace the original endothelium. Recent major innovations in corneal endothelial transplantation include replacement of diseased corneal endothelium with a thin lamellar posterior donor comprising a tissue-engineered endothelium carried or cultured on a thin substratum with an organized monolayer of cells. Repairing CECs is challenging because they have restricted proliferative ability in vivo. CECs can be cultivated in vitro and seeded successfully onto natural tissue materials or synthetic polymeric materials as grafts for transplantation. The optimal biomaterials for substrata of CEC growth are being investigated. Establishing a CEC culture system by tissue engineering might require multiple biomaterials to create a new scaffold that overcomes the disadvantages of single biomaterials. Chitosan and polycaprolactone are biodegradable biomaterials approved by the Food and Drug Administration that have superior biological, degradable, and mechanical properties for culturing substratum. We successfully hybridized chitosan and polycaprolactone into blended membranes, and demonstrated that CECs proliferated, developed normal morphology, and maintained their physiological phenotypes. The interaction between cells and biomaterials is important in tissue engineering of CECs. We are still optimizing culture methods for the maintenance and differentiation of CECs on biomaterials.

  11. Corneal Astigmatism Stability in Descemet Membrane Endothelial Keratoplasty for Fuchs Corneal Dystrophy.

    Science.gov (United States)

    Yokogawa, Hideaki; Sanchez, P James; Mayko, Zachary M; Straiko, Michael D; Terry, Mark A

    2016-07-01

    To calculate the magnitude and angle of the shift in corneal astigmatism associated with Descemet membrane endothelial keratoplasty (DMEK) surgery to determine the feasibility of concurrent astigmatism correction at the time of DMEK triple procedures. Retrospective study. Forty-seven eyes that previously underwent the DMEK procedure for Fuchs endothelial corneal dystrophy and that had more than 1.0 diopter (D) of front corneal astigmatism preoperatively were identified. All DMEK surgeries used a clear corneal temporal incision of 3.2 mm. Surgically induced astigmatism (SIA) was evaluated 6 months postsurgery with vector analysis using Scheimpflug image reading. We did not find a difference between pre- and postoperative magnitude of front astigmatism (P = 0.88; paired t test). The magnitude of the SIA front surface was 0.77 ± 0.63 D (range, 0.10-3.14 D). The centroid vector of the SIA front surface was 0.14 at 89.3°. A hyperopic corneal power shift was noted in both the front surface by 0.26 ± 0.74 D (range, 0.45-3.05 D) (P = 0.018; paired t test) and back surface by 0.56 ± 0.55 D (range, 0.25-2.40 D) (P astigmatism that is a with-the-rule shift associated with a temporal clear corneal incision. The stability of these data from preop to postop supports the plausibility of incorporating astigmatism correction with the cautious use of toric intraocular lenses for patients with Fuchs corneal dystrophy and cataract.

  12. In Vivo Confocal Microscopic Observation of Lamellar Corneal Transplantation in the Rabbit Using Xenogenic Acellular Corneal Scaffolds as a Substitute

    Science.gov (United States)

    Feng, Yun; Wang, Wei

    2015-01-01

    Background: The limiting factor to corneal transplantation is the availability of donors. Research has suggested that xenogenic acellular corneal scaffolds (XACS) may be a possible alternative to transplantation. This study aimed to investigate the viability of performing lamellar corneal transplantation (LCT) in rabbits using canine XACS. Methods: Fresh dog corneas were decellularized by serial digestion, and LCT was performed on rabbit eyes using xenogeneic decellularized corneal matrix. Cellular and morphological changes were observed by slit-lamp, light, and scanning electron microscopy at 7, 30 and 90 days postoperatively. Immunocytochemical staining for specific markers such as keratin 3, vimentin and MUC5AC, was used to identify cells in the graft. Results: Decellularized xenogenic corneal matrix remained transparent for about 1-month after LCT. The recipient cells were able to survive and proliferate into the grafts. Three months after transplantation, grafts had merged with host tissue, and graft epithelialization and vascularization had occurred. Corneal nerve fibers were able to grow into the graft in rabbits transplanted with XACS. Conclusions: Xenogenic acellular corneal scaffolds can maintain the transparency of corneal grafts about 1-month and permit growth of cells and nerve fibers, and is, therefore, a potential substitute or carrier for a replacement cornea. PMID:25836615

  13. Cellular Cytotoxicity of Antiglaucoma Drugs in Cultured Corneal Endothelial Cells

    Directory of Open Access Journals (Sweden)

    Kwou-Yeung Wu

    2007-03-01

    Full Text Available In this study, the various antiglaucoma drugs including betaxolol, timolol, levobunolol, carteolol, brimonidine, dipivefrin, dorzolamide, brinzolamide, latanoprost, unoprostone, and pilocarpine were used to investigate the effects of cellular cytotoxicity in cultured bovine corneal endothelial cells. After exposure to the drugs in three dilutions, 1/100, 1/1,000, and 1/10,000, for 100 minutes, cells were estimated based on the release assay of lactate dehydrogenase (LDH enzyme. It was found that cellular LDH was significantly released in the medium only at 1/100th dilution of betaxolol, brimonidine, dorzolamide, dipivefrin, latanoprost and unoprostone to 130%, 123%, 145%, 157%, 128% and 237%, respectively, compared with controls upon exposure to drugs for 100 minutes. Moreover, benzalkonium chloride preservative at the concentrations ranging from 0.001 to 0.00001mg/mL did not affect cellular LDH release in bovine corneal endothelial cells. These results indicate that high concentrations of antiglaucoma drugs may induce cytotoxicity in corneal endothelial cells.

  14. Extended incubation times improve corneal endothelial cell transplantation success.

    Science.gov (United States)

    Insler, M S; Lopez, J G

    1991-05-01

    To investigate the ability of extended incubation times to improve the success of endothelial cell transplantation, eight human donor corneas were denuded of their native endothelium, seeded twice during a 1-hr interval with a suspension of cultured infant human corneal endothelial cells, and then incubated for 144 hr under standard conditions. Subsequently the corneas were transplanted into African green monkeys using routine penetrating keratoplasty techniques. Rotational autografts and corneas devoid of endothelial cells served as controls. The seeded corneas appeared hazy at the time of surgery (mean pachymetry 48 hr postoperatively, 0.794 mm). Six corneas (75%) subsequently cleared, yielding a mean corneal thickness of 0.541 +/- 0.040 and 0.554 +/- 0.040 at 6 and 12 postoperative months, respectively. All control eyes showed advanced edema (thickness, greater than 1.0 mm) and developed extensive neovascularization. Clinically, the extended postseeding incubation corneas were observed to clear more rapidly and stabilize their thickness earlier than corneas incubated for only 24-48 hr. Scanning electron microscopy of extended postseeding incubation corneas revealed an intact monolayer of contact-inhibited cells with the hexagonal mosaic typical of corneal endothelium in vivo and improved intercellular contact compared with corneas incubated for only 24-48 hr.

  15. Corneal Backscatters as an Objective Index for Assessing Fuchs’ Endothelial Corneal Dystrophy: A Pilot Study

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    Hsueh-Yen Chu

    2017-01-01

    Full Text Available Purpose. To provide an objective, quantitative approach for monitoring Fuchs’ endothelial corneal dystrophy (FECD, with Scheimpflug imaging. Design. This is a retrospective case-control pilot study. Methods. The study group consisted of 53 eyes in 27 patients diagnosed with FECD, with normal subjects paired as control. Main outcome measures were corneal thickness, morphological patterns on densitograms, and indices of corneal density including the average area density (mean AD and the average ratio of Descemet’s membrane density versus area density (DM/AD in Pentacam Scheimpflug images. Results. There were no significant differences in age and corneal thickness between FECD and normal groups. Morphologically, hanging-hammock patterns were noted on the densitograms of FECD patients, which were different from the high-back chair patterns in normal subjects. Quantitatively, mean AD and DM/AD were both elevated in FECD patients as compared with normal subjects (P=0.01 and 0.025, resp.. In addition, FECD patients with corneal edema had significantly higher mean AD (P=0.018 than those without corneal edema. Conclusions. This pilot study shows that Pentacam system provides an objective, quantitative way to approach FECD corneas. It can assist ophthalmologists in detecting the early change and in monitoring disease progression of FECD. Further studies are needed to consolidate the findings.

  16. Contact lens-induced corneal endothelial polymegathism: functional significance and possible mechanisms.

    Science.gov (United States)

    Connor, C G; Zagrod, M E

    1986-07-01

    The corneal endothelium is principally responsible for maintenance of corneal deturgescence. Therefore, compromise of corneal endothelial functional integrity can result in corneal swelling and opacification. Contact lenses constitute a potential insult to the cornea because their wear reduces the oxygen available to that tissue. It has been reported that contact lens wear induces transient as well as permanent morphologic changes in the corneal endothelium. One of the permanent changes reported is referred to as polymegathism, which is a variation in cell size within the endothelial monolayer. Several investigators have suggested that polymegathism reflects a compromised endothelial functional status. Mechanisms proposed to explain contact lens-induced polymegathism include lactate accumulation, changes in pH, and elevation in CO2 content. We discuss these possibilities as well as speculate that these polymegathous shape changes may be a result of decreased endothelial ATP (adenosine triphosphate) levels and disturbed calcium homeostasis due to corneal endothelial hypoxia.

  17. Corneal endothelial cell density and morphology in Phramongkutklao Hospital

    Directory of Open Access Journals (Sweden)

    Narumon Sopapornamorn

    2008-03-01

    Full Text Available Narumon Sopapornamorn1, Manapon Lekskul1, Suthee Panichkul21Department of Ophthalmology, Phramongkutklao Hospital, Bangkok, Thailand; 2Department of Obstetrics and Gynecology, Phramongkutklao College of Medicine, Bangkok, ThailandObjective: To describe the corneal endothelial density and morphology in patients of Phramongkutklao Hospital and the relationship between endothelial cell parameters and other factors.Methods: Four hundred and four eyes of 202 volunteers were included. Noncontact specular microscopy was performed after taking a history and testing the visual acuity, intraocular pressure measurement, Schirmer’s test and routine eye examination by slit lamp microscope. The studied parameters included mean endothelial cell density (MCD, coefficient of variation (CV, and percentage of hexagonality.Results: The mean age of volunteers was 45.73 years; the range being 20 to 80 years old. Their MCD (SD, mean percentage of CV (SD and mean (SD percentage of hexagonality were 2623.49(325 cell/mm2, 39.43(8.23% and 51.50(10.99%, respectively. Statistically, MCD decreased significantly with age (p < 0.01. There was a significant difference in the percentage of CV between genders. There was no statistical significance between parameters and other factors.Conclusion: The normative data of the corneal endothelium of Thai eyes indicated that, statistically, MCD decreased significantly with age. Previous studies have reported no difference in MCD, percentage of CV, and percentage of hexagonality between gender. Nevertheless, significantly different percentages of CV between genders were presented in this study.Keywords: Corneal endothelial cell, parameters, age, gender, smoking, Thailand

  18. Effect of Mitomycin-C augmented trabeculectomy on corneal endothelial cells

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    Reza Zarei

    2015-01-01

    Conclusion: MMC application in trabeculectomy seems to cause a small but significant corneal endothelial loss. Most of the damage occurs intraoperatively, or in the early postoperative period, however progressive endothelial cell loss is not a major concern.

  19. Role of corneal epithelium in riboflavin/ultraviolet-A mediated corneal cross-linking treatment in rabbit eyes.

    Science.gov (United States)

    Tao, Xiangchen; Yu, Haiqun; Zhang, Yong; Li, Zhiwei; Jhanji, Vishal; Ni, Shouxiang; Wang, Ya; Mu, Guoying

    2013-01-01

    To evaluate the role of corneal epithelium in riboflavin/ultraviolet-A (UVA) mediated corneal collagen cross-linking treatment. Fifty New Zealand rabbits were divided into 5 groups: UVA treatment with or without corneal epithelium, UVA+riboflavin treatment with or without corneal epithelium, and control without any treatment. All rabbits were sacrificed after irradiation and subsequently 4 mm × 10 mm corneal strips were harvested for biomechanical evaluation. UVA irradiation alone did not enhance the maximal stress and Young's modulus of corneal specimens with (3.15 ± 0.56 mpa, 1.00 ± 0.09 mpa) or without (3.53 ± 0.85 mpa, 0.94 ± 0.21 mpa) the corneal epithelium, compared to specimens in the control group (4.30 ± 0.68 mpa, 1.03 ± 0.24 mpa). However, UVA irradiation combined with riboflavin significantly increased the maximal stress and Young's modulus of corneal specimens with (5.27 ± 1.09 mpa, 1.23 ± 0.23 mpa, P cornea in UVA+riboflavin and "epithelium-off" group were 35.9% and 15.4% higher compared to the UVA+riboflavin and "epithelium-on" group, respectively (P biomechanical properties of the cornea with and without epithelial removal. However, corneas without epithelium seem to benefit more compared to corneas with the epithelium.

  20. Discovery of Molecular Markers to Discriminate Corneal Endothelial Cells in the Human Body

    NARCIS (Netherlands)

    Yoshihara, Masahito; Ohmiya, Hiroko; Hara, Susumu; Kawasaki, Satoshi; Hayashizaki, Yoshihide; Itoh, Masayoshi; Kawaji, Hideya; Tsujikawa, Motokazu; Nishida, Kohji; Forrest, Alistair R. R.; Rehli, Michael; Baillie, J. Kenneth; de Hoon, Michiel J. L.; Haberle, Vanja; Lassmann, Timo; Kulakovskiy, Ivan V.; Lizio, Marina; Andersson, Robin; Mungall, Christopher J.; Meehan, Terrence F.; Schmeier, Sebastian; Bertin, Nicolas; Jørgensen, Mette; Dimont, Emmanuel; Arner, Erik; Schmidl, Christian; Schaefer, Ulf; Medvedeva, Yulia A.; Plessy, Charles; Vitezic, Morana; Severin, Jessica; Semple, Colin A.; Ishizu, Yuri; Francescatto, Margherita; Alam, Intikhab; Albanese, Davide; Altschuler, Gabriel M.; Archer, John A. C.; Arner, Peter; Babina, Magda; Baker, Sarah; Balwierz, Piotr J.; Beckhouse, Anthony G.; Pradhan-Bhatt, Swati; Blake, Judith A.; Blumenthal, Antje; Bodega, Beatrice; Bonetti, Alessandro; Briggs, James; Brombacher, Frank; Burroughs, A. Maxwell; Califano, Andrea; Cannistraci, Carlo V.; Carbajo, Daniel; Chen, Yun; Chierici, Marco; Ciani, Yari; Clevers, Hans C.; Dalla, Emiliano; Davis, Carrie A.; Detmar, Michael; Diehl, Alexander D.; Dohi, Taeko; Drabløs, Finn; Edge, Albert S. B.; Edinger, Matthias; Ekwall, Karl; Endoh, Mitsuhiro; Enomoto, Hideki; Fagiolini, Michela; Fairbairn, Lynsey; Fang, Hai; Farach-Carson, Mary C.; Faulkner, Geoffrey J.; Favorov, Alexander V.; Fisher, Malcolm E.; Frith, Martin C.; Fujita, Rie; Fukuda, Shiro; Furlanello, Cesare; Furuno, Masaaki; Furusawa, Jun-ichi; Geijtenbeek, Teunis B.; Gibson, Andrew; Gingeras, Thomas; Goldowitz, Daniel; Gough, Julian; Guhl, Sven; Guler, Reto; Gustincich, Stefano; Ha, Thomas J.; Hamaguchi, Masahide; Hara, Mitsuko; Harbers, Matthias; Harshbarger, Jayson; Hasegawa, Akira; Hasegawa, Yuki; Hashimoto, Takehiro; Herlyn, Meenhard; Hitchens, Kelly J.; Ho Sui, Shannan J.; Hofmann, Oliver M.; Hoof, Ilka; Hori, Fumi; Huminiecki, Lukasz; Iida, Kei; Ikawa, Tomokatsu; Jankovic, Boris R.; Jia, Hui; Joshi, Anagha; Jurman, Giuseppe; Kaczkowski, Bogumil; Kai, Chieko; Kaida, Kaoru; Kaiho, Ai; Kajiyama, Kazuhiro; Kanamori-Katayama, Mutsumi; Kasianov, Artem S.; Kasukawa, Takeya; Katayama, Shintaro; Kato, Sachi; Kawaguchi, Shuji; Kawamoto, Hiroshi; Kawamura, Yuki I.; Kawashima, Tsugumi; Kempfle, Judith S.; Kenna, Tony J.; Kere, Juha; Khachigian, Levon M.; Kitamura, Toshio; Klinken, S. Peter; Knox, Alan J.; Kojima, Miki; Kojima, Soichi; Kondo, Naoto; Koseki, Haruhiko; Koyasu, Shigeo; Krampitz, Sarah; Kubosaki, Atsutaka; Kwon, Andrew T.; Laros, Jeroen F. J.; Lee, Weonju; Lennartsson, Andreas; Li, Kang; Lilje, Berit; Lipovich, Leonard; Mackay-sim, Alan; Manabe, Ri-ichiroh; Mar, Jessica C.; Marchand, Benoit; Mathelier, Anthony; Mejhert, Niklas; Meynert, Alison; Mizuno, Yosuke; Morais, David A. de Lima; Morikawa, Hiromasa; Morimoto, Mitsuru; Moro, Kazuyo; Motakis, Efthymios; Motohashi, Hozumi; Mummery, Christine L.; Murata, Mitsuyoshi; Nagao-Sato, Sayaka; Nakachi, Yutaka; Nakahara, Fumio; Nakamura, Toshiyuki; Nakamura, Yukio; Nakazato, Kenichi; van Nimwegen, Erik; Ninomiya, Noriko; Nishiyori, Hiromi; Noma, Shohei; Nozaki, Tadasuke; Ogishima, Soichi; Ohkura, Naganari; Ohno, Hiroshi; Ohshima, Mitsuhiro; Okada-Hatakeyama, Mariko; Okazaki, Yasushi; Orlando, Valerio; Ovchinnikov, Dmitry A.; Pain, Arnab; Passier, Robert; Patrikakis, Margaret; Persson, Helena; Piazza, Silvano; Prendergast, James G. D.; Rackham, Owen J. L.; Ramilowski, Jordan A.; Rashid, Mamoon; Ravasi, Timothy; Rizzu, Patrizia; Roncador, Marco; Roy, Sugata; Rye, Morten B.; Saijyo, Eri; Sajantila, Antti; Saka, Akiko; Sakaguchi, Shimon; Sakai, Mizuho; Sato, Hiroki; Satoh, Hironori; Savvi, Suzana; Saxena, Alka; Schneider, Claudio; Schultes, Erik A.; Schulze-Tanzil, Gundula G.; Schwegmann, Anita; Sengstag, Thierry; Sheng, Guojun; Shimoji, Hisashi; Shimoni, Yishai; Shin, Jay W.; Simon, Christophe; Sugiyama, Daisuke; Sugiyama, Takaaki; Suzuki, Masanori; Swoboda, Rolf K.; 't Hoen, Peter A. C.; Tagami, Michihira; Takahashi, Naoko; Takai, Jun; Tanaka, Hiroshi; Tatsukawa, Hideki; Tatum, Zuotian; Thompson, Mark; Toyoda, Hiroo; Toyoda, Tetsuro; Valen, Eivind; van de Wetering, Marc; van den Berg, Linda M.; Verardo, Roberto; Vijayan, Dipti; Vorontsov, Ilya E.; Wasserman, Wyeth W.; Watanabe, Shoko; Wells, Christine A.; Winteringham, Louise N.; Wolvetang, Ernst; Wood, Emily J.; Yamaguchi, Yoko; Yamamoto, Masayuki; Yoneda, Misako; Yonekura, Yohei; Yoshida, Shigehiro; Zabierowski, Suzan E.; Zhang, Peter G.; Zhao, Xiaobei; Zucchelli, Silvia; Summers, Kim M.; Suzuki, Harukazu; Daub, Carsten O.; Kawai, Jun; Heutink, Peter; Hide, Winston; Freeman, Tom C.; Lenhard, Boris; Bajic, Vladimir B.; Taylor, Martin S.; Makeev, Vsevolod J.; Sandelin, Albin; Hume, David A.; Carninci, Piero

    2015-01-01

    The corneal endothelium is a monolayer of hexagonal corneal endothelial cells (CECs) on the inner surface of the cornea. CECs are critical in maintaining corneal transparency through their barrier and pump functions. CECs in vivo have a limited capacity in proliferation, and loss of a significant

  1. Corneal endothelial cell changes associated with cataract surgery in patients with type 2 diabetes mellitus

    DEFF Research Database (Denmark)

    Hugod, Mikkel; Storr-Paulsen, Allan; Norregaard, Jens Christian

    2011-01-01

    To investigate the corneal endothelial cell density and morphology in patients with and without diabetes after phacoemulsification with intraocular lens implantation.......To investigate the corneal endothelial cell density and morphology in patients with and without diabetes after phacoemulsification with intraocular lens implantation....

  2. Cytotoxicity of Voriconazole on Cultured Human Corneal Endothelial Cells▿

    OpenAIRE

    Han, Sang Beom; Shin, Young Joo; Hyon, Joon Young; Wee, Won Ryang

    2011-01-01

    The purpose of the present study was to evaluate the toxicity of voriconazole on cultured human corneal endothelial cells (HCECs). HCECs were cultured and exposed to various concentrations of voriconazole (5.0 to 1,000 μg/ml). Cell viability was measured using a Cell Counting Kit-8 (CCK-8) and live/dead viability/cytotoxicity assays. Cell damage was assessed using phase-contrast microscopy after 24 h of exposure to voriconazole. To analyze the effect of voriconazole on the intercellular barri...

  3. Cytotoxicity of Voriconazole on Cultured Human Corneal Endothelial Cells▿

    Science.gov (United States)

    Han, Sang Beom; Shin, Young Joo; Hyon, Joon Young; Wee, Won Ryang

    2011-01-01

    The purpose of the present study was to evaluate the toxicity of voriconazole on cultured human corneal endothelial cells (HCECs). HCECs were cultured and exposed to various concentrations of voriconazole (5.0 to 1,000 μg/ml). Cell viability was measured using a Cell Counting Kit-8 (CCK-8) and live/dead viability/cytotoxicity assays. Cell damage was assessed using phase-contrast microscopy after 24 h of exposure to voriconazole. To analyze the effect of voriconazole on the intercellular barrier, immunolocalization of zonula occludens 1 (ZO1) was performed. A flow cytometric assay was performed to evaluate the apoptotic and necrotic effects of voriconazole on HCECs. Cytotoxicity tests demonstrated the dose-dependent toxic effect of voriconazole on HCECs. Voriconazole concentrations of ≥100 μg/ml led to a significant reduction in cell viability. The morphological characteristics of HCECs also changed in a dose-dependent manner. Increasing concentrations of voriconazole resulted in fading staining for ZO1. Higher concentrations of voriconazole resulted in an increased number of propidium iodide (PI)-positive cells, indicating activation of the proapoptotic pathway. In conclusion, voriconazole may have a dose-dependent toxic effect on cultured HCECs. The results of this study suggest that although voriconazole concentrations of up to 50 μg/ml do not decrease cell viability, intracameral voriconazole concentrations of ≥100 μg/ml may increase the risk of corneal endothelial damage. PMID:21768517

  4. Biosynthetic corneal implants for replacement of pathologic corneal tissue: performance in a controlled rabbit alkali burn model.

    Science.gov (United States)

    Hackett, Joanne M; Lagali, Neil; Merrett, Kimberley; Edelhauser, Henry; Sun, Yifei; Gan, Lisha; Griffith, May; Fagerholm, Per

    2011-02-03

    To evaluate the performance of structurally reinforced, stabilized recombinant human collagen-phosphorylcholine (RHCIII-MPC) hydrogels as corneal substitutes in a rabbit model of severe corneal damage. One eye each of 12 rabbits received a deep corneal alkali wound. Four corneas were implanted with RHCIII-MPC hydrogels. The other eight control corneas were implanted with either allografts or a simple cross-linked RHCIII hydrogel. In all cases, 6.25 mm diameter, 350 μm thick buttons were implanted by anterior lamellar keratoplasty to replace damaged corneal tissue. Implants were followed for nine months by clinical examination and in vivo confocal microscopy, after which implanted corneas were removed and processed for histopathological and ultrastructural examination. Alkali exposure induced extensive central corneal scarring, ocular surface irregularity, and neovascularization in one case. All implants showed complete epithelial coverage by four weeks postoperative, but with accompanying suture-induced vascularization in 6 out of 12 cases. A stable, stratified epithelium with hemidesmosomal adhesion complexes regenerated over all implants, and subbasal nerve regeneration was observed in allograft and RHCIII-MPC implants. Initially acellular biosynthetic implants were populated with host-derived keratocytes as stromal haze subsided and stromal collagen was remodeled. Notably, RHCIII-MPC implants exhibited resistance to vascular ingrowth while supporting endogenous cell and nerve repopulation. Biosynthetic implants based on RHC promoted cell and nerve repopulation in alkali burned rabbit eyes. In RHCIII-MPC implants, evidence of an enhanced resistance to neovascularization was additionally noted.

  5. Corneal collagen cross-linking and liposomal amphotericin B combination therapy for fungal keratitis in rabbits

    Directory of Open Access Journals (Sweden)

    Zhao-Qin Hao

    2016-11-01

    Full Text Available AIM: To observe the therapeutic effect of corneal collagen cross-linking (CXL in combination with liposomal amphotericin B in fungal corneal ulcers. METHODS: New Zealand rabbits were induced fungal corneal ulcers by scratching and randomly divided into 3 groups, i.e. control, treated with CXL, and combined therapy of CXL with 0.25% liposomal amphotericin B (n=5 each. The corneal lesions were documented with slit-lamp and confocal microscopy on 3, 7, 14, 21 and 28d after treatment. The corneas were examined with transmission electron microscopy (TEM at 4wk. RESULTS: A rabbit corneal ulcer model of Fusarium was successfully established. The corneal epithelium defect areas in the two treatment groups were smaller than that in the control group on 3, 7, 14 and 21d (P<0.05. The corneal epithelium defect areas of the combined group was smaller than that of the CXL group (P<0.05 on 7 and 14d, but there were no statistical differences on 3, 21 and 28d. The corneal epithelium defects of the two treatment groups have been healed by day 21. The corneal epithelium defects of the control group were healed on 28d. The diameters of the corneal collagen fiber bundles (42.960±7.383 nm in the CXL group and 37.040±4.160 nm in the combined group were thicker than that of the control group (24.900±1.868 nm, but there was no difference between the two treatment groups. Some corneal collagen fiber bundles were distorted and with irregular arrangement, a large number of fibroblasts could be seen among them but no inflammatory cells in both treatment groups. CONCLUSION: CXL combined with liposomal amphotericin B have beneficial effects on fungal corneal ulcers. The combined therapy could alleviate corneal inflammattions, accelerate corneal repair, and shorten the course of disease.

  6. Establishment of a porcine corneal endothelial organ culture model for research purposes.

    Science.gov (United States)

    Kunzmann, Berenike C; Hellwinkel, Olaf J C; Klameth, Christian; Wenzel, Daniel; Bartz-Schmidt, Karl U; Spitzer, Martin S; Schultheiss, Maximilian

    2017-10-27

    Human corneas usually are not available for research, as they are used for transplantation only. At the same time, scientific studies on cultured human endothelial cells can produce misleading results due to inevitable dedifferentiation. Therefore, an organ-culture model of porcine corneas-displaying endothelial cell death rates comparable to those of cultured human corneas-would be very desirable. Fresh pig eyes were prepared under sterile conditions to obtain corneoscleral buttons, corneal buttons and so called "split corneal buttons" (new preparation method) and cultivated for 15 days. Morphology of the endothelial cell layer was observed by light microscopy on day 1, 8 and 15. On day 15 staining with trypan blue and alizarin red S was performed. Photographs were evaluated in a randomized, blinded manner. Here, the morphology of the corneal endothelium and the number of endothelial cells per mm(2) were analyzed. After 15 days of cultivation the endothelial cell layer was maintained only in corneal buttons and split corneal buttons. Alizarin red S stained areas and the existence of polymorphisms like rosette figures and reformation figures were significantly less frequent in split corneal buttons than in corneal buttons. Loss of endothelial cells was significantly greater in corneal buttons [575 ± 25/250 cells/mm(2) (median ± 25%/75%-quantile); 14.8%] than in split corneal buttons [417 ± 138/179 cells/mm(2) (median ± 25%/75%-quantile); 10.2%]. The new preparation method of split corneal buttons allows the cultivation of porcine corneas for 2 weeks with cell death rates comparable to those of the corresponding human tissue in cornea banks without the need to add de-swelling additives to the media. This is therefore a simple and highly reliable method model to be applied in intervention studies on corneal endothelial cells in their natural compound.

  7. Cornea stress test--evaluation of corneal endothelial function in vivo by contact lens induced stress

    Directory of Open Access Journals (Sweden)

    Saini Jagjit

    1997-01-01

    Full Text Available Reliable and valid assessment of corneal endothelial function is a critical input for diagnosing, prognosticating and monitoring progression of disorders affecting corneal endothelium. In 123 eyes, corneal endothelial function was assessed employing data from the corneal hydration recovery dynamics. Serial pachometric readings were recorded on Haag-Striet pachometer with Mishima-Hedbys modification before and after two hours of thick soft contact lens wear. Percentage Recovery Per Hour (PRPH was derived from raw data as an index of endothelial function. Assessed PRPH in pseudophakic corneal oedema and Fuchs′ endothelial dystrophy eyes (35.9 +/- 9.8% was significantly lower than normal controls (61.9 +/- 10.5%. On employing receiver operation characteristics curve analysis the tested results demonstrated high sensitivity (87% and specificity (92% for detection of low endothelial function at PRPH cut off of 47.5%. Using this PRPH cut off, 80% of Fuchs′ endothelial dystrophy and 93.3% of pseudophakic corneal oedema eyes could be demonstrated to have low endothelial function. A total of 66.7% of diabetic eyes also demonstrated PRPH of lower than 47.5%. Clear corneal grafts demonstrated PRPH values of 24.6% to 73.0%. Of 6 corneal grafts that demonstrated initial PRPH of lower than 47.5%, 4 failed within 4 to 6 months. Our data demonstrated high sensitivity and specificity of this corneal stress test. PRPH index was useful in quantifying endothelial function in clinical disorders including diabetes mellitus. The index PRPH was demonstrated to be useful in monitoring and prognosticating outcome of corneal grafts.

  8. Corneal endothelial cell density and morphology in normal Iranian eyes

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    Fallah Mohammad

    2006-03-01

    Full Text Available Abstract Background We describe corneal endothelial cell density and morphology in normal Iranian eyes and compare endothelial cell characteristics in the Iranian population with data available in the literature for American and Indian populations. Methods Specular microscopy was performed in 525 eyes of normal Iranian people aged 20 to 85 years old. The studied parameters including mean endothelial cell density (MCD, mean cell area (MCA and coefficient of variation (CV in cell area were analyzed in all of the 525 eyes. Results MCD was 1961 ± 457 cell/mm2 and MCA was 537.0 ± 137.4 μm2. There was no statistically significant difference in MCD, MCA and CV between genders (Student t-test, P = 0.85, P = 0.97 and P = 0.15 respectively. There was a statistically significant decrease in MCD with age (P r = -0.64. The rate of cell loss was 0.6% per year. There was also a statistically significant increase in MCA (P r = 0.56 and CV (P r = 0.30 from 20 to 85 years of age. Conclusion The first normative data for the endothelium of Iranian eyes seems to confirm that there are no differences in MCD, MCA and CV between genders. Nevertheless, the values obtained in Iranian eyes seem to be different to those reported by the literature in Indian and American populations.

  9. Inhibition of erythropoietin siRNA on corneal neovascularization of rabbit

    Directory of Open Access Journals (Sweden)

    Yu-Shun Xue

    2017-03-01

    Full Text Available AIM: To observe the expression of erythropoietin(EPOon the corneal of rabbit and evaluate the inhibition effect of EPO siRNA on corneal neovascularization(CNV. METHODS: Totally 22 healthy rabbits were randomly divided into 2 groups, which were experimental group and normal control group. Both eyes of rabbits in experimental group were chosen to establish corneal neovascularization model by alkali burn. The morphologic change of corneal was observed with slit lamp microscope and the area of CNV was calculated every day. After alkali burn, the right eye of the experimental group was accepted EPO siRNA injection under the conjunctiva, and the left eye was assigned to be experimental control group. The corneal with CNV was collected for immunohistochemistry at 3d, 7d, 14d, 21d after alkali burn, and the expression of EPO was measured. RESULTS: CNV began growing at the 3d after alkali burn in experimental group, and it was vigorous growing at 7d-14d period. The result of immunohistochemistry shows that the expression of EPO increased after the operation. Compared with experimental group, the rabbits who were treated by EPO siRNA was found with less neovascularization on their corneal, and the expression of EPO decreased. There were statistical significance between the two group at different time(PCONCLUSION: EPO is likely to play an important role on CNV growth, and EPO siRNA can inhibit the growth of CNV by restraining the expression of EPO.

  10. Concise Review: An Update on the Culture of Human Corneal Endothelial Cells for Transplantation.

    Science.gov (United States)

    Parekh, Mohit; Ferrari, Stefano; Sheridan, Carl; Kaye, Stephen; Ahmad, Sajjad

    2016-02-01

    The cornea forms the front window of the eye, enabling the transmission of light to the retina through a crystalline lens. Many disorders of the cornea lead to partial or total blindness, and therefore corneal transplantation becomes mandatory. Recently, selective corneal layer (as opposed to full thickness) transplantation has become popular because this leads to earlier rehabilitation and visual outcomes. Corneal endothelial disorders are a common cause of corneal disease and transplantation. Corneal endothelial transplantation is successful but limited worldwide because of lower donor corneal supply. Alternatives to corneal tissue for endothelial transplantation therefore require immediate attention. The field of human corneal endothelial culture for transplantation is rapidly emerging as a possible viable option. This manuscript provides an update regarding these developments. Significance: The cornea is the front clear window of the eye. It needs to be kept transparent for normal vision. It is formed of various layers of which the posterior layer (the endothelium) is responsible for the transparency of the cornea because it allows the transport of ions and solutes to and from the other layers of the cornea. Corneal blindness that results from the corneal endothelial dysfunction can be treated using healthy donor tissues. There is a huge demand for human donor corneas but limited supply, and therefore there is a need to identify alternatives that would reduce this demand. Research is underway to understand the isolation techniques for corneal endothelial cells, culturing these cells in the laboratory, and finding possible options to transplant these cells in the patients. This review article is an update on the recent developments in this field. ©AlphaMed Press.

  11. Development of Cell Analysis Software for Cultivated Corneal Endothelial Cells.

    Science.gov (United States)

    Okumura, Naoki; Ishida, Naoya; Kakutani, Kazuya; Hongo, Akane; Hiwa, Satoru; Hiroyasu, Tomoyuki; Koizumi, Noriko

    2017-11-01

    To develop analysis software for cultured human corneal endothelial cells (HCECs). Software was designed to recognize cell borders and to provide parameters such as cell density, coefficient of variation, and polygonality of cultured HCECs based on phase contrast images. Cultured HCECs with high or low cell density were incubated with Ca-free and Mg-free phosphate-buffered saline for 10 minutes to reveal the cell borders and were then analyzed with software (n = 50). Phase contrast images showed that cell borders were not distinctly outlined, but these borders became more distinctly outlined after phosphate-buffered saline treatment and were recognized by cell analysis software. The cell density value provided by software was similar to that obtained using manual cell counting by an experienced researcher. Morphometric parameters, such as the coefficient of variation and polygonality, were also produced by software, and these values were significantly correlated with cell density (Pearson correlation coefficients -0.62 and 0.63, respectively). The software described here provides morphometric information from phase contrast images, and it enables subjective and noninvasive quality assessment for tissue engineering therapy of the corneal endothelium.

  12. Corneal endothelial cell density and morphology and central corneal thickness in Guangxi Maonan and Han adolescent students of China

    Directory of Open Access Journals (Sweden)

    Hao Liang

    2015-06-01

    Full Text Available AIM:To investigate the corneal endothelial cell density and morphology and central corneal thickness in the Guangxi Maonan and Han adolescent students of China.METHODS:Noncontact specular microscope (Topcon SP3000P, Tokyo, Japan was performed in 133 adolescent students of Maonan nationality (M:F 54:79 and 105 adolescent students of Han nationality (M:F 50:55, 5 to 20y of age, who were randomly selected from 3 schools in Huanjiang Maonan Autonomous County of Guangxi Zhuang Autonomous Region of China. Parameters studied included endothelial cell density, mean cell area, coefficient of variation in cell size, percentage hexagonality and central corneal thickness.RESULTS:Endothelial cell density, mean cell area, coefficient of variation in cell size, percentage hexagonality and central corneal thickness in the study population were (2969.50±253.93 cells/mm2, (339.23±29.44 µm2, (29.96±4.07 %, (64.58±9.41 % and (523.71±32.82 µm in Maonan and (2998.26±262.65 cells/mm2, (336.11±30.07 µm2, (29.89±5.03 %, (64.91±11.64 % and (524.39±33.15 µm in Han, respectively. No significant differences were observed in endothelial cell density, mean cell area, coefficient of variation in cell size, percentage hexagonality and central corneal thickness between Maonan and Han (P=0.615, 0.659, 0.528, 0.551, 0.999. In Maonan and Han, we found age was negatively correlated with endothelial cell density and percentage hexagonality and positively correlated with mean cell area and coefficient of variation in cell size. Negative correlation was also found between central corneal thickness and age in Han, whereas no correlation was found in Maonan.CONCLUSION:There were no differences between Maonan and Han in corneal endothelial cell density and morphology and central corneal thickness. In these two nationalities, there were statistically significant decrease in endothelial cell density and percentage hexagonality with increasing age and statistically significant

  13. Intraocular pressure and corneal biomechanics in Fuchs' endothelial dystrophy and after posterior lamellar keratoplasty.

    Science.gov (United States)

    Clemmensen, Kåre; Hjortdal, Jesper

    2014-06-01

      To evaluate the precision of techniques for measuring intraocular pressure (IOP) in corneas with presumably altered biomechanical properties.   Intraocular pressure was measured with a Goldmann applanation tonometer (GAT), ocular response analyzer (ORA) and dynamic contour tonometer (DCT) in 70 eyes. Thirty-five eyes were normal corneas, 18 eyes had Fuchs' endothelial dystrophy, and 17 eyes had undergone Descemet's stripping automated endothelial keratoplasty (DSAEK) surgery. Corneal hysteresis (CH), corneal resistance factor (CRF) as well as central corneal thickness (CCT) were recorded with the ORA.   The measured cornea-corrected IOP using ORA was significantly higher than GAT in all three groups (pkeratoplasty. GAT and DCT seem to measure IOP correctly in patients with Fuchs' endothelial dystrophy as well as after posterior lamellar keratoplasty. Corneal-corrected IOP as measured with the ORA appears to overestimate IOP in patients with Fuchs' endothelial dystrophy as well as after posterior lamellar keratoplasty. © 2013 Acta Ophthalmologica Scandinavica Foundation. Published by Blackwell Publishing Ltd.

  14. Fourier transform method to determine human corneal endothelial morphology

    Science.gov (United States)

    Masters, Barry R.; Lee, Yim-Kul; Rhodes, William T.

    1991-08-01

    The statistical evaluation of the size, shape, density and regularity of the cells in the human corneal endothelium is an important diagnostic technique. A method based on the Fourier transform of the cell boundaries was developed which can yield these statistical properties. The development of a hybrid optical/digital technique to obtain these statistical perimeters is our goal. The input images were tracings of human endothelial cell patterns. The optical Fourier transform of each image was obtained, and the radial projection and the angular correlation function were plotted versus distance and angle respectively. The average size of the cells was obtained from the first peak of the radial projection. The width of this peak is related to the coefficient of variation of the average cell size. The separation of the peaks in the normalized angular correlation plot is related to cell shape. This method is suitable for rapid analysis of large numbers of endothelial cell images. This technique may have potential for diagnostic ophthalmology.

  15. Improvement In Rabbit Corneal Cell Suspension Viability After Freezing With Gingko Biloba Extrakt

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    Murad Aktan

    2007-01-01

    Full Text Available We investigated whether the addition of Gingko Biloba extract (EGb 761 to rabbit corneal epithelial medium before cell freezing improved cell viability after freezing then thawing. After removal of corneas, they were treated with enzymes and the corneal epithelium was prepared as a single cell suspension in freezing media with or without EGb 761. After freezing for two weeks then thawing, a higher cell viability was found in the cornea cell suspensions which had been frozen pretreated with EGb 761 in the media. The improvement with corneal cell viability with EGb 761 pretreatment is postulated to be based on the antioxidant capacity of the plant extract.

  16. Restoration of the rabbit corneal surface after total epithelial debridement and complete limbal excision

    Directory of Open Access Journals (Sweden)

    S.J. Faria-e-Sousa

    2012-05-01

    Full Text Available How is the corneal epithelium restored when all of it plus the limbus have been eliminated? This investigation explored the possibility that this may be achieved through the conjunctival epithelium. The corneal epithelium of the right eye of 12 rabbits (Oryctolagus cuniculus was totally scraped followed by surgical excision of the limbus plus 1.0-1.5 mm of the adjacent conjunctiva. Antibiotics and corticosteroids were applied for 1 week after surgery. Histological and immunohistochemical techniques were used to monitor the events taking place on the eye surface 2 weeks and 1, 3 and 6 months thereafter. Initially, the corneal surface was covered by conjunctival-like epithelium. After 1 month and more prominently at 3 and 6 months an epithelium displaying the morphological features of the cornea and reacting with the AE5 antibody was covering the central region. It is likely that the corneal epithelium originated from undifferentiated cells of the conjunctiva interacting with the corneal stroma.

  17. The corneal endothelium reflected: Studies on surgical damage tot the corneal endothelium and on endothelial specular microscopy

    NARCIS (Netherlands)

    B.T.H. van Dooren (Bart)

    2006-01-01

    textabstractThe endothelium is the innermost layer of the cornea. It is a mosaic of hexagonal cells that is only one cell thick. These endothelial cells actively maintain the corneal hydration equilibrium, and hence are very important for its transparency. However, they are vulnerable to trauma,

  18. Role of Corneal Epithelium in Riboflavin/Ultraviolet-A Mediated Corneal Cross-Linking Treatment in Rabbit Eyes

    Directory of Open Access Journals (Sweden)

    Xiangchen Tao

    2013-01-01

    Full Text Available Purpose. To evaluate the role of corneal epithelium in riboflavin/ultraviolet-A (UVA mediated corneal collagen cross-linking treatment. Methods. Fifty New Zealand rabbits were divided into 5 groups: UVA treatment with or without corneal epithelium, UVA+riboflavin treatment with or without corneal epithelium, and control without any treatment. All rabbits were sacrificed after irradiation and subsequently 4 mm × 10 mm corneal strips were harvested for biomechanical evaluation. Results. UVA irradiation alone did not enhance the maximal stress and Young’s modulus of corneal specimens with (3.15 ± 0.56 mpa, 1.00 ± 0.09 mpa or without (3.53 ± 0.85 mpa, 0.94 ± 0.21 mpa the corneal epithelium, compared to specimens in the control group (4.30 ± 0.68 mpa, 1.03 ± 0.24 mpa. However, UVA irradiation combined with riboflavin significantly increased the maximal stress and Young’s modulus of corneal specimens with (5.27 ± 1.09 mpa, 1.23 ± 0.23 mpa, P<0.05 or without (7.16 ± 1.88 mpa, 1.42 ± 0.16 mpa, P<0.05 corneal epithelium when compared to the control group. The maximal stress and Young’s modulus of cornea in UVA+riboflavin and “epithelium-off” group were 35.9% and 15.4% higher compared to the UVA+riboflavin and “epithelium-on” group, respectively (P<0.05. Conclusions. Our study shows that UVA+riboflavin treatment significantly affects the biomechanical properties of the cornea with and without epithelial removal. However, corneas without epithelium seem to benefit more compared to corneas with the epithelium.

  19. Paraoxonase-1 is associated with corneal endothelial cell alterations in patients with chronic obstructive pulmonary disease.

    Science.gov (United States)

    Soler, Núria; García-Heredia, Anabel; Marsillach, Judit; Mackness, Bharti; Mackness, Michael; Joven, Jorge; Romero, Pere; Camps, Jordi

    2013-08-28

    To investigate the relationships between the levels of the antioxidant enzyme paraoxonase-1 (PON1) and corneal endothelial alterations in patients with chronic obstructive pulmonary disease (COPD) undergoing cataract surgery. We studied 172 patients with cataract attending our ophthalmology clinic. Based on spirometric analysis, they were segregated into two groups, 110 (64%) with COPD and 62 (36%) without COPD. Corneal endothelial cell morphology was examined by widefield noncontact specular microscopy, which allows measurements of endothelial cell density (ECD), hexagonality, and endothelial cell size coefficient of variation (ECCV). Corneal thickness was measured by noncontact pachimetry. PON1 and plasma TNFα concentrations were analyzed by ELISA. Serum PON1 activity was analyzed by spectrophotometry. Patients with COPD had significant decreases in ECD, hexagonality, and corneal thickness, and a significant increase in ECCV. They also had significant decreases in serum PON1 activity but not in PON1 concentration. Serum PON1 activity showed a significant direct association with ECD, and an inverse association with corneal thickness. Results of the present study suggest that PON1 may be involved in the pathophysiology of corneal endothelial alterations in patients with COPD.

  20. Plastic compressed collagen as a novel carrier for expanded human corneal endothelial cells for transplantation.

    Directory of Open Access Journals (Sweden)

    Hannah J Levis

    Full Text Available Current treatments for reversible blindness caused by corneal endothelial cell failure involve replacing the failed endothelium with donor tissue using a one donor-one recipient strategy. Due to the increasing pressure of a worldwide donor cornea shortage there has been considerable interest in developing alternative strategies to treat endothelial disorders using expanded cell replacement therapy. Protocols have been developed which allow successful expansion of endothelial cells in vitro but this approach requires a supporting material that would allow easy transfer of cells to the recipient. We describe the first use of plastic compressed collagen as a highly effective, novel carrier for human corneal endothelial cells. A human corneal endothelial cell line and primary human corneal endothelial cells retained their characteristic cobblestone morphology and expression of tight junction protein ZO-1 and pump protein Na+/K+ ATPase α1 after culture on collagen constructs for up to 14 days. Additionally, ultrastructural analysis suggested a well-integrated endothelial layer with tightly opposed cells and apical microvilli. Plastic compressed collagen is a superior biomaterial in terms of its speed and ease of production and its ability to be manipulated in a clinically relevant manner without breakage. This method provides expanded endothelial cells with a substrate that could be suitable for transplantation allowing one donor cornea to potentially treat multiple patients.

  1. Plastic compressed collagen as a novel carrier for expanded human corneal endothelial cells for transplantation.

    Science.gov (United States)

    Levis, Hannah J; Peh, Gary S L; Toh, Kah-Peng; Poh, Rebekah; Shortt, Alex J; Drake, Rosemary A L; Mehta, Jodhbir S; Daniels, Julie T

    2012-01-01

    Current treatments for reversible blindness caused by corneal endothelial cell failure involve replacing the failed endothelium with donor tissue using a one donor-one recipient strategy. Due to the increasing pressure of a worldwide donor cornea shortage there has been considerable interest in developing alternative strategies to treat endothelial disorders using expanded cell replacement therapy. Protocols have been developed which allow successful expansion of endothelial cells in vitro but this approach requires a supporting material that would allow easy transfer of cells to the recipient. We describe the first use of plastic compressed collagen as a highly effective, novel carrier for human corneal endothelial cells. A human corneal endothelial cell line and primary human corneal endothelial cells retained their characteristic cobblestone morphology and expression of tight junction protein ZO-1 and pump protein Na+/K+ ATPase α1 after culture on collagen constructs for up to 14 days. Additionally, ultrastructural analysis suggested a well-integrated endothelial layer with tightly opposed cells and apical microvilli. Plastic compressed collagen is a superior biomaterial in terms of its speed and ease of production and its ability to be manipulated in a clinically relevant manner without breakage. This method provides expanded endothelial cells with a substrate that could be suitable for transplantation allowing one donor cornea to potentially treat multiple patients.

  2. Corneal epithelial and aqueous humor acidification during in vivo contact lens wear in rabbits.

    Science.gov (United States)

    Giasson, C; Bonanno, J A

    1994-03-01

    Based on contact lens-induced stromal acidification of the cornea, it has been suggested that the corneal epithelial and endothelial cells also become acidotic during contact lens wear. This alleged acidification may have a role in altered cell appearance and metabolism during contact lens wear. This study investigated the effects of anoxia, carbon dioxide retention, and contact lens gas transmissibility on the epithelial and aqueous humor pH in living rabbits. Epithelial intracellular pH (pHi) and aqueous humor pH were fluorophotometrically measured with a pH sensitive-dye (BCECF) during contact lens wear or exposure to various gas mixtures. Polymethylmethacrylate (PMMA) lens wear acidified epithelial cells by preventing CO2 efflux and by inducing hypoxia. Increasing lens oxygen transmissibility decreased epithelial acidification. After initiation of rigid, gas-permeable (RGP) lens wear or CO2-air exposure, pHi dropped transiently and then recovered partially. This recovery of pHi was not observed during anoxia, whether induced by PMMA lens wear or exposure to 100% N2. The aqueous humor also acidified during PMMA lens wear, a phenomenon not observed during RGP lens wear. Changes in aqueous pH were smaller, slower, and delayed when compared to their epithelial counterparts. Hypoxic contact lens wear acidifies the corneal epithelium and aqueous humor. The aqueous humor pH change indicates a probable endothelial acidification during hypoxic contact lens wear; the pH changes are caused by two separate and additive effects, CO2 retention and hypoxic acidosis. Increases in the oxygen transmissibility of the lens decrease the cellular acidosis, which might minimize cellular complications arising from contact lens wear. We estimate that a lens with an oxygen transmissibility (Dk/L) of 300 x 10(-11) (cm/sec)(ml O2/ml x mm Hg) is needed to prevent epithelial pHi changes in the open eye. In contrast, lenses with Dk/L as low as 18 x 10(-9) (cm/sec)(ml O2/ml x mm Hg) can

  3. Stability of the laser in situ keratomileusis corneal flap in rabbit eyes.

    Science.gov (United States)

    Laurent, John M; Schallhorn, Steven C; Spigelmire, Jennifer R; Tanzer, David J

    2006-06-01

    To quantify the force required to dislodge a laser in situ keratomileusis (LASIK) corneal flap in rabbits and correlate that force with sources of injury to LASIK corneal flaps in humans. Animal Vivarium and Department of Ophthalmology, Naval Medical Center, San Diego, California, USA. Corneal flaps were cut in 56 eyes of 30 rabbits. A -8.00 diopter excimer ablation was also performed in 32 of the eyes. Injury testing, using a burst of CO2 from a modified paintball gun, was conducted from 1 to 9 days postoperatively. Impact force was compared to that generated by mechanical trauma (fingers and a small tree branch) striking a force gauge. Of 43 eyes tested for injury, flaps could not be dislocated in 11 eyes (26%). There was no significant difference between eyes treated with flap only and those that also had ablation. When flaps were dislodged, the required force generally resulted in extensive intraocular injury. There was no significant difference between the force required for flap dislocation on postoperative days 1 and 9. In rabbits, LASIK corneal flaps were very resistant to high-speed wind trauma as early as 24 hours postoperatively. Flap stability was robust prior to the formation of collagen scar tissue, probably due to epithelial bridging at the edge of the flap and an osmotic gradient across the flap-stromal bed interface. Although this study used a rabbit model, it seems likely that both these flap adhesion mechanisms would also provide stability to the LASIK flap in humans.

  4. High resolution scanning electron microscopy of rabbit corneal endothelium to show effects of UV-visible irradiation in the presence of chlorpromazine

    Energy Technology Data Exchange (ETDEWEB)

    Lea, P.J.; Hollenberg, M.J.; Menon, I.A.; Temkin, R.J.; Persad, S.D.; Basu, P.K. (Univ. of Toronto, Ontario (Canada))

    1989-01-01

    The ultrastructure of rabbit cornea endothelial cells was examined by scanning electron microscopy (SEM) in freeze-cleaved corneas using a Hitachi S-570 scanning electron microscope in the high resolution mode (HRSEM). In order to study phototoxic effects in vitro, rabbit corneas (experimental) were cultured as organ culture in the presence of 5 micrograms/ml chlorpromazine (CPZ) and irradiated. For comparison, control 1 corneas were not irradiated but incubated in the dark without CPZ in the medium; control 2 corneas were also kept in the dark but in the presence of CPZ; control 3 corneas were irradiated with no CPZ in the medium. Cellular damage was not seen in the three types of control corneas, but in the experimental corneas the endothelial cells showed extensive disruption of the cell membrane and some deterioration of the intracellular components. Our study confirmed that HRSEM is a satisfactory new technique for visualizing damage of the intracellular organelles of corneal endothelium.

  5. Characterization Of Corneal Specular Endothelial Photomicrographs By Their Fourier Transforms

    Science.gov (United States)

    Masters, Barry R.

    1988-08-01

    The innermost layer of the cornea consists of a single layer of cells which are responsible for the active transport of fluid from the cornea to the aqueous humor. This cellular layer is about 5 pm thick and consist of cells of various shapes (pentagons, hexagons, hexagons, and octagons) although hexagons predominate. With aging and disease states, the cells become enlarged and the percent of hexagons decrease with the appearance of many five and seven side cells. The shape of the endothelial cells in the living cornea is usually evaluated from specular photomicrographs made in specularly reflected light which shows the cell borders in a square millimeter of corneal endothelium. The cell borders are traced out and the cells are analyzed with a digitizer pad, stylus, and computer software. This yields the cell area, density, coefficient of variation, and a histogram of n-sided cells. As an alternative approach to evaluate the cell shape and pattern optically, the following system was employed. The Fourier transform of the cell pattern was obtained on a digital image processor. This technique results in a fingerprint of the original cellular pattern which can be used to characterize the cellular arrangement.

  6. Proteomics of Fuchs' Endothelial Corneal Dystrophy Support That the Extracellular Matrix of Descemet's Membrane is Disordered

    DEFF Research Database (Denmark)

    Poulsen, Ebbe Toftgaard; Dyrlund, Thomas Franck; Runager, Kasper

    2014-01-01

    Fuchs' endothelial corneal dystrophy (FECD) is a major corneal disorder affecting the innermost part of the cornea, leading to visual impairment. As the morphological changes in FECD are mainly observed in the extracellular matrix of the Descemet's membrane/endothelial layer we determined...... that the morphological changes observed in FECD is caused in part by an aberrant assembly of the extracellular matrix within the Descemet's membrane/endothelial layer....... differentially regulated proteins, many of which are extracellular proteins known to be involved in proper assembly of the basement membrane in other tissues. In total 26 differentially regulated proteins were identified, of which 6 proteins were regulated by both methods. These results support...

  7. The effect of 193 nm excimer laser radiation on the human corneal endothelial cell density

    Energy Technology Data Exchange (ETDEWEB)

    Isager, P.; Hjortdal, J.Oe.; Ehlers, N. [Aarhus Univ. Hospital, Dept. of Ophthalmology, Aarhus (Denmark)

    1996-06-01

    The effect of 193 nm excimer laser radiation on human corneal endothelial cell density was examined. Fifty-five eyes from 35 patients underwent photorefractive keratectomy for myopia. Photomicrographs of the endothelium were taken a short time before the operation and on an average of 7 months postoperatively with a specular microscope. The average endothelial cell densities were preoperatively 3375 {+-} 266 cells/mm{sup 2} (means {+-} SD) and postoperatively 3348 {+-} 287 cells/mm{sup 2}, corresponding to a fall of 27 cells/mm{sup 2} (N = 55). This fall in endothelial cell density was not statistically significant. A significant correlation between the change in cell density and age of the patient was found, with older patients losing more cells (N = 35, 2p < 0.05). The magnification of the specular microscope was found to change with corneal thickness. The importance of correcting the endothelial cell densities for corneal thickness is discussed. (au) 14 refs.

  8. Generation of novel monoclonal antibodies for the enrichment and characterization of human corneal endothelial cells (hCENC) necessary for the treatment of corneal endothelial blindness.

    Science.gov (United States)

    Ding, Vanessa; Chin, Angela; Peh, Gary; Mehta, Jodhbir S; Choo, Andre

    2014-01-01

    Corneal transplantation is the primary treatment option to restore vision for patients with corneal endothelial blindness. Although the success rate of treatment is high, limited availability of transplant grade corneas is a major obstacle. Tissue-engineered corneal endothelial grafts constructed using cultivated human corneal endothelial cells (hCENC) isolated from cadaveric corneas may serve as a potential graft source. Currently, tools for the characterization of cultured hCENC and enrichment of hCENC from potential contaminating cells such as stromal fibroblasts are lacking. In this study, we describe the generation and characterization of novel cell surface monoclonal antibodies (mAbs) specific for hCENC. These mAbs could be used for enrichment and characterization of hCENC. Out of a total of 389 hybridomas, TAG-1A3 and TAG-2A12 were found to be specific to the corneal endothelial monolayer by immunostaining of frozen tissue sections. Both mAbs were able to clearly identify hCENC with good 'cobblestone-like' morphology from multiple donors. The antigen targets for TAG-1A3 and TAG-2A12 were found to be CD166/ALCAM and Peroxiredoxin-6 (Prdx-6), respectively, both of which have not been previously described as markers of hCENC. Additionally, unlike other Prdx-6 mAbs, TAG-2A12 was found to specifically bind cell surface Prdx-6, which was only expressed on hCENC and not on other cell types screened such as human corneal stromal fibroblasts (hCSF) and human pluripotent stem cells (hPSC). From our studies, we conclude that TAG-1A3 and TAG-2A12 are promising tools to quantitatively assess hCENC quality. It is also noteworthy that the binding specificity of TAG-2A12 could be used for the enrichment of hCENC from cell mixtures of hCSF and hPSC.

  9. Effects of edible bird's nest (EBN on cultured rabbit corneal keratocytes

    Directory of Open Access Journals (Sweden)

    Hun Lee

    2011-10-01

    Full Text Available Abstract Background There has been no effective treatment or agent that is available for corneal injury in promoting corneal wound healing. Previous studies on edible bird's nest extract (EBN had reported the presence of hormone-like substance; avian epidermal growth factor that could stimulate cell division and enhance regeneration. This study aimed to investigate the effects of EBN on corneal keratocytes proliferative capacity and phenotypical changes. Methods Corneal keratocytes from six New Zealand White Rabbits were isolated and cultured until Passage 1. The proliferative effects of EBN on corneal keratocytes were determined by MTT assay in serum-containing medium (FDS and serum-free medium (FD. Keratocytes phenotypical changes were morphologically assessed and gene expression of aldehyde dehydrogenase (ALDH, collagen type 1 and lumican were determined through RT-PCR. Results The highest cell proliferation was observed when both media were supplemented with 0.05% and 0.1% EBN. Cell proliferation was also consistently higher in FDS compared to FD. Both phase contrast micrographs and gene expression analysis confirmed the corneal keratocytes retained their phenotypes with the addition of EBN. Conclusions These results suggested that low concentration of EBN could synergistically induce cell proliferation, especially in serum-containing medium. This could be a novel breakthrough as both cell proliferation and functional maintenance are important during corneal wound healing. The in vitro test is considered as a crucial first step for nutri-pharmaceutical formation of EBN-based eye drops before in vivo application.

  10. Simultaneous bilensectomy and endothelial keratoplasty for angle-supported phakic intraocular lens-induced corneal decompensation

    Directory of Open Access Journals (Sweden)

    Vikas Mittal

    2011-01-01

    Full Text Available A 40-year-old lady presented with severe endothelial cell loss in both eyes 14 years after angle-supported phakic intraocular lens (AS PIOL implantation. The left eye had severe corneal edema with bullous keratopathy. The right eye had markedly reduced endothelial cell count (655 cells/mm 2 although the cornea was clear. She underwent simultaneous bilensectomy (AS PIOL explantation and phacoemulsification and Descemet′s stripping and endothelial keratoplasty (DSEK in the left eye. Explanted AS PIOL was identified as ZSAL-4 (Morcher, Stuttgart, Germany model. Corneal edema cleared completely in 2 months with a best corrected visual acuity (-2.25 D sph of 20/60. No intervention was done in the right eye. The present case illustrates that AS PIOL-induced endothelial decompensation can be effectively managed by simultaneous bilensectomy and endothelial keratoplasty.

  11. Changes in Anterior, Posterior, and Total Corneal Astigmatism after Descemet Membrane Endothelial Keratoplasty

    OpenAIRE

    Maged Alnawaiseh; Lars Zumhagen; André Rosentreter; Nicole Eter

    2017-01-01

    Purpose. To evaluate changes in anterior, posterior, and total corneal astigmatism in patients after Descemet membrane endothelial keratoplasty (DMEK). Methods. We retrospectively included 29 eyes of 23 patients (age 67.6???9.8 years, 13 female, 10 male) after DMEK surgery. The magnitude and axis orientation of anterior, posterior, and total corneal astigmatism before and after DMEK were determined using a rotating Scheimpflug system (Pentacam HR, Oculus). Results. The magnitude of anterior, ...

  12. Dynamic OCT measurements of corneal biomechanical properties after UV cross-linking in the rabbit

    Science.gov (United States)

    Twa, Michael D.; Li, Jiasong; Manapuram, Ravi K.; Menodiado, Floredes M.; Singh, Manmohan; Aglyamov, Salavat; Emelianov, Stanislav; Larin, Kirill V.

    2013-03-01

    Structural properties of the cornea determine the shape and optical quality of the eye. Keratoconus, a structural degeneration of the cornea, is often treated with UV-induced collagen cross-linking to increase tissue resistance to further deformation and degeneration. Optimal treatments would be customized to the individual and consider preexisting structural properties as well as the effects induced by treatment and this requires the capability to noninvasively measure tissue properties. The purpose of this study is to use novel methods of optical elastography to study the effects of UV-induced corneal collagen cross-linking in the rabbit eye. Low-amplitude (corneal stiffness.

  13. Descemet membrane endothelial keratoplasty with a stromal rim in the treatment of posterior polymorphous corneal dystrophy

    Directory of Open Access Journals (Sweden)

    Pavel Studeny

    2012-01-01

    Full Text Available A 20-year-old patient, diagnosed with posterior polymorphous corneal dystrophy, developed corneal edema for which he underwent Descemet membrane endothelial keratoplasty with a stromal rim (DMEK-S in the right eye. No intra- or postoperative complications were noted. At the last follow-up 2 years and 9 months after the procedure, the best corrected visual acuity was 1.0 and endothelial cell density declined from 3533 cells/mm 2 to 1012 cells/mm 2 . Despite the endothelial cell loss, DMEK-S appears to be a good alternative to other surgical techniques for the treatment of corneal endotheliopathies, and it may be of benefit to young patients.

  14. Donor Age and Corneal Endothelial Cell Loss 5 Years after Successful Corneal Transplantation: Specular Microscopy Ancillary Study Results

    Science.gov (United States)

    2010-01-01

    Objective To determine whether endothelial cell loss 5 years after successful corneal transplantation is related to the age of the donor. Design Multicenter, prospective, double-masked clinical trial. Participants Three hundred forty-seven subjects participating in the Cornea Donor Study who had not experienced graft failure 5 years after corneal transplantation for a moderate-risk condition (principally Fuchs’ dystrophy or pseudophakic corneal edema). Testing Specular microscopic images of donor corneas obtained before surgery and postoperatively at 6 months, 12 months, and then annually through 5 years were submitted to a central reading center to measure endothelial cell density (ECD). Main Outcome Measure Endothelial cell density at 5 years. Results At 5 years, there was a substantial decrease in ECD from baseline for all donor ages. Subjects who received a cornea from a donor 12 to 65 years old experienced a median cell loss of 69% in the study eye, resulting in a 5-year median ECD of 824 cells/mm2 (interquartile range, 613–1342), whereas subjects who received a cornea from a donor 66 to 75 years old experienced a cell loss of 75%, resulting in a median 5-year ECD of 654 cells/mm2 (interquartile range, 538–986) (P [adjusted for baseline ECD] = 0.04). Statistically, there was a weak negative association between ECD and donor age analyzed as a continuous variable (r [adjusted for baseline ECD] = −0.19; 95% confidence interval, −0.29 to −0.08). Conclusions Endothelial cell loss is substantial in the 5 years after corneal transplantation. There is a slight association between cell loss and donor age. This finding emphasizes the importance of longer-term follow-up of this cohort to determine if this relationship affects graft survival. PMID:18387408

  15. Inhibition of TGF-β signaling enables human corneal endothelial cell expansion in vitro for use in regenerative medicine.

    Directory of Open Access Journals (Sweden)

    Naoki Okumura

    Full Text Available Corneal endothelial dysfunctions occurring in patients with Fuchs' endothelial corneal dystrophy, pseudoexfoliation syndrome, corneal endotheliitis, and surgically induced corneal endothelial damage cause blindness due to the loss of endothelial function that maintains corneal transparency. Transplantation of cultivated corneal endothelial cells (CECs has been researched to repair endothelial dysfunction in animal models, though the in vitro expansion of human CECs (HCECs is a pivotal practical issue. In this study we established an optimum condition for the cultivation of HCECs. When exposed to culture conditions, both primate and human CECs showed two distinct phenotypes: contact-inhibited polygonal monolayer and fibroblastic phenotypes. The use of SB431542, a selective inhibitor of the transforming growth factor-beta (TGF-β receptor, counteracted the fibroblastic phenotypes to the normal contact-inhibited monolayer, and these polygonal cells maintained endothelial physiological functions. Expression of ZO-1 and Na(+/K(+-ATPase maintained their subcellular localization at the plasma membrane. Furthermore, expression of type I collagen and fibronectin was greatly reduced. This present study may prove to be the substantial protocol to provide the efficient in vitro expansion of HCECs with an inhibitor to the TGF-β receptor, and may ultimately provide clinicians with a new therapeutic modality in regenerative medicine for the treatment of corneal endothelial dysfunctions.

  16. Optimization of Human Corneal Endothelial Cells for Culture: The Removal of Corneal Stromal Fibroblast Contamination Using Magnetic Cell Separation

    Directory of Open Access Journals (Sweden)

    Gary S. L. Peh

    2012-01-01

    Full Text Available The culture of human corneal endothelial cells (CECs is critical for the development of suitable graft alternative on biodegradable material, specifically for endothelial keratoplasty, which can potentially alleviate the global shortage of transplant-grade donor corneas available. However, the propagation of slow proliferative CECs in vitro can be hindered by rapid growing stromal corneal fibroblasts (CSFs that may be coisolated in some cases. The purpose of this study was to evaluate a strategy using magnetic cell separation (MACS technique to deplete the contaminating CSFs from CEC cultures using antifibroblast magnetic microbeads. Separated “labeled” and “flow-through” cell fractions were collected separately, cultured, and morphologically assessed. Cells from the “flow-through” fraction displayed compact polygonal morphology and expressed Na+/K+ATPase indicative of corneal endothelial cells, whilst cells from the “labeled” fraction were mostly elongated and fibroblastic. A separation efficacy of 96.88% was observed. Hence, MACS technique can be useful in the depletion of contaminating CSFs from within a culture of CECs.

  17. Inhibition of Corneal Neovascularization by Hydrazinocurcumin ...

    African Journals Online (AJOL)

    Purpose: To investigate the effect of hydrazinocurcumin on a human vascular endothelial growth factor (VEGF)-induced corneal neovascularization in rabbit model. Methods: Murine corneal neovascularization (CorNV) was induced via two intrastromal implantations of VEGF polymer 2 mm from the limbus.

  18. Inhibition of Corneal Neovascularization by Hydrazinocurcumin

    African Journals Online (AJOL)

    Purpose: To investigate the effect of hydrazinocurcumin on a human vascular endothelial growth factor. (VEGF)-induced corneal neovascularization in rabbit model. Methods: Murine corneal neovascularization (CorNV) was induced via two intrastromal implantations of. VEGF polymer 2 mm from the limbus.

  19. [Morphometric changes of corneal endothelial cells in pseudoexfoliation syndrome and pseudoexfoliation glaucoma].

    Science.gov (United States)

    de Juan-Marcos, L; Cabrillo-Estévez, L; Escudero-Domínguez, F A; Sánchez-Jara, A; Hernández-Galilea, E

    2013-11-01

    To evaluate the corneal endothelial morphometry and central corneal thickness (CCT) in pseudoexfoliative (PEX) eyes with and without glaucoma and to compare with normal eyes and eyes with primary open-angle glaucoma (POAG). A total of 166 patients were included in this study: 36 eyes with pseudoexfoliation syndrome (PXS), 30 eyes with pseudoexfoliation glaucoma (PXG), 40 eyes with POAG, and 60 normal eyes. Corneal endothelial cell density (ECD), coefficient of variation (CV) in cell size, and percentage of hexagonal cells, were measured using a non-contact specular microscope, whereas CCT was measured with an ultrasonic pachymeter. ECD and percentage of hexagonal cells were lower in PEX groups and in the POAG group compared with normal eyes, while the CV in cell size was greater. There was a tendency for greater cell loss and morphological abnormalities of the corneal endothelial cells in PXG eyes compared to PXS eyes, when all pseudoexfoliative eyes were analyzed together. Changes in endothelial cells increased with age. There were no significant differences in mean CCT between the four groups. Endothelial cell density is significantly decreased, and pleomorphism and polymegathism of cells are increased in PEX eyes, particularly when intraocular pressure is high. Copyright © 2012 Sociedad Española de Oftalmología. Published by Elsevier Espana. All rights reserved.

  20. [Comparison of corneal wound healing of photorefractive keratectomy and laser in situ keratomileusis in rabbits].

    Science.gov (United States)

    Ma, Xiao-Hua; Li, Jing-Hai; Bi, Hong-Sheng; Zhou, Fang; Li, Yan

    2003-03-01

    To compare effects of photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) on the cornea and corneal wound healing and to investigate the possible mechanism of corneal haze and myopic regression histopathologically. Twenty-four New Zealand white rabbits were allocated randomly to correct -4.00 and -8.00 diopters and were operated with PRK on right eyes and LASIK on left eyes. At 10 days and 1, 3, and 6 months, corneal haze was observed, refraction was evaluated, and 3 rabbits were randomly selected for each time point to be enucleated, and corneas to be bisected. One half of each cornea was evaluated using transmission electron microscopy, and the other half was evaluated using either light microscopy or immunohistochemical staining for collagen type III, IV, fibronectin (FN) and transforming growth factor-beta(1) (TGF-beta(1)). Different degrees of corneal haze and myopic regression were observed after PRK: the higher the desired myopic correction, the heavier the haze. Corneal wound healing response was greater and lasted longer after PRK than after LASIK. Various pathological findings such as epithelial hyperplasia, basal membrane reforming and extracellular matrix deposits were found in the ablation zone and repair mechanisms were still active at 6 months after PRK. Whereas after LASIK, the interface between the flap and stromal bed was transparent except for growing epithelial plugs and lightly proliferating stroma coinciding with the flap margins. After both PRK and LASIK, all corneal cell types were consistently positive for TGF-beta(1) antibody during the corneal wound healing time. TGF-beta(1) antibody positivity decreased as the wound healing approached completion. The histopathological changes of corneal haze and myopic regression are as follows: epithelial hyperplasia, basal membrane immaturity, anterior stromal keratocyte increase and activity, new collagen III production and irregular arrangement thereof, and FN deposition in the

  1. Flex center method versus center method for endothelial corneal evaluation in eye banking: a comparative analysis.

    Science.gov (United States)

    Villalba, R; Jiménez, A; Fornés, G; Eisman, M; Villagrán, J L Gómez

    2014-12-01

    Specular microscopy can provide a non-invasive morphological analysis of the cornea endothelial cell layer. A variety of analysis programs are available to determine corneal endothelial quality. The flex-center endothelial analysis method (Konan Inc) is a newer technique including the outermost digitized cells and thus increases the number of cells for analysis. The aim of this study is to analyze whether the new flex-center method, increases the possibilities of corneal endothelium evaluation before implants. For this purpose 67 corneas were studied by both methods at the Eye Bank of the Tissue Establishment of Córdoba. Although we have found differences in the resulting of number of cells in the area analysed, no significant differences were found with respect to the endothelial cell count, coefficient of variation cell area, and the percentage of hexagonal cells recorded. Based on this data, both methods can be used satisfactorily in eye banking.

  2. Corneal endothelial morphology and central thickness in patients with type II diabetes mellitus

    DEFF Research Database (Denmark)

    Storr-Paulsen, Allan; Singh, Amardeep; Jeppesen, Helene

    2014-01-01

    PURPOSE: To investigate corneal endothelial cell density and morphology in type II diabetic and non-diabetic patients and to relate potential differences to the glycaemic status. METHODS: A prospective clinical study including 107 patients with type II diabetes and 128 non-diabetic patients. Sample...... blood tests. The endothelial cell density, the variation in endothelial cell size (CV), the percentage of hexagonal cells, and the central corneal thickness (CCT) were recorded. RESULTS: Type II diabetic subjects did not differ from the non-diabetic control subjects with regards to endothelial cell...... density, hexagonality or variation in CV, but showed a significant increase in CCT (538 versus 546 μm, p diabetic group, lower cell counts were associated with higher HbA1c values (p II diabetes has no impact...

  3. Proteomic Analyses of the Acute Tissue Response for Explant Rabbit Corneas and Engineered Corneal Tissue Models Following In Vitro Exposure to 1540 nm Laser Light

    National Research Council Canada - National Science Library

    Eurell, T. E; Johnson, T. E; Roach, W. P

    2005-01-01

    Two-dimensional electrophoresis and histomorphometry were used to determine if equivalent protein changes occurred within native rabbit corneas and engineered corneal tissue models following in vitro...

  4. Fully automated corneal endothelial morphometry of images captured by clinical specular microscopy

    Science.gov (United States)

    Bucht, Curry; Söderberg, Per; Manneberg, Göran

    2010-02-01

    The corneal endothelium serves as the posterior barrier of the cornea. Factors such as clarity and refractive properties of the cornea are in direct relationship to the quality of the endothelium. The endothelial cell density is considered the most important morphological factor of the corneal endothelium. Pathological conditions and physical trauma may threaten the endothelial cell density to such an extent that the optical property of the cornea and thus clear eyesight is threatened. Diagnosis of the corneal endothelium through morphometry is an important part of several clinical applications. Morphometry of the corneal endothelium is presently carried out by semi automated analysis of pictures captured by a Clinical Specular Microscope (CSM). Because of the occasional need of operator involvement, this process can be tedious, having a negative impact on sampling size. This study was dedicated to the development and use of fully automated analysis of a very large range of images of the corneal endothelium, captured by CSM, using Fourier analysis. Software was developed in the mathematical programming language Matlab. Pictures of the corneal endothelium, captured by CSM, were read into the analysis software. The software automatically performed digital enhancement of the images, normalizing lights and contrasts. The digitally enhanced images of the corneal endothelium were Fourier transformed, using the fast Fourier transform (FFT) and stored as new images. Tools were developed and applied for identification and analysis of relevant characteristics of the Fourier transformed images. The data obtained from each Fourier transformed image was used to calculate the mean cell density of its corresponding corneal endothelium. The calculation was based on well known diffraction theory. Results in form of estimated cell density of the corneal endothelium were obtained, using fully automated analysis software on 292 images captured by CSM. The cell density obtained by the

  5. Treatment Results of Corneal Collagen Cross-Linking Combined with Riboflavin and 440 Nm Blue Light for Bacterial Corneal Ulcer in Rabbits.

    Science.gov (United States)

    Wei, Shufang; Zhang, Cuiying; Zhang, Shaoru; Xu, Yanyun; Mu, Guoying

    2017-10-01

    To study the treatment effect of corneal collagen cross-linking (CXL) combined with 440 nm blue light and riboflavin on bacterial corneal ulcer using animal experiments. A total of 21 New Zealand white rabbits that developed Staphylococcus aureus corneal ulcer were randomly divided into three groups. Seven rabbits were used as blank control groups; seven rabbits were treated with CXL combined with riboflavin and 440 nm blue light; and seven rabbits were treated with CXL combined with riboflavin and 370 nm ultraviolet A light. Necrotic tissues or secretions from the ulcer surface, eye secretions, conjunctival hyperemia, hypopyon, corneal infiltration, and pathological changes of the cornea were all observed. The 1st, 3th, and 7th day after CXL treatment, a statistically significant difference was found among the inflammation scores of the three groups. The scores of 440 and 370 groups decreased gradually, significantly lower than that of the control group. Bacterial cultures of 440 and 370 groups turned to be negative while that of the control group remained positive. After 1 day of CXL treatment, pathology pictures of the three groups all showed loss of corneal epithelia with many inflammatory cells in deep stroma. After 7 days of CXL treatment, abscess formed in almost all corneal area in the control group, while in 440 and 370 groups, multilayer healing of corneal epithelia, neovascularization, and many inflammatory cells within ulcers and proliferation of a small amount of fibroblast were seen. CXL combined with riboflavin and 440 nm blue light is effective in treating S. aureus corneal ulcer.

  6. Actin filament organization during endothelial wound healing in the rabbit cornea: comparison between transcorneal freeze and mechanical scrape injuries.

    Science.gov (United States)

    Ichijima, H; Petroll, W M; Barry, P A; Andrews, P M; Dai, M; Cavanagh, H D; Jester, J V

    1993-08-01

    To compare and contrast the in vivo mechanism of wound healing after mechanical scrape and transcorneal freeze (TCF) injury in a rabbit eye model by examining changes in the cytoskeletal organization of contractile, filamentous actin (f-actin) microfilaments as relates to differences in cell migration or translocation during endothelial repair. Endothelial wound healing after mechanical scrape and transcorneal freeze injury was studied in rabbit eyes using laser scanning confocal microscopy (LSCM). Central corneal mechanical scrape injury was made using an olive tip cannula, and TCF injury was made using a 3-mm diameter stainless steel probe cooled with liquid nitrogen. Cytoskeletal changes in f-actin stained with phalloidin-FITC were observed during wound healing using LSCM. At 6 hours after mechanical scrape, the leading edge of the migrating sheet showed a decrease in the intensity of phalloidin-FITC staining, suggesting a decrease in cortical f-actin. Migrating endothelial cells in vivo did not appear to develop stress fibers after mechanical scrape, which is consistent with an in vitro cell spreading mechanism of endothelial wound healing. By 24 hours, the denuded area was almost fully resurfaced by migrating endothelial cells. On the other hand, TCF injury produced fibroblastic changes in the endothelial cells with extension and elongation of spindle-shaped endothelial cells at the leading edge by 24 hours after injury. Fibroblastic endothelial cells developed prominent actin stress-fibers, which is consistent with an in vitro cell migration mechanism of endothelial wound healing. Three days after TCF, the wounded area was resurfaced with two cell types: rough, fibroblast-like cells forming a retrocorneal fibrous membrane having prominent f-actin bundles or stress fibers with few cell-cell junctions, and smooth, polygonal-shaped endothelial cells having tight cell junctions with a cortical distribution of f-actin. After 28 days the retrocorneal fibrous

  7. Apatinib-loaded nanoparticles suppress vascular endothelial growth factor-induced angiogenesis and experimental corneal neovascularization.

    Science.gov (United States)

    Lee, Jung Eun; Kim, Koung Li; Kim, Danbi; Yeo, Yeongju; Han, Hyounkoo; Kim, Myung Goo; Kim, Sun Hwa; Kim, Hyuncheol; Jeong, Ji Hoon; Suh, Wonhee

    2017-01-01

    Pathological angiogenesis is one of the major symptoms of severe ocular diseases, including corneal neovascularization. The blockade of vascular endothelial growth factor (VEGF) action has been recognized as an efficient strategy for treating corneal neovascularization. In this study, we aimed to investigate whether nanoparticle-based delivery of apatinib, a novel and selective inhibitor of VEGF receptor 2, inhibits VEGF-mediated angiogenesis and suppresses experimental corneal neovascularization. Water-insoluble apatinib was encapsulated in nanoparticles composed of human serum albumin (HSA)-conjugated polyethylene glycol (PEG). In vitro angiogenesis assays showed that apatinib-loaded HSA-PEG (Apa-HSA-PEG) nanoparticles potently inhibited VEGF-induced tube formation, scratch wounding migration, and proliferation of human endothelial cells. In a rat model of alkali burn injury-induced corneal neovascularization, a subconjunctival injection of Apa-HSA-PEG nanoparticles induced a significant decrease in neovascularization compared to that observed with an injection of free apatinib solution or phosphate-buffered saline. An in vivo distribution study using HSA-PEG nanoparticles loaded with fluorescent hydrophobic model drugs revealed the presence of a substantial number of nanoparticles in the corneal stroma within 24 h after injection. These in vitro and in vivo results demonstrate that apatinib-loaded nanoparticles may be promising for the prevention and treatment of corneal neovascularization-related ocular disorders.

  8. Experimental study of plasmid TGF-beta 1 DNA gene transfer with lipofectamine into rabbit corneal epithelial cells in vitro.

    Science.gov (United States)

    Huang, Qiong; Hu, Yanhua; Jiang, Fagang; Chen, Hong

    2002-01-01

    To investigate whether the TGF-beta 1 plasmid DNA carried by lipofectamine could be introduced into cultured rabbit corneal epithelial cells, specific expression of the plasmid pMAM TGF-beta 1 in the cultured corneal epithelial cells was studied. Two days after 12 h of transfection of pMAMT-GF-beta 1 mediated by lipofectamine into the cultured corneal epithelial cells, the TGF-beta 1 protein expression specific for pMAMTGF-beta 1 in the cells was detected by means of immunohistochemical staining and the positive rate was 23.37%. The results suggested that foreign plasmid DNA could be effectively delivered into cultured rabbit corneal epithelial cells by means of lipofectamine, and this will provide a promising method of studying TGF-beta 1 on the mechanism of physiology and pathology concerned with corneal epithelial cells.

  9. Methods Development for the Isolation and Culture of Primary Corneal Endothelial Cells

    Science.gov (United States)

    2017-02-01

    stage component (C) incorporates a raised convex button for seating an inverted cornea. Panel D depicts the device as assembled for use. A...6. 21. Yue BY, Sugar J, Gilboy JE, Elvart JL. Growth of human corneal endothelial cells in culture. Invest Ophthalmol Vis Sci 1989; 30(2): 248-53

  10. Effects of phthalates on the human corneal endothelial cell line B4G12

    DEFF Research Database (Denmark)

    Krüger, Tanja; Cao, Yi; Kjærgaard, Søren K.

    2012-01-01

    Phthalates are industrial chemicals used in many cosmetics. We evaluated an in vitro model for eye irritancy testing using the human corneal endothelial cell line B4G12. Cell proliferation and toxicity were assessed after exposing to di-n-butyl phthalate (DBP), benzyl butyl phthalate (BBP), di-2...

  11. Hearing disability in patients with Fuchs' endothelial corneal dystrophy: unrecognized co-pathology?

    NARCIS (Netherlands)

    Stehouwer, Marilette; Bijlsma, Ward R.; van der Lelij, Allegonda

    2011-01-01

    To investigate a possible association between Fuchs' endothelial corneal dystrophy (FECD) and hearing disability. A cross-sectional observational study was performed at the University Medical Center Utrecht. Cases and controls were patients who were treated by a cornea specialist between 2004 and

  12. Collagen types in healing alkali-burned corneal stroma in rabbits.

    Science.gov (United States)

    Saika, S; Ooshima, A; Shima, K; Tanaka, S; Ohnishi, Y

    1996-01-01

    We evaluated the change in the type of collagen found during healing in the alkali-burned corneal stroma of rabbits. We estimated the relative proportions of alpha chains in pepsin-solubilized collagen, using sodium dodecylsulfate polyacrylamide gel electrophoresis. Bands of alpha 1(I), alpha 2(I), alpha 1(V), and alpha 1(III) chains stained with Coomassie Blue were separated. The alpha 1(III) and alpha 1(V) chains showed a transient proportional increase in healing corneal stroma in the area exposed to alkali. No significant alterations in collagen alpha chains were detected in peripheral corneal stroma that had not been directly exposed to alkali. Our results suggest that the keratocytes which repopulate the alkali-injured corneal stroma early in healing synthesize a higher proportion of collagen types III and V than type I, and then switch to synthesizing predominantly type I collagen as stromal healing progresses. Collagen types III and V thus appear to be of primary importance in the healing of the corneal stroma.

  13. Changes in Anterior, Posterior, and Total Corneal Astigmatism after Descemet Membrane Endothelial Keratoplasty

    Science.gov (United States)

    Zumhagen, Lars; Rosentreter, André; Eter, Nicole

    2017-01-01

    Purpose. To evaluate changes in anterior, posterior, and total corneal astigmatism in patients after Descemet membrane endothelial keratoplasty (DMEK). Methods. We retrospectively included 29 eyes of 23 patients (age 67.6 ± 9.8 years, 13 female, 10 male) after DMEK surgery. The magnitude and axis orientation of anterior, posterior, and total corneal astigmatism before and after DMEK were determined using a rotating Scheimpflug system (Pentacam HR, Oculus). Results. The magnitude of anterior, posterior, and total corneal astigmatism in the central cornea did not change significantly after surgery. Before surgery, we found a significant correlation between the magnitudes of anterior and posterior corneal astigmatism (Spearman's correlation coefficient (rS) = 0.526, P = 0.003), while after surgery this correlation was no longer significant (rS = 0.038, P = 0.843). There was a significant correlation between the vector difference between preoperative and postoperative posterior astigmatism and the change in corneal pachymetry (rP = 0.47, P = 0.010). Conclusions. Posterior corneal astigmatism (especially the orientation) and therefore the relationship between anterior and total corneal astigmatism may change after DMEK. This should be considered to improve the accuracy of toric IOL power calculations following phakic DMEK or in combined procedures. PMID:28553547

  14. Changes in Anterior, Posterior, and Total Corneal Astigmatism after Descemet Membrane Endothelial Keratoplasty

    Directory of Open Access Journals (Sweden)

    Maged Alnawaiseh

    2017-01-01

    Full Text Available Purpose. To evaluate changes in anterior, posterior, and total corneal astigmatism in patients after Descemet membrane endothelial keratoplasty (DMEK. Methods. We retrospectively included 29 eyes of 23 patients (age 67.6 ± 9.8 years, 13 female, 10 male after DMEK surgery. The magnitude and axis orientation of anterior, posterior, and total corneal astigmatism before and after DMEK were determined using a rotating Scheimpflug system (Pentacam HR, Oculus. Results. The magnitude of anterior, posterior, and total corneal astigmatism in the central cornea did not change significantly after surgery. Before surgery, we found a significant correlation between the magnitudes of anterior and posterior corneal astigmatism (Spearman’s correlation coefficient rS=0.526, P=0.003, while after surgery this correlation was no longer significant (rS=0.038, P=0.843. There was a significant correlation between the vector difference between preoperative and postoperative posterior astigmatism and the change in corneal pachymetry (rP=0.47, P=0.010. Conclusions. Posterior corneal astigmatism (especially the orientation and therefore the relationship between anterior and total corneal astigmatism may change after DMEK. This should be considered to improve the accuracy of toric IOL power calculations following phakic DMEK or in combined procedures.

  15. Corneal endothelial cell changes 5 years after laser in situ keratomileusis: femtosecond laser versus mechanical microkeratome.

    Science.gov (United States)

    Klingler, Kyle N; McLaren, Jay W; Bourne, William M; Patel, Sanjay V

    2012-12-01

    To compare corneal endothelial cell density (ECD) and morphology between flap creation with a femtosecond laser and flap creation with a mechanical microkeratome 5 years after laser in situ keratomileusis (LASIK). Mayo Clinic, Rochester, Minnesota, USA. Prospective randomized masked paired-eye study. In this study of LASIK for myopia or myopic astigmatism, fellow eyes were randomized by ocular dominance to flap creation by a femtosecond laser or by a mechanical microkeratome. Central endothelial images were analyzed before and 3 years and 5 years after LASIK; endothelial cell variables were compared between treatments at each examination. Relationships between endothelial cell loss and contact lens wear, residual bed thickness, and preoperative refractive error were evaluated. There were no differences in the ECD, percentage of hexagonal cells, or coefficient of variation of cell area between treatments at any examination (all P = .99); the smallest detectable differences were 120 cells/mm(2), 5%, and 2%, respectively. The mean annual rate of corneal endothelial cell loss was -0.1% ± 1.2% (SD) and -0.1% ± 1.0% for the femtosecond laser and the mechanical microkeratome, respectively. Endothelial cell loss was not associated with contact lens wear, residual bed thickness, or preoperative refractive error. The energy delivered to the cornea during femtosecond laser flap creation did not affect the corneal endothelium 5 years after LASIK when compared with flap creation with a mechanical microkeratome. Corneas that have had either method of flap creation could be accepted as donor tissue for endothelial keratoplasty from the standpoint of endothelial health. No author has a financial or proprietary interest in any material or method mentioned. Copyright © 2012 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

  16. The Effect of Ascorbic Acid (Vitamin C) on Transepithelial Corneal Cross-Linking in Rabbits.

    Science.gov (United States)

    Koc, Mustafa; Bostanci, Basak; Demirel, Ozlem Ozbas; Genc, Feyza; Tekin, Kemal; Koban, Yaran; Dincel, Aylin Sepici; Sen, Murat; Yilmazbas, Pelin

    2017-09-01

    To evaluate the effects of ascorbic acid (vitamin C), the main antioxidant agent in the cornea on transepithelial corneal cross-linking (CXL) where the main mechanism is oxidation. Twenty eyes of 20 rabbits were divided into 3 groups: Group 1 (7 eyes) had transepithelial corneal CXL after being fed with normal diet; Group 2 (7 eyes) had corneal CXL after once-daily subcutaneous injections of 200 mg of ascorbic acid in addition to normal diet; and the control group (6 eyes) was fed with normal diet but did not have corneal CXL performed. Ascorbic acid levels were measured in aqueous humor and plasma, and biomechanical measurements were applied to the cornea. There was a significant difference in ascorbic acid levels of plasma (P = 0.008) and aqueous humor (P = 0.006) between group 1 and 2. The Young's modulus values of group 1 and 2 were similar (P = 0.741) and were significantly higher than the control group (P = 0.02 and P = 0.01). The increase rate in Young's modulus values was 37.3% in group 1 and 43.9% in group 2 compared to control group. The ultimate strain values in group 1 and 2 were similar (P = 0.632) and were significantly higher than control group (P = 0.04, P = 0.03). The ultimate stress values in group 1 and 2 were similar (P = 0.836) and were significantly lower than control group (P = 0.001, P = 0.001). Systemic vitamin C does not appear to decrease effectiveness of transepithelial corneal CXL. Therefore, there is no reason to stop or reduce vitamin C supplementation before corneal CXL therapy.

  17. Disruption of zonula occludens-1 localization in the rabbit corneal epithelium by contact lens-induced hypoxia.

    Science.gov (United States)

    Yanai, Ryoji; Ko, Ji-Ae; Morishige, Naoyuki; Chikama, Tai-ichiro; Ichijima, Hideji; Nishida, Teruo

    2009-10-01

    Hypoxia impairs the barrier function of the corneal epithelium. This function depends on tight junctions, of which zonula occludens (ZO)-1 is a major component. The authors have investigated the effects of hypoxia on ZO-1 localization and expression in the rabbit corneal epithelium in vivo. A polymethylmethacrylate (PMMA) or rigid gas-permeable (RGP) lens was applied to one eye each of albino rabbits for 24 hours. The structure of the corneal epithelium was examined by in vivo confocal microscopy, and epithelial barrier function was evaluated by measurement of central corneal thickness. The distribution and expression of ZO-1 in the corneal epithelium were examined by immunofluorescence analysis and by immunoblot and reverse transcription-polymerase chain reaction analyses, respectively. Application of a PMMA lens, but not that of an RGP lens, resulted in a reduction in cell size at the surface of the corneal epithelium, compared with that in control eyes, and an increase in central corneal thickness. Immunofluorescence analysis revealed a continuous pattern of ZO-1 immunoreactivity around the perimeter of superficial corneal epithelial cells in control eyes or in eyes treated with an RGP lens. In contrast, the pattern of ZO-1 staining was discontinuous and patchy in eyes treated with a PMMA lens. Amounts of ZO-1 mRNA and protein in the corneal epithelium were reduced by application of a PMMA lens but not by that of an RGP lens. Hypoxia at the ocular surface induced the disruption of tight junctions between superficial cells in the rabbit corneal epithelium in vivo.

  18. Efficacy and safety of femtosecond laser-assisted corneal endothelial keratoplasty: a randomized multicenter clinical trial.

    Science.gov (United States)

    Cheng, Yanny Y Y; Schouten, Jan S A G; Tahzib, Nayyirih G; Wijdh, Robert-Jan; Pels, Elisabeth; van Cleynenbreugel, Hugo; Eggink, Catharina A; Rijneveld, Wilhelmina J; Nuijts, Rudy M M A

    2009-12-15

    To evaluate the efficacy and safety of femtosecond laser-assisted endothelial keratoplasty (FLEK) versus penetrating keratoplasty (PK) in patients with corneal endothelial disease. A randomized multicenter clinical trial of 80 eyes of 80 patients with corneal endothelial disease were randomized to FLEK or PK. Clinical outcomes (astigmatism and visual acuity) and incidence of postoperative complications were compared between the two groups. At 12 months, the percentage of eyes with a refractive astigmatism less than or equal to 3 diopters was higher in the FLEK group in comparison with the PK group (86.2% vs. 51.3%, P=0.004). The mean postoperative best corrected visual acuity was 20/70+/-2 lines in the FLEK group and 20/44+/-2 lines in the PK group (Pastigmatism and results in an absence of wound healing related problems in patients with endothelial disease. However, visual acuity is lower as compared with conventional PK, and the high level of endothelial cell loss warrants a modification of the insertion technique of the endothelial graft.

  19. [Corneal wound healing and immunohistological features of extracellular matrix following penetrating keratoplasty in rabbits].

    Science.gov (United States)

    Kato, T; Nakayasu, K; Kanai, A

    1999-07-01

    To analyze corneal wound healing after penetrating keratoplasty (PKP). We performed PKP on 20 white rabbit eyes, and applied immunohistochemical techniques. The distribution of type I, III and IV collagens, large proteoglycan, chondroitin 6-sulfate, chondroitin 4-sulfate, and vimentin was determined at postoperative intervals of 3 days, 1 week, 2 weeks, 1 month and 3 months. By day 3, staining for type IV collagen was observed along the host-graft junction. By day 7, staining for type III collagen, large proteoglycan and chondroitin 6-sulfate had increased in the repair region, and then diminished with increasing postoperative time. Epithelial wound healing required more than one month, whereas the remodeling of Descemet's membrane did not terminate at 3 months after PKP. The data from this study suggest that type III collagen, large proteoglycan and chondroitin 6-sulfate play a crucial role in the corneal wound healing after PKP.

  20. Preliminary study of rabbit model with corneal neovascularization after thermal burn under the constant temperature

    Directory of Open Access Journals (Sweden)

    Yong Jia

    2014-07-01

    Full Text Available AIM:To explore the suitable conditions in rapid model of corneal neovascularization(CNVafter thermal burn under different constant temperature in rabbit. METHODS: Total 45 New Zealand white rabbits were divided randomly into five groups(A, B, C, D, E. A groups: 100℃(n=10, B groups: 200℃(n=10, C groups: 300℃(n=10, D groups: 400℃(n=10, and E groups: control group(n=5. All left eyes of rabbits in A,B,C,D groups were induced corneal neovascularization by constant temperature burning device. The growth of CNV was observed by slit lamp microscope and the area of CNV were recorded on 4 th, 7 th, 14th, 30th days postoperatively. SPSS 19.0 statistical package was used for data analysis, and the data was recorded by mean±standard deviation. Comparison by analysis of variance was made by repeated measures in the area of neovascularization at each time point in groups. Statistical tests were considered significantly when P values were less than 0.05. RESULTS: On postoperative 4th, 7th, 14th, 30th days: no neovascularization was found after corneal thermal burn in A group, but only a few nebula left(n=2; the area of CNV were(9.16±1.45mm2,(37.73±5.49mm2,(62.44±7.54mm2,(40.28±7.39mm2 in B group respectively; and(11.45±1.04mm2,(44.51±4.64mm2,(66.13±4.13mm2,(43.04±2.33mm2 in C group respectively; and(13.23±0.86mm2,(47.26±4.59mm2,(67.57±4.56mm2,(45.59±4.44mm2 in D group respectively, and part corneal carbide(n=4was observed as well as corneal perforation(n=6were found on 3d in D group. No neovascularization was found in normal control group. Comparison of the areas of CNV at each time point between groups was statistically different, PPCOCLUSION: In 4 to 7d, the higher the temperature is, the more the neovascularization area of CNV are. It has no significant difference in 14 to 30d. But corneal carbide and corneal perforation are often found in 400℃ group, so its modeling failure rate is high. It is between 200℃ and 300℃ that

  1. Patch Grafting Using a Cryopreserved Descemet Stripping Automated Endothelial Keratoplasty Flap for Treating Corneal Perforation

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    Arisa Okada

    2016-04-01

    Full Text Available A 73-year-old woman with a corneal perforation of undetermined etiology was treated with corneal patch grafting. A residual partial-thickness corneal button obtained during a previous Descemet stripping automated endothelial keratoplasty (DSAEK surgery and stored at –80°C in Optisol GS for 3 months was used as a patch graft. Five days postoperatively, the anterior chamber was reformed and the perforation was masked by the donor cornea. During the next several weeks, gradual displacement of the anterior edge of the donor cornea in the limbal direction occurred. Seven weeks postoperatively, further displacement of the donor cornea resulted in unmasking of the perforated area. At this time, the corneal defect was closed by stromal scar tissue and corneal epithelium. Five months postoperatively, best corrected visual acuity was 1.0 without marked astigmatism and intraocular pressure was 9 mm Hg in the left eye. From this case, we learned that cryopreserved DSAEK flaps stored longer than reported previously can be used as patch grafts to treat emergency conditions. Scar tissue can fill a corneal stromal defect 1 mm in diameter during temporary patch grafting for less than 2 months.

  2. The effect of topical application of 0.15% ganciclovir gel on cytomegalovirus corneal endotheliitis.

    Science.gov (United States)

    Koizumi, Noriko; Miyazaki, Dai; Inoue, Tomoyuki; Ohtani, Fumie; Kandori-Inoue, Michiko; Inatomi, Tsutomu; Sotozono, Chie; Nakagawa, Hiroko; Horikiri, Tomoko; Ueta, Mayumi; Nakamura, Takahiro; Inoue, Yoshitsugu; Ohashi, Yuichi; Kinoshita, Shigeru

    2017-02-01

    The aim of this study was to evaluate the therapeutic efficacy and drug transfer of topical application of 0.15% ganciclovir (GCV) gel on cytomegalovirus (CMV) corneal endotheliitis. This study is a multicentre, prospective, interventional case series. Seven eyes of seven immunocompetent patients diagnosed with CMV corneal endotheliitis, based on clinical manifestations and qualitative PCR, were enrolled in this study. The patients were treated with topical applications of 0.15% GCV gel six times daily for 12 weeks without concomitant systemic GCV. Clinical evaluations and quantitative PCR of CMV were performed, and GCV concentrations in aqueous humour were measured by liquid chromatography/tandem mass spectrometry. Clinical improvement of coin-shaped lesions, other types of keratic precipitates, corneal oedema, and anterior chamber inflammation was confirmed at the 4-week visit in all seven eyes. The GCV treatment significantly decreased the CMV copy numbers (p<0.0001). After 12 weeks of treatment, six eyes recovered clear corneas with good vision, and endothelial function was well maintained. Detectable levels of GCV were confirmed in the aqueous humour of all the eyes. The mean GCV concentration in the anterior chamber was 162.0±202.4 ng/mL. The re-emergence of CMV without symptoms was observed in one eye with lower drug transfer. No side effects were observed. Clinical improvement and reduced CMV copy numbers in the aqueous humour were confirmed in the CMV corneal endotheliitis cases. Although the case numbers are limited and long-term follow-up is necessary, the topical application of 0.15% GCV gel appears to be a useful treatment option for CMV endotheliitis. UMIN000012435. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  3. Differing roles for TCF4 and COL8A2 in central corneal thickness and fuchs endothelial corneal dystrophy.

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    Robert P Igo

    Full Text Available Fuchs endothelial corneal dystrophy (FECD is the most common late-onset, vision-threatening corneal dystrophy in the United States, affecting about 4% of the population. Advanced FECD involves a thickening of the cornea from stromal edema and changes in Descemet membrane. To understand the relationship between FECD and central corneal thickness (CCT, we characterized common genetic variation in COL8A2 and TCF4, genes previously implicated in CCT and/or FECD. Other genes previously associated with FECD (PITX2, ZEB1, SLC4A11, and genes only known to affect CCT (COL5A1, FOXO1, AVGR8, ZNF469 were also interrogated. FECD probands, relatives and controls were recruited from 32 clinical sites; a total of 532 cases and 204 controls were genotyped and tested for association of FECD case/control status, a 7-step FECD severity scale and CCT, adjusting for age and sex. Association of FECD grade with TCF4 was highly significant (OR= 6.01 at rs613872; p = 4.8×10(-25, and remained significant when adjusted for changes in CCT (OR= 4.84; p = 2.2×10(-16. Association of CCT with TCF4 was also significant (p = 6.1×10(-7, but was abolished with adjustment for FECD grade (p = 0.92. After adjusting for FECD grade, markers in other genes examined were modestly associated (p ∼ 0.001 with FECD and/or CCT. Thus, common variants in TCF4 appear to influence FECD directly, and CCT secondarily via FECD. Additionally, changes in corneal thickness due to the effect of other loci may modify disease severity, age-at-onset, or other biomechanical characteristics.

  4. [Applying deep lamellar keratoplasty in corneal disorders without endothelial abnormalities].

    Science.gov (United States)

    Wylegała, Edward; Wróblewska, Ewa Marta; Tarnawska, Dorota; Mierzwa, Marzena

    2003-01-01

    To present own experiences in the surgery of deep lamellar keratoplasty. 25 eye of 25 patients suffering from various corneal pathologies with preserved endothelium: 22 with keratoconus eyes, 2 with post-inflammatory scar and one eye with leucoma in Stevens-Johnson syndrome. Mean follow-up was 25.5 months (from 6 to 32). Main outcome measures were Non-Corrected and Best Corrected Visual Acuity (NCVA, BCVA), astigmatism, refractive error, IOP and corneal transparency. The surgery was performed in general anesthesia. In majority of cases the recipient stroma was trephined to a depth of 70% with 7.5 mm diameter vacuum trephine. In cases of intraoperative perforation of Descemet membrane with shallow anterior chamber we converted the procedure into penetrating keratoplasty. The donor lenticule was trephined in the artificial chamber with the 0.5 mm oversized manual trephine to a depth of 90%. In two cases after DLK was finished, limbal stem cell grafting was additionally performed. Postoperative NCVA ranged from 0.01 to 0.8 and BCVA from 0.1 to 0.8. Astigmatism ranged from 0.5 to 10.0 D. Refractive error ranged from -8.0 to +1.0 D. In follow-up period all grafts maintained transparent. The complications were mainly intraoperative perforations of Descemet membrane (9 cases): in 7 cases we convert the procedure into penetrating keratoplasty. In 2 cases the double chamber-forming was observed: in one case on 7 day following surgery the penetrating keratoplasty was performed, in other a spontaneous attachment was observed. We also observed loose sutures in 2 cases, inflammatory infiltrates in one case. Two cases of ocular hypertension was successfully treated by medication. Deep lamellar keratoplasty is an effective procedure for treating various diseases of corneal stroma with unaffected endothelium.

  5. In vitro culture characteristics of corneal epithelial, endothelial, and keratocyte cells in a native collagen matrix.

    Science.gov (United States)

    Orwin, E J; Hubel, A

    2000-08-01

    The objective of this investigation was to demonstrate the effectiveness of a tissue-engineered collagen sponge as a substrate for the culture of human corneal cells. To that end, human kerotocyte, epithelial, and endothelial cells were cultured separately on collagen sponges composed of native fibrillar collagen with a pore size of approximately 0.1 mm. Co-culture experiments were also performed (epithelial/endothelial and epithelial/keratocyte cultures). Proliferation of keratocytes and matrix production was assessed. The morphology of the epithelial and endothelial cell cultures was characterized by histology and scanning electron microscopy. Keratocytes cultured on collagen sponges exhibited increased matrix synthesis over time as well as proliferation and repopulation of the matrix. Epithelial and endothelial cells showed the ability to migrate over the collagen sponge. The thickness of the epithelial layer was influenced by soluble factors produced by endothelial cells. The morphology of the bottom layer of epithelial cells was influenced by the presence of keratocytes in the culture. These studies indicate that human corneal cells exhibit normal cell phenotype when cultured individually on an engineered collagen sponge matrix and co-culture of different cell types in the cornea can influence cell behavior.

  6. Discovery of molecular markers to discriminate corneal endothelial cells in the human body.

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    Masahito Yoshihara

    Full Text Available The corneal endothelium is a monolayer of hexagonal corneal endothelial cells (CECs on the inner surface of the cornea. CECs are critical in maintaining corneal transparency through their barrier and pump functions. CECs in vivo have a limited capacity in proliferation, and loss of a significant number of CECs results in corneal edema called bullous keratopathy which can lead to severe visual loss. Corneal transplantation is the most effective method to treat corneal endothelial dysfunction, where it suffers from donor shortage. Therefore, regeneration of CECs from other cell types attracts increasing interests, and specific markers of CECs are crucial to identify actual CECs. However, the currently used markers are far from satisfactory because of their non-specific expression in other cell types. Here, we explored molecular markers to discriminate CECs from other cell types in the human body by integrating the published RNA-seq data of CECs and the FANTOM5 atlas representing diverse range of cell types based on expression patterns. We identified five genes, CLRN1, MRGPRX3, HTR1D, GRIP1 and ZP4 as novel markers of CECs, and the specificities of these genes were successfully confirmed by independent experiments at both the RNA and protein levels. Notably none of them have been documented in the context of CEC function. These markers could be useful for the purification of actual CECs, and also available for the evaluation of the products derived from other cell types. Our results demonstrate an effective approach to identify molecular markers for CECs and open the door for the regeneration of CECs in vitro.

  7. Discovery of molecular markers to discriminate corneal endothelial cells in the human body.

    Science.gov (United States)

    Yoshihara, Masahito; Ohmiya, Hiroko; Hara, Susumu; Kawasaki, Satoshi; Hayashizaki, Yoshihide; Itoh, Masayoshi; Kawaji, Hideya; Tsujikawa, Motokazu; Nishida, Kohji

    2015-01-01

    The corneal endothelium is a monolayer of hexagonal corneal endothelial cells (CECs) on the inner surface of the cornea. CECs are critical in maintaining corneal transparency through their barrier and pump functions. CECs in vivo have a limited capacity in proliferation, and loss of a significant number of CECs results in corneal edema called bullous keratopathy which can lead to severe visual loss. Corneal transplantation is the most effective method to treat corneal endothelial dysfunction, where it suffers from donor shortage. Therefore, regeneration of CECs from other cell types attracts increasing interests, and specific markers of CECs are crucial to identify actual CECs. However, the currently used markers are far from satisfactory because of their non-specific expression in other cell types. Here, we explored molecular markers to discriminate CECs from other cell types in the human body by integrating the published RNA-seq data of CECs and the FANTOM5 atlas representing diverse range of cell types based on expression patterns. We identified five genes, CLRN1, MRGPRX3, HTR1D, GRIP1 and ZP4 as novel markers of CECs, and the specificities of these genes were successfully confirmed by independent experiments at both the RNA and protein levels. Notably none of them have been documented in the context of CEC function. These markers could be useful for the purification of actual CECs, and also available for the evaluation of the products derived from other cell types. Our results demonstrate an effective approach to identify molecular markers for CECs and open the door for the regeneration of CECs in vitro.

  8. Cultivation of Human Microvascular Endothelial Cells on Topographical Substrates to Mimic the Human Corneal Endothelium

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    Jie Shi Chua

    2013-03-01

    Full Text Available Human corneal endothelial cells have a limited ability to replicate in vivo and in vitro. Allograft transplantation becomes necessary when an accident or trauma results in excessive cell loss. The reconstruction of the cornea endothelium using autologous cell sources is a promising alternative option for therapeutic or in vitro drug testing applications. The native corneal endothelium rests on the Descemet’s membrane, which has nanotopographies of fibers and pores. The use of synthetic topographies mimics the native environment, and it is hypothesized that this can direct the behavior and growth of human microvascular endothelial cells (HMVECs to resemble the corneal endothelium. In this study, HMVECs are cultivated on substrates with micron and nano-scaled pillar and well topographies. Closely packed HMVEC monolayers with polygonal cells and well-developed tight junctions were formed on the topographical substrates. Sodium/potassium (Na+/K+ adenine triphosphatase (ATPase expression was enhanced on the microwells substrate, which also promotes microvilli formation, while more hexagonal-like cells are found on the micropillars samples. The data obtained suggests that the use of optimized surface patterning, in particular, the microtopographies, can induce HMVECs to adopt a more corneal endothelium-like morphology with similar barrier and pump functions. The mechanism involved in cell contact guidance by the specific topographical features will be of interest for future studies.

  9. Complications associated with bovine corneal endothelial cell-lined homografts in the cat.

    Science.gov (United States)

    Bahn, C F; MacCallum, D K; Lillie, J H; Meyer, R F; Martonyi, C L

    1982-01-01

    Cultured bovine corneal endothelial cells were subcultured onto feline corneas from which the native endothelium had been mechanically removed, and transplanted into cats via penetrating keratoplasty. Although the transplants remained thin and clear in the immediate postoperative period, correlative clinical and morphologic analysis disclosed evidence of a host response directed against the heterologous endothelium by the ninth postoperative day. Eyes with rotational autografts or transplanted homografts did not disclose evidence of a similar host response.

  10. Hearing disability in patients with Fuchs’ endothelial corneal dystrophy: unrecognized co-pathology?

    OpenAIRE

    Stehouwer M; Bijlsma WR; Van der Lelij A

    2011-01-01

    Marilette Stehouwer, Ward R Bijlsma, Allegonda Van der LelijDepartment of Ophthalmology, University Medical Center Utrecht, Utrecht, The NetherlandsPurpose: To investigate a possible association between Fuchs’ endothelial corneal dystrophy (FECD) and hearing disability.Methods: A cross-sectional observational study was performed at the University Medical Center Utrecht. Cases and controls were patients who were treated by a cornea specialist between 2004 and 2008. FECD patients had ...

  11. Structural, Morphological, and Functional Correlates of Corneal Endothelial Toxicity Following Corneal Exposure to Sulfur Mustard Vapor

    Science.gov (United States)

    2013-10-01

    dehydrated in graded ethanol, and embedded in Poly/Bed 812 embedding resin (Polysciences, Inc., Warrington, PA) for transmission electron microscopy (TEM...osmium tetroxide, dehydrated in graded ethanol, and critical point dried. Mounted samples were ion beam coated using gold/palladium and imaged using...In Vivo at 24 Hours The ability of the endothelium to maintain corneal deturges- cence depends on osmotic pump activity and the integrity of the CEC

  12. Técnica de separação da membrana de Descemet para transplante de células endoteliais da córnea: estudo experimental em coelhos Technique for separating Descemet membrane for corneal endothelial cells transplantation: experimental study in rabbits

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    Daniel Wasilewski

    2010-02-01

    Full Text Available OBJETIVO: Avaliar a porcentagem de dano endotelial induzido por uma técnica cirúrgica para a separação da membrana de Descemet contendo endotélio sadio, analisar a viabilidade e eficácia desta técnica, e avaliar a porcentagem de dano endotelial causado pela inversão da córnea em câmara anterior artificial. MÉTODOS: As córneas de três grupos de 12 coelhos da linhagem Nova Zelândia foram avaliadas. O grupo 1 foi usado como controle; portanto, as córneas foram analisadas após coletadas e trepanadas. O grupo 2 foi analisado após a inversão da córnea (endotélio para cima na posição convexa, montada em câmara anterior artificial, para o cálculo da porcentagem do dano endotelial induzido por esta inversão. O grupo 3 foi avaliado após a separação entre a membrana de Descemet e o estroma com o uso de substância viscoelástica em córneas invertidas e montadas em câmara anterior artificial. O dano endotelial foi avaliado por meio de fotografias digitais tiradas no microscópio após impregnar o endotélio com vermelho de alizarina. Amostras do grupo 3 foram processadas para avaliação histopatológica. RESULTADOS: O grupo 3 (separação viscoelástica apresentou um índice de lesão celular endotelial de 10,06%, o grupo 2 apresentou um índice de 3,58% e o grupo controle um índice de 0,18% de lesão celular endotelial (pPURPOSE: To evaluate the percentage of endothelial cell damage induced during a surgical technique of Descemet's membrane separation containing healthy endothelium, analyze the viability and efficacy of this technique, and evaluate the percentage of endothelial cell damage caused by inversion of the cornea on an artificial anterior chamber. METHODS: The corneas from three groups of 12 New Zealand rabbits were evaluated. The Group one was used as the control, so the corneas were analyzed after collected and trephinated. The Group two was analyzed after inversion of the cornea (endothelial side up at a convex

  13. Short-term effect of topical brimonidine tartrate on intrastromal corneal pressure in rabbits.

    Science.gov (United States)

    Bolívar, Gema; Teus, Miguel A; Hernández-Verdejo, José L

    2010-07-01

    Because topical brimonidine tartrate has been reported to decrease flap adherence in the early postoperative period after LASIK, its effect on the intrastromal corneal pressure (ICP) was evaluated. An interventional, prospective, animal study was performed. Intrastromal corneal pressure was recorded for 45 minutes in eight eyes of rabbits treated with topical brimonidine tartrate three times daily for 3 consecutive days (study group); eight contralateral eyes were treated with artificial tears (control group). All measurements were performed by the same, masked investigator. Mean ICP was -4.00 +/- 2.90 mmHg, -6.70 +/- 3.00 mmHg, and -9.00 +/- 4.50 mmHg at 15, 30, and 45 minutes, respectively, in the control group. In the study group (brimonidine-treated eyes), the ICP readings were +2.75 +/- 2.90 mmHg, -2.50 +/- 13.00 mmHg, and -8.50 +/- 5.00 mmHg at the same time points, respectively. The differences in the ICP between both groups were statistically significant at 15 minutes (P = .01), but no significant difference was found at 30 or 45 minutes (P = .20 and P = .80, respectively). Topical treatment with brimonidine tartrate induces a significant decrease in ICP at 15 minutes. This effect on ICP may explain the decreased corneal flap adherence reported in eyes treated with this drug.

  14. Fuchs endothelial corneal dystrophy: clinical characteristics of surgical and nonsurgical patients

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    Goldberg RA

    2014-09-01

    Full Text Available Roger A Goldberg,1,2 Sabri Raza,1 Eric Walford,1 William J Feuer,1 Jeffrey L Goldberg1,3 1Bascom Palmer Eye Institute, University of Miami, Miami, FL, USA; 2Tufts-New England Eye Center/Ophthalmic Consultants of Boston, Boston, MA, USA; 3Shiley Eye Center, University of California San Diego, San Diego, CA, USA Purpose: To review the patient and clinical characteristics of patients with Fuchs endothelial corneal dystrophy (FECD. Methods: Review of records for every patient who presented to the Bascom Palmer Eye Institute between 2003 and 2009 whose visit was coded for endothelial corneal dystrophy (International Classification of Diseases, Ninth Revision [ICD9] 371.57, bullous keratopathy (ICD9 371.23, or who underwent a corneal surgery with or without cataract extraction. Demographic, clinical, and ancillary testing data were collected from the time of presentation, diagnosis, and follow-up, and the use, timing, and type of surgical interventions was documented, with 6-month and final visual acuities recorded. Results: A total of 2,370 charts were included in this study, of which 966 patients had a diagnosis of FECD. Of these, 197 patients (21% received a corneal transplantation procedure. The surgery most often performed was penetrating keratoplasty with or without cataract extraction (66%, followed by endothelial keratoplasty with or without cataract extraction (34%. The risk factors for surgery include worse visual acuity at presentation (20/60 Snellen visual acuity in surgical patients versus 20/40 Snellen visual acuity in nonsurgical patients, P<0.001, greater average central corneal thickness (635 µm versus 592 µm, P<0.001, loss of visual acuity over time (two lines lost versus zero lines lost, P<0.001, increasing age (P<0.001, and male sex (P=0.008. Over half of patients (52% did not receive surgery despite poor vision. Conclusion: During this time period, FECD did not have a consistent pattern for management or treatment, and despite

  15. Molecular evidence and functional expression of multidrug resistance associated protein (MRP) in rabbit corneal epithelial cells.

    Science.gov (United States)

    Karla, Pradeep K; Pal, Dananjay; Mitra, Ashim K

    2007-01-01

    Multidrug resistance associated protein (MRP) is a major family of efflux transporters involved in drug efflux leading to drug resistance. The objective of this study was to explore physical barriers for ocular drug absorption and to verify if the role of efflux transporters. MRP-2 is a major homologue of MRP family and found to express on the apical side of cell membrane. Cultured Rabbit Corneal Epithelial Cells (rCEC) were selected as an in vitro model for corneal epithelium. [14C]-erythromycin which is a proven substrate for MRP-2 was selected as a model drug for functional expression studies. MK-571, a known specific and potent inhibitor for MRP-2 was added to inhibit MRP mediated efflux. Membrane fraction of rCEC was used for western blot analysis. Polarized transport of [14C]-erythromycin was observed in rCEC and transport from B-->A was significantly high than from A-->B. Permeability's increased significantly from A-->B in the presence of MK-571 and ketoconozole. Uptake of [14C]-erythromycin in the presence of MK-571 was significantly higher than control in rCEC. RT-PCR analysis indicated a unique and distinct band at approximately 498 bp corresponding to MRP-2 in rCEC and MDCK11-MRP-2 cells. Immunoprecipitation followed by Western Blot analysis indicated a specific band at approximately 190 kDa in membrane fraction of rCEC and MDCK11-MRP-2 cells. For the first time we have demonstrated high expression of MRP-2 in rabbit corneal epithelium and its functional activity causing drug efflux. RT-PCR, immunoprecipitation followed by Western blot analysis further confirms the result.

  16. Abnormal Corneal Endothelial Maturation in Collagen XII and XIV Null Mice

    Science.gov (United States)

    Hemmavanh, Chinda; Koch, Manuel; Birk, David E.; Espana, Edgar M.

    2013-01-01

    Purpose. Maturation of the endothelium and the adjacent matrix was characterized in wild-type (WT) mice. The influence of FACIT collagen XII and XIV deficiency on the morphology, maturation, and function of the corneal endothelium was examined. Methods. Analysis of the endothelium and Descemet's membrane (DM) was performed using transmission electron microscopy at postnatal day (P)4, P14, and P30 in WT, Col12a1−/−, Col14a1−/−, and Col12a1−/−/Col14a1−/− mice. Endothelial junctions were analyzed using ZO-1. The presence of endothelial–stromal communications was evaluated with phalloidin staining as well as electron microscopy. Finally, corneal thickness was assessed. Results. A thin DM, clefts between endothelial cells and DM, and large “vacuole-like” structures were present in the endothelial cells of WT mice at P4 but not noted at P30. The endothelia of Col12a1−/−, Col14a1−/−, and compound Col12a1−/−/Col14a1−/− in the P30 cornea maintained the vacuole-like structures seen at P4. A mature endothelial junction pattern was delayed in the null corneas. Expression of ZO-1 in WT endothelia at P14 was diffuse and localized to the basolateral and apical cell membrane. At P30, staining was localized to intercellular junctions. ZO-1 reactivity was patchy in Col12a1−/−, Col14a1−/−, and compound Col12a1−/−/Col14a1−/− corneas at P14 and P30. Stromal thickness was increased in P30 null corneas. Endothelial cell processes were demonstrated penetrating the DM and into the underlying stroma, throughout the entire endothelial layer in the P4 cornea. Conclusions. Collagen XII and XIV null mice demonstrate delayed endothelial maturation. The structural alterations suggest functional changes in endothelial function resulting in increased corneal thickness. Endothelial–stromal interactions suggest a pathway for signal transduction. PMID:23599329

  17. New strategy to study corneal endothelial cell transplantation: the chick cornea model.

    Science.gov (United States)

    Mangioris, Georgios; Chiodini, Florence; Dosso, Andre

    2011-12-01

    To set up a culture assay of chick corneal endothelial cells (CECs) for transplantation into host corneas. Histology sections were performed at 6, 9, 12, 15, and 18 embryonic days of development of the chick embryo. Visualization of the gross morphology of CECs and of epithelium, stroma, and Descemet membrane was performed. Transplantation of CECs at 18 embryonic days of development into explanted, denuded from endothelial cell, host corneas of the same stage was attempted. The results from the histological sections clearly indicate that after embryonic day 12, the endothelial cells are well differentiated and the proliferation is complete. Transplanted CECs were able to migrate and integrate into the denuded host corneas. This study demonstrated its feasibility using an easy accessible model of chick cornea. With this technique, sufficient CECs may be obtained for biochemical and functional investigations using only nonhatched chickens that are easily accessible and easy to manipulate.

  18. [The expression level of MMP-2 and collagen of hydroxyapatite modified titanium for keratoprosthesis in the corneal stroma of rabbits].

    Science.gov (United States)

    Yang, Min; Du, Gai-ping; Wang, Li-qiang; Wang, Xiao-ping; Cui, Fu-zhai; Lu, Yu-jie; Huang, Yi-fei

    2013-10-01

    To investigate the expression level of metalloproteinases-2(MMP-2) and Collagen in a hydroxyapatite surfaced-modified of three Pan type titanium keratoprosthesis after that implanted into the corneal stroma of rabbits, further evaluate its biological compatibility. Experimental study. Twenty-four New Zealand white rabbits, 2.0-2.5 kg, were respectively divided into three groups. Surgery was performed in right eye of all animals. skirt of HA-Ti and Ti were respectively inserted into the corneal stroma of rabbit of experimental group A and group B; only a sack was made without implantation in control group C . Cornea edema and corneal neovascularization were observed at scheduled times after operation; animals were sacrificed 2, 4 and 16 weeks after operation and their cornea was removed and examined under light microscopy; the surface of skirt was observed under scanning electron microscope. During the study period, all skirts were stable without infected, dissolved and excluded. Different degree of cornea edema and neovascularization was revealed after surgery. MMP-2 were absent in the normal corneal matrix. The expression level of MMP-2 in group A was higher than group C at all time points (F = 6.083, P collagen and yellow red type I collagen, 16 weeks corneal mainly for bright red when within the collagen type I, still have a small amount of collagen type III. Rabbit cornea implanted HA-Ti skirts cause MMP-2 activation, continuous high expression didn't cause the cornea to dissolve; Collagen -III turned into collagen-I gradually in the extracellular matrix around the skirts. Hydroxyapatite modified titanium for Keratoprosthesis promoted the corneal neovascularization and improve the interfacial bio integration of skirt and host cornea.

  19. Study of Topical Human Umbilical Cord Blood Serum in the Treatment of Alkaline Corneal Epithelial Wounds in Rabbit Model

    Directory of Open Access Journals (Sweden)

    B Sharifi

    2011-04-01

    Full Text Available Introduction & Objective: One of the important functions of the cornea is to maintain normal vision by refracting light onto the lens and retina. This property is dependent in part on the ability of the corneal epithelium to undergo continuous renewal. Ocular surface failure which follows a variety of endogenous and exogenous precipitating factors, the most common being: chemical trauma, infection, alkaline burn, inflammation and hereditary conditions, lid or lash abnormalities, tear deficiency or reduced sensation. The core principal underpinning management strategy for ocular surface failure is establishing or promoting new growth of healthy conjunctiva and corneal epithelium. This process is mediated by many proteins that are inducers of corneal cell migration, proliferation, and differentiation. The current study was performed to investigate the efficacy of umbilical cord serum on alkaline corneal epithelial wound healing in the rabbit model. Materials & Methods: In this study conducted at Yasuj University of Medical Sciences in 2010, thirty two rabbits were randomly assigned into two equal groups. Central corneal alkali wound was formed in one eye of the rabbits by applying a 6-mm round filter paper, soaked in 1 N NaOH, for 60 seconds. Group one of animals received umbilical cord blood serum and group two received Sno*Tear in the eyes. The treatment was dosed 4 times a day with the eye drops, and epithelial wound closure was recorded using slit lamp. The data were analyzed to determine the rate of wound closure. Results: The mean wound radius closure rate was 0.77 mm/day (SD=0.013 for umbilical cord blood serum-treated eyes, 0.73 mm/day (SD=0.018 for artificial tear-treated eyes. Conclusion: This study shows that alkali-injured corneal epithelial wound heal faster when treated with umbilical cord blood serum than with artificial tear in rabbit model.

  20. Changes in corneal endothelial cell profile measurements after deep anterior lamellar keratoplasty for keratoconus.

    Science.gov (United States)

    Salouti, Ramin; Masoumpour, Masoumeh; Nowroozzadeh, Mohammad H; Zamani, Mohammad; Ghoreyshi, Maryam; Melles, Gerrit R J

    2013-06-01

    The primary objective was to evaluate whether postoperative alterations in corneal shape (as reflected by keratometry values) affect endothelial cell profile measurements after deep anterior lamellar keratoplasty (DALK) in a group of patients with keratoconus. Secondary objective was to describe the pattern of changes in corneal endothelial cell profile measurements during the first 3 years after DALK. In this prospective interventional case series, we enrolled patients who had significant keratoconus and were scheduled for DALK (Melles technique). Cases with concomitant intraocular surgeries, intra-/postoperative complications, and poor quality of images were excluded. Two hundred one eyes and 45 eyes (of the original 201) were enrolled for evaluating the primary and the secondary objectives, respectively. At 3 months post DALK, the mean endothelial cell density (ECD) had significantly increased and the mean cell area had decreased compared with preoperative measurements (2721 vs. 2823 cells/mm2, P = 0.015; and 378 vs. 362 μm2, P = 0.005, respectively). Regression analysis revealed a weak but significant nonlinear association between changes in mean keratometry and ECD changes at 3 months (R2 = 0.039, P = 0.02). Standard deviation of mean cell area had significantly decreased at 12 months after DALK compared with measurements taken at 3 months after surgery (P = 0.023) and remained stable thereafter. Apparent measurements of ECD may not show a decrease but instead even a slight increase in some cases after uncomplicated DALK (Melles technique) for keratoconus. This finding along with a later decrease in standard deviation of mean cell area suggests that notable postoperative changes in corneal biomechanical forces may affect endothelial cell profile measurements.

  1. Corneal endothelial rejection after penetrating keratoplasty treated with intravenous and topic corticosteroid: one year follow up

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    Ricardo Yuji Abe

    2013-02-01

    Full Text Available OBJECTIVE: To analyze the recovery of visual acuity (VA and graft survival after first episode of endothelial rejection in penetrating keratoplasty (PKP treated with intravenous (IV and topic corticosteroid. METHODS: Interventional, prospective, non-comparative case series study evolving 32 PKP patients in one year follow up, who presented first episode of corneal endothelial rejection. The patients were submitted to 500 mg IV injection of methylprednisolone in association with topical prednisolone. Main outcome measures included VA recovery and corneal edema regression. Second outcome included new rejections and graft failure. Multivariate analysis techniques were used to estimate rates of graft outcome events and the impact of risk factors. RESULTS: A total of 32 eyes from 32 patients (13 male and 19 female were included in the study. The mean VA (in number of letters before rejection was 48 (22 to 88 letters. Patients treated within 7 days or less of initial symptoms had better VA recovery, corneal edema regression and less graft failure (p<0.001. Patients with previous ocular surgery had worse VA recovery and more graft failure (p<0.047. CONCLUSION: The association between the other risk factors and the outcomes did not reach statistical significance in the multivariate model because of the small numbers of patients. Methylprednisolone in association with topical prednisolone is an alternative treatment for graft rejection. Our study showed that patients treated within 7 days of symptoms and no previous anterior segment surgery had better visual outcome and graft survival after treatment.

  2. A multicenter study to map genes for Fuchs endothelial corneal dystrophy: baseline characteristics and heritability.

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    Louttit, Megan D; Kopplin, Laura J; Igo, Robert P; Fondran, Jeremy R; Tagliaferri, Angela; Bardenstein, David; Aldave, Anthony J; Croasdale, Christopher R; Price, Marianne O; Rosenwasser, George O; Lass, Jonathan H; Iyengar, Sudha K

    2012-01-01

    To describe the methods for family and case-control recruitment for a multicenter genetic and associated heritability analyses of Fuchs endothelial corneal dystrophy (FECD). Twenty-nine enrolling sites with 62 trained investigators and coordinators gathered individual and family information, graded the phenotype, and collected blood and/or saliva for genetic analysis on all individuals with and without FECD. The degree of FECD was assessed in a 0 to 6 semiquantitative scale using standardized clinical methods with pathological verification of FECD on at least 1 member of each family. Central corneal thickness was measured by ultrasonic pachymetry. Three hundred twenty-two families with 330 affected sibling pairs with FECD were enrolled and included a total of 650 sibling pairs of all disease grades. Using the entire 7-step FECD grading scale or a dichotomous definition of severe disease, heritability was assessed in families via sib-sib correlations. Both binary indicators of severe disease and semiquantitative measures of disease severity were significantly heritable, with heritability estimates of 30% for severe disease, 37% to 39% for FECD score, and 47% for central corneal thickness. Genetic risk factors have a strong role in the severity of the FECD phenotype and corneal thickness. Genotyping this cohort with high-density genetic markers followed by appropriate statistical analyses should lead to novel loci for disease susceptibility.

  3. Corneal endothelial cell density and morphology in low and moderate myopic Chinese eyes

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    Jane Mei Chun

    2013-08-01

    Full Text Available AIM: To describe and compare the corneal endothelial cell density and morphology in young, low and moderate myopic Chinese adults in Malaysian Chinese population.METHODS: Non-contact specular microscopy (Topcon SP3000P, Tokyo, Japan was performed in low (n=78; 21.22±1.51 years and moderate (n=78; 21.82±1.40 years myopic subjects. The mean of three consecutive measurements of endothelial cell density (MCD, coefficient of variation (CV in the cell size, and hexagonal appearance of the cell were obtained.RESULTS: In low myopic eyes the MCD was 3 063.0±176.2/mm2, the mean CV was 33.4±4.0% and the mean hexagonal appearance of the cell was 57.9±2.7%. In moderate myopic eyes the MCD was 2961.6±159.0/mm2, the mean CV was 33.9±3.6% and mean hexagonal appearance of the cell was 56.2±4.7%. There were statistically significant differences in MCD (PPCONCLUSION:The corneal endothelial cell layer in more myopic eyes tends to have less MCD and cell hexagonality compared to lower myopic eyes. Nevertheless, there is no significant difference in CV between low and moderate myopic eyes.

  4. Cytotoxic effects of betaxolol on healthy corneal endothelial cells both in vitro and in vivo

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    Ying Miao

    2014-02-01

    Full Text Available AIM: To demonstrate the cytotoxic effect of betaxolol and its underlying mechanism on human corneal endothelial cells(HCE cells in vitro and cat corneal endothelial cells(CCE cells in vivo, providing experimental basis for safety anti-glaucoma drug usage in clinic of ophthalmology.METHODS: In vivo and in vitro experiments were conducted to explore whether and how betaxolol participates in corneal endothelial cell injury. The in vitro morphology, growth status, plasma membrane permeability, DNA fragmentation, and ultrastructure of HCE cells treated with 0.021875-0.28g/L betaxolol were examined by light microscope, 3-(4,5-dimethylthiahiazo (-z-y1-3,5-di-phenytetrazoliumromide (MTT assay, acridine orange (AO/ethidium bromide (EB double-fluorescent staining, DNA agarose gel electrophoresis, and transmission electron microscope (TEM. The in vivo density, morphology, and ultrastructure of CCE cells, corneal thickness, and eye pressure of cat eyes treated with 0.28g/L betaxolol were investigated by specular microscopy, applanation tonometer, alizarin red staining, scanning electron microscope (SEM, and TEM.RESULTS: Exposure to betaxolol at doses from 0.0875g/L to 2.8g/L induced morphological and ultrastructural changes of in vitro cultured HCE cells such as cytoplasmic vacuolation, cellular shrinkage, structural disorganization, chromatin condensation, and apoptotic body appearance. Simultaneously, betaxolol elevated plasma membrane permeability and induced DNA fragmentation of these cells in a dose-dependent manner in AO/EB staining. Furthermore, betaxolol at a dose of 2.8g/L also induced decrease of density of CCE cells in vivo, and non-hexagonal and shrunk apoptotic cells were also found in betaxolol-treated cat corneal endothelia.CONCLUSION: Betaxolol has significant cytotoxicity on HCE cells in vitro by inducing apoptosis of these cells, and induced apoptosis of CCE cells in vivo as well. The findings help provide new insight into the apoptosis

  5. Long-lasting corneal endothelial graft rejection successfully reversed after dexamethasone intravitreal implant

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    Giannaccare G

    2016-07-01

    Full Text Available Giuseppe Giannaccare, Michela Fresina, Alberto Pazzaglia, Piera Versura Ophthalmology Unit, Department of Experimental, Diagnostic and Specialty Medicine (DIMES, Alma Mater Studiorum University of Bologna, Sant’Orsola‑Malpighi Teaching Hospital, Bologna, Italy Abstract: Graft rejection is the most significant complication corneal transplantation and the leading indication for overall corneal transplantation. Corticosteroid therapy represents the mainstay of graft rejection treatment; however, the optimal route of administration of corticosteroid remains uncertain. We report herein for the first time the multimodal imaging of a case of long-lasting corneal endothelial graft rejection successfully reversed 3 months after dexamethasone intravitreal implant. A 29-year-old Asian female presented with a long-lasting corneal endothelial graft rejection in her left phakic eye. She underwent penetrating keratoplasty for advanced keratoconus 24 months before presentation. Hourly dexamethasone eyedrops, daily intravenous methylprednisolone, and one parabulbar injection of methylprednisolone acetate were administered during the 5 days of hospitalization. However, the clinical picture remained approximately unchanged despite therapy. By mutual agreement, we opted for the off-label injection of dexamethasone 0.7 mg intravitreal implant in order to provide therapeutic concentrations of steroid for a period of ~6 months. No other concomitant therapies were prescribed to the patient. Visual acuity measurement, slit lamp biomicroscopy, anterior segment photography, confocal microscopy, anterior segment optical coherence tomography, laser cell flare meter, intraocular pressure measurement, and ophthalmoscopy were performed monthly for the first postoperative 6 months. Three months after injection, both clinical and subclinical signs of rejection disappeared with a full recovery of visual acuity to 20/30 as before the episode. Currently, at the 12-month

  6. Biocompatibility of helicoidal multilamellar arginine-glycine-aspartic acid-functionalized silk biomaterials in a rabbit corneal model.

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    Wang, Liqiang; Ma, Ruijue; Du, Gaiping; Guo, Huiling; Huang, Yifei

    2015-01-01

    Silk proteins represent a unique choice in the selection of biomaterials that can be used for corneal tissue engineering and regenerative medical applications. We implanted helicoidal multilamellar arginine-glycine-aspartic acid-functionalized silk biomaterials into the corneal stroma of rabbits, and evaluated its biocompatibility. The corneal tissue was examined after routine hematoxylin-eosin staining, immunofluorescence for collagen I and III, and fibronectin, and scanning electron microscopy. The silk films maintained their integrity and transparency over the 180-day experimental period without causing immunogenic and neovascular responses or degradation of the rabbit corneal stroma. Collagen I increased, whereas Collagen III and fibronectin initially increased and then gradually decreased. The extracellular matrix deposited on the surface of the silk films, tightly adhered to the biomaterial. We have shown this kind of silk film graft has suitable biocompatibility with the corneal stroma and is an initial step for clinical trials to evaluate this material as a transplant biomaterial for keratoplasty tissue constructs. © 2014 Wiley Periodicals, Inc.

  7. Derivation of corneal endothelial cell-like cells from rat neural crest cells in vitro.

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    Chengqun Ju

    Full Text Available The aim of this study was to investigate the feasibility of inducing rat neural crest cells (NCC to differentiate to functional corneal endothelial cell (CEC-like cells in vitro. Rat NCC were induced with adult CEC-derived conditioned medium. Immunofluorescence, flow cytometry and real time RT-PCR assay were used to detect expression of the corneal endothelium differentiation marker N-cadherin and transcription factors FoxC1 and Pitx2. CFDA SE-labeled CEC-like cells were transplanted to the corneal endothelium of a rat corneal endothelium deficiency model, and an eye-down position was maintained for 24 hours to allow cell attachment. The animals were observed for as long as 2 months after surgery and underwent clinical and histological examination. Spindle-like NCC turned to polygonal CEC-like after induction and expressed N-cadherin, FoxC1, Pitx2, zonula occludens-1 and sodium-potassium pump Na(+/K(+ ATPase. The corneas of the experimental group were much clearer than those of the control group and the mean corneal thickness in the experimental group was significantly less than in the control group7, 14, 21 and 28 days after surgery. Confocal microscopy through focusing and histological analysis confirmed that green fluorescence-positive CEC-like cells formed a monolayer covering the Descemet's membrane in the experimental group. In conclusion, CEC-like cells derived from NCCs displayed characters of native CEC, and the induction protocol provides guidance for future human CEC induction from NCC.

  8. Changes in Corneal Biomechanical Properties After Descemet Stripping Automated Endothelial Keratoplasty for Pseudophakic Bullous Keratopathy.

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    Faramarzi, Amir; Feizi, Sepehr; Najdi, Danial; Ghiasian, Leila; Karimian, Farid

    2016-01-01

    To compare corneal biomechanical properties and intraocular pressure (IOP) in eyes with pseudophakic bullous keratopathy (PBK) before and after Descemet stripping automated endothelial keratoplasty (DSAEK). This prospective nonrandomized intrasubject comparative study was conducted on 44 eyes of 22 patients with the diagnosis of PBK who underwent DSAEK in one eye. IOP was measured by Goldmann applanation tonometer, and central corneal thickness was measured by ultrasound pachymetry. The ocular response analyzer was used to measure corneal hysteresis (CH), corneal resistance factor (CRF), Goldmann-related IOP, and cornea-compensated IOP. The same measurements were performed in the normal fellow eyes which served as controls. All measurements were performed preoperatively and 6 months postoperatively. The mean patient age was 67 ± 14 years. Mean preoperative CH and CRF values in the DSAEK group were 5.77 ± 2.94 and 6.39 ± 2.72 mm Hg, respectively, which were significantly lower than those measured in the control group (8.2 ± 2.47 and 8.43 ± 2.49 mm Hg, respectively, P = 0.001 for both comparisons). Postoperatively, CH and CRF demonstrated a significant increase (7.09 ± 3.68 mm Hg, P = 0.05 and 8.21 ± 3.84 mm Hg, P = 0.03, respectively) in operated eyes approaching the normal values measured in the control eyes. Corneal biomechanical parameters were significantly lower in PBK eyes than in the normal fellow eyes. These metrics significantly increased after DSAEK and reached values measured in the normal fellow eyes.

  9. Evaluation of the relationship of corneal biomechanical metrics with physical intraocular pressure and central corneal thickness in ex vivo rabbit eye globes.

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    Bao, FangJun; Deng, ManLi; Wang, QinMei; Huang, JinHai; Yang, Jing; Whitford, Charles; Geraghty, Brendan; Yu, Ayong; Elsheikh, Ahmed

    2015-08-01

    The relationship of corneal biomechanical metrics provided by the Ocular Response Analyzer (ORA) and Corvis ST (CVS) with physical intraocular pressure (IOPp) and central corneal thickness (CCT) was evaluated. Thirty fresh enucleated eyes of 30 rabbits were used in ex vivo whole globe inflation experiments. IOPp was measured with a pressure transducer and increased from 7.5 to 37.5 mmHg in steps of 7.5 mmHg while biomechanical data was acquired using the ORA and CVS. At least 3 examinations were performed at each pressure level, where CCT and twelve biomechanical metrics were recorded and analyzed as a function of IOPp. The biomechanical metrics included corneal hysteresis (CH) and corneal resistance factor (CRF), obtained by the ORA. They also included the applanation times (A1T, A2T), lengths (A1L, A2L) and velocities (A1V, A2V), in addition to the highest concavity time (HCT), peak distance (PD), radius (HR) and deformation amplitude (DA), obtained by the CVS. The variation of CCT and the twelve biomechanical metrics for the 30 rabbit eyes tested across the 5 pressure stages considered (inter-pressure differences) were statistically significant (P = 0.00). IOPp was highly to moderately correlated with most biomechanical metrics, especially CRF, A1T, A1V, A2V, PD and DA, while the relationships with CH, A2T, A1L and HCT were poor. IOP has important influences on most corneal biomechanical metrics provided by CVS and ORA. Two biomechanical metrics A1V and HR were influenced by CCT after correcting for the effect of IOP in most pressure stages, while the correlation with others were weak. Comparisons of research groups based on ORA and CVS with different IOPs and CCTs may lead to possible misinterpretations if both or one of which are not considered in the analysis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Corneal haze induced by excimer laser photoablation in rabbits is reduced by preserved human amniotic membrane graft

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    Wang, Ming X.; Gray, Trevor; Prabhasawat, Pinnita; Ma, Xiong; Culbertson, William; Forster, Richard; Hanna, Khalil; Tseng, Scheffer C. G.

    1998-06-01

    We conducted a study to determine if preserved human amniotic membrane can reduce corneal haze induced by excimer laser photoablation. Excimer photoablation was performed bilaterally on 40 New Zealand white rabbits with a 6 mm ablation zone and 120 micrometer depth (PTK) using the VISX Star. One eye was randomly covered with a preserved human amniotic membrane and secured using four interrupted 10 - 0 nylon sutures; the other eye served as control. The amniotic membranes were removed at one week, and the corneal haze was graded with a slit-lamp biomicroscopy by three masked corneal specialists (WC, KH and RF) biweekly for the ensuing 12 weeks. Histology and in situ TUNEL staining (for fragmented DNA as an index for apoptosis) was performed at days 1, 3 and 7 and at 12 weeks. One week after excimer photoablation, the amniotic membrane-covered corneas showed more anterior stromal edema, which resolved at the second week. A consistent grading of organized reticular corneal haze was noted among the three masked observers. Such corneal haze peaked at the seventh week in both groups. The amniotic membrane-covered group showed statistically significant less corneal haze (0.50 plus or minus 0.15) than the control groups (1.25 plus or minus 0.35) (p less than 0.001). The amniotic membrane-covered corneas had less inflammatory response at days 1 and 3, showing nearly nil DNA fragmentation on keratocytes on the ablated anterior stromal and less stromal fibroblast activation. There is less altered epithelial cell morphology and less epithelial hyperplasia at 1 week in these amniotic membrane-treated eyes. We concluded from this study that amniotic membrane matrix is effective in reducing corneal haze induced by excimer photoablation in rabbits and may have clinical applications.

  11. A case of corneal endotheliitis with mumps virus RNA in aqueous humor detected by rt-PCR.

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    Ando, Kiyomu; Ishihara, Mami; Kusumoto, Yoshifumi; Shibuya, Etsuko; Nakamura, Satoshi; Mizuki, Nobuhisa

    2013-04-01

    To report a case of corneal endotheliitis following the mumps parotitis. A 46-year-old man noticed ciliary infection and visual loss in the right eye after mumps parotitis. To determine the cause of the disease, reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify mumps virus RNA in samples from the aqueous humor. RT-PCR revealed mumps virus RNA in an aqueous humor sample. Although full recovery of vision was achieved within 1 month of the onset, corneal endothelial cells were severely damaged. The authors have detected mumps virus in the aqueous humor by RT-PCR for the first time.

  12. [Isolation, culture and identification of two types of endothelial progenitor cells from peripheral blood in rabbits].

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    Xiao, Fang-Yi; Zhang, Huai-Qin; Yu, Hua; Yang, De-Ye; Huang, Wei-Jian; Zhou, Hao

    2007-04-01

    To investigate how to isolate, culture and identify two types of endothelial progenitor cells from peripheral blood in rabbits. Mononuclear cells(MNCs) were isolated from rabbit peripheral blood. Endothelial progenitor cells (EPCs) and endothelial outgrowth cells (EOCs) were obtained from MNCs through different ways of isolation and culture. Two types of cells were assessed by DiI-ac-LDL uptake and lectin binding, and then they were identified by immunofluorescence of flk-1, immunocytochemistry of CD34 and VIII factor related antigen and vasculogenesis activity in vitro. Two types of endothelial progenitor cells were obtained from rabbit peripheral blood through different ways of isolation and culture. EPCs on the seventh day and EOCs on the sixteenth day were positive for ac-LDL uptake and lectin binding, and both of them expressed CD34, flk-1 and VIII factor related antigen. EOCs were assembled into primitive vascular tube-like structures when plated in matrigel. EPCs and EOCs could be obtained from rabbit peripheral blood when different ways of isolation and culture were performed. The system of cell culture can be applied to subsequent experiments in cell transplantation.

  13. Polymorphism of the Flap Endonuclease 1 Gene in Keratoconus and Fuchs Endothelial Corneal Dystrophy

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    Katarzyna A. Wojcik

    2014-08-01

    Full Text Available Oxidative stress is implicated in the pathogenesis of many diseases, including serious ocular diseases, keratoconus (KC and Fuchs endothelial corneal dystrophy (FECD. Flap endonuclease 1 (FEN1 plays an important role in the repair of oxidative DNA damage in the base excision repair pathway. We determined the association between two single nucleotide polymorphisms (SNPs, c.–441G>A (rs174538 and g.61564299G>T (rs4246215, in the FEN1 gene and the occurrence of KC and FECD. This study involved 279 patients with KC, 225 patients with FECD and 322 control individuals. Polymerase chain reaction (PCR and length polymorphism restriction fragment analysis (RFLP were applied. The T/T genotype of the g.61564299G>T polymorphism was associated with an increased occurrence of KC and FECD. There was no association between the c.–441G>A polymorphism and either disease. However, the GG haplotype of both polymorphisms was observed more frequently and the GT haplotype less frequently in the KC group than the control. The AG haplotype was associated with increased FECD occurrence. Our findings suggest that the g.61564299G>T and c.–441G>A polymorphisms in the FEN1 gene may modulate the risk of keratoconus and Fuchs endothelial corneal dystrophy.

  14. Effect of 1- and 6-Hour-Delayed Corneal Collagen Cross-Linking on Corneal Healing in a Rabbit Alkali-Burn Model: Clinical and Histological Observations.

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    Karti, Omer; Zengin, Mehmet Ozgur; Cinar, Esat; Tutuncu, Merve; Karahan, Eyyup; Celik, Asl; Kucukerdonmez, Cem

    2016-12-01

    To study the effect of 1- and 6-hour-delayed corneal collagen cross-linking (CXL) on wound-healing of experimental alkali burns of the cornea. Twenty-four albino rabbits were used. Alkali burns were created using 1 M NaOH. The animals were divided randomly into 2 groups: group 1 (control group, n = 6) and group 2 (experimental group, n = 18). The experimental group was further divided into 3 subgroups as follows: group 2A, untreated (non-CXL) subgroup; group 2B, 1-hour-delayed CXL treatment subgroup; and group 2C, 6-hour-delayed CXL treatment subgroup. All rabbits were examined periodically for 21 days after treatment and then killed. The corneas were excised and histologically examined. Corneal ulceration, edema, and opacity scores were 4.0 ± 1.64, 1.6 ± 0.65, and 3.5 ± 1.21 in group 2A, 1.5 ± 1.76, 1.3 ± 0.87, and 3.1 ± 1.12 in group 2B, and 2.0 ± 1.90, 1.5 ± 0.79, and 3.3 ± 1.09 in group 2C, respectively. These scores were significantly less in groups 2B and 2C than in group 2A (P = 0.023, P = 0.043, and P = 0.034, respectively). Corneal epithelialization, evident upon staining, was best in group 2B and worst in group 2A (P = 0.012). Histopathology revealed that destruction of corneal collagen fibers and infiltration of inflammatory cells into corneal tissue were reduced in groups 2B and 2C compared with group 2A. We found that CXL treatment exerted positive effects on severe alkali-induced corneal burns. However, the effects were more pronounced in the 1-hour treatment group. We believe that CXL treatment may be a possible treatment for corneal alkali burn.

  15. Assessment of corneal endothelial cell density in patients with keratoconus not using contact lenses.

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    Timucin, Ozgur Bulent; Karadag, Mehmet Fatih; Cinal, Adnan; Asker, Muntecep; Asker, Selvi; Timucin, Damla

    2013-04-01

    To assess the corneal endothelial cell density (ECD) in keratoconus patients with no history of contact lens use. Yuzuncu Yil University, School of Medicine and Van Training and Research Hospital, Department of Ophthalmology, Van, Turkey. Cross-sectional controlled study. The eyes of 65 patients with the diagnosis of keratoconus with no history of contact lens wear and the eyes of 40 healthy controls were prospectively examined using the Heidelberg Retinal Tomography Rostock Cornea Module (HRT3/RCM). The average ECD from the two groups were then compared. Of the cases with keratoconus, 44 (67.7%) were men and 21 (32.3%) were women. The mean age was 20.9±6.8 (range=10-41) years. Of the controls, 28 (70%) were men and 12 (30%) were women. The mean age was 23.9±5.8 (range=14-35) years. Of the 65 eyes with keratoconus, 19 (29.2%) had mild keratoconus, 21 (32.3%) had moderate keratoconus, and 25 (38.5%) had severe keratoconus. The mean ECD was 2731.6±303.2 cells/mm2 in cases with keratoconus and 2664.9±319.5 cells/mm2 in controls. There was no difference between the densities (unpaired t-test, P=0.4). No significant relationships were found between the ECD data and central corneal thickness or steepest keratometric. Endothelial cell density was unaltered in keratoconic patients without a history of contact lens use when compared with healthy controls. Change in ECD is independent from the central corneal thickness and the stage of keratoconus. Copyright © 2012 British Contact Lens Association. Published by Elsevier Ltd. All rights reserved.

  16. Dual-purpose corneal tissue for anterior lamellar keratoplasty and Descemet's membrane endothelial keratoplasty.

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    Menant-Tay, C Larena; Conlon, Ronan; Teja, Salina; Yeung, Season; Teichman, Joshua; Ziai, Setareh; Baig, Kashif

    2016-12-01

    To assess the intraoperative issues and surgical outcomes of preparing a single-donor corneal tissue for same-day use in both deep anterior lamellar keratoplasty (DALK) and Descemet's membrane endothelial keratoplasty (DMEK). Consecutive retrospective case series. Ten eyes of 10 patients who underwent DALK (5 patients) or DMEK (5 patients) surgery using dual-purpose corneal tissue. Five dual-purpose corneoscleral rims were used to prepare tissue for 5 DMEK and 5 DALK procedures. The submerged cornea using backgrounds away technique was first used to harvest the 5 DMEK grafts, and the remaining tissue was used for the 5 DALK grafts. Tissue preparation and operative use occurred on the same day. Tissue preparation challenges, intraoperative complications, and visual recovery were assessed. There were no difficulties in preparing the 5 dual-purpose tissues, and all 10 lamellar transplants were completed successfully. At the 6-month follow-up, the mean best-corrected distance visual acuity improved from 20/250 to 20/80 in the DALK patients, and from 20/300 to 20/25 in the DMEK patients. Postoperative complications after DALK included retained viscoelastic agent at the interface in 1 patient and a double anterior chamber managed with rebubbling in another. After DMEK, a peripheral partial graft detachment occurred in 1 patient and was managed successfully with rebubbling. All corneas demonstrated clarity on slit-lamp examination. Single-donor corneal tissue can be effectively used for both DALK and DMEK, and may represent a more efficient use of corneal tissue. Complications with the preparation of dual-purpose tissue were not encountered. Copyright © 2016 Canadian Ophthalmological Society. Published by Elsevier Inc. All rights reserved.

  17. Density and morphology of corneal endothelial cell after phacoemulsification using Ringer lactate versus balanced salt solution as irrigating solutions

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    Farahdina Rahmawati

    2018-02-01

    Full Text Available AIM: To compare the difference in corneal endothelial cell density and morphology after phacoemulsification using Ringer lactate(RLand balanced salt solution(BSSirrigating solutions.METHODS: The prospective randomized controlled trial study was conducted between February 2017 and April 2017 in Dr. YAP Eye Hospital, Yogyakarta, Indonesia. There were a total of 52 subjects(52 eyeswho were senile cataract patients further grouped into two, 26 patients undergoing the phacoemulsification procedure using RL irrigating solution and the other 26 patients with BSS irrigating solution, both conducted by one operator. On the 1, 7, and 28d post operative, an evaluation was done to measure the density and corneal endothelial cell morphology, as well as the variable of inflammation in the two groups.RESULTS: Fifty-two eyes had undergone phacoemulsification with posterior intraocular lens implantation. Both groups were evaluated for the endothelial cell reduction and corneal endothelial cell morphology change, along with post-operative inflammation. On the 28d post-operative, endothelial cell reduction in the BSS group(173.96 cell/mm2, 8.12%was lower than the RL group(253.20 cell/mm2, 10.25%, percentage of corneal endothelial cell variation coefficient increase in the BSS group(2.92%, 8.36%was lower compared to the RL group(3.42%, 9.96%, decrease of hexagonal cells of corneal endothelium cells presentation percentage in the BSS group(4.30%, 8.17%was lower compared to the RL group(4.84%, 8.97%, and the percentage increase of central corneal thickness in the BSS group(4.69 μm, 0.89%was almost equal to the RL group(4.53 μm, 0.90%. All of the results regarding difference in density and corneal cell endothelium morphology between the two groups did not reveal any statistically significant difference(P>0.05. Inflammatory variable in the two groups were even.CONCLUSION: BSS and RL were equal in their capability of maintaining endothelial cell loss and endothelial

  18. Correlation of corneal thickness, endothelial cell density and anterior chamber depth with ocular surface temperature in normal subjects.

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    Pattmöller, Johanna; Wang, Jiong; Zemova, Elena; Seitz, Berthold; Eppig, Timo; Langenbucher, Achim; Szentmáry, Nóra

    2015-09-01

    To analyze corneal surface temperature profile in a young and healthy study population and to determine the impact of corneal thickness (CT), anterior chamber depth (ACD), and endothelial cell density (ECD) on surface temperature. In this prospective, single-center study 61 healthy right eyes of 61 subjects without tear film pathologies (mean age 24.9 ± 6.7 years) were recruited. Ocular surface temperature (OST) was measured with the Ocular Surface Thermographer TG-1000. From Pentacam HR CT and ACD, and from specular microscopy ECD and central corneal thickness (CCT) were acquired. From the raw measurement data (OST, CT and ACD) we extracted a) local OST the corneal center and 3mm away from the center at the 3, 6, and 9 o'clock positions, and b) Zernike parameters Z1, Z2 and Z3 to evaluate the general temperature profile within a 6mm circular area around the center. Overall, there was no correlation between OST and CT, ACD or ECD. Local OST did not correlate with CT at any measurement position. On average local OST was highest at measurement positions where CT was lowest, but without reaching statistical significance. Baseline OST was highest at thin corneal regions and temperature decay over time was smallest in those regions. Z1, Z2 and Z3 correlated well with CT. In healthy subjects corneal thickness, endothelial cell density and anterior chamber depth have no effect on corneal surface temperature. The general temperature profile seems to be influenced by the corneal thickness profile effecting a higher temperature and lower decay at thinner corneal regions. Copyright © 2014. Published by Elsevier GmbH.

  19. The effect of subconjunctival bevacizumab on corneal neovascularization, inflammation and re-epithelization in a rabbit model

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    Glauco Reggiani Mello

    2011-01-01

    Full Text Available PURPOSE: To evaluate the use of subconjunctival bevacizumab on corneal neovascularization in an experimental rabbit model for its effect on vessel extension, inflammation, and corneal epithelialization. METHODS: In this prospective, randomized, blinded, experimental study, 20 rabbits were submitted to a chemical trauma with sodium hydroxide and subsequently divided into two groups. The experimental group received a subconjunctival injection of bevacizumab (0.15 m; 3.75 mg, and the control group received an injection of 0.15 ml saline solution. After 14 days, two blinded digital photograph analyses were conducted to evaluate the inflammation/diameter of the vessels according to pre-established criteria. A histopathological analysis of the cornea evaluated the state of the epithelium and the number of polymorphonuclear cells. RESULTS: A concordance analysis using Kappa's statistic showed a satisfactory level of agreement between the two blinded digital photography analyses. The neovascular vessel length was greater in the control group (p<0.01 than in the study group. However, the histopathological examination revealed no statistically significant differences between the groups in terms of the state of the epithelium and the number of polymorphonuclear cells. CONCLUSIONS: Subconjunctival bevacizumab inhibited neovascularization in the rabbit cornea. However, this drug was not effective at reducing inflammation. The drug did not induce persistent corneal epithelial defects.

  20. Long-term changes in corneal endothelial morphology after discontinuation of low gas-permeable contact lens wear

    NARCIS (Netherlands)

    Odenthal, M.T.; Gan, I.M.; Oosting, J.; Kijlstra, A.; Beekhuis, W.H.

    2005-01-01

    Low gas-permeable contact lens wear of polymethyl methacrylate or hydroxyethyl methacrylate material is known to cause morphologic abnormalities in the corneal endothelial cell layer. These lenses were widely prescribed and successfully worn until their use was actively discouraged in the late 1980s

  1. Effects of laser peripheral iridotomy on corneal endothelial cell density and cell morphology in primary angle closure suspect subjects

    Directory of Open Access Journals (Sweden)

    Hossein Jamali

    2016-01-01

    Conclusion: In PACS eyes, we did not find a decline in corneal endothelial cell density or a change in cell morphological characteristics, including cell hexagonality and CV, in the central, nasal, and temporal regions of the cornea in any of our subjects over a one-year follow-up period.

  2. Serum-free corneal organ culture medium (SFM) but not conventional minimal essential organ culture medium (MEM) protects human corneal endothelial cells from apoptotic and necrotic cell death.

    Science.gov (United States)

    Jäckel, Thekla; Knels, Lilla; Valtink, Monika; Funk, Richard H W; Engelmann, Katrin

    2011-01-01

    To evaluate the influence of organ culture media on corneal endothelial cell survival. The human corneal endothelial cell line HCEC-12 was cultured in five different media: human corneal endothelial cell (HCEC) growth medium (F99(HCEC)), standard minimal essential corneal organ culture medium (MEM)+2% fetal calf serum (FCS), MEM+5% FCS, and humanised, endothelial serum-free medium (SFM) (with and without antibiotics). A portion of the cells was treated with 0.5 μmol/l staurosporine and examined for signs of apoptosis by assessing mitochondrial membrane polarisation state (intravital JC-1 staining), by YO-PRO-1 and propidium iodide staining, by determining fragmentation of nuclei by sub-G1 DNA content, by immunocytochemistry for cleaved caspase-3, cleaved caspase-8, Bcl2-associated X protein (Bax) and B-cell lymphoma 2 (Bcl-2), and by western blotting for cleaved caspase-3 and cleaved poly (ADP-ribose) polymerase (PARP). The number of apoptotic cells in untreated control cultures was significantly higher in MEM compared with F99(HCEC) and SFM. Staurosporine treatment induced apoptosis in all tested cultures to varying degrees. Cells cultured in MEM showed stronger staining for cleaved caspase-3, cleaved caspase-8, Bax, Bcl-2 and cleaved PARP, increased sub-G1 DNA content, more propidium iodide- and YO-PRO-1-positive cells, and more mitochondria with depolarised membranes. All parameters were significantly higher in MEM compared with F99(HCEC) and SFM. SFM cultures were significantly less susceptible to cell stress. SFM is superior to MEM in promoting HCEC survival.

  3. Nonlinear optical corneal collagen crosslinking of ex vivo rabbit eyes.

    Science.gov (United States)

    Bradford, Samantha M; Brown, Donald J; Juhasz, Tibor; Mikula, Eric; Jester, James V

    2016-11-01

    To determine whether riboflavin-induced collagen crosslinking (CXL) could be precisely achieved in the corneal stroma of ex vivo rabbit eyes using nonlinear optical excitation with a low numerical aperture lens and enlarged focal volume. Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, USA. Experimental study. The corneal epithelium was removed and the corneas were soaked in 0.5% riboflavin solution. Using a 0.1 numerical aperture objective, a theoretical excitation volume of 150 μm × 3 μm was generated using 1 W of 760 nm femtosecond laser light and raster scanned with 4.4 μm line separation at varying effective speeds over a 4.50 mm × 2.25 mm area. Corneal sections were examined for collagen autofluorescence. Collagen autofluorescence was enhanced 2.9 times compared with ultraviolet-A (UVA) CXL. Also, increasing speed was linearly associated with decreasing autofluorescence intensity. The slowest speed of 2.69 mm/s showed a mean of 182.97 μm ± 52.35 (SD) long autofluorescent scan lines axially in the central cornea compared with 147.84 ± 4.35 μm for UVA CXL. Decreasing dwell time was linearly associated with decreasing autofluorescence intensity, approaching that of UVA CXL at a speed of 8.9 mm/s. Using an effective speed of 8.9 mm/s, nonlinear optical CXL could be achieved over a 3.0 mm diameter area in fewer than 4 minutes. Further development of nonlinear optical CXL might result in safer, faster, and more effective CXL treatments. None of the authors has a financial or proprietary interest in any material or method mentioned. Copyright © 2016 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

  4. Wound healing in the rabbit cornea after corneal collagen cross-linking with riboflavin and UVA.

    Science.gov (United States)

    Wollensak, Gregor; Iomdina, Elena; Dittert, Dag-Daniel; Herbst, Hermann

    2007-06-01

    This study was undertaken to investigate the wound healing process of the first 6 weeks after photodynamic cross-linking treatment in the rabbit cornea, using the photosensitizer riboflavin and UVA. After removal of the central epithelium, the right corneas of 8 Chinchilla rabbits were cross-linked with a photosensitizing 0.1% riboflavin solution and UVA light (370 nm; irradiance, 3 mW/cm(2); dose, 5.4 J/cm(2)) for 30 minutes. Two animals were euthanized 3 days, 7 days, 4 weeks, and 6 weeks postoperatively. The corneas of the enucleated eyes were evaluated using 4-microm light microscopic sections with routine stains and avidin-biotin complex immunostaining with anti-alpha-smooth muscle actin. By day 3 after treatment, complete apoptotic damage and loss of the endothelial cells and the stromal keratocytes were found in the irradiated area through the entire thickness of the stroma. There was marked stromal edema (850 +/- 66 vs. 332 +/- 43 microm in the untreated controls; P neutrophils. By day 7, the endothelium was already intact again, and keratocyte repopulation of the posterior stroma was noted. By week 4, the keratocyte repopulation of the anterior stroma was observed with some acellular areas between. By week 6, the cytoarchitecture of the cornea seemed normal again. By weeks 4 and 6, alpha-actin-positive keratocytes were identified, especially in the periphery of the irradiated area. After riboflavin/UVA cross-linking of rabbit cornea, a complete cell loss occurs in the irradiation area with an irradiance of 3 mW/cm(2). The cytotoxic damage is repaired by repopulation after approximately 4-6 weeks. A combination of cross-linking with other procedures such as the implantation of intracorneal rings should be performed only after a sufficient time interval of approximately 2 months, allowing cellular regeneration.

  5. Microbiologic, Pharmacokinetic, and Clinical Effects of Corneal Collagen Cross-Linking on Experimentally Induced Pseudomonas Keratitis in Rabbits.

    Science.gov (United States)

    Cosar, C Banu; Kucuk, Mutlu; Celik, Ekrem; Gonen, Tansu; Akyar, Isin; Serteser, Mustafa; Tokat, Fatma; Ince, Umit

    2015-10-01

    To determine the effects of corneal collagen cross-linking (CXL) on the penetration of topical 0.5% moxifloxacin, on the number of colony-forming units (CFUs) in the cornea, and on the clinical course in a rabbit eye model of experimentally induced Pseudomonas aeruginosa keratitis. In this prospective animal study, experimental Pseudomonas corneal ulcers were induced in 56 corneas of 28 albino New Zealand rabbits. The corneas were randomly divided into the following 4 groups: the control group (14 eyes), the MOX group (moxifloxacin) (14 eyes), the MOX + CXL group (14 eyes), and the CXL group (14 eyes). On day 4 of the experiment, the eyes in the control group were enucleated and CFU counting was performed. On day 10 of the experiment, all eyes were enucleated and CFU counting was performed. In the MOX and MOX + CXL groups, the moxifloxacin level in the cornea, aqueous humor, iris, plasma, and serum was measured by reverse-phase high-performance liquid chromatography. The difference in the corneal CFU count between the MOX group and the MOX + CXL group was not significant (P = 0.317). Clinical improvement was greatest in the MOX + CXL group (P < 0.001). The mean corneal moxifloxacin level was 0.391 ± 0.09 μg·mg in the MOX group versus 0.291 ± 0.09 μg·mg in the MOX + CXL group; as such, CXL did not have a significant effect on antibiotic penetrance (P = 0.386). Clinical improvement was greatest in the MOX + CXL group. The synergistic effect of CXL on corneal ulcer treatment is not through antibiotic penetrance.

  6. Effects of Antiglaucoma Drugs on Calcium Mobility in Cultured Corneal Endothelial Cells

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    Kwou-Yeung Wu

    2006-02-01

    Full Text Available The aim of this study was to estimate the effects of various antiglaucoma drugs including betaxolol, timolol, levobunolol, brimonidine, carteolol, dipivefrin, dorzolamide, brinzolamide, latanoprost, unoprostone, and pilocarpine on intracellular free Ca2+ ([Ca2+]i mobility in cultured bovine corneal endothelial cells. Various antiglaucoma drugs were diluted from original concentrations to 1/100, 1/1,000, and 1/10,000. The [Ca2+]i mobility was studied by spectrofluorophotometry after loading with the ester of fura-2 (fura-2/AM. It was found that timolol (58 μM and 5.8 μM, levobunolol (171 μM, 17.1 μM, and 1.71 μM, betaxolol (162 μM, 16.2 μM, and 1.62 μM, carteolol (680 μM and 68 μM, dipivefrin (28 μM and 2.8 μM, dorzolamide (616 μM and 61.6 μM, brinzolamide (260 μM, latanoprost (1.1 μM, unoprostone (28.2 μM, 2.82 μM, and 0.282 μM, and pilocarpine (408 μM and 40.8 μM induced a significant increase in [Ca2+]i. Nevertheless, only brimonidine (68 μM and 6.8 μM decreased [Ca2+]i concentration significantly. Benzalkonium chloride preservative did not affect [Ca2+]i after addition of 0.001, 0.0001 and 0.00001 mg/mL to cells. These results indicate that all antiglaucoma drugs may affect the physiologic function of corneal endothelial cells through change of [Ca2+]i.mobility.

  7. [Corneal power after descemet stripping automated endothelial keratoplasty (DSAEK) - Modeling and concept for calculation of intraocular lenses].

    Science.gov (United States)

    Langenbucher, Achim; Szentmáry, Nóra; Spira, Corinna; Seitz, Berthold; Eppig, Timo

    2016-06-01

    Descemet stripping automated endothelial keratoplasty (DSAEK) is an established treatment option for pathologies of the corneal endothelium. It is typically accompanied with a hyperopic shift in refraction. The purpose of this work is to predict corneal geometry after DSAEK based on model data and to present a concept how to determine corneal power, e.g. for intraocular power calculation to prevent a refractive surprise with a subsequent cataract surgery. Based on data of the Kooijman schematic model eye we simulated a microkeratome cut parallel to the corneal front surface for donor trephination to determine the radial thickness profile of the posterior corneal donor lamella. This donor lamella was tension-neutrally adapted to the back surface of the host and the profile of the cornea after DSAEK was derived and characterized by a quadric surface. Comparison with the curvature of the host without and with donor could resample hyperopic shift which was published in literature. A method was shown how to determine corneal power after DSAEK. From the data of the Kooijman schematic model eye and the donor characteristics central / peripheral corneal thickness was increased by 150 / 250μm due to adaptation of the donor lamella. Geometry of corneal back surface showed a reduced radius of curvature (by about 0.9mm) and a change in conic constant (by about -0.13). Persistent clinically observed hyperopic shift correlates to the change in geometry of the cornea due to adaptation of the donor lamella, which reduces corneal power by 0.88 D. DSAEK leads to a hyperopic shift in refraction, which can be explained by a change in corneal back surface geometry. In case of subsequent cataract surgery, the intraocular lens power should be calculated with consideration of both corneal surfaces rather than keratometry or corneal topography in order to minimize a systematic hyperopic shift due to misinterpretation of corneal power after DSAEK. In case of a Triple-DSAEK, a target

  8. A study of corneal endothelial changes in soft contact lens wearers using non-contact specular microscopy

    Directory of Open Access Journals (Sweden)

    Renu M Magdum

    2013-01-01

    Full Text Available Aim: To study the corneal endothelial changes after soft contact lens wear, to correlate these changes with the duration of soft contact lens wear, and to study the pattern of use and preferences of contact lens among young adults. Materials and Methods: This observational study was carried out in 100 eyes of 50 soft contact lens users aged between 19 and 27 years. Both eyes of 50 medical students who had never worn contact lenses served as controls. Data from each subject were collected using a structured questionnaire of 24 items that included demographic profile, pattern of contact lens use, symptoms, brand name, number of years worn, and hours of daily wear. These data were analyzed using Chi square for association. Specular microscopy was done using TOPCON SP-3000P. Computerized morphometry was used to evaluate central corneal thickness, size, shape, mean cellular density, hexagonality, coefficient of variation, and polymegathism of the corneal cells . Results: It was found that central corneal thickness was 0.532 ± 0.0309 mm in lens users and 0.514 ± 0.03 mm in controls, cell density was 2570.91 ± 432.06 cells/mm 2 in lens users and 2723.17 ± 327.64 cells/mm 2 in controls, while hexagonality was 54.81 ± 39.72% in lens users and 67.65 ± 36.49% in controls. Conclusion: Despite the known effects of long duration of soft contact lens use on corneal endothelial cell morphology, this study could not draw a significant correlation between them. However, a significant difference was found in the corneal endothelial thickness, cell density, and hexagonality. Among the soft contact lens users, 62% used soft disposable type while 38% used soft extended wear contact lens. Contact lenses were preferred over spectacles for better cosmetic appearance, comfort, and wider visual field.

  9. Corneal thickness and intraocular pressure in edematous corneas before and after Descemet stripping with automated endothelial keratoplasty.

    Science.gov (United States)

    Chang, Diane T W; Pantcheva, Mina B; Noecker, Robert J

    2010-10-01

    It is important to accurately measure intraocular pressure (IOP) in eyes with corneal endothelial dysfunction both before and after Descemet stripping with automated endothelial keratoplasty (DSAEK). Glaucoma is a common comorbidity in this population, and IOP elevation can worsen corneal edema. Additionally, preexisting glaucoma and steroid-responsive ocular hypertension are significant risk factors for graft rejection after DSAEK. Accurate tonometry is limited by variations in central corneal thickness (CCT) and corneal hydration that may affect corneal biomechanical properties. We analyzed CCT and IOP in eyes before and after DSAEK to determine whether changes in corneal biomechanics because of edema, grafted tissue, and subsequent stromal deturgescence affect IOP measurement. A retrospective chart review was performed on 32 eyes from 31 patients with corneal edema secondary to Fuchs endothelial dystrophy, bullous keratopathy, or prior graft failure, or rejection that received uncomplicated DSAEK with no evidence of persistent corneal edema or steroid-induced ocular hypertension. IOP was measured by Tono-Pen XL, and CCT was measured by ultrasound pachymetry before and approximately 3 months after surgery. We used paired t tests to evaluate changes in CCT and IOP after DSAEK and linear regression to determine the relationship between CCT and IOP before and after surgery. CCT significantly decreased from 703 ± 82 to 650 ± 52 μm after DSAEK (P = 0.0026), but there was no significant change in measured IOP (16.7 ± 3.4 mm Hg preoperatively and 16.3 ± 4.1 mm Hg postoperatively; P = 0.61). There was no significant relationship between CCT and IOP before (slope = 0.10 ± 0.07 mm Hg/10 μm; r = 0.062; P = 0.17) or after (slope = 0.21 ± 0.14 mm Hg/10 μm; r = 0.072; P = 0.14) DSAEK. CCT is significantly reduced by DSAEK but remains well above the normal range. IOP remains near the preoperative level 3 months after DSAEK. Furthermore, no correction is required for

  10. Ex vivo expansion of bovine corneal endothelial cells in xeno-free medium supplemented with platelet releasate.

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    Ming-Li Chou

    Full Text Available Clinical-grade ex vivo expansion of corneal endothelial cells can increase the availability of corneal tissues for transplantation and treatment of corneal blindness. However, these cells have very limited proliferative capacity. Successful propagation has required so far to use very complex growth media supplemented with fetal bovine serum and other xenocomponents. We hypothesized that human platelet releasates rich in multiple growth factors, and in particular neurotrophins, could potentially be a useful supplement for ex vivo expansion of corneal endothelium cells due to their neural crest origin. Platelet releasates were prepared by calcium salt activation of apheresis platelet concentrates, subjected or not to complement inactivation by heat treatment at 56°C for 30 minutes. Platelet releasates were characterized for their content in proteins and were found to contain high amount of growth factors including platelet-derived growth factor-AB (30.56 to 39.08 ng/ml and brain-derived neurotrophic factor (30.57 to 37.11 ng/ml neurotrophins. We compared the growth and viability of corneal endothelium cells in DMEM-F12 medium supplemented with different combinations of components, including 2.5%∼10% of the platelet releasates. Corneal endothelium cells expanded in platelet releasates exhibited good adhesion and a typical hexagonal morphology. Their growth and viability were enhanced when using the complement-inactivated platelet releasate at a concentration of 10%. Immunostaining and Western blots showed that CECs maintained the expressions of four important membrane markers: Na-K ATPase α1, zona occludens-1, phospho-connexin 43 and N-cadherin. In conclusion, our study provides the first proof-of-concept that human platelet releasates can be used for ex vivo expansion of corneal endothelium cells. These findings open a new paradigm for ex vivo propagation protocols of corneal endothelium cells in compliance with good tissue culture practices

  11. Evaluation of standardized endodontic paper point tear test in New Zealand white rabbits and comparison between corneal sensitivity followed tear tests.

    Science.gov (United States)

    Lima, Leandro; Lange, Rogério Ribas; Turner-Giannico, Amália; Montiani-Ferreira, Fabiano

    2015-01-01

    To evaluate endodontic paper point tear test (PPTT) in rabbits and compare changes in corneal touch threshold (CTT) induced by Schirmer tear test (STT) and PPTT. Normal corneal sensitivity recovery time after both tear tests was also measured. Also, mean PPTT and STT values were obtained. Tear production of 20 eyes, from 10 rabbits, was evaluated using STT and the PPTT. Central corneal touch threshold was measured by a Cochet-Bonnet esthesiometer before any tear test was performed (zero time), immediately after the test (1 min), and consecutively at 6, 11, 16, and 26 min. Tests were conducted on three consecutive days: Day 1 - control condition, no tear tests performed only the CTT; Day 2 - CTT before and after PPTT; and Day 3 - CTT before and after STT. CTT values were compared using repeated measures ANOVA. Corneal touch threshold was significantly increased for at least 16 min after STT, indicating STT causes corneal discomfort. No difference was found between CTT following PPTT and controls, indicating PPTT caused minimal corneal discomfort. The mean (±SD) value for STT was 5.2 ± 1.0 mm/min and for PPTT was 13.8 ± 1.5 mm/min. The aqueous fraction of rabbit's tears can be successfully measured by PPTT. This report established reference values for PPTT in rabbits. Additionally, the absence of a significant difference in CTT after PPTT compared with controls shows that PPTT is well tolerated by rabbits. Considering the improved comfort (compared with STT), accuracy, and low cost, PPTT is a bona fide method of measuring aqueous tear production in rabbits. © 2014 American College of Veterinary Ophthalmologists.

  12. Effect of different culture media and deswelling agents on survival of human corneal endothelial and epithelial cells in vitro.

    Science.gov (United States)

    Valtink, Monika; Donath, Patricia; Engelmann, Katrin; Knels, Lilla

    2016-02-01

    To examine the effects of media and deswelling agents on human corneal endothelial and epithelial cell viability using a previously developed screening system. The human corneal endothelial cell line HCEC-12 and the human corneal epithelial cell line HCE-T were cultured in four different corneal organ culture media (serum-supplemented: MEM +2 % FCS, CorneaMax®/CorneaJet®, serum-free: Human Endothelial-SFM, Stemalpha-2 and -3) with and without 6 % dextran T500 or 7 % HES 130/0.4. Standard growth media F99HCEC and DMEM/F12HCE-T served as controls. In additional controls, the stress inducers staurosporine or hydrogen peroxide were added. After 5 days in the test media, cell viability was assessed by flow cytometrically quantifying apoptotic and necrotic cells (sub-G1 DNA content, vital staining with YO-PRO-1® and propidium iodide) and intracellular reactive oxygen species (ROS). The MEM-based media were unable to support HCEC-12 and HCE-T survival under stress conditions, resulting in significantly increased numbers of apoptotic and necrotic cells. HCEC-12 survival was markedly improved in SFM-based media even under staurosporine or hydrogen peroxide. Likewise, HCE-T survival was improved in SFM with or without dextran. The media CorneaMax®, CorneaJet®, and CorneaMax® with HES supported HCEC-12 survival better than MEM-based media, but less well than SFM-based media. HCE-T viability was also supported by CorneaJet®, but not by CorneaMax® with or without HES. Stemalpha-based media were not suitable for maintaining viability of HCEC-12 or HCE-T in the applied cell culture system. The use of serum-supplemented MEM-based media for corneal organ culture should be discontinued in favour of serum-free media like SFM.

  13. Functional significance of thermosensitive transient receptor potential melastatin channel 8 (TRPM8) expression in immortalized human corneal endothelial cells.

    Science.gov (United States)

    Mergler, Stefan; Mertens, Charlotte; Valtink, Monika; Reinach, Peter S; Székely, Violeta Castelo; Slavi, Nefeli; Garreis, Fabian; Abdelmessih, Suzette; Türker, Ersal; Fels, Gabriele; Pleyer, Uwe

    2013-11-01

    Human corneal endothelial cells (HCEC) maintain appropriate tissue hydration and transparency by eliciting net ion transport coupled to fluid egress from the stroma into the anterior chamber. Such activity offsets tissue swelling caused by stromal imbibition of fluid. As corneal endothelial (HCE) transport function is modulated by temperature changes, we probed for thermosensitive transient receptor potential melastatin 8 (TRPM8) functional activity in immortalized human corneal endothelial cells (HCEC-12) and freshly isolated human corneal endothelial cells (HCEC) as a control. This channel is either activated upon lowering to 28 °C or by menthol, eucalyptol and icilin. RT-PCR and quantitative real-time PCR (qPCR) verified TRPM8 gene expression. Ca(2+) transients induced by either menthol (500 μmol/l), eucalyptol (3 mmol/l), or icilin (2-60 μmol/l) were identified using cell fluorescence imaging. The TRP channel blocker lanthanum III chloride (La(3+), 100 μmol/l) as well as the TRPM8 blockers BCTC (10 μmol/l) and capsazepine (CPZ, 10 μmol/l) suppressed icilin-induced Ca(2+) increases. In and outward currents induced by application of menthol (500 μmol/l) or icilin (50 μmol/l) were detected using the planar patch-clamp technique. A thermal transition from room temperature to ≈ 18 °C led to Ca(2+) increases that were inhibited by a TRPM8 blocker BCTC (10 μmol/l). Other thermosensitive TRP pathways whose heterogeneous Ca(2+) response patterns are suggestive of other Ca(2+) handling pathways were also detected upon strong cooling (≈10 °C). Taken together, functional TRPM8 expression in HCEC-12 and freshly dissociated HCEC suggests that HCE function can adapt to thermal variations through activation of this channel subtype. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Real-time assessment of corneal endothelial cell damage following graft preparation and donor insertion for DMEK.

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    Maninder Bhogal

    Full Text Available To establish a method for assessing graft viability, in-vivo, following corneal transplantation.Optimization of calcein AM fluorescence and toxicity assessment was performed in cultured human corneal endothelial cells and ex-vivo corneal tissue. Descemet membrane endothelial keratoplasty grafts were incubated with calcein AM and imaged pre and post preparation, and in-situ after insertion and unfolding in a pig eye model. Global, macroscopic images of the entire graft and individual cell resolution could be attained by altering the magnification of a clinical confocal scanning laser microscope. Patterns of cell loss observed in situ were compared to those seen using standard ex-vivo techniques.Calcein AM showed a positive dose-fluorescence relationship. A dose of 2.67μmol was sufficient to allow clear discrimination between viable and non-viable areas (sensitivity of 96.6% with a specificity of 96.1% and was not toxic to cultured endothelial cells or ex-vivo corneal tissue. Patterns of cell loss seen in-situ closely matched those seen on ex-vivo assessment with fluorescence viability imaging, trypan blue/alizarin red staining or scanning electron microscopy. Iatrogenic graft damage from preparation and insertion varied between 7-35% and incarceration of the graft tissue within surgical wounds was identified as a significant cause of endothelial damage.In-situ graft viability assessment using clinical imaging devices provides comparable information to ex-vivo methods. This method shows high sensitivity and specificity, is non-toxic and can be used to evaluate immediate cell viability in new grafting techniques in-vivo.

  15. Corneal endothelial autocrine trophic factor VIP in a mechanism-based strategy to enhance human donor cornea preservation for transplantation.

    Science.gov (United States)

    Koh, Shay-Whey Margaret

    2012-02-01

    Vasoactive intestinal peptide (VIP) and ciliary neurotrophic factor (CNTF) are identified as autocrines of human corneal endothelial (CE) cells working in concert to maintain the differentiated state and promote the survival of the corneal endothelium. From VIP gene knockdown study, endogenous VIP is shown to maintain the level of the differentiation marker, the adhesion molecule N-cadherin, CE cell size, shape, and retention, in situ in the human donor corneoscleral explants. Exogenous VIP protects the corneal endothelium against the killing effect of oxidative stress, in part by upholding ATP levels in CE cells dying of oxidative stress-induced injury, allowing them to die of an apoptotic death instead of an acute necrotic one. The switch from the acute necrosis to the programmed cell death (apoptosis) may have allowed the injured CE cell to be rescued by the VIP-upregulated pathways, including those of Bcl-2 and N-cadherin, and resulted in long-term CE cell survival. The endogenous VIP in CE cells is upregulated by CNTF, which is released by CE cells surviving the oxidative stress. The CNTF receptor (CNTFRα) is expressed in CE cells in human donor corneoscleral explant and gradually becomes lost during corneal storage. VIP treatment (10(-8) M, 37 °C, 30 min) prior to storage of freshly dissected human donor corneoscleral explants increases their CE cell CNTFRα level and responsiveness to CNTF in upregulating the gap junctional protein connexin-43 expression. VIP treatment of both fresh and preserved corneoscleral explants reduces CE damage in the corneoscleral explants and in the corneal buttons trephined from them. CE cell loss is a critical risk factor in corneal graft failure at any time in the life of the graft, which can be as late as 5-10 years after an initially successful transplant. A new procedure, Descemet's stripping automated endothelial keratoplasty (DSAEK), which is superior to the traditional full thickness transplantation in many aspects

  16. Expression of MMP-2, MT1-MMP, and TIMP-2 by cultured rabbit corneal fibroblasts under mechanical stretch.

    Science.gov (United States)

    Liu, Chengxing; Feng, Pengfei; Li, Xiaona; Song, Jie; Chen, Weiyi

    2014-08-01

    Refractive surgery not only leads to tissue injury but also evokes mechanical stress increase of the cornea. How the mechanical stress affects the corneal matrix remodeling, specifically, matrix metalloproteinases (MMPs) and their inhibitors (tissue inhibitors of metalloproteinases; TIMPs) is not well understood. In this study, cultured rabbit corneal fibroblasts in vitro were subjected to regimen of 5%, 10%, or 15% equibiaxial stretch at 0.1 Hz for 3 or 24 h. MMP-2 protein level was measured by gelatin zymography and Western blotting. MMP-2, membrane type 1 MMP (MT1-MMP), and TIMP-2 mRNA levels were quantified by real-time quantitative PCR. Extracellular regulated protein kinase (ERK) phosphorylation protein levels were assessed by Western blotting. Our results showed that a 15% stretch resulted in increases in MMP-2 protein, MMP-2 mRNA, and MT1-MMP mRNA levels, but a decrease in TIMP-2 mRNA level. However, a 5% stretch caused decreases in MMP-2 protein and mRNA level, but an increase in TIMP-2 mRNA level, and no change in MT1-MMP mRNA level. A 15% stretch also caused a significant increase in ERK1/2 phosphorylation. Inhibition of the mitogenactivated protein kinase (MEK) pathway with PD98059 attenuated stretch-induced increase in MMP-2 production and ERK activity. These results suggest that small-magnitude stretching may promote corneal matrix synthetic events, whereas large-magnitude stretching promotes corneal matrix degradation by changing the balance between MMPs and TIMPs in corneal fibroblasts. Large-magnitude stretch-induced increase in pro-MMP-2 production was in an ERK-dependent manner. © 2014 by the Society for Experimental Biology and Medicine.

  17. A new nanosecond UV laser at 355 nm: early results of corneal flap cutting in a rabbit model.

    Science.gov (United States)

    Trost, Andrea; Schrödl, Falk; Strohmaier, Clemens; Bogner, Barbara; Runge, Christian; Kaser-Eichberger, Alexandra; Krefft, Karolina; Vogel, Alfred; Linz, Norbert; Freidank, Sebastian; Hilpert, Andrea; Zimmermann, Inge; Grabner, Günther; Reitsamer, Herbert A

    2013-12-03

    A new 355 nm UV laser was used for corneal flap cutting in an animal model and tested for clinical and morphologic alterations. Corneal flaps were created (Chinchilla Bastards; n = 25) with an UV nanosecond laser at 355 nm (150 kHz, pulse duration 850 ps, spot-size 1 μm, spot spacing 6 × 6 μm, side cut Δz 1 μm; cutting depth 130 μm) and pulse energies of 2.2 or 2.5 μJ, respectively. Following slit-lamp examination, animals were killed at 6, 12, and 24 hours after treatment. Corneas were prepared for histology (hematoxylin and eosin [HE], TUNEL-assay) and evaluated statistically, followed by ultrastructural investigations. Laser treatment was tolerated well, flap lift was easier at 2.5 μJ compared with 2.2 μJ. Standard HE at 24 hours revealed intact epithelium in the horizontal cut, with similar increase in corneal thickness at both energies. Irrespective of energy levels, TUNEL assay revealed comparable numbers of apoptotic cells in the horizontal and vertical cut at 6, 12, and 24 hours, becoming detectable in the horizontal cut as an acellular stromal band at 24 hours. Ultrastructural analysis revealed regular morphology in the epi- and endothelium, while in the stroma, disorganized collagen lamellae were detectable representing the horizontal cut, again irrespective of energy levels applied. This new UV laser revealed no epi- nor endothelial damage at energies feasible for corneal flap cutting. Observed corneal swelling was lower compared with existing UV laser studies, albeit total energy applied here was much higher. Observed loss of stromal keratinocytes is comparable with available laser systems. Therefore, this new laser is suitable for refractive surgery, awaiting its test in a chronic environment.

  18. A Rabbit Model of Acanthamoeba Keratitis: Use of Infected Soft Contact Lenses After Corneal Epithelium Debridement With a Diamond Burr.

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    Ortillés, Ángel; Goñi, Pilar; Rubio, Encarnación; Sierra, Marta; Gámez, Ekaterina; Fernández, María T; Benito, María; Cristóbal, José Á; Calvo, Begoña

    2017-02-01

    To develop a rabbit model of Acanthamoeba keratitis (AK) as the best method to reproduce the natural course of this disease. To induce AK, infected contact lenses (1000 amoebae/mm2, 90% trophozoites) were placed over the previously debrided corneal surface, in combination with a temporary tarsorrhaphy. Environmental and clinical strains of Acanthamoeba spp. (genotype T4) were used. Three groups (1L, n = 32; 2L-21d, n = 5; 2L-3d, n = 23) were established according to the number of contact lenses used (1L, 1 lens; 2L-21d and 2L-3d, 2 lenses) and the placement day of these (1L, day 1; 2L-21d, days 1 and 21; 2L-3d, days 1 and 3). The infection was quantified by a clinical score system and confirmed using corneal cytology and culture, polymerase chain reaction and histopathologic analysis. The infection rate obtained was high (1L, 87.5%; 2L-21d, 100%; 2L-3d, 82.6%), although no clinical signs were observed in the 50% of the infected animals in group 1L. Among groups, group 2L-3d showed more cases of moderate and severe infection. Among strains, no statistically significant differences were found in the infection rate. In the control eyes, cross infection was confirmed when a sterile contact lens was placed in the previously debrided corneas but not if the eye remained intact. The combination of two infected contact lenses after corneal debridement seems to be an alternative model, clinically and histopathologically similar to its human counterpart, to induce the different AK stages and reproduce the course of the disease in rabbits.

  19. Polymorphism of the Transferrin Gene in Eye Diseases: Keratoconus and Fuchs Endothelial Corneal Dystrophy

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    Katarzyna A. Wójcik

    2013-01-01

    Full Text Available Oxidative stress may play a role in the pathogenesis of keratoconus (KC and Fuchs endothelial corneal dystrophy (FECD. Iron may promote the stress by the Fenton reaction, so its homeostasis should be strictly controlled. Transferrin is essential for iron homeostasis because it transports iron from plasma into cells. The malfunction of transferrin, which may be caused by variation in its gene (TF variation, may contribute to oxidative stress and change KC and FECD risk. To verify this hypothesis we investigated the association between three polymorphisms of the TF gene, g.3296G>A (rs8177178, g.3481A>G (rs8177179, and c.–2G>A (rs1130459, and KC and FECD occurrence. Genotyping was performed in blood lymphocytes in 216 patients with KC, 130 patients with FECD and 228 controls by PCR-RFLP. We studied also the influence of other risk factors. The A/A genotype and the A allele of the g.3296G>A polymorphism were associated with KC occurrence, while the G allele was negatively correlated with it. We observed a decrease in KC occurrence associated with the A/G genotype of the g.3481A>G polymorphism. We did not find any association between the c.–2G>A polymorphism and KC. No association was found between all three polymorphisms and FECD occurrence.

  20. Transforming growth factor-β2 induces morphological alteration of human corneal endothelial cells in vitro

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    Jing Wang

    2014-10-01

    Full Text Available AIM:To investigate the morphological altering effect of transforming growth factor-β2 (TGF-β2 on untransfected human corneal endothelial cells (HCECs in vitro.METHODS: After untransfected HCECs were treated with TGF-β2 at different concentrations, the morphology, cytoskeleton distribution, and type IV collagen expression of the cells were examined with inverted contrast light microscopy, fluorescence microscopy, immunofluorescence or Western Blot.RESULTS:TGF-β2 at the concentration of 3-15 μg/L had obviously alterative effects on HCECs morphology in dose and time-dependent manner, and 9 μg/L was the peak concentration. TGF-β2 (9 μg/L altered HCE cell morphology after treatment for 36h, increased the mean optical density (P<0.01 and the length of F-actin, reduced the mean optical density (P<0.01 of the collagen type IV in extracellular matrix (ECM and induced the rearrangement of F-actin, microtubule in cytoplasm and collagen type IV in ECM after treatment for 72h. CONCLUTION:TGF-β2 has obviously alterative effect on the morphology of HCECs from polygonal phenotype to enlarged spindle-shaped phenotype, in dose and time-dependence manner by inducing more, elongation and alignment of F-actin, rearrangement of microtubule and larger spread area of collagen type IV.

  1. Effect of Intraocular Forward Scattering and Corneal Higher-Order Aberrations on Visual Acuity after Descemet's Stripping Automated Endothelial Keratoplasty.

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    Kazutaka Kamiya

    Full Text Available To assess the relationship of intraocular forward scattering and corneal higher-order aberrations (HOAs with best spectacle corrected visual acuity (BSCVA after Descemet's stripping automated endothelial keratoplasty (DSAEK, and to compare these parameters between DSAEK and non-Descemet's stripping automated endothelial keratoplasty (n-DSAEK groups.This retrospective study enrolled thirty eyes of 30 consecutive patients who underwent standard DSAEK, and who underwent successful phacoemulsification with intraocular lens implantation before DSAEK. The mean age at the time of surgery was 71.7 ± 10.4 years. We quantitatively evaluated the objective scattering index (OSI using the double-pass instrument (OQAS II, Visiometrics and corneal HOAs using Hartmann-Shack aberrometry (KR-9000PW, Topcon 3 months postoperatively.The mean OSI, corneal HOAs, and logMAR BSCVA 3 months after DSAEK were 7.91 ± 3.58, 0.43 ± 0.27 μm, and 0.32 ± 0.25, respectively. We found a significant correlation between the OSI and logMAR BSCVA (Spearman correlation coefficient r=0.714, p0.05.Our pilot study demonstrated that the postoperative corrected visual acuity was significantly correlated with intraocular forward scattering, but not with corneal HOAs in post-DSAEK eyes, suggesting that intraocular forward scattering plays a more essential role in postoperative visual performance than corneal aberrations after DSAEK. The detailed visual performance, such as HOAs and intraocular scattering, after n-DSAEK appears to be essentially equivalent to that after DSAEK.

  2. Preservation of vascular DDAH activity contributes to the protection of captopril against endothelial dysfunction in hyperlipidemic rabbits.

    Science.gov (United States)

    Lin, Yuan; Feng, Mei; Lu, Chang-Wu; Lei, Yan-Ping; He, Zhi-Min; Xiong, Yan

    2017-03-05

    Endothelial dysfunction plays a pivotal role in the pathogenesis of atherosclerosis. Endogenous inhibitor of nitric oxide synthase (NOS) asymmetric dimethylarginine (ADMA) has been recognized as an independent risk factor of endothelial dysfunction and the biomarker of atherosclerosis. This study was to investigate whether endogenous ADMA and its metabolic enzyme dimethylarginine dimethylaminohydrolase (DDAH) were involved in mechanisms of captopril protection against endothelial dysfunction in high fat diet feeding rabbits. Half of model rabbits were treated with captopril (10mg/kg/d, i.g.) for 12w. Vascular morphology and serum lipid profiles were detected. Serum ADMA concentration were assayed by high performance liquid chromatography. Recombinant DDAH2 gene adenoviruses were ex vivo transferred to thoracic aortas of high fat diet feeding rabbits. Endothelium-dependent relaxation of aortas response to acetylcholine and DDAH activity were measured. Atherosclerosis was confirmed in high fat diet feeding rabbits by increased serum lipid profiles and morphologic changes of vascular wall. Serum ADMA levels were significantly increased in hyperlipidemic rabbits accompanied with impairment of endothelium-dependent relaxation and inhibition of DDAH activity in thoracic aortas. Captopril treatment not only decreased vascular intima thickening and serum ADMA concentration but also preserved vascular DDAH activity and endothelium-dependent relaxation in hyperlipidemic rabbits without influence on serum lipid profiles. Similar beneficial effects on endothelial function and DDAH activity could be achieved by DDAH2 gene transfection. These results indicated that captopril could protect against injuries of vascular morphology and endothelial function in hyperlipidemic rabbits, the mechanisms may be related to the preservation of DDAH activity and decrease of ADMA accumulation in vascular endothelium. Copyright © 2017. Published by Elsevier B.V.

  3. Comparison of the changes in corneal endothelial cells after pars plana and anterior chamber ahmed valve implant.

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    Seo, Ji Won; Lee, Jong Yeon; Nam, Dong Heun; Lee, Dae Yeong

    2015-01-01

    Purpose. To compare the changes in corneal endothelial cells after pars plana Ahmed glaucoma valve (AGV) implantation with those after the anterior chamber AGV implantation for refractory glaucoma. Methods. The medical records of 18 eyes with pars plana implantation of AGV (ppAGV) were reviewed retrospectively and were compared with 18 eyes with the anterior chamber AGV (acAGV) implant. The preoperative and postoperative endothelial cells, intraocular pressure (IOP), and postoperative complications during the follow-up in both groups were compared. Results. The average follow-up was 18 months. The postoperative endothelial cells in the ppAGV and acAGV groups were 2044 ± 303 and 1904 ± 324, respectively (P = 0.25). The average percentage decrease in the endothelial cells in the ppAGV and acAGV groups at 18 months was 12.5% and 18.4%, respectively, and showed significant difference between the 2 groups (P = 0.01). No difference in IOP control and the number of postoperative glaucoma medications was observed between the 2 groups. Conclusions. Endothelial cell damage in the ppAGV group for refractory glaucoma appeared to be lower than that in the acAGV group. Therefore, pars plana implantation of AGV may be preferred as it may have lower level of endothelial cell damage while maintaining similar level of IOP control.

  4. Comparison of the Changes in Corneal Endothelial Cells after Pars Plana and Anterior Chamber Ahmed Valve Implant

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    Ji Won Seo

    2015-01-01

    Full Text Available Purpose. To compare the changes in corneal endothelial cells after pars plana Ahmed glaucoma valve (AGV implantation with those after the anterior chamber AGV implantation for refractory glaucoma. Methods. The medical records of 18 eyes with pars plana implantation of AGV (ppAGV were reviewed retrospectively and were compared with 18 eyes with the anterior chamber AGV (acAGV implant. The preoperative and postoperative endothelial cells, intraocular pressure (IOP, and postoperative complications during the follow-up in both groups were compared. Results. The average follow-up was 18 months. The postoperative endothelial cells in the ppAGV and acAGV groups were 2044 ± 303 and 1904 ± 324, respectively (P=0.25. The average percentage decrease in the endothelial cells in the ppAGV and acAGV groups at 18 months was 12.5% and 18.4%, respectively, and showed significant difference between the 2 groups (P=0.01. No difference in IOP control and the number of postoperative glaucoma medications was observed between the 2 groups. Conclusions. Endothelial cell damage in the ppAGV group for refractory glaucoma appeared to be lower than that in the acAGV group. Therefore, pars plana implantation of AGV may be preferred as it may have lower level of endothelial cell damage while maintaining similar level of IOP control.

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

    Science.gov (United States)

    Kim, Min Su; Kim, Kyoung Nam; Kim, Chang-Sik

    2016-12-01

    To compare changes in corneal endothelial cell density (CECD) after Ahmed glaucoma valve (AGV) implantation and trabeculectomy. 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. 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 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 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 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.

  6. Attenuation of oxidative stress, inflammation, and endothelial dysfunction in hypercholesterolemic rabbits by allicin.

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    El-Sheakh, Ahmed R; Ghoneim, Hamdy A; Suddek, Ghada M; Ammar, El Sayed M

    2015-08-14

    Allicin, the active substance of garlic, exerts a broad spectrum of pharmacological activities and is considered to have potential therapeutic applications. The present study was designed to investigate the possible beneficial effects of allicin against oxidative stress, inflammation, and endothelial dysfunction in hypercholesterolemic rabbits. Male New Zealand white rabbits were used in this study. Rabbits randomly received 1 of the following treatments: normal chow diet for 4 weeks, 1% high cholesterol diet (HCD), HCD plus allicin (10 mg/kg/day), or HCD plus atorvastatin (10 mg/kg/day). Blood samples were collected at the end of experimental diets for measurement of serum total cholesterol (TC), triglycerides (TGs), high-density lipoprotein cholesterol (HDL-C), C-reactive protein (CRP), malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (SOD). In addition, the aorta was removed for measurement of vascular reactivity, histopathological changes, intima/media (I/M) ratio, and immunohistochemical staining of both tumor necrosis-alpha (TNF-α) and nuclear factor (NF)-κB. HCD induced significant increases in serum TC, TGs, low-density lipoprotein cholesterol (LDL-C), CRP, and MDA. Moreover, HCD caused significant decrease in serum GSH and SOD. In addition, aortic relaxation response to acetylcholine (ACh) was impaired. Immunohistochemical staining of aortic specimens from HCD-fed rabbits revealed high expression levels of both TNF-α and the oxidant-induced transcription factor, NF-κB. Allicin supplementation significantly decreased serum MDA and CRP, increased serum HDL-C, GSH, and SOD levels while nonsignificantly affecting HCD-induced elevations in serum TC and LDL-C. Additionally, allicin significantly protected against HCD-induced attenuation of rabbit aortic endothelium-dependent relaxation to ACh and elevation in I/M ratio. This effect was confirmed by histopathological examination of the aorta. Moreover, allicin has substantially

  7. Surgical Management of Limbal Dermoids Using Anterior Corneal Buttons From Descemet Stripping Automated Endothelial Keratoplasty Donor Tissue as Patch Grafts.

    Science.gov (United States)

    Wu, Kuan-I; Chu, Hsiao-Sang; Pai, Amy Shih-I; Hou, Yu-Chih; Lin, Szu-Yuan; Chen, Wei-Li; Hu, Fung-Rong

    2017-01-01

    To assess the surgical and clinical outcomes of anterior lamellar keratoplasty using anterior corneal buttons from Descemet stripping automated endothelial keratoplasty (DSAEK) donor tissue. Retrospective data from 8 patients with unilateral limbal dermoids, treated between February 2011 and January 2016 at National Taiwan University Hospital, were analyzed. Donor corneas for DSAEK were divided into anterior and posterior lamellae using a 350-μm microkeratome. Anterior corneal buttons were stored for up to 4 weeks in storage media before being used as patch grafts for anterior lamellar keratoplasty. Corneoscleral integrity was preserved in all cases. Three of the 8 patients showed improved best-corrected visual acuity after surgery. Three patients' astigmatism reduced by more than 0.75 diopters. All 8 patients had satisfactory cosmesis after surgery. Neovascularization at the graft-host junction and graft edema was noted in 1 patient and was treated using bevacizumab injection and topical steroid. Anterior corneal buttons obtained from DSAEK can be used as patch grafts for surgical management of limbal dermoids. This procedure achieved satisfactory cosmetic and visual outcomes in our study. This procedure may potentially allow one corneal tissue to be received by multiple patients.

  8. Endothelial keratoplasty combined with cataract surgery or alone using polyethylene glycol hydrogel sealant for closure of corneal incisions.

    Science.gov (United States)

    Spierer, Oriel; O'Brien, Terrence P

    2015-03-01

    We present a surgical technique in which polyethylene glycol hydrogel is used to seal corneal incisions and maintain air in the anterior chamber. This technique was used in 11 eyes that had Descemet-stripping endothelial keratoplasty (DSEK). In 2 cases, DSEK was combined with cataract extraction and intraocular lens implantation. In all cases, the anterior chamber was well formed with no leakage and the donor graft remained attached following surgery. The use of polyethylene glycol hydrogel in DSEK and combined DSEK-cataract surgery may shorten surgery, decrease suture-induced corneal astigmatism, and prevent the need for suture removal. Neither author has a financial or proprietary interest in any material or method mentioned. Copyright © 2015. Published by Elsevier Inc.

  9. Polymorphisms of the Homologous Recombination Gene RAD51 in Keratoconus and Fuchs Endothelial Corneal Dystrophy

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    Ewelina Synowiec

    2013-01-01

    Full Text Available Purpose. We investigated the association between genotypes and haplotypes of the c.-61G>T (rs 1801320 and c.-98G>C (rs 1801321 polymorphisms of the RAD51 gene and the occurrence of keratoconus (KC and Fuchs endothelial corneal dystrophy (FECD in dependence on some environmental factors. Methods. The polymorphisms were genotyped in peripheral blood lymphocytes of 100 KC and 100 FECD patients as well as 150 controls with PCR-RFLP. Results. The G/T genotype of the c.-61G>T polymorphism was associated with significantly increased frequency occurrence of KC (crude OR 2.99, 95% CI 1.75–5.13. On the other hand, the G/G genotype of this polymorphism was positively correlated with a decreased occurrence of this disease (crude OR 0.52, 95% CI 0.31–0.88. We did not find any correlation between genotypes/alleles of the c.-98G>C polymorphism and the occurrence of KC. We also found that the G/G genotype and G allele of the c.-98G>C polymorphism had a protective effect against FECD (crude OR 0.51, 95% CI 0.28–0.92; crude OR 0.53, 95% CI 0.30–0.92, resp., while the G/C genotype and the C allele increased FECD occurrence (crude OR 1.85, 95% CI 1.01–3.36; crude OR 1.90, 95% CI 1.09–3.29, resp.. Conclusions. The c.-61T/T and c.-98G>C polymorphisms of the RAD51 gene may have a role in the KC and FECD pathogenesis and can be considered as markers in these diseases.

  10. Comparison of manual & automated analysis methods for corneal endothelial cell density measurements by specular microscopy.

    Science.gov (United States)

    Huang, Jianyan; Maram, Jyotsna; Tepelus, Tudor C; Modak, Cristina; Marion, Ken; Sadda, SriniVas R; Chopra, Vikas; Lee, Olivia L

    2017-08-07

    To determine the reliability of corneal endothelial cell density (ECD) obtained by automated specular microscopy versus that of validated manual methods and factors that predict such reliability. Sharp central images from 94 control and 106 glaucomatous eyes were captured with Konan specular microscope NSP-9900. All images were analyzed by trained graders using Konan CellChek Software, employing the fully- and semi-automated methods as well as Center Method. Images with low cell count (input cells number <100) and/or guttata were compared with the Center and Flex-Center Methods. ECDs were compared and absolute error was used to assess variation. The effect on ECD of age, cell count, cell size, and cell size variation was evaluated. No significant difference was observed between the Center and Flex-Center Methods in corneas with guttata (p=0.48) or low ECD (p=0.11). No difference (p=0.32) was observed in ECD of normal controls <40 yrs old between the fully-automated method and manual Center Method. However, in older controls and glaucomatous eyes, ECD was overestimated by the fully-automated method (p=0.034) and semi-automated method (p=0.025) as compared to manual method. Our findings show that automated analysis significantly overestimates ECD in the eyes with high polymegathism and/or large cell size, compared to the manual method. Therefore, we discourage reliance upon the fully-automated method alone to perform specular microscopy analysis, particularly if an accurate ECD value is imperative. Copyright © 2017. Published by Elsevier España, S.L.U.

  11. A histological study of rabbit corneas after transepithelial corneal crosslinking using partial epithelial photoablation or ethanol treatment

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    Mehmet Cuneyt Ozmen

    2014-12-01

    Full Text Available AIM: To evaluate the histological changes after transepithelial corneal crosslinking (CXL using partial thickness excimer laser ablation or epithelial ethanol application in an experimental rabbit study.METHODS: Right eyes of twenty-four rabbits were studied. Four eyes received total epithelial debridement (group I. Four eyes received partial thickness epithelial ablation with excimer laser (group II. Twelve eyes were treated with different durations (30s and 60s and concentrations (18% to 48% of ethanol (group III. Riboflavin was applied for 30min intervals along with topical proparacaine drops with benzalkonium chloride, and 370 nm irradiation was performed for 30min, while riboflavin was instilled every 3min. Four eyes (group IV received 48% ethanol for 30s without riboflavin and irradiation. Eyes were collected after 24h and examined histologically.RESULTS: All eyes in group I showed keratocyte loss in the superficial 300 µ of corneal storma. In group II, 1-4 layers of epithelium were preserved and no keratocyte loss occurred. In group III, CXL after treatment with ethanol up to 24% concentration and up to 60s revealed no keratocyte loss. CXL after treatment with 48% and higher ethanol concentrations yielded keratocyte loss in the superficial 200 µ to 300 µ of cornea.CONCLUSION: Incomplete excimer laser ablation of the epithelium or treatment with ethanol up to 24% concentration and up to 60s duration yielded no stromal keratocyte loss. To get the same histological appearance seen in epithelial debridement group, partial thickness excimer laser epithelial ablation or ethanol application is not adequate for transepithelial CXL.

  12. Construction of tissue-engineered full-thickness cornea substitute using limbal epithelial cell-like and corneal endothelial cell-like cells derived from human embryonic stem cells.

    Science.gov (United States)

    Zhang, Canwei; Du, Liqun; Sun, Peng; Shen, Lin; Zhu, Jing; Pang, Kunpeng; Wu, Xinyi

    2017-04-01

    The aim of this study was to construct a full-thickness artificial cornea substitute in vitro by coculturing limbal epithelial cell-like (LEC-like) cells and corneal endothelial cell-like (CEC-like) cells derived from human embryonic stem cells (hESCs) on APCM scaffold. A 400 μm thickness, 11 mm diameter APCM lamella containing Bowman's membrane was prepared as the scaffold using trephine and a special apparatus made by ourselves. LEC-like cells and CEC-like cells, derived from hESCs as our previously described, were cocultured on the scaffold using a special insert of 24-well plates that enabled seeding both sides of the scaffold. Three or four layers of epithelium-like cells and a uniform monolayer of CEC-like cells could be observed by H&E staining. The thickness, endothelial cell density, and mechanical properties of the construct were similar to that of native rabbit corneas. Immunofluorescence analysis showed expression of ABCG2 and CK3 in the epithelium-like cell layers and expression of N-cadherin, ZO-1 and Na+/K + ATPase in the CEC-like cells. The corneal substitutes were well integrated within the host corneas, and the transparency increased gradually in 8-week follow-up after transplantation in the rabbits. These results suggest that the strategy we developed is feasible and effective for construction of tissue-engineered full-thickness cornea substitute with critical properties of native cornea. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Near infra-red light attenuates corneal endothelial cell dysfunction in situ and in vitro.

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    Núñez-Álvarez, Claudia; Del Olmo-Aguado, Susana; Merayo-Lloves, Jesús; Osborne, Neville N

    2017-08-01

    In the present study mechanical damage to the corneal endothelium was induced by elevation of intraocular pressure (IOP, 140 mmHg, 60 min) to one eye of rats, delivered either in complete darkness or in the presence of red light (16.5 W/m2, 3000 lx, 625-635 nm). IOP raised in the dark revealed the endothelium to be damaged as staining for the gap junction protein ZO-1 was irregular in appearance with some cells displaced in position or lost to leave gaps or holes. This damage was clearly attenuated when red light was focused through the pupil during the insult of raised IOP. Moreover, staining of endothelium with JC-1 dye showed mitochondria to be activated by both elevated IOP and red light but the activation of mitochondria persisted longer for red light. We interpret this finding to suggest that raised IOP causes apoptosis of endothelial cells and that their mitochondria are activated in the initial stages of the process. In contrast, red light activates mitochondria to induce a protective mechanism to counteract the negative influence of raised IOP on endothelial cells. Evidence is provided to support this notion by the finding that red light stimulates mitochondrial cytochrome oxidase IV (COX IV). Moreover, mitochondria in corneal endothelial cell cultures are activated by red light, revealed by staining with JC-1, that results in an increased rate of proliferation and are also able to counteract toxic insults (sodium azide or cobalt chloride) to the cultures. The present studies therefore show that a non-toxic level of red light attenuates damage to the corneal endothelium both in situ and in vitro through action on COX IV located in mitochondria that results in an enhancement of a cell's survival mechanisms. The study provides proof of principle for the non-invasive use of red-light therapy to attenuate any dysfunctions associated with the corneal endothelium and so preserve maximum visual acuity. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Spontaneous Corneal Clearance in the Presence of a Partially Detached Graft after Non-Descemet Stripping Automated Endothelial Keratoplasty

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    Konstantinos Droutsas

    2016-05-01

    Full Text Available Purpose: To report the explantation of a detached and opaque donor disc as an alternative to secondary keratoplasty in a case of persistent graft detachment followed by spontaneous clearance of the recipient cornea after non-Descemet stripping automated endothelial keratoplasty (non-DSAEK. Methods: A 57-year-old man with cataract and bullous keratopathy after herpes simplex virus endotheliitis of the right eye and best spectacle-corrected visual acuity (BSCVA of 0.1 underwent simultaneous phacoemulsification and non-DSAEK. Due to early detachment of the donor disc, two additional intracameral air injections were necessary in order to achieve graft attachment. However, the donor disc gradually detached and became fibrotic while the recipient cornea anterior to the detached graft became transparent and without any edema. Therefore, a mere explantation of the DSAEK graft was performed. Results: Four months after graft explantation, BSCVA was 0.5 and endothelial cell density (ECD was 1,221 cells/mm2. After 13 months, BSCVA was still 0.6 while ECD had fell to 800, and 2 years later, the endothelium decompensated. BSCVA was 0.3 and ECD was not measurable. Conclusions: To our knowledge this is the first report of explantation of an endothelial graft as an alternative to re-keratoplasty in a case of spontaneous corneal clearance. This minimally invasive treatment may be considered in similar cases. However, due to the ongoing loss of endothelial cells after endothelial keratoplasty, a re-keratoplasty may still be needed in the long term.

  15. Replacing the endothelium without corneal surface incisions or sutures: the first United States clinical series using the deep lamellar endothelial keratoplasty procedure.

    Science.gov (United States)

    Terry, Mark A; Ousley, Paula J

    2003-04-01

    To report the 6- and 12-month results of the first United States clinical series of deep lamellar endothelial keratoplasty (DLEK) in the treatment of endothelial dysfunction. Prospective, noncomparative, interventional case series. Eight eyes of eight patients with corneal edema from Fuchs' dystrophy and pseudophakia. A 9.0-mm limbal, scleral, partial-depth incision provided access for a deep lamellar corneal pocket dissection. A 7.5- to 8.0-mm posterior lamellar disc of recipient tissue was then excised and replaced through the pocket with a same size donor disc containing healthy endothelium. A temporary air bubble in the anterior chamber was used for donor tissue adherence, and no surface corneal incisions or sutures were necessary. Preoperative and postoperative best spectacle-corrected visual acuity (BSCVA), manifest refraction astigmatism, TMS-1 topography, ultrasonic pachymetry, Orbscan topography, and endothelial cell density were evaluated. Intraoperative and postoperative complications are reported. At 6 and 12 months after surgery, all eight corneas were clear and the grafts were healed in good position. At 6 months, the BSCVA varied between 20/30 and 20/70, the average change in astigmatism from before surgery was +1.13 diopters (D; +/-1.50 D), the average change in corneal power was -0.4 D (+/-1.7 D), the average pachymetry was 648 micro m (+/-134 micro m), and the average endothelial cell count was 2290 cells/mm(2) (+/-372 cells/mm(2)). At 12 months, three of the four eyes reaching this time gate were 20/40 or better, with a change in astigmatism from before surgery of only +0.81 D (+/- 0.55 D), a corneal power change of -1.3 D (+/- 0.4 D), and an endothelial density of 2409 cells/mm(2) (+/- 154 cells/mm(2)). One of the original nine eyes entered into this study required conversion to standard penetrating keratoplasty as a result of a microperforation during recipient pocket dissection and has experienced no ill effects. The DLEK procedure, with its

  16. Menadione-Induced DNA Damage Leads to Mitochondrial Dysfunction and Fragmentation During Rosette Formation in Fuchs Endothelial Corneal Dystrophy.

    Science.gov (United States)

    Halilovic, Adna; Schmedt, Thore; Benischke, Anne-Sophie; Hamill, Cecily; Chen, Yuming; Santos, Janine Hertzog; Jurkunas, Ula V

    2016-06-20

    Fuchs endothelial corneal dystrophy (FECD), a leading cause of age-related corneal edema requiring transplantation, is characterized by rosette formation of corneal endothelium with ensuing apoptosis. We sought to determine whether excess of mitochondrial reactive oxygen species leads to chronic accumulation of oxidative DNA damage and mitochondrial dysfunction, instigating cell death. We modeled the pathognomonic rosette formation of postmitotic corneal cells by increasing endogenous cellular oxidative stress with menadione (MN) and performed a temporal analysis of its effect in normal (HCEnC, HCECi) and FECD (FECDi) cells and ex vivo specimens. FECDi and FECD ex vivo specimens exhibited extensive mtDNA and nDNA damage as detected by quantitative PCR. Exposure to MN triggered an increase in mitochondrial superoxide levels and led to mtDNA and nDNA damage, while DNA amplification was restored with NAC pretreatment. Furthermore, MN exposure led to a decrease in ΔΨm and adenosine triphosphate levels in normal cells, while FECDi exhibited mitochondrial dysfunction at baseline. Mitochondrial fragmentation and cytochrome c release were detected in FECD tissue and after MN treatment of HCEnCs. Furthermore, cleavage of caspase-9 and caspase-3 followed MN-induced cytochrome c release in HCEnCs. This study provides the first line of evidence that accumulation of oxidative DNA damage leads to rosette formation, loss of functionally intact mitochondria via fragmentation, and subsequent cell death during postmitotic cell degeneration of ocular tissue. MN induced rosette formation, along with mtDNA and nDNA damage, mitochondrial dysfunction, and fragmentation, leading to activation of the intrinsic apoptosis via caspase cleavage and cytochrome c release. Antioxid. Redox Signal. 24, 1072-1083.

  17. REACTION OF THE RABBIT CORNEAL ENDOTHELIUM TO NYLON SUTURES - A SEM STUDY

    NARCIS (Netherlands)

    JONGEBLOED, WL; VANDERVEEN, G; KALICHARAN, D; RIJNEVELD, WJ; HOUTMAN, WA; WORST, JGF

    1990-01-01

    Nylon and stainless steel sutures separately placed deeply into rabbit corneas by splitting the stroma for a few millimeters, without closing sutures, remained in the cornea for two, four and six weeks respectively. In contrast to the stainless steel sutures an extensive tissue reaction could be

  18. Synergistic action of heparin and serum on basic fibroblast growth factor-modulated DNA synthesis and mitochondrial activity of cultured bovine corneal endothelial cells

    NARCIS (Netherlands)

    Hoppenreijs, V. P.; Pels, E.; Felten, P. C.; Ruijter, J. M.; Vrensen, G. F.; Treffers, W. F.

    1996-01-01

    Basic fibroblast growth factor (bFGF) is a major mitogen and chemoattractant for many cell types. The synergistic role of fetal bovine serum (FBS) and heparin on the modulation of tissue-cultured bovine corneal endothelial cells by bFGF was studied. Cell modulation was assessed by DNA synthesis

  19. Corneal dystrophies

    Directory of Open Access Journals (Sweden)

    Klintworth Gordon K

    2009-02-01

    diseases (mucopolysaccharidoses, lipidoses, mucolipidoses, and several skin diseases (X-linked ichthyosis, keratosis follicularis spinolosa decalvans. The management of the corneal dystrophies varies with the specific disease. Some are treated medically or with methods that excise or ablate the abnormal corneal tissue, such as deep lamellar endothelial keratoplasty (DLEK and phototherapeutic keratectomy (PTK. Other less debilitating or asymptomatic dystrophies do not warrant treatment. The prognosis varies from minimal effect on the vision to corneal blindness, with marked phenotypic variability.

  20. Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty

    OpenAIRE

    Bhogal, M.; Matter, K.; Balda, M. S.; Allan, B D

    2016-01-01

    Purpose To evaluate the effect of media composition and storage method on pre-prepared Descemet's membrane endothelial keratoplasty (DMEK) grafts. Methods 50 corneas were used. Endothelial wound healing and proliferation in different media were assessed using a standard injury model. DMEK grafts were stored using three methods: peeling with free scroll storage; partial peeling with storage on the stroma and fluid bubble separation with storage on the stroma. Endothelial cell (EC) phenotype an...

  1. Evaluation of statin therapy on endothelial function in hypercholesterolemic rabbits by automatic measurement of arterial wall movement using ultrasound images.

    Science.gov (United States)

    Rahmani-Cherati, Tavoos; Mokhtari-Dizaji, Manijhe; Vajhi, Alireza; Rostami, Abdorrazzagh

    2014-10-01

    The aim of this study was to evaluate arterial endothelial function, assessed as acetylcholine-mediated dilation (AMD), in a hypercholesterolemic atherosclerotic rabbit model to investigate the effects of atorvastatin in the atherosclerotic process, using a new computerized analysis model and ultrasound images. Twenty-seven rabbits were fed a high-cholesterol (2%) diet for 6 wk and then divided into three groups for an additional 9 wk: Group A received regular chow food, group B received a 2% cholesterol-rich diet plus atorvastatin drug, and group C received regular chow food plus atorvastatin. Ultrasound examinations of endothelial function of the rabbit abdominal aorta artery were performed immediately after the 6 weeks (0 wk) and then 3, 6 and 9 wk after that. For off-line analysis, a computerized analysis method for evaluating instantaneous changes in the wall of the rabbit abdominal aorta was used. As parameters of improvement resulting from treatment, endothelium-dependent acetylcholine-induced dilation and endothelium-independent nitroglycerin-induced dilation were evaluated in treated rabbits. Differences among groups were tested using analysis of variance. On histopathology, intima-media thickness decreased after treatment in all groups. There were no significant differences in arterial diameter and blood velocity changes among treated rabbits at 0, 3, 6 and 9 wk of treatment in all groups, except in end-diastolic velocity, radial strain percentage, pulse index and resistance index in group C. In group A, AMD did not significantly improve after 3, 6 and 9 wk, as compared with 0 wk. Atorvastatin treatment significantly increased AMD (18%) at 3 wk in group B, compared with week 0. AMD significantly increased after 3 (26%), 6 (124%) and 9 (182%) wk in group C, compared with 0 wk. It is concluded that the new automatic method enables accurate and repeated evaluation of endothelial function during the progression and regression of atherosclerosis. Also, the

  2. Evaluation of intracameral injection of ranibizumab and bevacizumab on the corneal endothelium by scanning electron microscopy.

    Science.gov (United States)

    Ari, Seyhmus; Nergiz, Yusuf; Aksit, Ihsan; Sahin, Alparslan; Cingu, Kursat; Caca, Ihsan

    2015-03-01

    To evaluate the effects of intracameral injection of ranibizumab and bevacizumab on the corneal endothelium by scanning electron microscopy (SEM). Twenty-eight female rabbits were randomly divided into four equal groups. Rabbits in groups 1 and 2 underwent intracameral injection of 1 mg/0.1 mL and 0.5 mg/0.05 mL ranibizumab, respectively; group 3 was injected with 1.25 mg/0.05 mL bevacizumab. All three groups were injected with a balanced salt solution (BSS) into the anterior chamber of the left (fellow) eye. None of the rabbits in group 4 underwent an injection. Corneal thickness and intraocular pressure were measured before the injections, on the first day, and in the first month after injection. The rabbits were sacrificed and corneal tissues were excised in the first month after injection. Specular microscopy was used for the corneal endothelial cell count. Endothelial cell density was assessed and comparisons drawn between the groups and the control. Micrographs were recorded for SEM examination. The structure of the corneal endothelial cells, the junctional area of the cell membrane, the distribution of microvillus, and the cell morphology of the eyes that underwent intracameral injection of vascular endothelial growth factor (VEGF), BSS, and the control group were compared. Corneal thickness and intraocular pressure were not significantly different between the groups that underwent anti-VEGF or BSS injection and the control group on the first day and in the first month of injection. The corneal endothelial cell count was significantly diminished in all three groups; predominantly in group 1 and 2 (Pendothelium and intercellular junctions were normal. However, a relative reduction was observed in the microvillus density of endothelial cells. Although eyes in group 3 were morphologically similar to fellow eyes and the control group, disarrangement in endothelial cell borders was evident. The SEM examination pointed out deterioration in endothelial

  3. The effect of riboflavin/UVA collagen cross-linking therapy on the structure and hydrodynamic behaviour of the ungulate and rabbit corneal stroma.

    Directory of Open Access Journals (Sweden)

    Sally Hayes

    Full Text Available PURPOSE: To examine the effect of riboflavin/UVA corneal crosslinking on stromal ultrastructure and hydrodynamic behaviour. METHODS: One hundred and seventeen enucleated ungulate eyes (112 pig and 5 sheep and 3 pairs of rabbit eyes, with corneal epithelium removed, were divided into four treatment groups: Group 1 (28 pig, 2 sheep and 3 rabbits were untreated; Group 2 (24 pig were exposed to UVA light (3.04 mW/cm(2 for 30 minutes and Group 3 (29 pig and Group 4 (31 pig, 3 sheep and 3 rabbits had riboflavin eye drops applied to the corneal surface every 5 minutes for 35 minutes. Five minutes after the initial riboflavin instillation, the corneas in Group 4 experienced a 30 minute exposure to UVA light (3.04 mW/cm(2. X-ray scattering was used to obtain measurements of collagen interfibrillar spacing, spatial order, fibril diameter, D-periodicity and intermolecular spacing throughout the whole tissue thickness and as a function of tissue depth in the treated and untreated corneas. The effect of each treatment on the hydrodynamic behaviour of the cornea (its ability to swell in saline solution and its resistance to enzymatic digestion were assessed using in vitro laboratory techniques. RESULTS: Corneal thickness decreased significantly following riboflavin application (p<0.01 and also to a lesser extent after UVA exposure (p<0.05. With the exception of the spatial order factor, which was higher in Group 4 than Group 1 (p<0.01, all other measured collagen parameters were unaltered by cross-linking, even within the most anterior 300 microns of the cornea. The cross-linking treatment had no effect on the hydrodynamic behaviour of the cornea but did cause a significant increase in its resistance to enzymatic digestion. CONCLUSIONS: It seems likely that cross-links formed during riboflavin/UVA therapy occur predominantly at the collagen fibril surface and in the protein network surrounding the collagen.

  4. The Effect of Riboflavin/UVA Collagen Cross-linking Therapy on the Structure and Hydrodynamic Behaviour of the Ungulate and Rabbit Corneal Stroma

    Science.gov (United States)

    Hayes, Sally; Kamma-Lorger, Christina S.; Boote, Craig; Young, Robert D.; Quantock, Andrew J.; Rost, Anika; Khatib, Yasmeen; Harris, Jonathan; Yagi, Naoto; Terrill, Nicholas; Meek, Keith M.

    2013-01-01

    Purpose To examine the effect of riboflavin/UVA corneal crosslinking on stromal ultrastructure and hydrodynamic behaviour. Methods One hundred and seventeen enucleated ungulate eyes (112 pig and 5 sheep) and 3 pairs of rabbit eyes, with corneal epithelium removed, were divided into four treatment groups: Group 1 (28 pig, 2 sheep and 3 rabbits) were untreated; Group 2 (24 pig) were exposed to UVA light (3.04 mW/cm2) for 30 minutes and Group 3 (29 pig) and Group 4 (31 pig, 3 sheep and 3 rabbits) had riboflavin eye drops applied to the corneal surface every 5 minutes for 35 minutes. Five minutes after the initial riboflavin instillation, the corneas in Group 4 experienced a 30 minute exposure to UVA light (3.04 mW/cm2). X-ray scattering was used to obtain measurements of collagen interfibrillar spacing, spatial order, fibril diameter, D-periodicity and intermolecular spacing throughout the whole tissue thickness and as a function of tissue depth in the treated and untreated corneas. The effect of each treatment on the hydrodynamic behaviour of the cornea (its ability to swell in saline solution) and its resistance to enzymatic digestion were assessed using in vitro laboratory techniques. Results Corneal thickness decreased significantly following riboflavin application (pcollagen parameters were unaltered by cross-linking, even within the most anterior 300 microns of the cornea. The cross-linking treatment had no effect on the hydrodynamic behaviour of the cornea but did cause a significant increase in its resistance to enzymatic digestion. Conclusions It seems likely that cross-links formed during riboflavin/UVA therapy occur predominantly at the collagen fibril surface and in the protein network surrounding the collagen. PMID:23349690

  5. Polyvinylidene fluoride for proliferation and preservation of bovine corneal endothelial cells by enhancing type IV collagen production and deposition.

    Science.gov (United States)

    Wang, Tsung-Jen; Wang, I-Jong; Chen, Yi-Hsin; Lu, Jui-Nan; Young, Tai-Horng

    2012-01-01

    In this study, biomaterials with different hydrophobic properties including polyvinyl alcohol (PVA), poly(ethylene-co-vinyl alcohol) (EVAL), tissue culture polystyrene (TCPS), and polyvinylidene fluoride (PVDF) were examined in the bovine corneal endothelial cells (BCECs) culture system to elucidate their possible impact on clinical demand and scientific interest. It was found that BCECs were inhibited to attach onto the PVA surface. Conversely, relatively more hydrophobic biomaterials EVAL, TCPS, and PVDF successfully initiate BCEC adhesion. Compared to EVAL, cultured BCECs on TCPS and PVDF exhibited higher viability. Furthermore, fibroblastic transformation on EVAL and TCPS was observed at day 17, but BCECs maintained typical hexagonal shape on the PVDF surface at day 21. This phenomenon can be rescued by previously coating type IV collagen on TCPS but not on EVAL. In addition, when BCECs were cultured on PVDF, the expressions of gap junction connexin-43, differentiation marker N-cadherin, and tight junction ZO-1 were well-developed, resembling the physiological phenotypes. After examining the type IV collagen expression by Western blot analysis and protein absorption test, a possible explanation for the better proliferation and preservation of BCECs on the PVDF substrate is that PVDF is a bioactive substratum which enables BCECs to synthesize and reserve more extracellular matrix type IV collagen, paving an important way to provide a more preferential environment for BCEC cultures. Accordingly, promoting CEC growth effects after cell-biomaterial association may be applied to the tissue engineering of corneal endothelium. Copyright © 2011 Wiley Periodicals, Inc.

  6. Increased proliferation and replicative lifespan of isolated human corneal endothelial cells with L-ascorbic acid 2-phosphate.

    Science.gov (United States)

    Shima, Nobuyuki; Kimoto, Miwa; Yamaguchi, Masahiro; Yamagami, Satoru

    2011-11-07

    To explore an alternative culture method for human corneal endothelial cells (HCECs) and to examine the effect of l-ascorbic acid 2-phosphate (Asc-2P) on the growth of these cells. The influence of various mitogens, extracellular matrices (ECMs), and Asc-2P on growth of cultured HCECs was examined. HCECs were obtained from donors ranging in age from 12 to 74 years, and primary cultures and subcultures were performed with or without Asc-2P. Expanded HCECs were characterized with immunostaining and reverse transcription polymerase chain reaction (RT-PCR) and evaluated for generation of 8-hydroxy-2-deoxyguanosine (8-OHdG) with immunostaining and an enzyme-linked immunosorbent assay (ELISA). Culture with Asc-2P and bFGF on atelocollagen promoted the proliferation of HCECs in both primary cultures and subcultures as efficiently as conventional culture using ECM derived from bovine corneal endothelial cells. Zonula occludens-1, N-cadherin, connexin 43, and Na+/K+-ATPase were localized at plasma membranes of cultured HCECs. mRNAs of the voltage-dependent anion channels (VDAC2 and VDAC3), sodium bicarbonate cotransporter member 4 (SLC4A4), and chloride channel proteins (CLCN2 and CLCN3) were detected by RT-PCR. During multiple passages, cultures without Asc-2P showed a decrease in growth and irregular cell morphology, whereas cultures with Asc-2P sustained cell growth and maintained the characteristic polygonal morphology. ELISA for 8-OHdG showed that the levels in mitochondrial DNA significantly decreased when HCECs were subcultured with Asc-2P. Combination of Asc-2P and bFGF on atelocollagen allows successful culture for HCECs. Asc-2P extends the lifespan of cultured HCECs, partly due to protection against oxidative DNA damage.

  7. Effects of low and hyper Dk rigid gas permeable contact lenses on Bcl-2 expression and apoptosis in the rabbit corneal epithelium.

    Science.gov (United States)

    Yamamoto, K; Ladage, P M; Ren, D H; Li, L; Petroll, W M; Jester, J V; Cavanagh, H D

    2001-07-01

    To study Bcl-2 expression and apoptotic cell shedding of the rabbit corneal epithelium during extended wear of low and hyper Dk rigid gas permeable (RGP) contact lenses. Rabbits were fit with either a low or a hyper Dk RGP lens (Dk/Ltotal= 10 and 97). The rabbits wore the lenses for either 24 hours, 3 days, or 1 week at which point they were humanely sacrificed. Immunocytochemistry and western blot analyses were performed to detect Bcl-2 in the corneal epithelium; TUNEL assay (TdT-mediated dUTP nick-end labeling) was used to identify apoptotic epithelial cells. 1) Immunocytochemistry: In the normal cornea, antibodies to Bcl-2 uniformly stained nuclei of all epithelial cell layers. Occasional surface epithelial cells, however, showed no anti-Bcl-2 nuclear staining; concomitant TUNEL assay revealed that all TUNEL-labeled-surface cells were Bcl-2 negative. By contrast, RGP contact lens wear, regardless of test lens oxygen transmissibility or lens wearing interval, significantly decreased both the total number of Bcl-2 negative and TUNEL-labeled cells on the epithelial surface (P Dk lens test group but was similar to control values in the hyper Dk lens test group. Bcl-2 protein seems to play an important role in the regulation of apoptotic cell shedding in the normal rabbit corneal epithelium. The identical staining pattern was seen in previous studies of the normal human cornea. RGP contact lens wear, however, appears to block the changes in Bcl-2 protein prior to apoptotic surface cell shedding, suggesting a lens-related anti-apoptotic effect. Taken together, these findings may explain why contact lens wear reduces surface cell exfoliation as previously reported in human studies.

  8. In vitro biomimetic platforms featuring a perfusion system and 3D spheroid culture promote the construction of tissue-engineered corneal endothelial layers.

    Science.gov (United States)

    Li, Shanyi; Han, Yuting; Lei, Hao; Zeng, Yingxin; Cui, Zekai; Zeng, Qiaolang; Zhu, Deliang; Lian, Ruiling; Zhang, Jun; Chen, Zhe; Chen, Jiansu

    2017-04-10

    Corneal endothelial cells (CECs) are very important for the maintenance of corneal transparency. However, in vitro, CECs display limited proliferation and loss of phenotype via endothelial to mesenchymal transformation (EMT) and cellular senescence. In this study, we demonstrate that continuous supplementary nutrition using a perfusion culture bioreactor and three-dimensional (3D) spheroid culture can be used to improve CEC expansion in culture and to construct a tissue-engineered CEC layer. Compared with static culture, perfusion-derived CECs exhibited an increased proliferative ability as well as formed close cell-cell contact junctions and numerous surface microvilli. We also demonstrated that the CEC spheroid culture significantly down-regulated gene expression of the proliferation marker Ki67 and EMT-related markers Vimentin and α-SMA, whereas the gene expression level of the CEC marker ATP1A1 was significantly up-regulated. Furthermore, use of the perfusion system in conjunction with a spheroid culture on decellularized corneal scaffolds and collagen sheets promoted the generation of CEC monolayers as well as neo-synthesized ECM formation. This study also confirmed that a CEC spheroid culture on a curved collagen sheet with controlled physiological intraocular pressure could generate a CEC monolayer. Thus, our results show that the use of a perfusion system and 3D spheroid culture can promote CEC expansion and the construction of tissue-engineered corneal endothelial layers in vitro.

  9. Risk factors for donor endothelial loss in eye bank-prepared posterior lamellar corneal tissue for descemet stripping automated endothelial keratoplasty.

    Science.gov (United States)

    Liu, Yu-Chi; Alvarez Paraz, Carisa M; Cajucom-Uy, Howard Yu; Agahari, Djoni; Sethuraman, Selvam; Tan, Donald T-H; Mehta, Jodhbir S

    2014-07-01

    The aim of this study was to investigate donor, tissue, and precut procedure risk factors for endothelial cell density (ECD) loss in posterior lamellar corneal tissue preparation by an eye bank for Descemet stripping automated endothelial keratoplasty. A total of 259 corneoscleral rims precut by the Singapore Eye Bank from October 2011 to August 2013 were evaluated. Donor characteristics, tissue characteristics, and precut procedure parameters were analyzed. The mean donor age was 57.18 ± 11.35 years, and the mean cutting transition time was 4.16 ± 0.75 seconds. The mean ECD was 2826 ± 225 and 2787 ± 224 cells per square millimeter before and after precutting, respectively, with an average ECD change of -1.38% ± 3.28%. The precutting procedure failure rate was 1.2%. Mutivariate regression analysis showed that an older donor age, a higher ECD before cutting, and a slower cutting transition speed were significant factors. Corneas with an ECD >2800 cells per square millimeter before precutting, cutting transition time >5.5 seconds, and corneas with donor age >65 years were significantly more likely to have greater than 5% ECD loss after precutting (odds ratio, 6.42, 1.66, and 1.62; 95% confidence interval, 1.44-29.43, 1.45-2.72, and 1.66-5.82, respectively). Donor source, death-to-preservation time (range, 0.67-10.88 hours), death-to-precutting time (range, 0-7 days), and graft thickness (range, 43-232 μm) were not statistically significant factors. The ECD loss in the precut tissue prepared by the eye bank was very low. The risk factors identified provide better understanding of how to improve the quality and safety profiles when preparing graft tissue for Descemet stripping automated endothelial keratoplasty.

  10. Increased Hepatic Expression of Endothelial Lipase Inhibits Cholesterol Diet-Induced Hypercholesterolemia and Atherosclerosis in Transgenic Rabbits.

    Science.gov (United States)

    Wang, Chuan; Nishijima, Kazutoshi; Kitajima, Shuji; Niimi, Manabu; Yan, Haizhao; Chen, Yajie; Ning, Bo; Matsuhisa, Fumikazu; Liu, Enqi; Zhang, Jifeng; Chen, Y Eugene; Fan, Jianglin

    2017-07-01

    Endothelial lipase (EL) is a key determinant in plasma high-density lipoprotein-cholesterol. However, functional roles of EL on the development of atherosclerosis have not been clarified. We investigated whether hepatic expression of EL affects plasma lipoprotein metabolism and cholesterol diet-induced atherosclerosis. We generated transgenic (Tg) rabbits expressing the human EL gene in the liver and then examined the effects of EL expression on plasma lipids and lipoproteins and compared the susceptibility of Tg rabbits with cholesterol diet-induced atherosclerosis with non-Tg littermates. On a chow diet, hepatic expression of human EL in Tg rabbits led to remarkable reductions in plasma levels of total cholesterol, phospholipids, and high-density lipoprotein-cholesterol compared with non-Tg controls. On a cholesterol-rich diet for 16 weeks, Tg rabbits exhibited significantly lower hypercholesterolemia and less atherosclerosis than non-Tg littermates. In Tg rabbits, gross lesion area of aortic atherosclerosis was reduced by 52%, and the lesions were characterized by fewer macrophages and smooth muscle cells compared with non-Tg littermates. Increased hepatic expression of EL attenuates cholesterol diet-induced hypercholesterolemia and protects against atherosclerosis. © 2017 American Heart Association, Inc.

  11. Methods Development for the Isolation and Culture of Primary Corneal Endothelial Cells

    Science.gov (United States)

    2017-02-01

    widespread stockpiling and the lack of definitive medical treatment. The vast majority of SM exposure cases involve corneal injury. At low-exposure doses...10 2 5 8 11 3 6 9 12 Phase II testing involved further evaluation of individual Proulx medium constituents. Opti-MEM I (a...coating (Figure 2A) or over FNC-coated TCP (Figure 2B). No difference in morphology or growth rate was observed between Nunclon Delta or Falcon Primaria

  12. Transfection of the Human Heme Oxygenase Gene Into Rabbit Coronary Microvessel Endothelial Cells: Protective Effect Against Heme and Hemoglobin Toxicity

    Science.gov (United States)

    Abraham, N. G.; Lavrovsky, Y.; Schwartzman, M. L.; Stoltz, R. A.; Levere, R. D.; Gerritsen, M. E.

    1995-07-01

    Heme oxygenase (HO) is a stress protein and has been suggested to participate in defense mechanisms against agents that may induce oxidative injury such as metals, endotoxin, heme/hemoglobin, and various cytokines. Overexpression of HO in cells might therefore protect against oxidative stress produced by certain of these agents, specifically heme and hemoglobin, by catalyzing their degradation to bilirubin, which itself has antioxidant properties. We report here the successful in vitro transfection of rabbit coronary microvessel endothelial cells with a functioning gene encoding the human HO enzyme. A plasmid containing the cytomegalovirus promoter and the human HO cDNA complexed to cationic liposomes (Lipofectin) was used to transfect rabbit endothelial cells. Cells transfected with human HO exhibited an ≈3.0-fold increase in enzyme activity and expressed a severalfold induction of human HO mRNA as compared with endogenous rabbit HO mRNA. Transfected and nontransfected cells expressed factor VIII antigen and exhibited similar acetylated low-density lipoprotein uptake (two important features that characterize endothelial cells) with >85% of cells staining positive for each marker. Moreover, cells transfected with the human HO gene acquired substantial resistance to toxicity produced by exposure to recombinant hemoglobin and heme as compared with nontransfected cells. The protective effect of HO overexpression against heme/hemoglobin toxicity in endothelial cells shown in these studies provides direct evidence that the inductive response of human HO to such injurious stimuli represents an important tissue adaptive mechanism for moderating the severity of cell damage produced by these blood components.

  13. Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty.

    Science.gov (United States)

    Bhogal, Maninder; Matter, Karl; Balda, Maria S; Allan, Bruce D

    2016-11-01

    To evaluate the effect of media composition and storage method on pre-prepared Descemet's membrane endothelial keratoplasty (DMEK) grafts. 50 corneas were used. Endothelial wound healing and proliferation in different media were assessed using a standard injury model. DMEK grafts were stored using three methods: peeling with free scroll storage; partial peeling with storage on the stroma and fluid bubble separation with storage on the stroma. Endothelial cell (EC) phenotype and the extent of endothelial overgrowth were examined. Global cell viability was assessed for storage methods that maintained a normal cell phenotype. 1 mm wounds healed within 4 days. Enhanced media did not increase EC proliferation but may have increased EC migration into the wounded area. Grafts that had been trephined showed evidence of EC overgrowth, whereas preservation of a physical barrier in the bubble group prevented this. In grafts stored in enhanced media or reapposed to the stroma after trephination, endothelial migration occurred sooner and cells underwent endothelial-mesenchymal transformation. Ongoing cell loss, with new patterns of cell death, was observed after returning grafts to storage. Grafts stored as free scrolls retained more viable ECs than grafts prepared with the fluid bubble method (74.2± 3% vs 60.3±6%, p=0.04 (n=8). Free scroll storage is superior to liquid bubble and partial peeling techniques. Free scrolls only showed overgrowth of ECs after 4 days in organ culture, indicating a viable time window for the clinical use of pre-prepared DMEK donor material using this method. Methods for tissue preparation and storage media developed for whole corneas should not be used in pre-prepared DMEK grafts without prior evaluation. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  14. Pseudotyping and culture conditions affect efficiency and cytotoxicity of retroviral gene transfer to human corneal endothelial cells.

    Science.gov (United States)

    Valtink, Monika; Stanke, Nicole; Knels, Lilla; Engelmann, Katrin; Funk, Richard H W; Lindemann, Dirk

    2011-08-29

    To evaluate retroviral vectors as a tool to transduce normal human corneal endothelial cells (HCECs) and to optimize transduction to increase gene transfer efficiency. Enhanced green fluorescent protein (EGFP) encoding retroviral vectors based on HIV-1 or murine leukemia virus (MLV), pseudotyped with either vesicular stomatitis virus glycoprotein (VSV-G) or a modified foamy virus envelope protein (FV Env), and prototype foamy virus (PFV) were produced. Transduction was performed in four HCEC culture media that were previously described for specific cultivation of HCECs or organ culture of donor corneas, namely enriched HCEC growth medium F99(HCEC), its unsupplemented basal medium F99, MEM + 2% fetal calf serum (FCS) (MEM), and Human Endothelial-SFM (SFM). Transduction efficiency was evaluated by marker gene transfer assay, and cytotoxic effects of virus infection were evaluated by means of resazurin conversion assay. PFV- and HIV-1-based vectors showed superior transduction efficiency compared with MLV-based vectors. Pseudotyping with a modified FV Env increased transduction efficiency compared with pseudotyping with VSV-G. In medium SFM, transduction efficiency of PFV, HIV-1-/FV Env, and MLV-based vectors was markedly reduced compared with the other culture media. When cells were cultured in F99-based media, cell viability was reduced by retroviral transduction compared with uninfected or mock infected controls, but remained unaffected when cells were cultured in SFM and was even increased when cells were cultured in MEM. HIV-1-based vectors pseudotyped with FV Env can efficiently be used to transduce primary HCECs in vitro. However, transduction efficiency is dependent on culture conditions and impairs metabolic activity and viability of HCECs in vitro.

  15. Cytotoxicity of atropine to human corneal endothelial cells by inducing mitochondrion-dependent apoptosis.

    Science.gov (United States)

    Wen, Qian; Fan, Ting-Jun; Tian, Cheng-Lei

    2016-07-01

    Atropine, a widely used topical anticholinergic drug, might have adverse effects on human corneas in vivo. However, its cytotoxic effect on human corneal endothelium (HCE) and its possible mechanisms are unclear. Here, we investigated the cytotoxicity of atropine and its underlying cellular and molecular mechanisms using an in vitro model of HCE cells and verified the cytotoxicity using cat corneal endothelium (CCE) in vivo. Our results showed that atropine at concentrations above 0.3125 g/L could induce abnormal morphology and viability decline in a dose- and time-dependent manner in vitro. The cytotoxicity of atropine was proven by the induced density decrease and abnormality of morphology and ultrastructure of CCE cells in vivo. Meanwhile, atropine could also induce dose- and time-dependent elevation of plasma membrane permeability, G1 phase arrest, phosphatidylserine externalization, DNA fragmentation, and apoptotic body formation of HCE cells. Moreover, 2.5 g/L atropine could also induce caspase-2/-3/-9 activation, mitochondrial transmembrane potential disruption, downregulation of anti-apoptotic Bcl-2 and Bcl-xL, upregulation of pro-apoptotic Bax and Bad, and upregulation of cytoplasmic cytochrome c and apoptosis-inducing factor. In conclusion, atropine above 1/128 of its clinical therapeutic dosage has a dose- and time-dependent cytotoxicity to HCE cells in vitro which is confirmed by CCE cells in vivo, and its cytotoxicity is achieved by inducing HCE cell apoptosis via a death receptor-mediated mitochondrion-dependent signaling pathway. Our findings provide new insights into the cytotoxicity and apoptosis-inducing effect of atropine which should be used with great caution in eye clinic. © 2016 by the Society for Experimental Biology and Medicine.

  16. Evaluation of the Endothelial Cell Density and the Central Corneal Thickness in Pseudoexfoliation Syndrome and Pseudoexfoliation Glaucoma

    Directory of Open Access Journals (Sweden)

    Bożydar T. Tomaszewski

    2014-01-01

    Full Text Available Purpose. Evaluation of central corneal thickness (CCT and endothelial cell density (ECD in patients with senile cataract and coexisting pseudoexfoliation (PEX syndrome with glaucoma (PEXG and without glaucoma using specular microscopy. Participants and Methods. The study included 122 patients (217 eyes. In this group of patients we identified 133 eyes with PEX syndrome (65 with glaucoma, 68 without glaucoma and 84 eyes without PEX syndrome. ECD and CCT were measured in each eye by specular microscopy. Results. ECD in eyes with PEX syndrome without glaucoma (2297 ± 359 cell/mm2 and in eyes with PEXG (2241 ± 363 cell/mm2 was lower than in the control group (2503 ± 262 cell/mm2 (P<0.001. CCT in eyes with PEXG (508.2 ± 32.6 μm was thinner than in eyes with PEX syndrome without glaucoma (529.7 ± 30.3 μm and control group (527.7 ± 29.4 μm (P<0.001. Conclusions. This research shows that in eyes with PEX syndrome, both with and without glaucoma, ECD was statistically significantly lower than in the control group. In patients with PEXG, CCT was statistically significantly thinner than in the PEX syndrome and control group.

  17. Impact of photodynamic inactivation (PDI) using the photosensitizer chlorin e6 on viability, apoptosis, and proliferation of human corneal endothelial cells.

    Science.gov (United States)

    Wang, Jiong; Stachon, Tanja; Eppig, Timo; Langenbucher, Achim; Seitz, Berthold; Szentmáry, Nóra

    2013-04-01

    Photodynamic inactivation (PDI) may be a potential alternative in case of therapy-resistant infectious keratitis. PDI using the photosensitizer chlorin e6 (Ce6) with high photosensitizing efficacy offers a valuable option, also for keratitis. The purpose of our study was to determine the impact of PDI with the photosensitizer Ce6 on viability, apoptosis, and proliferation of human corneal endothelial cells (HCECs), in vitro. Human corneal endothelial cell line was cultured in DMEM/Ham's F12 medium supplemented with 5 % fetal calf serum. HCECs cultures underwent illumination using red (670 nm) light for 13 min following exposure to 50-500 nM concentrations of Ce6 in the culture medium. Twenty-four hours after PDI, cell viability was evaluated by the Alamar blue assay, total DNA content of the cells and apoptosis using the APO-DIRECT Kit, and cell proliferation by the BrdU Cell Proliferation Assay Kit. Using Ce6 or illumination only, we did not detect significant changes of cell viability, apoptosis, and proliferation. Following PDI, viability and total DNA content of HCECs decreased significantly above 150 nM Ce6 concentration (P proliferation of endothelial cells decreased significantly (P proliferation, and also triggers apoptosis of HCECs in vitro. PDI using the photosensitizer Ce6 may be a potential treatment alternative in infectious keratitis. However, to avoid endothelial cell damage, the photosensitizer must not penetrate the endothelium.

  18. Long-Term Biomechanical and Histologic Results of WST-D/NIR Corneal Stiffening in Rabbits, Up to 8 Months Follow-up.

    Science.gov (United States)

    Brekelmans, Jurriaan; Goz, Alexandra; Dickman, Mor M; Brandis, Alexander; Sui, Xiaomeng; Wagner, H Daniel; Nuijts, Rudy M M A; Scherz, Avigdor; Marcovich, Arie L

    2017-08-01

    To determine the long-term safety and efficacy of WST-D/near-infrared (NIR) corneal stiffening. One eye of 23 New Zealand White rabbits was de-epithelialized mechanically followed by topical application of 2.5 mg/mL WST11, combined with dextran-500 (WST-D) for 20 minutes. Subsequently, samples were irradiated with a NIR (755 nm) laser at 10 mW/cm2 for 30 minutes. Untreated fellow eyes served as controls. One week (n = 4), 1 month (n = 6), 4 months (n = 9), or 8 months (n = 4) after treatment rabbits were euthanized. Corneal strips were cut in superior-inferior direction for extensiometry testing (1, 4, and 8 months), and histologic sections were prepared for evaluation of keratocyte distribution (1 week and 8 months). Elastic modulus after treatment was significantly higher than in paired controls (16.0 ± 2.3 MPa versus 9.6 ± 3.6 MPa [P = 0.008], 18.1 ± 4.5 MPa versus 12.6 ± 2.3 MPa [P = 0.003], and 18.6 ± 3.6 MPa versus 14.2 ± 3.6 MPa [P = 0.010], at 1, 4, and 8 months, respectively). A significant decrease in keratocyte count at the anterior stroma was observed directly after treatment (1.5 ± 1.7 vs. 19.0 ± 4.1 [P = 0.002]). At 8 months keratocyte repopulation appeared completed, with similar distribution in treated and untreated corneas (15.9 ± 1.1 vs. 14.5 ± 2.5 [P = 0.562]). Corneal thickness was comparable between treated and untreated corneas at all time points. WST-D/NIR treatment resulted in significant and persistent long-term increase in corneal stiffness. Initial keratocyte apoptosis in the anterior stroma is followed by repopulation to normal level at 8 months after treatment. The safe nature of NIR light allows treatment of corneas of any thickness without endangering corneal endothelium or deeper ocular structures, potentially benefiting patients deemed unsuitable for riboflavin/UV-A cross-linking.

  19. [Combined rapamycin eye drop in nanometer vector and poly (lactic acid) wafers of cyclosporine A effectively prevents high-risk corneal allograft rejection in rabbits].

    Science.gov (United States)

    Jiang, Wei; Sun, Hui-Min; Li, Xiao-Rong; Yuan, Xu-Bo; Wang, Yu-Qing; Zhang, Shu-Xian; Tian, En-Jiang; Yuan, Jia-Qin

    2009-06-01

    To evaluate the combined effect of topical rapamycin (RAPA) eye drop in nanometer vector and poly (lactic acid) (PLA) wafers of cyclosporine A (CsA) in the prevention of acute allograft rejection after rabbit corneal transplantation. Methods It was an experimental study. RAPA was incorporated into the nanometer particles and CsA was incorporated into PLA wafers. A was syngeneic control whose both donor and recipient are New Zealand rabbit. Gray donor corneas were implanted into the 102 recipients of New Zealand albino rabbits with corneal neovascularization who were randomly divided into B, C, D, E, F, G 6 groups to receive the different types of therapy: B was no therapy control; C was eye drop of nanometer vector but no RAPA twice a day, 28 days; D was PLA wafers in the anterior chamber of rabbit eyes but no drugs; E was 0.5% RAPA eye drop of nanometer vector twice a day, 28 days; F was PLA wafers of CsA in the anterior chamber of rabbit eyes; G was PLA wafers of CsA in the anterior chamber of rabbit eyes and 0.5% RAPA eye drop of nanometer vector eye drop twice a day for 28 days together. Postoperative evaluation included slit-lamp biomicroscopy, histopathology and immunohistology, Cytokines related with neovascularization and immunosuppression in the corneal tissue by RT-PCR. The graft survival was assessed by One-Way ANOVA and q test. Corneal allograft survival time: A (100.00 +/- 0.00), B (8.44 +/- 1.24), C (8.89 +/- 2.57), D (8.56 +/- 2.30), E (43.11 +/- 5.58), F (43.67 +/- 9.54), G (72.00 +/- 15.34) d. Group G led to a statistically significant prolongation of transplant survival and was superior than group E and F which was a statistical prolongation compared with group B, C and D (qGE = 11.42, qGF = 11.24, qEB = 13.64, qEC = 13.38, qED = 13.46, qFB = 13.82, qFC = 13.56, qFD = 13.64; P < 0.01). Immunohistopathologically, the grafts were subjected to an immune response contained a dense infiltrate of neutrophils, CD4+ and CD8+ T lymphocytes in the group B

  20. Autologous bone-marrow mesenchymal stem cell implantation and endothelial function in a rabbit ischemic limb model.

    Directory of Open Access Journals (Sweden)

    Shinsuke Mikami

    Full Text Available BACKGROUND: The purpose of this study was to determine whether autologous mesenchymal stem cells (MSCs implantation improves endothelial dysfunction in a rabbit ischemic limb model. METHODS: We evaluated the effect of MSC implantation on limb blood flow (LBF responses to acetylcholine (ACh, an endothelium-dependent vasodilator, and sodium nitroprusside (SNP, an endothelium-independent vasodilator, in rabbits with limb ischemia in which cultured MSCs were implanted (n = 20 or saline was injected as a control group (n = 20. LBF was measured using an electromagnetic flowmeter. A total of 10(6 MSCs were implanted into each ischemic limb. RESULTS: Histological sections of ischemic muscle showed that capillary index (capillary/muscle fiber was greater in the MSC implantation group than in the control group. Laser Doppler blood perfusion index was significantly increased in the MSC implantation group compared with that in the control group. LBF response to ACh was greater in the MSC group than in the control group. After administration of N(G-nitro-L-arginine, a nitric oxide synthase inhibitor, LBF response to ACh was similar in the MSC implantation group and control group. Vasodilatory effects of SNP in the two groups were similar. CONCLUSIONS: These findings suggest that MSC implantation induces angiogenesis and augments endothelium-dependent vasodilation in a rabbit ischemic model through an increase in nitric oxide production.

  1. Targeted corneal transplantation.

    Science.gov (United States)

    Jhanji, Vishal; Mehta, Jod S; Sharma, Namrata; Sharma, Bhavana; Vajpayee, Rasik B

    2012-07-01

    Corneal transplantation surgery has moved from an era of conventional penetrating keratoplasty to selective replacement of the diseased corneal layer with complementary healthy donor corneal tissue. Anterior lamellar transplantation surgeries do not involve replacement of corneal endothelium, consequently eliminating the occurrence of endothelial rejection. Similarly, in diseases affecting the corneal endothelium, selective replacement with a lamellar lenticule bearing healthy endothelium provides better outcomes in terms of ocular surface, lesser astigmatism and quick visual recovery. In addition to the advantages of enhanced surgical outcomes, targeted corneal transplantation allows the use of one donor cornea for more than one recipient, thereby offering a viable solution to the problem of paucity of donor corneas. Evolving techniques of corneal transplantation have enabled better utilization of donor corneal tissue. Anterior lamellar as well as endothelial keratoplasty surgeries have become first-choice surgeries in appropriately selected cases. This review briefly discusses some of these novel surgical techniques. A better understanding of targeted corneal transplantation would lead to adaptation of the concept of component corneal surgery. This would further enable the corneal surgeons to circumvent the problem of donor corneal shortage especially in the developing world.

  2. Topical 1% Nalbuphine on corneal sensivity and epitheilization after experimental lamellar keratectomy in rabbits Nalbufina 1% tópica sobre a sensibilidade e a epitelização corneal após ceratectomia lamelar experimental

    Directory of Open Access Journals (Sweden)

    Miguel Ladino Silva

    2012-04-01

    Full Text Available The present study was aimed to evaluate the effects of topical 1% nalbuphine on corneal sensitivity and re-epithelialization, after lamellar keratectomy in rabbits. All protocols were approved by the Animal Care Comission of São Paulo State University (Protocol 028793-08 and were conducted in accordance with the Institutional Animal Committee and the Association for Research in Vision and Ophthalmology (ARVO statement for the use of animals in research. Surgeries were performed on the left eye (Nalbuphine Group and on the right eye (Control Group. Two groups were formed (n=10 and corneas received either 30µl of 1% nalbuphine (NG or 30µl of 0,9% saline (CG. Treatments occurred at 7, 11, 15 and 19 hours. After the surgery, the corneas were stained with fluorescein and photographed daily; corneal touch threshold (CTT was assessed with Cochet-Bonnet aesthesiometer, at 7 and 19 hours, 20 minutes after treatments. Data were statistically compared with repeated measures ANOVA and Bonferroni post-hoc test, and T test (P0.05; however, a higher area under the curve for both parameters was observed in the NG (2771, in comparison to CG (2164. Topical 1% nalbuphine did not change significantly corneal sensitivity and re-epithelialization, after experimental lamellar keratectomy in rabbits.Avaliaram-se os efeitos da nalbufina 1% sobre o limiar de sensibilidade corneal (LSC e a epitelização corneal em coelhos submetidos à ceratectomia lamelar unilateral. Os procedimentos foram aprovados pela Comissão de Ética no Uso de Animais da Faculdade de Ciências Agrarias e Veterinárias da Universidade Estadual Paulista (Protocolo no 028793-08, de acordo com as normas do Institutional Animal Committee and the Association for Research in Vision and Ophthalmology (ARVO. Conceberam-se dois grupos (n=10 e os olhos foram tratados com 30µl de Nalbufina 1% (Olho esquerdo - GN ou com 30µl de solução salina (Olho direito - GC, às 7, 11, 15 e 19 horas das

  3. Corneal endothelial cell loss and corneal biomechanical characteristics after two-step sequential or combined phaco-vitrectomy surgery for idiopathic epiretinal membrane

    DEFF Research Database (Denmark)

    Hamoudi, Hassan; Christensen, Ulrik Correll; La Cour, Morten

    2017-01-01

    PURPOSE: To assess the impact of sequential and combined surgery [cataract surgery and 23-gauge pars plana vitrectomy (PPV) with peeling] on corneal endothelium cell density (CED) and corneal biomechanical characteristics. METHODS: Phakic eyes with epiretinal membrane (ERM) were prospectively...... allocated to (i) cataract surgery and subsequent PPV (CAT group), (ii) PPV and subsequent cataract surgery (VIT group) or (iii) phacovitrectomy (COMBI group). Eyes were examined at baseline, 1 month after each surgery, and at 3 and 12 months follow-up. Corneal endothelium cell density (CED) was assessed...... with non-contact specular microscopy. Pachymetry [central cornea thickness (CCT)], keratometry and cornea volume (CV) were measured with Pentacam Scheimpflug camera. Primary outcome was change in CED after 12 months; secondary outcomes were changes in CCT and CV after 12 months. RESULTS: Sixty-two eyes...

  4. Alignment and cell-matrix interactions of human corneal endothelial cells on nanostructured collagen type I matrices.

    Science.gov (United States)

    Gruschwitz, Rita; Friedrichs, Jens; Valtink, Monika; Franz, Clemens M; Müller, Daniel J; Funk, Richard H W; Engelmann, Katrin

    2010-12-01

    To use nanoscopically defined, two-dimensional matrices assembled from aligned collagen type I fibrils as a sheet substratum for in vitro cultivation of human corneal endothelial cells (HCECs). To assess the effect of matrix architecture on HCEC morphology and to characterize integrin-mediated HCEC-matrix interaction. Cell alignment and cell-matrix interactions of primary HCECs and three different immortalized HCEC populations on native and UV-cross-linked collagen type I matrices were examined by time-lapse microscopy. Specific integrin α(2)β(1) binding to the collagen matrix was demonstrated using a function-blocking α(2) antibody. Integrin α(2) subunit expression levels of the four HCEC populations were analyzed by Western blot analysis. All HCEC populations aligned along the oriented collagen fibrils. Primary HCECs and, to a lesser extent, the other tested HCEC populations exerted high traction forces, leading to progressive matrix destruction. Cross-linking of the collagen matrices considerably increased matrix stability. Integrin subunit α(2) expression levels of the four cell types correlated with the degree of cell alignment and exertion of traction forces. In turn, blocking integrin subunit α(2) reduced cell alignment and prevented matrix destruction. HCECs align directionally along parallel arrays of collagen type I fibrils. The interactions of HCECs with collagen type I are primarily mediated by integrin α(2)β(1). Integrin subunit α(2) levels correlate with matrix contraction and subsequent destruction. Sustained cultivation of HCECs on ultrathin collagen matrices thus requires matrix cross-linking and moderate integrin α(2)β(1) expression levels.

  5. Preparation of arginine-glycine-aspartic acid-modified biopolymeric nanoparticles containing epigalloccatechin-3-gallate for targeting vascular endothelial cells to inhibit corneal neovascularization.

    Science.gov (United States)

    Chang, Che-Yi; Wang, Ming-Chen; Miyagawa, Takuya; Chen, Zhi-Yu; Lin, Feng-Huei; Chen, Ko-Hua; Liu, Guei-Sheung; Tseng, Ching-Li

    2017-01-01

    Neovascularization (NV) of the cornea can disrupt visual function, causing ocular diseases, including blindness. Therefore, treatment of corneal NV has a high public health impact. Epigalloccatechin-3-gallate (EGCG), presenting antiangiogenesis effects, was chosen as an inhibitor to treat human vascular endothelial cells for corneal NV treatment. An arginine-glycine-aspartic acid (RGD) peptide-hyaluronic acid (HA)-conjugated complex coating on the gelatin/EGCG self-assembly nanoparticles (GEH-RGD NPs) was synthesized for targeting the α v β 3 integrin on human umbilical vein endothelial cells (HUVECs) in this study, and a corneal NV mouse model was used to evaluate the therapeutic effect of this nanomedicine used as eyedrops. HA-RGD conjugation via COOH and amine groups was confirmed by 1 H-nuclear magnetic resonance and Fourier-transform infrared spectroscopy. The average diameter of GEH-RGD NPs was 168.87±22.5 nm with positive charge (19.7±2 mV), with an EGCG-loading efficiency up to 95%. Images of GEH-RGD NPs acquired from transmission electron microscopy showed a spherical shape and shell structure of about 200 nm. A slow-release pattern was observed in the nanoformulation at about 30% after 30 hours. Surface plasmon resonance confirmed that GEH-RGD NPs specifically bound to the integrin α v β 3 . In vitro cell-viability assay showed that GEH-RGD efficiently inhibited HUVEC proliferation at low EGCG concentrations (20 μg/mL) when compared with EGCG or non-RGD-modified NPs. Furthermore, GEH-RGD NPs significantly inhibited HUVEC migration down to 58%, lasting for 24 hours. In the corneal NV mouse model, fewer and thinner vessels were observed in the alkali-burned cornea after treatment with GEH-RGD NP eyedrops. Overall, this study indicates that GEH-RGD NPs were successfully developed and synthesized as an inhibitor of vascular endothelial cells with specific targeting capacity. Moreover, they can be used in eyedrops to inhibit angiogenesis in corneal NV

  6. The Effect of a p38 Mitogen-Activated Protein Kinase Inhibitor on Cellular Senescence of Cultivated Human Corneal Endothelial Cells.

    Science.gov (United States)

    Hongo, Akane; Okumura, Naoki; Nakahara, Makiko; Kay, EunDuck P; Koizumi, Noriko

    2017-07-01

    We have begun a clinical trial of a cell-based therapy for corneal endothelial dysfunction in Japan. The purpose of this study was to investigate the usefulness of a p38 MAPK inhibitor for prevention cellular senescence in cultivated human corneal endothelial cells (HCECs). HCECs of 10 donor corneas were divided and cultured with or without SB203580 (a p38 MAPK inhibitor). Cell density and morphology were evaluated by phase-contrast microscopy. Expression of function-related proteins was examined by immunofluorescent staining. Cellular senescence was evaluated by SA-β-gal staining and Western blotting for p16 and p21. Senescence-associated factors were evaluated by membrane blotting array, quantitative PCR, and ELISA. Phase-contrast microscopy showed a significantly higher cell density for HCECs cultured with SB203580 than without SB203580 (2623 ± 657 cells/mm2 and 1752 ± 628 cells/mm2, respectively). The HCECs cultured with SB203580 maintained a hexagonal morphology and expressed ZO-1, N-cadherin, and Na+/K+-ATPase in the plasma membrane, whereas the control HCECs showed an altered staining pattern for these marker proteins. HCECs cultured without SB203580 showed high positive SA-β-gal staining, a low nuclear/cytoplasm ratio, and expression of p16 and p21. IL-6, IL-8, CCL2, and CXCL1 were observed at high levels in low cell density HCECs cultured without SB203580. Activation of p38 MAPK signaling due to culture stress might be a causative factor that induces cellular senescence; therefore, the use of p38 MAPK inhibitor to counteract senescence may achieve sufficient numbers of HCECs for tissue engineering therapy for corneal endothelial dysfunction.

  7. Optimized human platelet lysate as novel basis for a serum-, xeno-, and additive-free corneal endothelial cell and tissue culture.

    Science.gov (United States)

    Thieme, Daniel; Reuland, Lynn; Lindl, Toni; Kruse, Friedrich; Fuchsluger, Thomas

    2017-09-21

    The expansion of donor-derived corneal endothelial cells (ECs) is a promising approach for regenerative therapies in corneal diseases. To achieve the best Good Manufacturing Practice standard the entire cultivation process should be devoid of nonhuman components. However, so far, there is no suitable xeno-free protocol for clinical applications. We therefore introduce a processed variant of a platelet lysate for the use in corneal cell and tissue culture based on a Good Manufacturing Practice-grade thrombocyte concentrate. This processed human platelet lysate (phPL), free of any animal components and of anticoagulants such as heparin with a physiological ionic composition, was used to cultivate corneal ECs in vitro and ex vivo in comparison to standard cultivation with fetal calf serum (FCS). Human donor corneas were cut in quarters while 2 quarters of each cornea were incubated with the respective medium supplement. Three fields of view per quarter were taken into account for the analysis. Evaluation of phPL as a medium supplement in cell culture of immortalized EC showed a superior viability compared with FCS control with reduced cell proliferation. Furthermore, the viability during the expansion of primary cells is significantly (3-fold ±0.5) increased with phPL compared with FCS standard medium. Quartering donor corneas was traumatic for the endothelium and therefore resulted in increased EC loss. Interestingly, however, cultivation of the quartered pieces for 2 weeks in 0.1-mg/ml pHPL in Biochrome I showed a 21 (±10) % EC loss compared with 67 (±12) % EC loss when cultivated in 2% FCS in Biochrome I. The cell culture protocol with pHPL as FCS replacement seems to be superior to the standard FCS protocols with respect to EC survival. It offers a xeno-free and physiological environment for corneal endothelial cells. This alternative cultivation protocol could facilitate the use of EC for human corneal cell therapy. Copyright © 2017 John Wiley & Sons, Ltd.

  8. Corneal Laceration

    Medline Plus

    Full Text Available ... What Is Corneal Laceration? Corneal Laceration Symptoms What Causes Corneal Laceration? Corneal Laceration Diagnosis Corneal Laceration Treatment ... the corneal laceration is deep enough it can cause a full thickness laceration. This is when the ...

  9. Corneal Laceration

    Medline Plus

    Full Text Available ... What Is Corneal Laceration? Corneal Laceration Symptoms What Causes Corneal Laceration? Corneal Laceration Diagnosis Corneal Laceration Treatment What Is Corneal Laceration? Leer en Español: ¿Qué ...

  10. Corneal and Aqueous Humor Concentrations of Amphotericin B Using Three Different Routes of Administration in a Rabbit Model

    National Research Council Canada - National Science Library

    Qu, Linghui; Li, Liangmao; Xie, Hanping

    2010-01-01

    ...), the current drug of choice in fungal keratitis. Concentrations of AMB were investigated in the cornea and aqueous humor of rabbits after using one of three different routes of administration: topical 0.25...

  11. Effect of valsartan on ACAT-1 and PPAR-γ expression in intima with carotid artery endothelial balloon injury in rabbit.

    Science.gov (United States)

    Ma, Tao; Ma, Zhi-Qiang; Du, Xiao-Hui; Yu, Qiu-Shi; Wang, Rong; Liu, Li

    2015-01-01

    To study the effect of valsartan on ACAT-1 and PPAR-γ expression after vascular endothelial balloon injury in intimal hyperplasia process. 24 male New Zealand white rabbits were randomly divided into three groups with 8 in each group. rabbits were fed with normal diet; Balloon injury group: rabbits were fed with 0.5% cholesterol, 5% lard rabbit feed; balloon injury + valsartan group, rabbits were fed with 0.5% cholesterol, 5% lard rabbit feed added with 10 mg/(kg.d) valsartan gavage. RT-PCR and Western blotting method were used to detect the carotid ACAT-1, PPAR-γ mRNA and protein expression after 8 weeks of feeding. In carotid artery balloon injury group, vascular smooth muscle cells (VSMC) proliferation and intimal hyperplasia were significantly higher 14 d after endothelial injury. In 14 days valsartan treatment group VSMC proliferation and intimal hyperplasia were lighter than the surgery group. Compared with the control group, ACAT-1, PPAR-γ mRNA and protein were significantly increased in balloon injury group and valsartan group (P ACAT-1 mRNA and protein were significantly lower in valsartan group and balloon injury group (P ACAT-1 and PPAR-γ mRNA in balloon injury group and valsartan group showed negative correlation (P ACAT-1, PPAR-γ mRNA and protein content were significantly increased in intimal hyperplasia process after vascular endothelial balloon injury. The effect of valsartan suppressed intimal hyperplasia correlated with the expression of down-regulated ACAT-1 and up-regulated PPAR-γ.

  12. Overexpression of human HMW FGF-2 but not LMW FGF-2 reduces the cytotoxic effect of lentiviral gene transfer in human corneal endothelial cells.

    Science.gov (United States)

    Valtink, Monika; Knels, Lilla; Stanke, Nicole; Engelmann, Katrin; Funk, Richard H W; Lindemann, Dirk

    2012-05-31

    Recently, insertion of immuno-modulatory or anti-apoptotic genes into corneal endothelial cells (HCECs) came into focus. Basic FGF-2 occurs in one secreted (low molecular weight, LMW, 18 kD) and four nuclear (high molecular weight, HMW, 22-34 kD) isoforms. HMW isoforms are known differentiation and survival factors, while LMW FGF-2 is a known mitogen. The effect of FGF-2 overexpression of each of the five known isoforms on HCEC cell survival after lentiviral gene transfer in different culture media was investigated. Cells were transduced with lentiviral vectors encoding for each of the five FGF-2 isoforms. Transduction efficiency and expression of individual FGF-2 isoforms was assessed by marker gene transfer and western blotting. Primary HCECs were cultured and transduced in four different media previously described for HCEC cultivation or corneal organ cultivation. Cytotoxic effect of virus infection and a possible rescue effect of FGF-2 overexpression were determined by resazurin conversion assay. Transduction with FGF-2 encoding lentiviral vectors resulted in overexpression of the respective isoform in all tested cell populations. Western blotting after total cell lysis proved nuclear localization of transgenic HMW isoforms. Overexpression of HMW FGF-2-especially 34 kD FGF-2-reduced lentiviral cytotoxicity, while overexpression of LMW FGF-2 aggravated viral cytotoxicity. Cytotoxicity of lentiviral gene transfer in corneal endothelial cells may be reduced by using bicistronic vectors that encode for the target gene and the 34-kD isoform of human FGF-2. Such cotransduction of a survival factor may increase cell survival after gene transfer, thereby improving gene therapeutic approaches.

  13. Research progress of corneal transplantation

    OpenAIRE

    Bing-Jie Zhang; Heng Sun; Yuan-Ping Zhang; Lin-Kun Ma

    2015-01-01

    Corneal transplantation is an ophthalmology treatment technique for corneal disease to help restore vision or control the development of corneal diseases by removing a scarred or damaged host cornea and replacing it with a clear and healthy donor cornea. Traditional corneal transplantation includes penetrating keratoplasty and lamellar keratoplasty. In recent ten years, deep lamellar keratoplasty and endothelial keratoplasty have gradually developed. At present, the development of keratoprost...

  14. Corneal endothelial cell loss and corneal biomechanical characteristics after two-step sequential or combined phaco-vitrectomy surgery for idiopathic epiretinal membrane.

    Science.gov (United States)

    Hamoudi, Hassan; Christensen, Ulrik Correll; La Cour, Morten

    2017-08-01

    To assess the impact of sequential and combined surgery [cataract surgery and 23-gauge pars plana vitrectomy (PPV) with peeling] on corneal endothelium cell density (CED) and corneal biomechanical characteristics. Phakic eyes with epiretinal membrane (ERM) were prospectively allocated to (i) cataract surgery and subsequent PPV (CAT group), (ii) PPV and subsequent cataract surgery (VIT group) or (iii) phacovitrectomy (COMBI group). Eyes were examined at baseline, 1 month after each surgery, and at 3 and 12 months follow-up. Corneal endothelium cell density (CED) was assessed with non-contact specular microscopy. Pachymetry [central cornea thickness (CCT)], keratometry and cornea volume (CV) were measured with Pentacam Scheimpflug camera. Primary outcome was change in CED after 12 months; secondary outcomes were changes in CCT and CV after 12 months. Sixty-two eyes were enrolled and allocated to the three groups. The mean preoperative CED was 2776, 2794 and 2653 cells/mm(2,) which decreased significantly at 12 months by 15.3, 20.0 and 19.3% in the CAT, VIT and COMBI group. There was no significant difference in percentage cell loss between the groups at final follow-up. The CED decreased significantly after cataract surgery, but was unaffected by PPV. Central cornea thickness (CCT) increased by 10 μm (p = 0.005) and CV by 1.38 mm(3) (2.3%, p < 0.001) in the COMBI group. There were no significant differences in CCT or CV between the groups at final follow-up. Combined and sequential surgery in ERM leads to a small decrease in CED. Performing cataract surgery before, after or in combination with vitrectomy did not make any significant difference with respect to final CED, CCT or CV. © 2017 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

  15. Descemet's stripping automated endothelial keratoplasty (DSAEK) using corneal donor tissue not acceptable for use in penetrating keratoplasty as a result of anterior stromal scars, pterygia, and previous corneal refractive surgical procedures.

    Science.gov (United States)

    Phillips, Paul M; Terry, Mark A; Shamie, Neda; Chen, Edwin S; Hoar, Karen L; Stoeger, Chris; Friend, Daniel J; Saad, Hisham A

    2009-09-01

    The purpose of this study was to evaluate outcomes of Descemet's stripping automated endothelial keratoplasty (DSAEK) using anterior stromal flawed (ASF) donor corneas that were unsuitable for use in full-thickness penetrating keratoplasty as a result of stromal scars, pterygia, or previous corneal refractive surgery and to compare results with DSAEK using standard tissue. We conducted a review of our initial 42 (19 with 6-month follow up) consecutive DSAEK surgeries using ASF tissue compared with 357 (199 with 6-month follow up) time-matched controls using standard tissue. Intraoperative and perioperative complications, including dislocations and primary graft failures, were compared. Six-month best spectacle-corrected vision, incidence of rejection episodes, postoperative refractive astigmatism, keratometric values, pre- and postoperative topography-derived surface asymmetry index, and surface regularity index were compared. One surgeon-cut ASF tissue was perforated before surgery and was discarded. No surgeon-cut standard tissue was perforated. No intraoperative complications and no episodes of primary graft failure or pupillary block glaucoma occurred in either group. One (2.4%) postoperative graft dislocation and one (5.2%) graft rejection episode occurred in the study group. There were 10 (2.8%) dislocations and 8 (2.2%) graft rejection in the controls. A statistically similar significant improvement in best spectacle-corrected vision occurred in both groups. Corneal topography, pachymetry, and manifest astigmatism were not significantly different between groups. Postoperative results of DSAEK using donor tissue excluded from use in penetrating keratoplasty as a result of stromal flaws are equivalent to results using standard donor tissue. Central corneal thickness measurements should be performed before cutting to avoid tissue perforation. The use of ASF tissue for DSAEK will expand the cornea donor pool.

  16. Ultraviolet-visible light spectral transmittance of rabbit corneas after riboflavin/ultraviolet-A (365 nm) corneal collagen cross-linking.

    Science.gov (United States)

    Hwang, Ho Sik; Kim, Man Soo

    2013-01-01

    To determine the effect of riboflavin/ultraviolet-A (365 nm) corneal collagen cross-linking on the transmission of the ultraviolet-visible (UV-VIS) light spectrum through the cornea. Twelve New Zealand white male rabbits were used in this research. Cross-linking was performed unilaterally on the right eyes of the animals while only the epithelium was removed on the left eyes as the control. Seven weeks after cross-linking, the animals were euthanized, and the enucleated eyes were processed for transmission spectroscopy. To confirm that the cross-linking procedures was done successfully on the right corneas, the tensile force-extension relationship was measured using six corneas from three of the rabbits after the transmission spectrum was determined. Seven weeks after cross-linking, ten of the 12 rabbits had clear corneas in the cross-linked and control eyes. The two rabbits with neovascularization and granular opacities in the right corneas were not included in subsequent measurements. In the cross-linked corneas, transmittance was 87.57% at 650 nm, and decreased continuously as the wavelength shortened. From 315 nm, the transmittance rapidly decreased and was 35.52% at 300 nm. In the control corneas, transmittance was 95.95% at 650 nm and decreased continuously as the wavelength shortened. Below 315 nm, the transmittance rapidly decreased, to 40.29% at 300 nm. The transmittance of the cross-linking corneas was 10%-20% lower than that of the control corneas. The difference was 8.38% at 650 nm and increased as the wavelength shortened, reaching a maximum of 20.59% at 320 nm, and decreased rapidly to 4.77% at 300 nm. The tensile force-extension relationship showed that a greater force was necessary to extend the cross-linking corneas over 500 µm than that of the control corneas. The transmittance of the cross-linked corneas was 10%-20% lower than that of the control corneas. The difference increased as the wavelength decrease, reaching a maximum at 320 nm and then

  17. Ultraviolet-visible light spectral transmittance of rabbit corneas after riboflavin/ultraviolet-A (365 nm) corneal collagen cross-linking

    Science.gov (United States)

    Hwang, Ho Sik

    2013-01-01

    Purpose To determine the effect of riboflavin/ultraviolet-A (365 nm) corneal collagen cross-linking on the transmission of the ultraviolet-visible (UV-VIS) light spectrum through the cornea. Methods Twelve New Zealand white male rabbits were used in this research. Cross-linking was performed unilaterally on the right eyes of the animals while only the epithelium was removed on the left eyes as the control. Seven weeks after cross-linking, the animals were euthanized, and the enucleated eyes were processed for transmission spectroscopy. To confirm that the cross-linking procedures was done successfully on the right corneas, the tensile force-extension relationship was measured using six corneas from three of the rabbits after the transmission spectrum was determined. Results Seven weeks after cross-linking, ten of the 12 rabbits had clear corneas in the cross-linked and control eyes. The two rabbits with neovascularization and granular opacities in the right corneas were not included in subsequent measurements. In the cross-linked corneas, transmittance was 87.57% at 650 nm, and decreased continuously as the wavelength shortened. From 315 nm, the transmittance rapidly decreased and was 35.52% at 300 nm. In the control corneas, transmittance was 95.95% at 650 nm and decreased continuously as the wavelength shortened. Below 315 nm, the transmittance rapidly decreased, to 40.29% at 300 nm. The transmittance of the cross-linking corneas was 10%–20% lower than that of the control corneas. The difference was 8.38% at 650 nm and increased as the wavelength shortened, reaching a maximum of 20.59% at 320 nm, and decreased rapidly to 4.77% at 300 nm. The tensile force-extension relationship showed that a greater force was necessary to extend the cross-linking corneas over 500 µm than that of the control corneas. Conclusions The transmittance of the cross-linked corneas was 10%–20% lower than that of the control corneas. The difference increased as the wavelength decrease

  18. Baculovirus-mediated vascular endothelial growth factor-D(ΔNΔC) gene transfer induces angiogenesis in rabbit skeletal muscle.

    Science.gov (United States)

    Heikura, Tommi; Nieminen, Tiina; Roschier, Miia M; Karvinen, Henna; Kaikkonen, Minna U; Mähönen, Anssi J; Lesch, Hanna P; Rissanen, Tuomas T; Laitinen, Olli H; Airenne, Kari J; Ylä-Herttuala, Seppo

    2012-01-01

    Occluded arteries and ischemic tissues cannot always be treated by angioplasty, stenting or by-pass-surgery. Under such circumstances, viral gene therapy may be useful in inducing increased blood supply to ischemic area. There is evidence of improved blood flow in ischemic skeletal muscle and myocardium in both animal and human studies using adenoviral vascular endothelial growth factor (VEGF) gene therapy. However, the expression is transient and repeated gene transfers with the same vector are inefficient due to immune responses. Different baculoviral vectors pseudotyped with or without vesicular stomatitis virus glycoprotein (VSV-G) and/or carrying woodchuck hepatitis virus post-transcriptional regulatory element (Wpre) were tested both in vitro and in vivo. VEGF-D(ΔNΔC) was used as therapeutic transgene and lacZ as a control. In vivo efficacy was evaluated as capillary enlargement and transgene expression in New Zealand White (NZW) rabbit skeletal muscle. A statistically significant capillary enlargement was detected 6 days after gene transfer in transduced areas compared to the control gene transfers with baculovirus and adenovirus encoding β-galactosidase (lacZ). Substantially improved gene transfer efficiency was achieved with a modified baculovirus pseudotyped with VSV-G and carrying Wpre. Dose escalation experiments revealed that either too large volume or too many virus particles caused inflammation and necrosis in the target tissue, whereas 10(9) plaque forming units injected in multiple aliquots resulted in transgene expression with only mild immune reactions. We show the first evidence of biologically significant baculoviral gene transfer in skeletal muscle of NZW rabbits using VEGF-D(ΔNΔC) as a therapeutic transgene. Copyright © 2012 John Wiley & Sons, Ltd.

  19. Soluble CD163 and interleukin-6 are increased in aqueous humour from patients with endothelial rejection of corneal grafts

    DEFF Research Database (Denmark)

    Funding, Mikkel; Vorum, Henrik; Nexø, Ebba

    2005-01-01

    PURPOSE: To evaluate soluble CD163 (sCD163) as a new marker of macrophage activity in aqueous humour from patients with corneal rejection and to investigate correlations between sCD163, the CD163 inducing interleukin-6 (IL-6), and albumin; to investigate whether increases in sCD163 and IL-6 levels...

  20. [Phosphatase and tensin homolog deleted on chromosome ten/phosphatidyl Inositol 3-kinase/vascular endothelial growth factor signaling pathway changes in the rabbit Kawasaki disease model].

    Science.gov (United States)

    Niu, L; Fu, M Y; Tian, J; He, X H; Zhang, H N; Wang, Q W; Wang, Y; Li, C L; Wang, Z Z; An, X J

    2016-03-01

    To observe the changes of phosphatase and tensin homolog deleted on chromosome ten(PTEN)/ phosphatidyl Inositol 3-kinase(PI3K)/ vascular endothelial growth factor(VEGF)signaling pathway in a rabbit Kawasaki disease model. Model of Kawasaki disease was established in weanling Japanese big-eared rabbits with 10% bovine serum venous injection (2.5 ml/kg, 2 times, and 2 week's interval) through the ear. Twenty four rabbits were divided into 4 groups: control group (without injection of 10% bovine serum albumin, six rabbits); 1 day group (sacrificed a the second day after the establishment of Kawasaki disease models, six rabbits); 7 day group (sacrificed at the seventh day after establishment of Kawasaki disease model, six rabbits); 30 day group (sacrificed at the thirtieth day after establishment of Kawasaki disease model, six rabbits). Pathological analysis was performed on coronary artery tissue samples. The express of PTEN and PI3K were detected by immunohistochemistry. The levels of VEGF and CK were also examined with ELISA and white blood cells were counted. (1) Coronary artery of model groups was thinner, distorted and had enlarged lumen. (2) PTEN expression in 1 d group, 7 d group and 30 d group were 58.5 ± 12.9, 73.2±9.9 and 109.6 ± 24.4, respectively, significantly higher than in the control group (25.5 ± 6.9, P0.05) and significantly lower than 1 d and 7 d group (both P0.05). (6)White blood cell count were significantly higher in 1 d group, 7 d group and 30 d group than in control group (all PKawasaki disease model and the signaling pathway might be involved in this model.

  1. Research progress of corneal transplantation

    Directory of Open Access Journals (Sweden)

    Bing-Jie Zhang

    2015-06-01

    Full Text Available Corneal transplantation is an ophthalmology treatment technique for corneal disease to help restore vision or control the development of corneal diseases by removing a scarred or damaged host cornea and replacing it with a clear and healthy donor cornea. Traditional corneal transplantation includes penetrating keratoplasty and lamellar keratoplasty. In recent ten years, deep lamellar keratoplasty and endothelial keratoplasty have gradually developed. At present, the development of keratoprosthesis provides a new choice for the patients no suitable for traditional. The review describes current surgical techniques in the field of corneal transplantation about indications, postoperative complications, and so on.

  2. Corneal Laceration

    Medline Plus

    Full Text Available ... Corneal Laceration? Corneal Laceration Diagnosis Corneal Laceration Treatment What Is Corneal Laceration? Leer en Español: ¿Qué Es una Laceración de la Córnea? Written By: Daniel ...

  3. Sustained-released mixture of vascular endothelial growth factor 165 and fibrin glue strengthens healing of ileal anastomoses in a rabbit model with intraperitoneal infection.

    Science.gov (United States)

    Li, Zhanwu; Wang, Wenjun; Wang, Xiaozhou; Jiang, Lei; Wang, Fengyi; Liu, Qiang

    2017-09-01

    To investigate the effects of a sustained-released mixture of vascular endothelial growth factor 165 (VEGF165) and fibrin glue (FG) local administration on postoperative rabbit ileal anastomoses. One hundred twenty-eight male and female New Zealand white rabbits underwent intraperitoneal infection subsequent ileal anastomosis surgery were divided randomly into 4 groups, including 32 animals in each, applied with saline solution, FG, rhVEGF165 and a mixture of rhVEGF165 with FG (VEGF + FG) on the anastomoses, respectively. The incidences of anastomotic leakage were observed. Histopathological examination for inflammatory infiltration, fibroblast proliferation, and capillary vascular proliferation were performed. Then, bursting pressure and hydroxyproline concentrations were assessed in anastomoses sits on postoperative days 3, 5, 7, and 14. Rabbits in VEGF + FG group had the lowest incidence of leakage (P < 0.05). Histological evaluations revealed that granulation tissue was formed on days 5 after anastomosis; fibroblast proliferation and capillary vascular proliferation were significantly increased on days 7 and 14 in VEGF + FG group. Furthermore, there was a statistically significant difference in the mean bursting pressures between VEGF + FG group and other groups on days 7 and 14 (P < 0.05), and rabbits in VEGF + FG group exhibited a higher concentration than VEGF group (P < 0.05) and FG group (P < 0.05) on day 14. Administration of VEGF165 mixed with FG to ileal anastomosis accelerates wound healing and enhances the anastomosis by increased angiogenesis.

  4. Localization of type XII collagen in normal and healing rabbit cornea by in situ hybridization.

    Science.gov (United States)

    Zhan, Q; Burrows, R; Cintron, C

    1995-05-01

    To identify the cell types responsible for type XII collagen synthesis in normal and healing rabbit cornea, a partial cDNA sequence of rabbit type XII collagen, obtained from an adult rabbit cornea cDNA library, was used to develop highly specific oligonucleotide probes for Northern blot analysis and in situ hybridization. Approximately 2000 bases of a type XII collagen 2.2 kb cDNA clone were sequenced. Comparative sequence analysis of the bases showed a 74% identity with chick alpha 1 (XII) chain of type XII collagen. The deduced amino acid sequence indicated a 72% identity with chick type XII collagen. Northern blot analysis showed that cultures of cornea stromal and endothelial cells each contain two RNA species, greater than 10 kb, that hybridize to rabbit type XII collagen oligonucleotide probes. Although normal stromal cells failed to show type XII collagen mRNA, normal endothelial cells contain mRNA for this collagen. These results indicate that endothelium of normal rabbit cornea has a potential to synthesize type XII collagen. During corneal wound healing, both endothelium-derived and stroma-derived cells in the developing scar tissue contained type XII mRNA. In view of the known presence of type XII collagen in corneal stromas from chick and mouse, the distribution of mRNA in normal cornea is puzzling.

  5. Quantitative evaluation of the corneal endothelium in the mouse after grafting

    OpenAIRE

    Plskova, J; Kuffova, L; Filipec, M; Holan, V; Forrester, J V

    2004-01-01

    Background/aim: Corneal graft survival depends critically on the quality of the endothelium. In this study the authors aimed to evaluate corneal endothelium in mice at different times after transplantation and to correlate endothelial integrity with corneal graft survival.

  6. [Corneal toxicity due to amantadine].

    Science.gov (United States)

    Avendaño-Cantos, E M; Celis-Sánchez, J; Mesa-Varona, D; Gálvez-Martínez, J; López-Arroquia, E; González Del Valle, F

    2012-09-01

    A 64 year-old female with Parkinson disease treated with amantadine for two years who suddenly suffered bilateral corneal oedema. It was initially treated as herpetic endotheliitis without improvement as we lacked information on her chronic treatment. The corneal oedema finally resolved after withdrawing the drug. Amantadine hydrochloride may produce endothelial dysfunction. Once the amantadine treatment is stopped, the corneal oedema may be reversible but endothelial density remains low. An ophthalmologist examination should be performed before the initiation of amantadine treatment in order to establish a risk: benefit ratio, especially in those patients with low endothelial density or any endothelial anomaly. Copyright © 2011 Sociedad Española de Oftalmología. Published by Elsevier Espana. All rights reserved.

  7. Corneal Ulcer

    Science.gov (United States)

    ... Español Eye Health / Eye Health A-Z Corneal Ulcer Sections What Is a Corneal Ulcer? Corneal Ulcer ... Diagnosis Corneal Ulcer Treatment What Is a Corneal Ulcer? Leer en Español: ¿Qué es una Úlcera de ...

  8. Toxicidade da mitomicina C ao endotélio da córnea de coelhos Mitomycin C toxicity in rabbit corneal endothelium

    Directory of Open Access Journals (Sweden)

    Maria Rosa Bet de Moraes Silva

    2009-04-01

    transmissão.PURPOSE: To evaluate corneal endothelium alterations after applying mitomycin C to the sclera using transmission and scanning electron microscopy, correlating alterations with time, concentration, and evaluation methods. METHODS: The corneal endothelium of both eyes of 32 albino rabbits was evaluated and distributed into four groups of 8. Mitomycin C was applied under a scleral flap in the right eye for 5 minutes. Mitomycin C concentrations were 0.5 mg/ml for G1 and G2 and 0.2 mg/ml for G3 and G4. Examinations were performed 15 days after application to G1 and G3, and 30 days after application to G2 and G4. Four cornea in each group were prepared for transmission electron microscopy and four for scanning electron microscopy. Left eyes of all animals were used as controls. RESULTS: Transmission electron microscopy showed corneal endothelium alterations in all groups: rarefied cytoplasm, dilation and fragmentation of rough endoplasmic reticulum cisternae, Golgi apparatus with cisternal dilation, reduced vacuoles, and irregularities of internal membrane more noticeable in G1 and G2. Scanning electron microscopy revealed alterations in all groups except G1: changes in the shape and size of cells and longer filopodial projections. CONCLUSIONS: 1 - Corneal endothelium alterations were seen at both 0.5 and 0.2 mg/ml concentrations and at 15 and 30 days after mytomicin C application; 2 - Alterations were more intense with higher mytomicin C concentration by transmission electron but not by scanning electron microscopy; 3 - The alterations correlated with time by scanning electron microscopy but not by transmission electron microscopy.

  9. Tissue Engineering of Corneal Endothelium

    Directory of Open Access Journals (Sweden)

    Satoru Yamagami

    2012-10-01

    Full Text Available Human corneal endothelial cells (HCECs do not replicate after wounding. Therefore, corneal endothelial deficiency can result in irreversible corneal edema. Descemet stripping automated endothelial keratoplasty (DSAEK allows selective replacement of the diseased corneal endothelium. However, DSAEK requires a donor cornea and the worldwide shortage of corneas limits its application. This review presents current knowledge on the tissue engineering of corneal endothelium using cultured HCECs. We also provide our recent work on tissue engineering for DSAEK grafts using cultured HCECs. We reconstructed DSAEK grafts by seeding cultured DiI-labelled HCECs on collagen sheets. Then HCEC sheets were transplanted onto the posterior stroma after descemetorhexis in the DSAEK group. Severe stromal edema was detected in the control group, but not in the DSAEK group throughout the observation period. Fluorescein microscopy one month after surgery showed numerous DiI-labelled cells on the posterior corneal surface in the DSAEK group. Frozen sections showed a monolayer of DiI-labelled cells on Descemet’s membrane. These findings indicate that cultured adult HCECs, transplanted with DSAEK surgery, maintain corneal transparency after transplantation and suggest the feasibility of performing DSAEK with HCECs to treat endothelial dysfunction.

  10. The inhibitory effect of different concentrations of KH902 eye drops on corneal neovascularization induced by alkali burn

    Directory of Open Access Journals (Sweden)

    Yan Wu

    2017-01-01

    Full Text Available Purpose: The aim of this study was to evaluate the inhibitory effect of different concentrations of KH902 eye drops on rabbit corneal neovascularization (CNV induced by alkali burn. Methods: Forty-eight adult rabbits were randomized into four groups after alkali burning: Group A (2.5 mg/ml, Group B (5 mg/ml, and Group C (10 mg/ml by different concentrations of KH902 eye drops and Group D by saline solution as control with three times a day for 2 weeks. At days 7, 14, and 28, the anterior segment photographs, confocal microscopy, and histopathology were performed to evaluate corneal opacity, neovascularization, inflammatory cell density, vessel size, and edema. Immunohistochemistry was applied to analyze the vascular endothelial growth factor (VEGF level. Results: (1 The CNV in the medicine-treated groups showed a reduction without obvious corneal side effects histologically. (2 Compared to the control group, the three medicine-treated groups showed a reduction in the VEGF levels and CNV areas on days 7, 14, and 28 and in the inflammatory cell density on days 14 and 28 (P 0.05. Conclusion: KH902 eye drops in lower concentration showed an obvious reduction of the CNV growing for rabbit corneal alkali burn without side effects.

  11. Effects of corneal irregular astigmatism on visual acuity after conventional and femtosecond laser-assisted Descemet's stripping automated endothelial keratoplasty.

    Science.gov (United States)

    Tomida, Daisuke; Yamaguchi, Takefumi; Ogawa, Akiko; Hirayama, Yumiko; Shimazaki-Den, Seika; Satake, Yoshiyuki; Shimazaki, Jun

    2015-07-01

    To compare short-term outcomes of Descemet's stripping automated endothelial keratoplasty (DSAEK) using a graft prepared with either a femtosecond laser or a microkeratome. Thirty-eight patients underwent DSAEK with grafts prepared with either a femtosecond laser (f-DSAEK; 21 eyes) or a microkeratome (m-DSAEK; 17 eyes). Visual acuity, endothelial cell density, regular astigmatism and irregular astigmatism were compared between the two groups preoperatively and at 1, 3, and 6 months post-operatively. Fourier analysis was conducted to calculate astigmatism of the anterior and posterior surfaces, and total cornea, using anterior segment optical coherence tomography (AS-OCT). Visual acuity (logMAR) improved from 1.20 ± 0.60 to 0.43 ± 0.25 after m-DSAEK (P astigmatism of the total cornea and the posterior surface was significantly larger after f-DSAEK than after m-DSAEK, although there was no significant difference in irregular astigmatism of the anterior surface at 6 months. Postoperative visual acuity was significantly correlated with the postoperative irregular astigmatism of the total cornea (r = 0.6657 and P astigmatism caused by posterior surface irregularities.

  12. L450W and Q455K Col8a2 knock-in mouse models of Fuchs endothelial corneal dystrophy show distinct phenotypes and evidence for altered autophagy.

    Science.gov (United States)

    Meng, Huan; Matthaei, Mario; Ramanan, Narendrakumar; Grebe, Rhonda; Chakravarti, Shukti; Speck, Caroline L; Kimos, Martha; Vij, Neeraj; Eberhart, Charles G; Jun, Albert S

    2013-03-28

    We compared the cellular phenotypes and studied the role of autophagy in the pathogenesis of Fuchs endothelial corneal dystrophy (FECD) using two α2 collagen VIII (Col8a2) knock-in mouse models and human FECD tissues. In vivo corneal endothelial cell (CEC) counts and morphology were analyzed by clinical confocal microscopy. Ultrastructural analysis of CECs was performed by transmission electron microscopy. Real-time PCR and Western blotting were performed using total RNA, and protein extracted from mouse CECs and human CECs obtained from FECD and autopsy patients. Both Col8a2 mouse models exhibited hallmarks of FECD; however, the Col8a2(L450W/L450W) mice exhibited a milder phenotype compared to the Col8a2(Q455K/Q455K) mice. Both models exhibited upregulation of the unfolded protein response (UPR) as evidenced by dilated rough endoplasmic reticulum (RER), and upregulation of UPR-associated genes and proteins. Real-time PCR of Col8a2(L450W/L450W) and Col8a2(Q455K/Q455K) CECs at 40 weeks revealed a 2.1-fold (P human FECD endothelium revealed a 10.4-fold upregulation of DRAM1 (P human FECD endothelial cells suggested a role for altered autophagy in this disease.

  13. Clinical evaluation and induced corneal vascularization study by native and anionic collagen membranes in rabbits corneas Avaliação clínica e estudo da vascularização corneal induzida pelas membranas de colágeno nativo e aniônico em córneas de coelhos

    Directory of Open Access Journals (Sweden)

    Thaís Eliane Binotto

    2009-12-01

    Full Text Available PURPOSE: To evaluate the corneal vascularization (CV and the clinical aspects induced by interlamellar graft with native (NCM and anionic (ACM collagen membranes in rabbits corneas. METHODS: An interlamellar graft with a 0.25 x 0.25 cm NCM (group 1 or ACM (group 2 fragment was performed in the right eye (treated eye. In the left eye, an estromal tunnel was done (control eye. Sixteen rabbits were used, and they were subdivided into two experimental groups of eight animals each. The clinical evaluation was performed at the 1st, 3rd, 7th, 15th and 30th postoperative days. Corneal vascularization analysis was performed after 30 days by the Images Analizator System Leica Qwin-550®. RESULTS: After 7 days, corneal vascularization was observed at about 2.25 ± 0.71 mm (NCM and at about 1.0 ± 1.69 mm (ACM, respectively, from the limbus in direction to the central cornea. After 15 days, CV increased in both groups (5.25 ± 1.03 mm - NCM; 2.0 ± 2.39 mm - ACM and then progressively decreased until day 30 (2.25 ± 2.10 mm - NCM; 0.75 ± 2.12 mm - ACM. The statistical analysis indicated that the averages of the distances from the limb vessels to the grafts observed after 7 and 15 days had not differed statistically (p=0.17, and after 15 and 30 postoperative days had a tendency to differ statistically (p=0.09. The control eyes did not present any changes. CONCLUSION: The interlamellar graft with native and anionic collagen membranes induced corneal vascularization when applied to rabbit corneas, but anionic collagen membrane induced a smaller corneal vascularization when compared to native collagen membrane. Although further studies are required, the results found in this study demonstrated the usefulness of interlamellar graft with native and anionic collagen membranes in keratoplasties. These membranes consists in one more graft option for the surgical treatment of corneal repair in rabbits and others animals, when other forms of medical and surgical

  14. Acellular ostrich corneal stroma used as scaffold for construction of tissue-engineered cornea

    Directory of Open Access Journals (Sweden)

    Xian-Ning Liu

    2016-03-01

    Full Text Available AIM: To assess acellular ostrich corneal matrix used as a scaffold to reconstruct a damaged cornea. METHODS: A hypertonic saline solution combined with a digestion method was used to decellularize the ostrich cornea. The microstructure of the acellular corneal matrix was observed by transmission electron microscopy (TEM and hematoxylin and eosin (H&E staining. The mechanical properties were detected by a rheometer and a tension machine. The acellular corneal matrix was also transplanted into a rabbit cornea and cytokeratin 3 was used to check the immune phenotype. RESULTS: The microstructure and mechanical properties of the ostrich cornea were well preserved after the decellularization process. In vitro, the methyl thiazolyl tetrazolium results revealed that extracts of the acellular ostrich corneas (AOCs had no inhibitory effects on the proliferation of the corneal epithelial or endothelial cells or on the keratocytes. The rabbit lamellar keratoplasty showed that the transplanted AOCs were transparent and completely incorporated into the host cornea while corneal turbidity and graft dissolution occurred in the acellular porcine cornea (APC transplantation. The phenotype of the reconstructed cornea was similar to a normal rabbit cornea with a high expression of cytokeratin 3 in the superficial epithelial cell layer. CONCLUSION: We first used AOCs as scaffolds to reconstruct damaged corneas. Compared with porcine corneas, the anatomical structures of ostrich corneas are closer to those of human corneas. In accordance with the principle that structure determines function, a xenograft lamellar keratoplasty also confirmed that the AOC transplantation generated a superior outcome compared to that of the APC graft.

  15. Paradigm shifts in corneal transplantation.

    Science.gov (United States)

    Tan, Donald T H; Anshu, Arundhati; Mehta, Jodhbir S

    2009-04-01

    Conventional corneal transplantation, in the form of penetrating keratoplasty (PK), involves full-thickness replacement of the cornea, and is a highly successful procedure. However, the cornea is anatomically a multi-layered structure. Pathology may only affect individual layers of the cornea, hence selective lamellar surgical replacement of only the diseased corneal layers whilst retaining unaffected layers represents a new paradigm shift in the field. Recent advancements in surgical techniques and instrumentation have resulted in several forms of manual, microkeratome and femto-second laser-assisted lamellar transplantation procedures. Anterior lamellar keratoplasty (ALK) aims at replacing only diseased or scarred corneal stroma, whilst retaining the unaffected corneal endothelial layer, thus obviating the risk of endothelial allograft rejection. Posterior lamellar keratoplasty/endothelial keratoplasty (PLK/EK) involves the replacement of the dysfunctional endothelial cell layer only. Whilst significant technical and surgical challenges are involved in performing lamellar micro-dissection of a tissue which is only 0.5 mm thick, the benefits of a more controlled surgical procedure and improved graft survival rates have resulted in a shift away from conventional PK. This review details the current advances in emerging lamellar corneal surgical procedures and highlights the main advantages and disadvantages of these new lamellar corneal procedures.

  16. CORNEAL BLINDNESS AND XENOTRANSPLANTATION

    Science.gov (United States)

    Lamm, Vladimir; Hara, Hidetaka; Mammen, Alex; Dhaliwal, Deepinder; Cooper, David K.C.

    2014-01-01

    Approximately 39 million people are blind worldwide, with an estimated 285 million visually impaired. The developing world shoulders 90% of the world’s blindness, with 80% of causative diseases being preventable or treatable. Blindness has a major detrimental impact on the patient, community, and healthcare spending. Corneal diseases are significant causes of blindness, affecting at least 4 million people worldwide. The prevalence of corneal disease varies among parts of the world. Trachoma, for instance, is the second leading cause of blindness in Africa, after cataracts, but is rarely found today in developed nations. When preventive strategies have failed, corneal transplantation is the most effective treatment for advanced corneal disease. The major surgical techniques for corneal transplantation include penetrating keratoplasty (PK), anterior lamellar keratoplasty (ALK), and endothelial keratoplasty (EK). Indications for corneal transplantation vary among countries, with Fuchs’ dystrophy being the leading indication in the U.S. and keratoconus in Australia. With the exception of the US, where EK will soon overtake PK as the most common surgical procedure, PK is the overwhelming procedure of choice. Success using corneal grafts in developing nations, such as Nepal, demonstrates the feasibility of corneal transplantation on a global scale. The number of suitable corneas from deceased human donors that becomes available will never be sufficient, and so research into various alternatives, e.g., stem cells, amniotic membrane transplantation, synthetic and biosynthetic corneas, and xenotransplantation, is progressing. While each of these has potential, we suggest that xenotransplantation holds the greatest potential for a corneal replacement. With the increasing availability of genetically-engineered pigs, pig corneas may alleviate the global shortage of corneas in the near future. PMID:25268248

  17. The Use of Fiber-Reinforced Scaffolds Cocultured with Schwann Cells and Vascular Endothelial Cells to Repair Rabbit Sciatic Nerve Defect with Vascularization

    Directory of Open Access Journals (Sweden)

    Hongyang Gao

    2013-01-01

    Full Text Available To explore the feasibility of biodegradable fiber-reinforced 3D scaffolds with satisfactory mechanical properties for the repair of long-distance sciatic nerve defect in rabbits and effects of vascularized graft in early stage on the recovery of neurological function, Schwann cells and vascular endothelial cells were cocultured in the fiber-reinforced 3D scaffolds. Experiment group which used prevascularized nerve complex for the repair of sciatic nerve defect and control group which only cultured with Schwann cells were set. The animals in both groups underwent electromyography to show the status of the neurological function recovery at 4, 8, and 16 weeks after the surgery. Sciatic nerve regeneration and myelination were observed under the light microscope and electron microscope. Myelin sheath thickness, axonal diameter, and number of myelinated nerve fiber were quantitatively analyzed using image analysis system. The recovery of foot ulcer, the velocity of nerve conduction, the number of regenerating nerve fiber, and the recovery of ultrastructure were increased in the experimental group than those in the control group. Prevascularized tissue engineered fiber-reinforced 3D scaffolds for the repair of sciatic nerve defects in rabbits can effectively promote the recovery of neurological function.

  18. The induction of an angiogenic response in corneal myofibroblasts by platelet-activating factor (PAF).

    Science.gov (United States)

    He, Jiucheng; Eastlack, Jason P; Bazan, Haydee E P

    2010-12-01

    Although the exact mechanisms underlying corneal neovascularization remain unclear, cytokines and growth factors play an important role in their development. We have shown previously that the inflammatory mediator platelet-activating factor (PAF) is a potent inducer of corneal neovascularization in vivo. In this study, we investigate the role of stromal myofibroblasts in neovascularization and the effect of PAF on this process. Myofibroblasts were obtained from rabbit corneal keratocytes and identified with anti-α-SMA antibody. Cells were treated with PAF (100 nM) for 24 hr. In some experiments, cells were pre-treated with the PAF antagonist LAU-0901 (150 nM). Expression of vascular endothelial growth factor (VEGF) and thrombospondin-1 (TSP-1) was examined by immunofluorescence and immunoblotting. To study the effect of myofibroblasts on vessel formation in vitro, Vybrant(®) CM-DiI labeled human umbilical vein endothelial cells (HUVECs) were cultured on myofibroblasts in a thin layer of collagen gel. CD31 was used as the cell marker of HUVEC. VEGF and TSP-1 were not detectable in keratocytes, but they were positively stained in myofibroblasts. PAF induced a significant increase in VEGF expression and a decrease in TSP-1 expression. These changes were inhibited in the presence of LAU-0901. HUVECs co-cultured with corneal myofibroblasts formed a typical structure of vessel-like tubes within 1 week. The addition of PAF to the medium increased HUVEC-induced vessel-like tube formation, which was abolished by LAU-0901. Addition of anti-VEGF antibody to the medium completely prevented the formation of vessel-like tubes. We provide evidence for the role of stromal myofibroblasts in the corneal neovascularization process. By enhancing VEGF production and decreasing TSP-1 production in myofibroblasts, PAF augments the angiogenic response. The PAF antagonist LAU-0901 could represent a new therapeutic venue for inhibiting corneal neovascularization.

  19. Corneal Laceration

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    Full Text Available ... or apply pressure to eye avoid giving aspirin, ibuprofen or other non-steroidal, anti-inflammatory drugs. These ... lost sight from a corneal scar as a child. Now that I’m older, will a corneal ...

  20. Corneal Laceration

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    Full Text Available ... rub or apply pressure to eye avoid giving aspirin, ibuprofen or other non-steroidal, anti-inflammatory drugs. ... lost sight from a corneal scar as a child. Now that I’m older, will a corneal ...

  1. Corneal Laceration

    Medline Plus

    Full Text Available ... Now that I’m older, will a corneal transplant help me? May 15, 2015 Why Do My ... May Have A Future Alternative to Full Corneal Transplantation Nov 29, 2016 Combating Eye Injuries from Air ...

  2. Femtosecond-laser-assisted Descemet's stripping endothelial keratoplasty.

    Science.gov (United States)

    Cheng, Yanny Y Y; Pels, Elisabeth; Nuijts, Rudy M M A

    2007-01-01

    To our knowledge, we describe the first patient with pseudophakic bullous keratoplasty treated with femtosecond-laser-assisted endothelial keratoplasty. A 5.5 mm corneoscleral tunnel incision was made; after Descemet's membrane was stripped, an 8.0 mm posterior lamellar corneal disk prepared with a femtosecond laser was inserted into the anterior chamber against the recipient cornea without the use of corneal sutures. Four months postoperatively, the posterior corneal disk was clear and the induced astigmatism was 2.1 diopters, demonstrating a functional corneal endothelial layer. The femtosecond laser offers a new surgical approach for minimally invasive endothelial keratoplasty in corneal endothelial disorders.

  3. Efeito do colírio de 5-fluorouracil sobre o epitélio corneano íntegro de coelhos Effects of 5-fluorouracil eye drops on intact rabbit corneal epithelium

    Directory of Open Access Journals (Sweden)

    Lucieni Cristina Barbarini Ferraz

    2003-08-01

    Full Text Available OBJETIVO: Observar os efeitos da aplicação tópica de 5-fluorouracil (5-FU sobre o epitélio corneano íntegro. MÉTODOS: Foram utilizados 10 coelhos albinos (14 olhos, divididos em: grupo controle (GC, 4 olhos nos quais não se administraram drogas, grupo 1 (G1, 5 olhos que receberam 5-fluorouracil na concentração 1,25% e grupo 2 (G2, 5 olhos que receberam 5-fluorouracil na concentração de 2,5%. A droga foi instilada 4 vezes por dia, durante 7 dias, quando os animais foram sacrificados, os olhos removidos, separando-se a córnea que foi preparada de modo convencional para estudo em microscópico eletrônico de varredura. RESULTADOS: GC: observaram-se células de formato hexagonal, claras, escuras e intermediárias, compondo o epitélio corneano de coelhos. Presença de numerosas microplicas, principalmente nas células claras. Cada célula possui cerca de 1 a 3 criptas. Nos animais do G1, observou-se maior número de células escuras, regiões com diminuição no número de criptas; alterações da superfície celular, protusão na região do núcleo e descamação de células epiteliais. Os do G2 tiveram aumento de microprojeções na superfície celular, modificações nas junções intercelulares até separação de células adjacentes; diminuição do número e variabilidade no tamanho das criptas. As alterações mais freqüentes ocorreram nas células da periferia da córnea. CONCLUSÃO: O 5-fluorouracil teve efeitos deletérios no epitélio íntegro corneano de coelhos. As alterações observadas foram mais importantes nos animais que receberam a droga mais concentrada (G2 e mais freqüentes na periferia da córnea.PURPOSE: To assess the influence of the antiproliferative agent (5-FU on the intact rabbit corneal epithelium. METHODS: 10 rabbits (14 eyes,were divided into: control group (CG, 4 eyes without drug administration; G1, 5 eyes underwent treatment with topical 12.5 mg/ml 5-fluorouracil and G2, 5 eyes received 5-fluorouracil

  4. Influence of corneal hydration on optical coherence elastography

    Science.gov (United States)

    Twa, Michael D.; Vantipalli, Srilatha; Singh, Manmohan; Li, Jiasong; Larin, Kirill V.

    2016-03-01

    Corneal biomechanical properties are influenced by several factors, including intraocular pressure, corneal thickness, and viscoelastic responses. Corneal thickness is directly proportional to tissue hydration and can influence corneal stiffness, but there is no consensus on the magnitude or direction of this effect. We evaluated the influence of corneal hydration on dynamic surface deformation responses using optical coherence elastography (OCE). Fresh rabbit eyes (n=10) were prepared by removing the corneal epithelium and dropping with 0.9% saline every 5 minutes for 1 hour, followed by 20% dextran solution every 5 minutes for one hour. Corneal thickness was determined from structural OCT imaging and OCE measurements were performed at baseline and every 20 minutes thereafter. Micron-scale deformations were induced at the apex of the corneal tissue using a spatially-focused (150μm) short-duration (corneal thickness due to hydration process. Corneal thickness rapidly increased and remained constant following epithelium removal and changed little thereafter. Likewise, corneal stiffness changed little over the first hour and then decreased sharply after Dextran application (thickness: -46% [-315/682 μm] RR: - 24% [-0.7/2.88 ms-1]; GV: -19% [-0.6/3.2 m/s]). Corneal thickness and corneal stiffness (RR) were well correlated (R2 = .66). Corneal biomechanical properties are highly correlated with tissue hydration over a wide range of corneal thickness and these changes in corneal stiffness are quantifiable using OCE.

  5. Genetics of corneal endothelial dystrophies

    Indian Academy of Sciences (India)

    2009-12-31

    Dec 31, 2009 ... additional extracellular matrix known as the posterior col- lagenous layer, posterior to the normal DM, as a response to different types of diseases including ... clude clusters of vesicles or blister-like lesions, band-shaped irregularities, or more extensive irregularity involving the en- tire DM and the variable ...

  6. Combined effects of interleukin-1? and cyclic stretching on metalloproteinase expression in corneal fibroblasts in vitro

    OpenAIRE

    Feng, Pengfei; Li, Xiaona; Chen, Weiyi; Liu, Chengxing; Rong, Shuo; Wang, Xiaojun; Du, Genlai

    2016-01-01

    Background Corneal tensile strain increases if the cornea becomes thin or if intraocular pressure increases. However, the effects of mechanical stress on extracellular matrix (ECM) remodelling in the corneal repair process and the corneal anomalies are unknown. Methods In this study, the combined effects of interleukin-1? (IL-1?) on matrix metalloproteinases (MMPs) in corneal fibroblasts under cyclic stretching were investigated in vitro. Cultured rabbit corneal fibroblasts were subjected to ...

  7. Corneal endothelium of the Magellanic penguin (Spheniscus magellanicus) by scanning electron microscopy.

    Science.gov (United States)

    Pigatto, João A T; Laus, José L; Santos, Jaime M; Cerva, Cristine; Cunha, Luciana S; Ruoppolo, Valéria; Barros, Paulo S M

    2005-12-01

    The corneal endothelium is essential for the maintenance of the corneal transparency. The aim of this study was to examine the morphology of the endothelial surface and perform morphometric analysis of the normal corneal endothelial cells of the Magellanic penguin (Spheniscus magellanicus) using scanning electron microscopy. The present work demonstrates that the corneal endothelium of the Magellanic penguin is similar to those described in other vertebrates.

  8. Progress in corneal wound healing

    Science.gov (United States)

    Ljubimov, Alexander V.; Saghizadeh, Mehrnoosh

    2015-01-01

    Corneal wound healing is a complex process involving cell death, migration, proliferation, differentiation, and extracellular matrix remodeling. Many similarities are observed in the healing processes of corneal epithelial, stromal and endothelial cells, as well as cell-specific differences. Corneal epithelial healing largely depends on limbal stem cells and remodeling of the basement membrane. During stromal healing, keratocytes get transformed to motile and contractile myofibroblasts largely due to activation of transforming growth factor-β system. Endothelial cells heal mostly by migration and spreading, with cell proliferation playing a secondary role. In the last decade, many aspects of wound healing process in different parts of the cornea have been elucidated, and some new therapeutic approaches have emerged. The concept of limbal stem cells received rigorous experimental corroboration, with new markers uncovered and new treatment options including gene and microRNA therapy tested in experimental systems. Transplantation of limbal stem cell-enriched cultures for efficient re-epithelialization in stem cell deficiency and corneal injuries has become reality in clinical setting. Mediators and course of events during stromal healing have been detailed, and new treatment regimens including gene (decorin) and stem cell therapy for excessive healing have been designed. This is a very important advance given the popularity of various refractive surgeries entailing stromal wound healing. Successful surgical ways of replacing the diseased endothelium have been clinically tested, and new approaches to accelerate endothelial healing and suppress endothelial-mesenchymal transformation have been proposed including Rho kinase (ROCK) inhibitor eye drops and gene therapy to activate TGF-β inhibitor SMAD7. Promising new technologies with potential for corneal wound healing manipulation including microRNA, induced pluripotent stem cells to generate corneal epithelium, and

  9. [Corneal biomechanics].

    Science.gov (United States)

    Torres, R M; Merayo-Lloves, J; Jaramillo, M A; Galvis, V

    2005-04-01

    To review the corneal biomechanic concepts and to analyse, clarify and understand their relevance in refractive surgery. A literature review has been done using different databases. Corneal biomechanic concepts are not new and are applied implicitly in numerous surgical procedures. Their origin is related to tonometry studies, but they gained in popularity when they were linked to the treatment of keratoconus, a pathology in which the mechanical properties of the cornea are altered. Factors determining corneal stability were thus defined. Corneal biomechanics have also been used following refractive surgery to study post-operative keratectasia and to improve ablation patterns, which ignores the corneal response. The new ablation systems need to include the biomechanical factors, which motivate research conducted in physical-mathematical models and in corneal wound healing, improving our knowledge about the corneal biomechanical response. The corneal biomechanic concepts have gained in popularity with the advent of refractive surgery, although they did exist previously. Their relevance is linked to improvements in the ablation systems used in an attempt to obtain more accurate and reliable results.

  10. Corneal Laceration

    Medline Plus

    Full Text Available ... Answers How often and for how long should I use a hair dryer to treat my Fuchs’ dystrophy? May 06, 2017 I lost sight from a corneal scar as a child. Now that I’m older, will a corneal transplant help me? ...

  11. Corneal Laceration

    Medline Plus

    Full Text Available ... Tips & Prevention News Ask an Ophthalmologist Patient Stories Español Eye Health / Eye Health A-Z Corneal Laceration ... Laceration Treatment What Is Corneal Laceration? Leer en Español: ¿Qué Es una Laceración de la Córnea? Written ...

  12. Corneal Laceration

    Medline Plus

    Full Text Available ... Health Find an Ophthalmologist Academy Store Eye Health A-Z Symptoms Glasses & Contacts Tips & Prevention News Ask ... Ophthalmologist Patient Stories Español Eye Health / Eye Health A-Z Corneal Laceration Sections What Is Corneal Laceration? ...

  13. Keratan sulfate and dermatan sulfate proteoglycans associate with type VI collagen in fetal rabbit cornea

    National Research Council Canada - National Science Library

    Takahashi, T; Cho, HI; Kublin, CL; Cintron, C

    1993-01-01

    .... Because certain cytochemical data suggested that proteoglycans are associated with type VI collagen in the fetal rabbit cornea, we developed polyclonal antibodies specific to the core proteins of rabbit corneal KSPG...

  14. Clinical applications of corneal confocal microscopy

    Directory of Open Access Journals (Sweden)

    Mitra Tavakoli

    2008-06-01

    Full Text Available Mitra Tavakoli1, Parwez Hossain2, Rayaz A Malik11Division of Cardiovascular Medicine, University of Manchester and Manchester Royal Infirmary, Manchester, UK; 2University of Southampton, Southampton Eye Unit, Southampton General Hospital, Southampton, UKAbstract: Corneal confocal microscopy is a novel clinical technique for the study of corneal cellular structure. It provides images which are comparable to in-vitro histochemical techniques delineating corneal epithelium, Bowman’s layer, stroma, Descemet’s membrane and the corneal endothelium. Because, corneal confocal microscopy is a non invasive technique for in vivo imaging of the living cornea it has huge clinical potential to investigate numerous corneal diseases. Thus far it has been used in the detection and management of pathologic and infectious conditions, corneal dystrophies and ecstasies, monitoring contact lens induced corneal changes and for pre and post surgical evaluation (PRK, LASIK and LASEK, flap evaluations and Radial Keratotomy, and penetrating keratoplasty. Most recently it has been used as a surrogate for peripheral nerve damage in a variety of peripheral neuropathies and may have potential in acting as a surrogate marker for endothelial abnormalities.Keywords: corneal confocal microscopy, cornea, infective keratitis, corneal dystrophy, neuropathy

  15. Corneal topography.

    Science.gov (United States)

    Seitz, B; Behrens, A; Langenbucher, A

    1997-08-01

    In the review period, limitations of individual Placido disk-based topography systems have been studied and new principles, such as raster photogrammetry, pancorneal slit topography, laser holographic interferometry, and confocal laser scanning topography, have been introduced for laboratory or clinical work. Both Fourier analysis and Zernike decomposition of topographic height data seem to be powerful new tools for cross-sectional analysis of complex topographic corneal images, such as after cataract surgery, penetrating keratoplasty, and refractive surgery, as well as for longitudinal studies of corneal changes, such as in schoolchildren. Subdividing into rational optical components may improve consistency and standardization of topography data from different systems. Topography-based flying-spot-mode excimer laser photoablation after Zernike decomposition of topography height data has been proposed for correction of irregular corneal astigmatism. Topography-based central power measurements are of increasing value for intraocular lens power calculation before cataract surgery in eyes with irregular corneal surfaces, such as in keratoconus or after refractive surgery procedures. Quantitative and qualitative classification of corneal topography maps after corneal transplantation following conventional mechanical and nonmechanical trephination or after refractive surgery may lead to a better understanding of impaired visual acuity despite a clear graft or despite significantly reduced ametropia or corneal astigmatism.

  16. Corneal Regeneration by Deep Anterior Lamellar Keratoplasty (DALK Using Decellularized Corneal Matrix.

    Directory of Open Access Journals (Sweden)

    Yoshihide Hashimoto

    Full Text Available The purpose of this study is to demonstrate the feasibility of DALK using a decellularized corneal matrix obtained by HHP methodology. Porcine corneas were hydrostatically pressurized at 980 MPa at 10°C for 10 minutes to destroy the cells, followed by washing with EGM-2 medium to remove the cell debris. The HHP-treated corneas were stained with H-E to assess the efficacy of decellularization. The decellularized corneal matrix of 300 μm thickness and 6.0 mm diameter was transplanted onto a 6.0 mm diameter keratectomy wound. The time course of regeneration on the decellularized corneal matrix was evaluated by haze grading score, fluorescein staining, and immunohistochemistry. H-E staining revealed that no cell nuclei were observed in the decellularized corneal matrix. The decellularized corneal matrices were opaque immediately after transplantation, but became completely transparent after 4 months. Fluorescein staining revealed that initial migration of epithelial cells over the grafts was slow, taking 3 months to completely cover the implant. Histological sections revealed that the implanted decellularized corneal matrix was completely integrated with the receptive rabbit cornea, and keratocytes infiltrated into the decellularized corneal matrix 6 months after transplantation. No inflammatory cells such as macrophages, or neovascularization, were observed during the implantation period. The decellularized corneal matrix improved corneal transparency, and remodelled the graft after being transplanted, demonstrating that the matrix obtained by HHP was a useful graft for corneal tissue regeneration.

  17. Corneal Laceration

    Medline Plus

    Full Text Available ... by something sharp flying into the eye. It can also be caused by something striking the eye ... If the corneal laceration is deep enough it can cause a full thickness laceration. This is when ...

  18. Corneal Laceration

    Medline Plus

    Full Text Available ... itself. A corneal laceration is a very serious injury and requires immediate medical attention to avoid severe ... 27, 2015 Dark Spot in Vision After Blunt Trauma Dec 21, 2014 Pain a Year After Eyelid ...

  19. Corneal Laceration

    Medline Plus

    Full Text Available ... the blood and may increase bleeding after you have finished protecting the eye, see a physician immediately ... Jun 30, 2017 People with Advanced Keratoconus May Have A Future Alternative to Full Corneal Transplantation Nov ...

  20. Corneal Laceration

    Medline Plus

    Full Text Available ... People with Advanced Keratoconus May Have A Future Alternative to Full Corneal Transplantation Nov 29, 2016 Combating Eye Injuries from ... of Service For Advertisers For Media Ophthalmology Job Center © American Academy of Ophthalmology 2017 ...

  1. Corneal Laceration

    Medline Plus

    Full Text Available ... inflammatory drugs. These drugs thin the blood and may increase bleeding after you have finished protecting the ... a hair dryer to treat my Fuchs’ dystrophy? May 06, 2017 I lost sight from a corneal ...

  2. Corneal Laceration

    Medline Plus

    Full Text Available ... By: Devin A Harrison MD Sep. 01, 2017 The cornea is the clear front window of the eye . A corneal laceration is a cut on the cornea. It is usually caused by something sharp ...

  3. Corneal Laceration

    Medline Plus

    Full Text Available ... itself. A corneal laceration is a very serious injury and requires immediate medical attention to avoid severe ... Ask an Ophthalmologist Answers Did my traumatic brain injury cause early cataracts? Jan 21, 2018 Did I ...

  4. Corneal Laceration

    Medline Plus

    Full Text Available ... caused by something striking the eye with significant force, like a metallic hand tool. A corneal laceration ... and preserving your vision. Privacy Policy Related Top 5 Eye Health Stories of 2017 Dec 21, 2017 ...

  5. Corneal Laceration

    Medline Plus

    Full Text Available ... itself. A corneal laceration is a very serious injury and requires immediate medical attention to avoid severe ... Dangerous for Your Eyes Sep 20, 2017 Eye Injuries from Laundry Packets On the Rise Jun 30, ...

  6. Corneal collagens.

    Science.gov (United States)

    Robert, L; Legeais, J M; Robert, A M; Renard, G

    2001-05-01

    Cornea is a highly differentiated tissue rich in extracellular matrix (ECM) specifically distributed in space in order to insure its dual role--transparency and protection of inner eye-tissues. Corneal ECM is especially rich in collagens. Since the characterisation of a number of distinct collagen types it appeared that most of them are present in the cornea. Their synthesis follows a specific program of sequential expression of the different collagen types to be synthesised during the development and maturation of the cornea. The precise regulation of the diameter and orientation of fibers, and of the interfibrillar spaces is partially at least attributed to interactions between glycosaminoglycans and collagens. The 'program' of vectorial collagen synthesis and GAG-collagen interactions changes also with age and in several pathological conditions as corneal dystrophies and wound healing. The Maillard reaction, especially in diabetes, is one of these important factors involved in age-dependent modifications of corneal structure and function. Far from being inert, corneal collagens were shown to have relatively short half-lives. The biosynthesis of corneal collagens was studied also during wound healing. The refibrillation of wounded corneas does not follow the original 'program' of ECM-synthesis as shown by the comparative study of wound healing using biochemical and morphometric methods. This review recapitulates briefly previous and recent studies on corneal collagens in order to present to clinicians and scientists an overview of the state of the art of this important field at the intersection of eye research and matrix biology.

  7. Effects of ethyl-cyanoacrylate and octyl-cyanoacrylate on experimental corneal lesions in rabbits Efeitos do etil-cianoacrilato ou do octil-cianoacrilato sobre lesões corneais experimentais em coelhos

    Directory of Open Access Journals (Sweden)

    V.T. Barbosa

    2009-10-01

    Full Text Available The use of ethyl-cyanoacrylate and octyl-cyanoacrylate were clinically and histopathologically compared on the corneas of 36 rabbits after lamellar keratectomy (standardized diameter and depth. The animals were distributed into two groups, one for each type of adhesive. From each group, six subgroups were histopathologically evaluated on the 3rd, 7th, 14th, 21st, 30th, and 60th day post-operative. General (daily and ophthalmic (days 0, 1, 3, 5, 7, 14, 21, 30, 44, and 60 evaluations clinically indicated that there were significant differences for the variables water intake, attitude, blepharitis, corneal edema, and fluorescein test. The adhesive permanence time for octyl-cyanoacrylate (17.22 days was greater than that for ethyl-cyanoacrylate (7.66 days. With respect to the histopathological evaluation, corneal epithelization and collagen organization occurred without severe complications. However, treatment with ethyl-cyanoacrylate led to a moderate inflammatory reaction in the initial phases. With octyl-cyanoacrylate, re-epithelization and collagen organization proceeded more slowly with a discrete inflammatory reaction in the initial phases. From clinical and histopathologic points of view, octyl-cyanoacrylate showed advantages over ethyl-cyanoacrylate, whereas wound healing was achieved in both groups without major complications.Comparou-se o uso do etil-cianoacrilato e do octil-cianoacrilato em córneas de 36 coelhos após ceratectomia lamelar (diâmetro e profundidade padronizados. Os animais foram distribuídos em dois grupos, segundo o tipo de adesivo, e redistribuídos em seis subgrupos com três animais cada, para as avaliações histológicas aos 3, 7, 14, 21, 30 e 60 dias de pós-operatório. As avaliações clínicas gerais (diárias e as oftálmicas (dias 0, 1, 3, 5, 7, 14, 21, 30, 44 e 60, indicaram diferença entre os dois grupos, quanto ao consumo de água, atitude, blefarite, edema da córnea e teste da fluoresceína. O Tempo de

  8. Femtosecond laser corneal refractive surgery

    Science.gov (United States)

    Kurtz, Ron M.; Spooner, Greg J. R.; Sletten, Karin R.; Yen, Kimberly G.; Sayegh, Samir I.; Loesel, Frieder H.; Horvath, Christopher; Liu, HsiaoHua; Elner, Victor; Cabrera, Delia; Muenier, Marie-Helene; Sacks, Zachary S.; Juhasz, Tibor

    1999-06-01

    We evaluated the efficacy, safety, and stability of femtosecond laser intrastromal refractive procedures in ex vivo and in vivo models. When compared with longer pulsewidth nanosecond or picosecond laser pulses, femtosecond laser-tissue interactions are characterized by significantly smaller and more deterministic photodisruptive energy thresholds, as well as reduced shock waves and smaller cavitation bubbles. We utilized a highly reliable, all-solid-state femtosecond laser system for all studies to demonstrate clinical practicality. Contiguous tissue effects were achieved by scanning a 5 μm focused laser spot below the corneal surface at pulse energies of approximately 2 - 4 microjoules. A variety of scanning patterns was used to perform three prototype procedures in animal eyes; corneal flap cutting, keratomileusis, and intrastromal vision correction. Superior dissection and surface quality results were obtained for lamellar procedures (corneal flap cutting and keratomileusis). Preliminary in vivo evaluation of intrastromal vision correction in a rabbit model revealed consistent and stable pachymetry changes, without significant inflammation or loss of corneal transparency. We conclude that femtosecond laser technology may be able to perform a variety of corneal refractive procedures with high precision, offering advantages over current mechanical and laser devices and techniques.

  9. Technology needs for corneal transplant surgery

    Science.gov (United States)

    Vaddavalli, Pravin K.; Yoo, Sonia H.

    2011-03-01

    Corneal transplant surgery has undergone numerous modifications over the years with improvements in technique, instrumentation and eye banking. The main goals of corneal transplantation are achieving excellent optical clarity with long-term graft survival. Penetrating, anterior and posterior lamellar surgery along with femtosecond laser technology have partially met these goals, but outcomes are often unpredictable and surgeon dependent. Technology to predictably separate stroma from Descemet's membrane, techniques to minimize endothelial cell loss, improvements in imaging technology and emerging techniques like laser welding that might replace suturing, eventually making corneal transplantation a refractively predictable procedure are on the wish list of the cornea surgeon.

  10. Functional evaluation of corneal endothelium by combined measurement of corneal volume alteration and cell density after phacoemulsification.

    Science.gov (United States)

    Suzuki, Hisaharu; Oki, Kotaro; Takahashi, Keizo; Shiwa, Toshihiko; Takahashi, Hiroshi

    2007-12-01

    To assess a new method of functional evaluation of corneal endothelium using combined measurement of corneal volume alteration and cell density (CD) after phacoemulsification. Nippon Medical School Hospital, Tokyo, Japan. Phacoemulsification was performed in 28 cases. The corneal volume within 3.0 mm and 10.0 mm circles of the cornea was measured using Scheimpflug imaging (Pentacam, Oculus, Inc.) and the cell density, using specular microscopy (Noncon Robo, Konan Medical). The volume stress index (VSI), a parameter for measuring functional evaluation, was calculated by dividing the volume increase within the 3.0 mm circle (preoperative corneal volume - corneal volume at time of measurement) by the central cell density value. The resulting value is indicative of the amount of postoperative increase in corneal volume with which each corneal endothelial cell should be able to cope. Three months after surgery, the 3.0 mm corneal volume returned to preoperative values, while the 10.0 mm corneal volume remained significantly higher than preoperatively. The cell density values did not significantly change throughout the study. The VSI showed a significant decrease by 7 days after surgery followed by a gradual decrease until the end of the study. At 3 months, the VSI value was significantly smaller than at 1 day. The corneal volume increase after surgery may be indicative of the degree of the endothelial damage in the area; thus, the VSI may be useful in the functional assessment of the corneal endothelium.

  11. Corneal decompensation after intraocular ophthalmic surgery--Missouri, 1998.

    Science.gov (United States)

    1998-04-24

    During January 8-14, 1998, six of eight patients undergoing elective intraocular surgery at a Veterans Affairs medical center (VAMC) in St. Louis, Missouri, developed corneal endothelial decompensation (corneal edema and opacification) instruments sterilized by the Abtox Plazlyte system (Abtox, Inc., Chicago, Illinois). This report summarizes the results of the investigation of these cases and indicates that using the Abtox Plazlyte system to sterilize opthalmologic surgical equipment led to corneal decompensation.

  12. Corneal Transplantation

    DEFF Research Database (Denmark)

    Hjortdal, Jesper Østergaard

    Corneal transplantation has been performed for more than 100 years. Until 15 years ago the state-of-the art type of transplantation was penetrating keratoplasty, but since the start of this millennium, newly designed surgical techniques have developed considerably. Today, the vast majority...... with less risk of rejection episodes. Besides covering updated chapters on penetrating keratoplasty, and anterior and posterior lamellar procedures, this textbook also gives a thorough overview of the history of corneal transplantation and a detailed presentation of the microstructural components....... Economic considerations on cost and benefi t of medical treatment and surgical procedures are today an integrated part of the health system in many countries, and a chapter covers these aspects of corneal transplantation. This textbook is aimed at presenting an updated review of the new techniques...

  13. The influence of contact lens wear on the corneal response to ultraviolet radiation.

    Science.gov (United States)

    Ahmedbhai, N; Cullen, A P

    1988-01-01

    One eye of each of 15 male pigmented rabbits was fitted with a gas-permeable rigid contact lens. Lenses were worn on a daily basis for an initial period of 7 days with the other eye serving as a control. After this initial period the lenses were removed and both eyes were irradiated with UV-B (305 nm +/- 9 full width half maximum) using either 0.03 J cm-2 or 0.12 J cm-2 which were approximately the epithelial and endothelial damage thresholds for this waveband respectively. The responses of the cornea were followed by microscopy and pachometry. Biomicroscopically there was a dramatic difference in response between contact lens wearing and non-wearing eyes for the higher radiant exposure, whereas little difference was observed at the lower level. For both levels of radiant exposure there was a significantly less corneal thickness increase in contact lens wearing eyes after UV-B irradiation. Recovery patterns were similar.

  14. Corneal Intelligence

    African Journals Online (AJOL)

    Murdoch3

    the damping effect of the tissue to an applied force) or corneal compliance being the major risk factor. Figure 1. The percentage of participants in the observation group who developed primary open-angle glaucoma (median follow-up, 72 months) ...

  15. Corneal ulcers

    African Journals Online (AJOL)

    visual acuity chart. • fluorescein strips. • topical anaesthetic eye drops. • direct ophthalmoscope. Examination. Assessing vision with a Snellen visual acuity chart gives a clue as to the extent of the problem (e.g. a corneal abrasion with good vision is unlikely to require specialist intervention). Each eye should be tested ...

  16. Corneal Intelligence

    African Journals Online (AJOL)

    Murdoch3

    Corneal Intelligence. Ian Murdoch. Institute of Ophthalmology, Bath Street, London. In 2002, the ocular hypertension treatment study (OHTS) published their results. This study had taken 1636 ocular hypertensives. 1, 2. (IOP 24-32mmHg) and randomized them to receive therapy or no therapy. The primary outcome of the ...

  17. Trends in corneal transplantation: indications and techniques.

    Science.gov (United States)

    Ple-Plakon, Patricia A; Shtein, Roni M

    2014-07-01

    To describe trends in corneal transplantation surgery, including indications for surgery, evolution of lamellar keratoplasty, current surgical techniques, and future directions. Over the past decade, anterior and posterior lamellar keratoplasty have begun to supplant penetrating keratoplasty. Surgical techniques continue to change and improve outcomes. In recent years, Descemet membrane endothelial keratoplasty (DMEK) has gained interest as it eliminates the corneal stromal interface, which may limit visual acuity after Descemet stripping automated endothelial keratoplasty. Despite the promising results with improved visual acuity and decreased rejection, the technical challenges associated with DMEK have limited widespread acceptance. With technical refinements and more eye banks providing precut tissue for both Descemet stripping automated endothelial keratoplasty and DMEK, it is likely both procedures will continue to increase over time. Corneal transplantation has evolved rapidly over the past decade, from full-thickness penetrating keratoplasty towards lamellar keratoplasty to only remove and replace damaged layers of the cornea. Achieving minimal induced astigmatism with excellent visual acuity remains a challenge in corneal transplantation. Further refinements in surgical technique may help improve technical challenges and visual outcomes. In this article, we review changing trends in corneal transplantation and highlight developing medical treatments that may be available in the future.

  18. Quantitative evaluation of corneal epithelial injury caused by n-heptanol using a corneal resistance measuring device in vivo

    Directory of Open Access Journals (Sweden)

    Fukuda M

    2012-04-01

    Full Text Available Masamichi Fukuda1, Hiroshi Sasaki11Department of Ophthalmology, Kanazawa Medical University, Uchinada, JapanPurpose: We attempted to develop a device for measuring electrical corneal resistance (CR using corneal contact lens electrodes to quantitatively evaluate corneal injury in vivo. In the present study, full-thickness detachment of the corneal epithelium was induced by n-heptanol, and the feasibility of the quantitative evaluation of this injury by corneal contact lens electrodes was evaluated in vivo.Methods: The central area of an albino rabbit's cornea was exposed to a filter paper pre-immersed in n-heptanol for 1 minute to induce injury of the corneal epithelium. After induction of injury, the electrical CR was measured and the percentage of CR (%CR was calculated. Fluorescein solution (3 µL was applied to the wound/affected area of the corneal epithelium for photography with a slit-lamp biomicroscope. The wound/affected area was measured using an image analysis system. The correlation between the %CR and the wound/affected area was analyzed.Results: As the size of the wound/affected area of the corneal epithelium increased, the %CR decreased after corneal epithelium detachment. Thus, a close correlation was found between the area of corneal epithelium detachment and the %CR.Conclusion: The corneal resistance device that we developed was capable of quantitatively evaluating n-heptanol-induced full-thickness injuries of the corneal epithelium.Keywords: eyes in vivo, corneal injury, corneal contact lens electrode corneal resistance device, n-heptanol

  19. Polysaccharide coating of human corneal endothelium

    DEFF Research Database (Denmark)

    Schroder, H D; Sperling, S

    1977-01-01

    Electron microscopy revealed the presence of a 600-1500 A thick layer of polysaccharide on the surface of human corneal endothelial cells. The surface layer was visualized by combined fixation and staining in a mixture of ruthenium red and osmium tetroxide. The coating material was stable...

  20. Corneal biomechanical properties from two-dimensional corneal flap extensiometry: application to UV-riboflavin cross-linking.

    Science.gov (United States)

    Kling, Sabine; Ginis, Harilaos; Marcos, Susana

    2012-07-27

    Corneal biomechanical properties are usually measured by strip extensiometry or inflation methods. We developed a two-dimensional (2D) flap extensiometry technique, combining the advantages of both methods, and applied it to measure the effect of UV-Riboflavin cross-linking (CXL). Corneal flaps (13 pig/8 rabbit) from the de-epithelialized anterior stroma (96 μm) were mounted on a custom chamber, consisting of a BK7 lens, a reflective retina, and two reservoirs (filled with Riboflavin and silicone oil). Stretching the corneal flap during five pressure increase/decrease cycles (0-30 mm Hg) changed the refractive power of the system, whose Zernike aberrations were monitored with a ray-tracing aberrometer. Porcine flaps were used to test the system. Rabbits were treated with CXL unilaterally in vivo following standard clinical procedures. Flaps were measured 1 month postoperatively. An analytical model allowed estimating Young's modulus from the change in surface (strain) and pressure (stress). Confocal microscopy examination was performed before, and at different times after CXL. Flap curvature changed with increased function of IOP in pig flaps (23.4 × 10⁻³ D/mm Hg). In rabbit flaps curvature changed significantly less in 1 month post CXL (P = 0.026) than in untreated corneas [17.0 vs. 6.36 millidiopter (mD)/mm Hg]. Young's modulus was 2.29 megapascals (MPa) in porcine corneas, 1.98 MPa in untreated rabbit corneas, and 4.83 MPa in 1 month post CXL rabbit corneas. At the same time, highly reflective structures were observed in the rabbit midstroma after treatment. 2D flap extensiometry allows estimating corneal elasticity in vitro. The measurements are spatially resolved in depth, minimize the effects of corneal hydration, and preserve the integrity of the cornea. The method proved the efficacy of CXL in increasing corneal rigidity after 1 month in rabbits.

  1. Corneal Thickness as a Predictor of Corneal Transplant Outcome

    Science.gov (United States)

    Verdier, David D.; Sugar, Alan; Baratz, Keith; Beck, Roy; Dontchev, Mariya; Dunn, Steven; Gal, Robin L.; Holland, Edward J.; Kollman, Craig; Lass, Jonathan H.; Mannis, Mark J.; Penta, Jeffrey

    2013-01-01

    Purpose Assess corneal thickness (CT) and correlation with graft outcome after penetrating keratoplasty in the Cornea Donor Study. Methods 887 subjects with a corneal transplant for a moderate risk condition (principally Fuchs or pseudophakic corneal edema) had post-operative CT measurements throughout a 5 year follow up time. Relationships between baseline (recipient, donor, and operative) factors and CT were explored. Proportional hazards models were used to assess association between CT and graft failure. Relationship between CT and cell density was assessed with a longitudinal repeated measures model and Spearman correlation estimates. Results Higher longitudinal CT measurements were associated with diagnosis of pseudophakic or aphakic corneal edema (P 25mmHg during the first post-operative month (P=0.003), white (non-Hispanic) donor race (P=0.002) and respiratory causes of donor death (P600μm. In multivariate analysis, both 1 year CT and cell density were associated with subsequent graft failure (P=0.002 and 0.009). CT increase was modestly associated with endothelial cell loss during follow up (r=-0.29). Conclusion During the first 5 years following penetrating keratoplasty, CT can serve as a predictor of graft survival. However, CT is not a substitute for cell density measurement as both measures were independently predictive of graft failure. PMID:23343949

  2. Corneal endothelium in patients with diabetes mellitus: a historical cohort study.

    Science.gov (United States)

    Shenoy, Radha; Khandekar, Rajeev; Bialasiewicz, Alexander; Al Muniri, Abdullah

    2009-01-01

    Retinopathy is the major cause of ocular morbidity in patients with diabetes mellitus. Chronic hyperglycemia spares no organ and can affect the morphology and function of the various corneal layers, compromising its transparency. This study was conducted to associate the status of corneal endothelium to diabetes mellitus (DM) and identify risk factors of compromised corneal endothelium. A total of 220 eyes of randomly selected patients (110 diabetic and 110 nondiabetic) were subjected to detailed slitlamp and fundus evaluation. Corneal endothelial status was evaluated using the Nidek Confoscan 2. Cell density, percentage polymegathism, and pleomorphism were calculated. The findings in diabetic patients were compared to those without disease. The outcome was correlated to diabetic retinopathy (DR). The effects of hypertension, hyperlipidemia, age, gender, type, duration, glycemic control, and grades of DR was also considered. The mean corneal endothelial cell density was -175 cells/mm2 (95% CI -317 to -33 cells/mm2) less in eyes of diabetic patients. The number of endothelial cells with polymegathism was significantly greater among eyes of diabetic patients. There were less corneal endothelial cells with pleomorphism in nondiabetic patients. Polymegathism and pleomorphism of corneal endothelial cells seems to be positively associated with DM type II. Cell density was significantly lower in eyes with DR than those without DR. Corneal endothelium in diabetic patients seems to be compromised. Evaluation of corneal endothelium should be part of protocol for eye care of diabetic patients.

  3. Glaucoma and Corneal Transplant Procedures

    Directory of Open Access Journals (Sweden)

    Ammar M. Al-Mahmood

    2012-01-01

    Full Text Available Glaucoma after corneal transplantation is a leading cause of ocular morbidity after penetrating keratoplasty. The incidence reported is highly variable and a number of etiologic factors have been identified. A number of treatment options are available; surgical intervention for IOP control is associated with a high incidence of graft failure. IOP elevation is less frequently seen following deep anterior lamellar keratoplasty. Descemet's striping-automated endothelial keratoplasty is also associated with postprocedure intraocular pressure elevation and secondary glaucoma and presents unique surgical challenges in patients with preexisting glaucoma surgeries. Glaucoma exists in up to three-quarters of patients who undergo keratoprosthesis surgery and the management if often challenging. The aim of this paper is to highlight the incidence, etiology, and management of glaucoma following different corneal transplant procedures. It also focuses on the challenges in the diagnosis of glaucoma and intraocular pressure monitoring in this group of patients.

  4. A case of CMV endotheliitis treated with intravitreal ganciclovir injection.

    Science.gov (United States)

    Choi, Won Seok; Cho, Joon Hee; Kim, Ha Kyoung; Kim, Hyun Soo; Shin, Young Joo

    2013-04-01

    We report a case of CMV corneal endotheliitis that was treated with intravitreal ganciclovir injection. A 56-year-old man who has suffered from uveitis was referred to our clinic due to corneal endothelial abnormality. Slit lamp examination showed a localized sectoral corneal edema and linear keratic precipitates along the boundary of edema. In spite of treatment with oral steroid and acyclovir, the disease progressed and two new coin-like lesions were developed. After topical ganciclovir and intavitreal injection of ganciclovir, the corneal lesions disappeared.

  5. Evaluation of the effects of circular Descemet's membrane incision on the biomechanical, topographic and optical properties of rabbit corneas.

    Science.gov (United States)

    Hindman, Holly B; McCally, Russell L; Kim, Alisa; D'Anna, Salvatore E; Eberhart, Charles G; Jun, Albert S

    2011-01-01

      Prospective interventional animal case series to investigate quantitatively changes in corneal light-scattering, corneal hysteresis, keratometry and pachymetry induced by circular Descemet's membrane incision.   Thirty mature New Zealand White rabbits were divided into three study groups: (i) surgical intervention with circular Descemet's incision; (ii) surgical control; and (iii) medical control. Group 1 eyes had two paracenteses placed 120 degrees apart and an 8.5-mm-diameter Descemetorhexis was created with a reverse Sinskey hook. Group 2 eyes had two paracenteses placed 120 degrees apart. The main outcome measures were scatterometry, corneal hysteresis, pachymetry and keratometry measurements, which were performed prior to and 2 weeks following the interventions. Histology and transmission electron microscopy were performed post-mortem in representative eyes.   Eyes that had undergone circular Descemet's incision had significantly decreased mean keratometry (43.9 ± 0.7 dioptres [mean ± standard deviation] preoperatively vs. 43.5 ± 0.9 dioptres postoperatively, P = 0.007). Circular Descemet's membrane incision did not significantly change corneal hysteresis (4.4 ± 1.1 mmHg preoperatively vs. 4.6 ± 0.9 mmHg postoperatively, P = 0.664). Corneal light scattering decreased after Descemet's scoring (0.00254 ± 0.00059 preoperatively vs. 0.00206 ± 0.00031 postoperatively, P = 0.0025). Pachymetry measurements remained relatively stable (341.3 ± 18.6 µm preoperatively vs. 330.6 ± 20.0 µm postoperatively) without postoperative oedema.   Circular Descemet's scoring flattened the corneal curvature by a mean of 0.4 dioptres without affecting corneal hysteresis in rabbit corneas. These findings may have important implications for ongoing developments in endothelial keratoplasty. © 2011 The Authors. Clinical and Experimental Ophthalmology © 2011 Royal Australian and New Zealand College of

  6. Current concepts and techniques in corneal transplantation.

    Science.gov (United States)

    Laibson, Peter R

    2002-08-01

    The corneal endothelium is the most important single layer in corneal transplantation. In his Castroviejo Lecture, William Bourne, MD, summarizes his work on the corneal endothelium and its importance to corneal transplantation. Almost half the corneal transplants performed in the United States are done so because of malfunctioning, diseased, or absent endothelial cells. If just this layer could be transplanted, the long wait for better vision after keratoplasty (up to two years in some countries) can be eliminated, as well as the problems of epithelial and subepithelial graft rejection. The significant astigmatism after keratoplasty could also be reduced. Transplantation of the endothelium in deep lamellar keratoplasty is being done in limited fashion throughout the world and the first patients have now been done in the United States. In many countries where corneal tissue is difficult to obtain, keratoplasty is only performed on one eye, even though both eyes may need it. One article this year discusses binocular vision recovery in bilateral keratoplasty and the objective and subjective improvements after bilateral keratoplasty. Patients who are bilaterally blind from diseases such as Stevens-Johnson syndrome, and ocular pemphigoid have little hope of visual recovery from conventional corneal transplants. The use of a keratoprosthesis to bypass the totally abnormal conjunctival surface has helped many people in the past. The preoperative prognostic categories of patients who may benefit from keratoprosthesis has been carefully reviewed in a large number of keratoprosthesis patients and this information will help ophthalmologists decide who will benefit the most benefit from keratoprosthesis. This year, articles on corneal transplantation after conjunctival flaps, suture-related complications in keratoplasty, the implantation of an intraocular lens after penetrating keratoplasty, and long-term results of penetrating keratoplasty with glaucoma drainage tube

  7. Endothelial replacement: the limbal pocket approach.

    Science.gov (United States)

    Terry, Mark A

    2003-03-01

    The limbal pocket approach to endothelial replacement shows extraordinary promise in fulfilling the ideal goals of surgical treatment for endothelial dysfunction. From its inception, DLEK surgery has demonstrated that, by preserving the limbus and eliminating surface corneal incisions and sutures, the corneal topography can be stabilized with little change from preoperative measurements of astigmatism and corneal power. The surgical dissection planes of DLEK surgery are inherently stronger than that of PK or any other form of endothelial replacement, allowing early stability of the refraction and making the eye safer from blunt trauma over the long term. Further work with DLEK must be performed in perfecting the optical properties of the stromal interface before this technique can be used widely; however, as technology and techniques improve, DLEK surgery holds the promise of being the ideal method for endothelial replacement in the 21st century.

  8. Inflammatory cytokine TNF-α promotes corneal endothelium apoptosis via upregulating TIPE2 transcription during corneal graft rejection.

    Science.gov (United States)

    Wang, Qun; Wei, Chao; Ma, Li; Wang, Xin; Li, Lin; Zhou, Qingjun; Shi, Weiyun

    2018-02-26

    Endothelial dysfunction accounts for 50% of total corneal transplantation failures, suggesting that corneal endothelial damage is the leading cause of graft failure. Tumor necrosis factor-α (TNF-α) is known to contribute to the negative regulation of corneal transplantation, but how it does so remains unclear. Here, we report a regulatory loop involving TNF-α, TNF-α-induced protein 8 like 2 (TNFAIP8L2 or TIPE2), and apoptosis during corneal graft rejection. We established mice models of penetrating keratoplasty to verify whether the quantification of TNF-α in allogeneic corneas is enhanced through ELISA assay and immunofluorescence staining. In cornea tissues, we obtained corneal endothelium and measured apoptosis of the removed cells. Meanwhile, quantitative real-time PCR and Western blotting were used to detect the mRNA and protein expression of TIPE2. In human corneal endothelial cells, we verified the conclusions through some experiments. By specifically knocking down TIPE2, we detected the importance of TIPE2 in TNF-α-triggered apoptosis. In mice models, TNF-α was higher in the cornea and aqueous humor in allograft group and TNF-α elevation increased the apoptosis of the corneal endothelium. In addition, high levels of TIPE2 were found in allograft rejection models following TNF-α elevation. In human corneal endothelial cells (HCECs), TNF-α clearly augments TIPE2 expression and promotes cell apoptosis through upregulating TIPE2 transcription. Knocking down markedly decreased cell apoptosis. Our study identifies the molecular mechanisms underlying the interplay of TNF-α, TIPE2, and apoptosis during allograft rejection, and it suggests that both TNF-α and TIPE2 might be potential targets for the successfully grafted corneal endothelium.

  9. Corneal Transplantation in Children

    OpenAIRE

    Gabrić, N.; Dekaris, I.; Vojniković, B.; Karaman, Ž.; Mravičić, I.; Katušić, J.

    2001-01-01

    The main purpose of the study was to describe the surgical success rate and visual results of penetrating keratoplasty in children. This retrospective study included children that underwent corneal transplantation at the Department of Ophthalmology, General Hospital »Sveti Duh«, in the period 1994–1999. Patients’ age ranged from 6 to 16 years. Twenty-five corneal transplants were performed in 24 eyes. Corneal pathologies were corneal leucoma, congenital dystrophy, corneal combu...

  10. [Nanostructured bioplastic material for traumatic corneal injuries].

    Science.gov (United States)

    Kanyukov, V N; Stadnikov, A A; Trubina, O M; Yakhina, O M

    2015-01-01

    To substantiate the use of nanostructured bioplastic material for the treatment of traumatic eye injuries. The study enrolled 96 eyes of 48 rabbits and was carried out in 3 series of experiments, different in the type of induced corneal trauma: mechanical erosion, alkaline or acid burn. The animals were clinically monitored and sacrificed for morphological investigation at days 3, 7, 14, 30, and 90. The size of mechanical corneal erosions was repeatedly evaluated with fluorescein eye stain test. In the experimental group, Hyamatrix biomaterial was topically administered according to an original technique. In the controls, soft contact lenses were inserted and sutured. Complete closure of the epithelial defect with no impact on corneal properties was achieved in 3 days in the experimental group and in4 days in the control group. As for alkaline and acid corneal burns, experimental and control groups received Hyamatrix biomaterial and Solcoseryl eye gel correspondingly. In the experimental group of alkaline burn the defect closed by day 7, in the controls--by day 10-11. Acid-induced corneal edema also resolved by day 7 in the experimental group and by day 14 in the control group. 1. The results of this experimental and morphological study prove the hyaluronic acid-derived nanostructured bioplastic material effective in accelerating corneal re-epithelialization after mechanical erosions as compared with the controls. 2. Topical application of the hyaluronic acid-derived nanostructured bioplastic material shortens the exudative phase of inflammation, promotes corneal defect closure with formation of a more subtle opacification, and stimulates corneal restoration after chemical burns.

  11. Influence of corneal astigmatism, corneal curvature and meridional differences on corneal hysteresis and corneal resistance factor.

    Science.gov (United States)

    Wong, Yin-zhi; Lam, Andrew K C

    2011-09-01

    This study investigated the influence of corneal astigmatism, corneal curvature and meridional differences on corneal hysteresis (CH) and the corneal resistance factor (CRF) in a group of normal Chinese persons. Ninety-five participants were recruited and data from the eye with higher corneal astigmatism were analysed. The anterior corneal curvature was measured by corneal topography. The Goldmann-correlated intraocular pressure (IOPg), corneal-compensated intraocular pressure (IOPcc), CH and CRF at different meridians (default horizontal position, 10°, 20° and 30° along the superotemporal and inferonasal meridians) were obtained from an ocular response analyser. The corneal powers at these specific meridians also were calculated. At the default position, the IOPg and CRF had weak correlations with corneal astigmatism, while the IOPcc and CH were not significantly correlated with corneal astigmatism. Both the IOPg and IOPcc were measured significantly higher at the default position. The CH and CRF were lower at the default position but the difference in the CRF from obliquity could not reach statistical significance. The CH was not significantly correlated with the corneal power at all meridians. The CRF correlated with the corneal power only at 30° superotemporal. Corneal astigmatism and head tilt did not have much effect on the measurement of CH and the CRF, both of which were lowest along the horizontal meridian. Clinically, the difference was small. The influence of corneal power on CH and the CRF was minimal. © 2011 The Authors. Clinical and Experimental Optometry © 2011 Optometrists Association Australia.

  12. Quantitative evaluation of the corneal endothelium in the mouse after grafting.

    Science.gov (United States)

    Plskova, J; Kuffova, L; Filipec, M; Holan, V; Forrester, J V

    2004-09-01

    Corneal graft survival depends critically on the quality of the endothelium. In this study the authors aimed to evaluate corneal endothelium in mice at different times after transplantation and to correlate endothelial integrity with corneal graft survival. Syngeneic and allogeneic corneal grafts at various times (days 0-60) after engraftment were examined in flat mount preparation by confocal microscopy, by evaluating the hexagonal pattern of the endothelial monolayer using actin staining of the cell cortex. Corneas from untreated mice and from mice, who were grafted after removal of draining lymph nodes served as controls. In control corneas, more than 90% of the posterior surface was covered by endothelium. Syngeneic grafts were always covered by 54-99% of endothelium. In contrast, the posterior surface of corneal allografts showed great variation in the degree of endothelial cell coverage (0-98%). In addition, clinical opacity grading measure was not a reliable predictor of endothelial coverage. In corneal allografts there is progressive loss of endothelium over time, unlike with syngeneic grafts. However, in the early stages of allograft rejection, the grade of graft opacity does not accurately reflect the degree of endothelial cell coverage. Although corneal opacity grade is considered the main determinant of graft rejection, the data suggest that both the grade of corneal opacity plus a sufficient post-graft time duration (>8 weeks in the mouse) are required for the diagnosis of irreversible graft rejection.

  13. Reliability of the Effect of Artificial Anterior Chamber Pressure and Corneal Drying on Corneal Graft Thickness.

    Science.gov (United States)

    Romano, Vito; Steger, Bernhard; Chen, Jern Y; Hassaan, Sherif; Batterbury, Mark; Willoughby, Colin E; Ahmad, Sajjad; Elsheikh, Ahmed; Kaye, Stephen B

    2015-08-01

    To investigate the effect of artificial anterior chamber (AAC) pressure and corneal drying on the graft thickness in preparation for Descemet stripping automated endothelial keratoplasty. Twenty-seven corneoscleral discs were placed in an AAC. The AAC pressure (15, 45, 92, 109, and 198 mm Hg) was controlled using the height of an infusion bottle and a roller clamp. The endothelium was removed in 1 subgroup. Corneas were exposed to room air or repeatedly dried using cellulose spears. Central corneal thickness was measured every 90 seconds for the first 15 minutes and again at 20 minutes using an ultrasound pachymeter (SP-100, Tomey). There was a significant linear relationship between the corneal thickness and both AAC pressure and corneal drying. Very high coefficients of determination and narrow 95% confidence intervals were present, in particular for high pressures and drying. The rate of thinning increased with increasing pressure and drying to 1.6% per minute. At the maximum rate of thinning, a 10% reduction in corneal thickness occurred in 6 minutes or 100 μm in 8.8 minutes. Removal of the corneal endothelium reduced the rate of thinning to 0.3% per minute (R = 0.72). Increasing AAC pressure and corneal drying reduced the graft thickness at a very predictable rate. Adequate corneal thinning can be achieved by increasing the pressure in the AAC by closing the clamp followed by removal of the residual corneal epithelium and repeated drying with a cellulose spear for 5 to 10 minutes, depending on the initial corneal thickness. This method is simple and is both suitable for use in the eye bank and by the surgeon.

  14. Riboflavin concentration in corneal stroma after intracameral injection

    Directory of Open Access Journals (Sweden)

    Na Li

    2015-06-01

    Full Text Available AIM:To evaluate the enrichment of riboflavin in the corneal stroma after intracameral injection to research the barrier ability of the corneal endothelium to riboflavin in vivo.METHODS:The right eyes of 30 New Zealand white rabbits were divided into three groups. Different concentrations riboflavin-balanced salt solutions (BSS were injected into the anterior chamber (10 with 0.5%, 10 with 1%, and 10 with 2%. Eight corneal buttons of 8.5 mm in diameter from each group were dissected at 30min after injection and the riboflavin concentrations in the corneal stroma were determined using high-performance liquid chromatography (HPLC after removing the epithelium and endothelium. The other two rabbits in every group were observed for 24h and sacrificed. As a comparison, the riboflavin concentrations from 16 corneal stromal samples were determined using HPLC after instillation of 0.1% riboflavin-BSS solution for 30min on the corneal surface (8 without epithelium and 8 with intact epithelium.RESULTS: The mean riboflavin concentrations were 11.19, 18.97, 25.08, 20.18, and 1.13 µg/g for 0.5%, 1%, 2%, de-epithelialzed samples, and the transepithelial groups, respectively. The color change of the corneal stroma and the HPLC results showed that enrichment with riboflavin similar to classical de-epithelialized corneal collagen crosslinking (CXL could be achieved by intracameral1% riboflavin-BSS solution after 30min; the effect appeared to be continuous for at least 30min.CONCLUSION:Riboflavin can effectively penetrate the corneal stroma through the endothelium after an intracameral injection in vivo, so it could be an enhancing method that could improve the corneal riboflavin concentration in transepithelial CXL.

  15. Value of recombinant human epidermal growth factor in corneal wound repair after corneal foreign body elimination

    Directory of Open Access Journals (Sweden)

    Hong-Jie Han

    2013-11-01

    Full Text Available AIM: To investigate the repair efficacy of recombinant human epidermal growth factor on corneal epithelium after corneal foreign body eliminating operation. METHODS: There were 102 patients with corneal foreign body(188 affected eyeschosen for the study. All patients were divided into treatment group and control group according to the random number table. Both groups received corneal foreign body elimination by slit lamp. Postoperatively, the treatment group was given eye drops containing epidermal growth factor(JinYinShucombined with tobramycin while the control group was only administrated with tobramycin. Treatment effects were compared 3d after treatment. RESULTS: Three days after treatment, the cure rate in the treatment group(93.7%, was significantly higher than that in the control group(76.6%(PPCONCLUSION: The recombinant human epidermal growth factor is capable of integrating with corneal epithelial cells and endothelial cell receptor, shortening healing time of corneal epithelial wound, thus making it an effective treatment of traumatic corneal epithelial defect.

  16. Kv3.3 potassium channels in lens epithelium and corneal endothelium.

    Science.gov (United States)

    Rae, J L; Shepard, A R

    2000-03-01

    Human Kv3.3/KCNC3 is a Shaw-type potassium channel that has been mapped to chromosome 19q13.3-13.4. Complete mouse and rat Kv3.3 cDNA coding sequences have been published, yet the human Kv3.3 cDNA has remained incomplete for years. We report here for the first time the amino acid sequence for hKv3.3 and the electrophysiological behavior of the encoded channel in transiently transfected mammalian cells. In addition, we report the occurrence of Kv3.3 message in rabbit corneal endothelial cells and the properties of the currents when the corneal channel is expressed. The hKv3.3 gene is highly GC-rich (69%) and contains numerous GC runs which made DNA sequencing and PCR amplification especially problematic. The full-length sequence contains two possible start codons. The encoded 757 amino acid hKv3. 3 protein is about 93% identical to mouse and rat Kv3.3 in the first 659 amino acids before the C-terminal domains diverge greatly as a result of alternative splicing. The rabbit cornea Kv3.3 is a close sequence match for hKv3.3 even in the C-terminal domain. However, we have not yet found a cornea sequence which contains the first potential start codon from hKv3.3. Electrophysiologically, the hKv3. 3 channel produces an A-current although expression of constructs which lack the 5' region of the first start codon inactivate much more slowly than full-length constructs. This short hKv3.3 construct also shows changes in activation. Copyright 2000 Academic Press.

  17. Suppression of inflammatory corneal lymphangiogenesis by application of topical corticosteroids.

    Science.gov (United States)

    Hos, Deniz; Saban, Daniel R; Bock, Felix; Regenfuss, Birgit; Onderka, Jasmine; Masli, Sharmila; Cursiefen, Claus

    2011-04-01

    To analyze whether topical application of corticosteroids inhibits inflammatory corneal lymphangiogenesis and to study the potential underlying antilymphangiogenic mechanisms. Inflammatory corneal neovascularization was induced by suture placement, and the corneas were then treated with topical fluorometholone, prednisolone acetate, or dexamethasone sodium phosphate. After 1 week, the corneas were stained with lymphatic vessel endothelial hyaluronan receptor 1 for detection of pathological corneal lymphangiogenesis. The effect of these corticosteroids on macrophage recruitment was assessed via fluorescence-activated cell sorting analysis. The effect of these corticosteroids on proinflammatory cytokine expression by peritoneal exudate cells was tested via real-time polymerase chain reaction. Furthermore, the effect of steroid treatment on the proliferation of lymphatic endothelial cells was assessed via enzyme-linked immunosorbent assay. Treatment with corticosteroids resulted in a significant reduction of inflammatory corneal lymphangiogenesis. The antilymphangiogenic effect of fluorometholone was significantly weaker than that of prednisolone and dexamethasone. Corneal macrophage recruitment was also significantly inhibited by the application of topical steroids. Treatment of peritoneal exudate cells with corticosteroids led to a significant downregulation of the RNA expression levels of tumor necrosis factor and interleukin 1β. Additionally, proliferation of lymphatic endothelial cells was also inhibited. Corticosteroids are strong inhibitors of inflammatory corneal lymphangiogenesis, with significant differences between various corticosteroids in terms of their antilymphangiogenic potency. The main mechanism of the antilymphangiogenic effect seems to be through the suppression of macrophage infiltration, proinflammatory cytokine expression, and direct inhibition of proliferation of lymphatic endothelial cells. Steroids block corneal lymphangiogenesis, the main

  18. Endothelial keratoplasty: evolution and horizons

    Directory of Open Access Journals (Sweden)

    Gustavo Teixeira Grottone

    2012-12-01

    Full Text Available Endothelial keratoplasty has been adopted by corneal surgeons worldwide as an alternative to penetrating keratoplasty (PK in the treatment of corneal endothelial disorders. Since the first surgeries in 1998, different surgical techniques have been used to replace the diseased endothelium. Compared with penetrating keratoplasty, all these techniques may provide faster and better visual rehabilitation with minimal change in refractive power of the transplanted cornea, minimal induced astigmatism, elimination of suture-induced complications and late wound dehiscence, and a reduced demand for postoperative care. Translational research involving cell-based therapy is the next step in work on endothelial keratoplasty. The present review updates information on comparisons among different techniques and predicts the direction of future treatment.

  19. Corneal Biomechanical Properties after FS-LASIK with Residual Bed Thickness Less Than 50% of the Original Corneal Thickness

    Directory of Open Access Journals (Sweden)

    Haixia Zhang

    2018-01-01

    Full Text Available Background. The changes in corneal biomechanical properties after LASIK remain an unknown but important topic for surgical design and prognostic evaluation. This study aims to observe the postoperative corneal biomechanical properties one month after LASIK with amount of corneal cutting (ACC greater than 50% of the central corneal thickness (CCT. Methods. FS-LASIK was performed in 10 left rabbit eyes with ACC being 60% (L60 and 65% (L65 of the CCT, while the right eyes (R were the control. After 4 weeks, rabbits were executed and corneal strip samples were prepared for uniaxial tensile tests. Results. At the same strain, the stresses of L65 and L60 were larger than those of R. The elastic moduli of L60 and L65 were larger than those of R when the stress was 0.02 MPa, while they began to be less than those of R when stress exceeds the low-stress region. After 10 s relaxation, the stress of specimens L65, L60, and R increased in turn. Conclusion. The elastic moduli of the cornea after FS-LASIK with ACC greater than 50% of the CCT do not become less under normal rabbit IOP. The limit stress grows with the rise of ACC when relaxation becomes stable.

  20. Damage Threshold of In Vivo Rabbit Cornea by 2 micron Laser Irradiation

    National Research Council Canada - National Science Library

    Chen, Bo; Oliver, Jeffery; Dutta, Soumak; Rylander, III, Grady H; Thomsen, Sharon L; Welch, Ashley J

    2007-01-01

    To support refinement of the Maximum Permissible Exposure (MPE) safety limits, a series of experiments were conducted in vivo on Dutch Belted rabbit corneas to determine corneal minimum visible lesion thresholds...

  1. Graft Biomechanics Following Three Corneal Transplantation Techniques

    OpenAIRE

    Sepehr Feizi; Talieh Montahai; Hamidreza Moein

    2015-01-01

    Purpose: To compare corneal biomechanical properties following three different transplantation techniques, including Descemet stripping automated endothelial keratoplasty (DSAEK), deep anterior lamellar keratoplasty (DALK) and penetrating keratoplasty (PK) in comparison to normal eyes. Methods: This cross-sectional comparative study included 118 eyes: 17 eyes of 17 patients received DSAEK, 23 eyes of 21 patients underwent DALK using Anwar's big bubble technique, and 45 eyes of 36 patients ...

  2. Biocompatibility of poly(ethylene glycol) and poly(acrylic acid) interpenetrating network hydrogel by intrastromal implantation in rabbit cornea

    OpenAIRE

    Zheng, Luo Luo; Vanchinathan, Vijay; Dalal, Roopa; Noolandi, Jaan; Waters, Dale J.; Hartmann, Laura; Cochran, Jennifer R.; Frank, Curtis W.; Yu, Charles Q.; Ta, Christopher N.

    2015-01-01

    We evaluated the biocompatibility of a poly(ethylene glycol) and poly(acrylic acid) (PEG/PAA) interpenetrating network hydrogel designed for artificial cornea in a rabbit model. PEG/PAA hydrogel measuring 6 mm in diameter was implanted in the corneal stroma of twelve rabbits. Stromal flaps were created with a microkeratome. Randomly, six rabbits were assigned to bear the implant for 2 months, two rabbits for 6 months, two rabbits for 9 months, one rabbit for 12 months, and one rabbit for 16 m...

  3. Adherens junction proteins are expressed in collagen corneal equivalents produced in vitro with human cells

    OpenAIRE

    Giasson, Claude J.; Deschambeault, Alexandre; Carrier, Patrick; Germain, Lucie

    2014-01-01

    Purpose To test whether adherens junction proteins are present in the epithelium and the endothelium of corneal equivalents. Methods Corneal cell types were harvested from human eyes and grown separately. Stromal equivalents were constructed by seeding fibroblasts into a collagen gel on which epithelial and endothelial cells were added on each side. Alternatively, bovine endothelial cells were used. At maturity, sections of stromal equivalents were processed for Masson's trichrome or indirect...

  4. Comparison of confocal biomicroscopy and noncontact specular microscopy for evaluation of the corneal endothelium.

    Science.gov (United States)

    Hara, Makiko; Morishige, Naoyuki; Chikama, Tai-Ichiro; Nishida, Teruo

    2003-08-01

    To compare the clinical efficacy of confocal biomicroscopy with that of noncontact specular microscopy for the evaluation of the corneal endothelium. The corneal endothelium was examined in 14 normal subjects (28 eyes) and in 6 patients (11 eyes) with Fuchs corneal endothelial dystrophy using a noncontact specular microscope (SP-2000P, Topcon, Japan) and a confocal biomicroscope (ConfoScan, Tomey, Japan). The images and the calculated densities of corneal endothelial cells obtained by the 2 techniques were compared. For normal subjects, the images of corneal endothelial cells obtained by the 2 techniques were almost identical, although the density of these cells determined by confocal biomicroscopy (2916 +/- 334 cells/mm2) was slightly higher than that determined by noncontact specular microscopy (2765 +/- 323 cells/mm2). In contrast, whereas clear images of corneal endothelial cells, allowing the determination of cell density, were obtained for all 11 eyes of the patient group by confocal biomicroscopy, clear images were obtained for only 4 of these 11 eyes (36.4%) by noncontact specular microscopy. Both noncontact specular microscopy and confocal biomicroscopy revealed the shapes and number of endothelial cells in the normal cornea. However, for corneas with Fuchs dystrophy, clear images were obtained only by confocal biomicroscopy. Confocal biomicroscopy is thus an effective tool for evaluation of the diseased corneal endothelium.

  5. Corneal collagen cross-linking outcomes: review.

    Science.gov (United States)

    Jankov Ii, Mirko R; Jovanovic, Vesna; Delevic, Sladjana; Coskunseven, Efekan

    2011-02-11

    Keratoconus is a condition characterized by biomechanical instability of the cornea, presenting in a progressive, asymmetric and bilateral way. Corneal collagen cross-linking with riboflavin and UVA (CXL) is a new technique of corneal tissue strengthening that combines the use of riboflavin as a photo sensitizer and UVA irradiation. The studies showed that CXL was effective in halting the progression of keratoconus over a period of up to four years. The published studies also revealed a reduction of max K readings by more than 2 D, while the postoperative SEQ was reduced by an average of more than 1 D, and refractive cylinder decreased by about 1 D. No eyes lost any line of BCDVA. Moreover, there was no significant decrease in endothelial cell density. It was also found that CXL treatment was effective with reducing corneal and total wavefront aberrations. Corneal cross-linking has also led to an arrest and/or even a partial reversal of keratectasia in the treatment of iatrogenic ectasia after excimer laser ablation. A primary intervention such as CXL should be considered to potentially increase the biomechanical stability of the corneal tissue and postpone the need of lamellar or penetrating keratoplasty.

  6. MicroRNA-184 Regulates Corneal Lymphangiogenesis.

    Science.gov (United States)

    Grimaldo, Sammy; Yuen, Don; Theis, Jaci; Ng, Melissa; Ecoiffier, Tatiana; Chen, Lu

    2015-11-01

    MicroRNAs are a class of small noncoding RNAs that negatively regulate gene expression by binding to complimentary sequences of target messenger RNA. Their roles in corneal lymphangiogenesis are largely unknown. This study was to investigate the specific role of microRNA-184 (mir-184) in corneal lymphangiogenesis (LG) in vivo and lymphatic endothelial cells (LECs) in vitro. Standard murine suture placement model was used to study the expressional change of mir-184 in corneal inflammatory LG and the effect of synthetic mir-184 mimic on this process. Additionally, a human LEC culture system was used to assess the effect of mir-184 overexpression on cell functions in vitro. Expression of mir-184 was significantly downregulated in corneal LG and, accordingly, its synthetic mimic suppressed corneal lymphatic growth in vivo. Furthermore, mir-184 overexpression in LECs inhibited their functions of adhesion, migration, and tube formation in vitro. These novel findings indicate that mir-184 is involved critically in LG and potentially could be used as an inhibitor of the process. Further investigation holds the promise for divulging new therapies for LG disorders, which occur inside and outside the eye.

  7. In-vitro corneal transparency measuring system

    Science.gov (United States)

    Ventura, Liliane; da Costa Vieira, Marcelo A.; Isaac, Flavio; Chiaradia, Caio; Faria de Sousa, Sidney J.

    2001-06-01

    A system for measuring the average corneal transparency of preserved corneas has been developed in order to provide a more accurate and standard report of the corneal tissue. The donated cornea transparency is one of the features to be analyzed previously to its indication for the transplant. The small portable system consists of two main parts: the optical and the electronic parts. The optical system consists of a white light, lenses and pin-holes that collimate white light beams that illuminates the cornea in its preservative medium. The light that passes through the cornea is detected by a resistive detector and the average corneal transparency is shown in a display. In order to obtain just the tissue transparency, the electronic circuit was built in a way that there is a baseline input of the preservative medium, previous to the measurement of the corneal transparency. Manipulating the system consists of three steps: (1) Adjusting the zero percentage in the absence of light (at this time the detectors in the dark); (2) Placing the preservative medium in the system and adjusting the 100% value (this is the baseline input); (3) Preserving the cornea and placing it in the system. The system provides the tissue transparency. The system is connected to an endothelium evaluation system for Slit Lamp, that we have developed, and statistics about the relationship of the corneal transparency and density of the endothelial cells will be provided in the next years. The system is being used in a public Eye Bank in Brasil.

  8. Human tears reveal insights into corneal neovascularization.

    Science.gov (United States)

    Zakaria, Nadia; Van Grasdorff, Sigi; Wouters, Kristien; Rozema, Jos; Koppen, Carina; Lion, Eva; Cools, Nathalie; Berneman, Zwi; Tassignon, Marie-José

    2012-01-01

    Corneal neovascularization results from the encroachment of blood vessels from the surrounding conjunctiva onto the normally avascular cornea. The aim of this study is to identify factors in human tears that are involved in development and/or maintenance of corneal neovascularization in humans. This could allow development of diagnostic tools for monitoring corneal neovascularization and combination monoclonal antibody therapies for its treatment. In an observational case-control study we enrolled a total of 12 patients with corneal neovascularization and 10 healthy volunteers. Basal tears along with reflex tears from the inferior fornix, superior fornix and using a corneal bath were collected along with blood serum samples. From all patients, ocular surface photographs were taken. Concentrations of the pro-angiogenic cytokines interleukin (IL)-6, IL-8, Vascular Endothelial Growth Factor (VEGF), Monocyte Chemoattractant Protein 1 (MCP-1) and Fas Ligand (FasL) were determined in blood and tear samples using a flow cytometric multiplex assay. Our results show that the concentration of pro-angiogenic cytokines in human tears are significantly higher compared to their concentrations in serum, with highest levels found in basal tears. Interestingly, we could detect a significantly higher concentration of IL- 6, IL-8 and VEGF in localized corneal tears of patients with neovascularized corneas when compared to the control group. This is the first study of its kind demonstrating a significant difference of defined factors in tears from patients with neovascularized corneas as compared to healthy controls. These results provide the basis for future research using animal models to further substantiate the role of these cytokines in the establishment and maintenance of corneal neovascularization.

  9. Ocular tolerance in rabbits after intracameral administration of a fixed combination of tropicamide, phenylephrine, and lidocaine with and without rinsing.

    Science.gov (United States)

    Nuijts, Rudy M M A; Mencucci, Rita; Viaud-Quentric, Karen; Elena, Pierre-Paul; Olmière, Céline; Behndig, Anders

    2017-05-01

    To evaluate the safety and tolerability of a single intracameral administration of a combined mydriatic (tropicamide and phenylephrine) and anesthetic (lidocaine) formulation (Mydrane) with or without rinsing. Iris Pharma, La Gaude, France. Experimental study. Sixty pigmented rabbits received 100 μL or 200 μL of the combination product or a placebo (sodium chloride 0.9%) by intracameral injection. For the combination product, separate groups were included with and without rinsing after administration. From day 1 day to day 7, assessments included general clinical and ocular observations, pupil diameter measurements, corneal assessments, confocal microscopy, and electroretinography (ERG). Necropsy examinations were performed at study completion at day 8. Rapid mydriasis, stable 24 minutes after injection and returning to baseline levels by day 1, was induced in all groups that received the combination mydriatic and anesthetic drug. Rinsing had no effect. The combination product induced no adverse effects on the anterior or posterior segment of the eye (ie, no increased corneal thickness and endothelial cell loss, no abnormalities in ERG). Slitlamp examination showed slightly increased anterior chamber inflammation with rinsing in both the study group and placebo group. This observation was not confirmed by aqueous flare examination. No toxic effects of the products were found on histological evaluation. The combination mydriatic and anesthetic drug administered to pigmented rabbits as a single intracameral injection at volumes of 100 μL and 200 μL was well tolerated with no ocular adverse effects and no effect on the corneal endothelium. Copyright © 2017 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

  10. Equine corneal stromal abscesses

    DEFF Research Database (Denmark)

    Henriksen, M. D. L.; Andersen, P. H.; Plummer, C. E.

    2013-01-01

    The last 30 years have seen many changes in the understanding of the pathogenesis and treatment of equine corneal stromal abscesses (SAs). Stromal abscesses were previously considered an eye problem related to corneal bacterial infection, equine recurrent uveitis, corneal microtrauma and corneal...... foreign bodies in horses. They were more commonly diagnosed in horses living in subtropical climatic areas of the world. Therapeutic recommendations to treat equine SAs were historically nearly always a medical approach directed at bacteria and the often associated severe iridocyclitis. Today...... the pathogenesis of most equine SAs appears to be more often related to fungal inoculation of the anterior corneal stroma followed by posterior migration of the fungi deeper into the corneal stroma. There is also now an increased incidence of diagnosis of corneal SAs in horses living in more temperate climates...

  11. Corneal transplantation in children.

    Science.gov (United States)

    Gabrić, N; Dekaris, I; Vojniković, B; Karaman, Z; Mravicić, I; Katusić, J

    2001-01-01

    The main purpose of the study was to describe the surgical success rate and visual results of penetrating keratoplasty in children. This retrospective study included children that underwent corneal transplantation at the Department of Ophthalmology, General Hospital "Sveti Duh", in the period 1994-1999. Patients' age ranged from 6 to 16 years. Twenty-five corneal transplants were performed in 24 eyes. Corneal pathologies were corneal leucoma, congenital dystrophy, corneal combustion, corneal scar after perforating injury, keratoconus, corneal melting, hematocornea and rekeratoplasty. The follow-up period was at least 6 months. The rate of graft survival was 1 year in 75% of eyes with congenital dystrophy and keratoconus. Hematocornea and rekeratoplasty ended with graft failure. Postoperative visual acuity improvement was recorded in 14 out of 25 eyes. Penetrating keratoplasty in children showed very good surgical success. The final visual outcome was affected by irreversible amblyopia.

  12. Ways to Improve (Visual) Outcome in Corneal Transplantation, Corneal Pathology

    NARCIS (Netherlands)

    M.C. Bartels (Marjolijn)

    2005-01-01

    textabstractThe normally transparent cornea can lose its ability to refract light regularly from various conditions. Among these conditions are corneal opacities and corneal diseases leading to a distortion of the corneal contour. Vision might be restored by a corneal transplantation. Corneal

  13. Intraoperative corneal thickness change and clinical outcomes after corneal collagen crosslinking: Standard crosslinking versus hypotonic riboflavin.

    Science.gov (United States)

    Rosenblat, Elan; Hersh, Peter S

    2016-04-01

    To determine intraoperative changes in corneal thickness and outcomes of corneal collagen crosslinking (CXL) using 2 intraoperative regimens: riboflavin-dextran or hypotonic riboflavin. Cornea and refractive surgery practice, Teaneck, New Jersey, USA. Prospective randomized case series. Eyes with keratoconus or corneal ectasia were treated. All eyes received preloading with riboflavin 0.1% in 20% dextran. During ultraviolet-A (UVA) exposure, patients were randomly assigned to 1 of 2 study arms; that is, riboflavin-dextran or hypotonic riboflavin. Intraoperative pachymetry was measured before and after the corneal epithelium was removed, after initial riboflavin loading, and after UVA light exposure. Patients were evaluated for maximum keratometry (K), uncorrected distance visual acuity (UDVA), corrected distance visual acuity, corneal thickness, and endothelial cell count (ECC). Forty-eight eyes were treated. After removal of the epithelium and riboflavin loading, the mean pachymetry was 430 μm and 432 μm in the standard group and hypotonic group, respectively. Immediately after 30-minute UVA administration, the mean pachymetry was 302 μm and 342 μm, respectively. There was no statistically significant difference in the postoperative maximum K change, UDVA, corneal thickness, or ECC between the 2 groups. The cornea thinned substantially during the CXL procedure. The use of hypotonic riboflavin rather than riboflavin-dextran during UVA administration decreased the amount of corneal thinning during the procedure by 30%, from 128 μm to 90 μm. However, there were no significant differences in clinical efficacy or changes in ECC or function between groups postoperatively. In general, corneal thinning during CXL did not seem to compromise the safety of the endothelium. Dr. Hersh is a consultant to Avedro, Inc. Dr. Rosenblat has no financial or proprietary interest in any material or method mentioned. Copyright © 2016 ASCRS and ESCRS. Published by Elsevier

  14. Corneal collagen crosslinking for keratoconus. A review

    Directory of Open Access Journals (Sweden)

    M. M. Bikbov

    2014-01-01

    Full Text Available Photochemical crosslinking is widely applied in ophthalmology. Its biochemical effect is due to the release of singlet oxygen that promotes anaerobic photochemical reaction. Keratoconus is one of the most common corneal ectasia affecting 1 in 250 to 250 000 persons. Currently, the rate of iatrogenic ectasia following eximer laser refractive surgery increases due to biomechanical weakening of the cornea. Morphologically and biochemically, ectasia is characterized by corneal layers thinning, contact between the stroma and epithelium resulting from Bowman’s membrane rupture, chromatin fragmentation in keratocyte nuclei, phagocytosis, abnormal staining and arrangement of collagen fibers, enzyme system disorders, and keratocyte apoptosis. In corneal ectasia, altered enzymatic processes result in the synthesis of abnormal collagen. Collagen packing is determined by the activity of various extracellular matrix enzymes which bind amines and aldehydes of collagen fiber amino acids. In the late stage, morphological changes of Descemet’s membrane (i.e., rupture and detachment develop. Abnormal hexagonal-shaped keratocytes and their apoptosis are the signs of endothelial dystrophy. The lack of analogs in domestic ophthalmology encouraged the scientists of Ufa Eye Research Institute to develop a device for corneal collagen crosslinking. The parameters of ultraviolet (i.e., wavelength, exposure time, power to achieve the desired effect were identified. The specifics of some photosensitizers in the course of the procedure were studied. UFalink, a device for UV irradiation of cornea, and photosensitizer Dextralink were developed and adopted. Due to the high risk of endothelial damage, this treatment is contraindicated in severe keratoconus (CCT less than 400 microns. Major effects of corneal collagen crosslinking are the following: Young’s modulus (modulus of elasticity increase by 328.9 % (on average, temperature tolerance increase by 5

  15. Corneal collagen crosslinking for keratoconus. A review

    Directory of Open Access Journals (Sweden)

    M. M. Bikbov

    2014-10-01

    Full Text Available Photochemical crosslinking is widely applied in ophthalmology. Its biochemical effect is due to the release of singlet oxygen that promotes anaerobic photochemical reaction. Keratoconus is one of the most common corneal ectasia affecting 1 in 250 to 250 000 persons. Currently, the rate of iatrogenic ectasia following eximer laser refractive surgery increases due to biomechanical weakening of the cornea. Morphologically and biochemically, ectasia is characterized by corneal layers thinning, contact between the stroma and epithelium resulting from Bowman’s membrane rupture, chromatin fragmentation in keratocyte nuclei, phagocytosis, abnormal staining and arrangement of collagen fibers, enzyme system disorders, and keratocyte apoptosis. In corneal ectasia, altered enzymatic processes result in the synthesis of abnormal collagen. Collagen packing is determined by the activity of various extracellular matrix enzymes which bind amines and aldehydes of collagen fiber amino acids. In the late stage, morphological changes of Descemet’s membrane (i.e., rupture and detachment develop. Abnormal hexagonal-shaped keratocytes and their apoptosis are the signs of endothelial dystrophy. The lack of analogs in domestic ophthalmology encouraged the scientists of Ufa Eye Research Institute to develop a device for corneal collagen crosslinking. The parameters of ultraviolet (i.e., wavelength, exposure time, power to achieve the desired effect were identified. The specifics of some photosensitizers in the course of the procedure were studied. UFalink, a device for UV irradiation of cornea, and photosensitizer Dextralink were developed and adopted. Due to the high risk of endothelial damage, this treatment is contraindicated in severe keratoconus (CCT less than 400 microns. Major effects of corneal collagen crosslinking are the following: Young’s modulus (modulus of elasticity increase by 328.9 % (on average, temperature tolerance increase by 5

  16. Deep lamellar endothelial keratoplasty (DLEK): pursuing the ideal goals of endothelial replacement.

    Science.gov (United States)

    Terry, M A

    2003-11-01

    Endothelial dysfunction is a leading cause of corneal vision loss and treatment requires surgical replacement with donor endothelium. Standard penetrating keratoplasty (PK) suffers from the inherent problems of surface corneal incisions and sutures and poor wound healing of vertical stromal wounds. This often results in high irregular astigmatism, unpredictable corneal power, and the risk of long-term visual loss from suture-induced vascularization, ulceration, rejection, and late wound rupture. This paper delineates five ideal goals of endothelial replacement, which include: (1) a smooth surface topography without significant change in astigmatism from preoperative to postoperative; (2) a highly predictable and stable corneal power; (3) a healthy donor endothelium that resolves all oedema; (4) a tectonically stable globe, safe from injury and infection; and (5) an optically pure cornea. Deep lamellar endothelial keratoplasty (DLEK) is a surgical method of endothelial replacement that is performed through a limbal scleral incision that leaves the surface of the recipient cornea untouched. The early results of this innovative surgery are discussed and compared to the results of PK in terms of fulfillment of the five ideal goals of endothelial replacement. With further refinement of interface creation, DLEK surgery may be the ideal method for endothelial replacement.

  17. [Corneal trophism and sensitivity changes after penetrating keratoplasty].

    Science.gov (United States)

    Javaloy Estañ J; Aracil Marco A; Belmonte Martínez J; Gallar, J

    2000-09-01

    To evaluate in the rabbit eye the changes in mechanical sensitivity and trophic function of the corneal epithelium after the denervation consecutive to circular trephinations of the peripheral cornea, similar to those produced by penetrating keratoplasty. Complete trephination (8 mm diameter) with rotation of corneal button (PK) was performed in 12 eyes. Non-penetrating trephination, affecting 2/3 of the corneal depth, was made in a group of 12 eyes (NPK). A separate group of 16 nonoperated eyes served as control. Mechanical corneal sensitivity was measured with the Cochet-Bonnet esthesiometer (No. 12 filament). Trophic function of the corneal epithelium was assessed by determining the healing rate of epithelial wounds performed with n-heptanol. Three months after surgery, sensitivity in the center of the cornea was significantly reduced in both PK and NPK eyes (240+/-0 mg/S and 179+/-20 mg/S, respectively) when compared with control (88+/-10 mg/S, pPK and NPK eyes. Results suggest that three months after corneal graft, throphic maintenance of the epithelium remains unaffected in spite of the significant hypoesthesia recorded at this time. Further experiments are needed to analyze the interaction between corneal epithelial cells and the trigeminal neurons innervating the cornea

  18. Preparation and Biomechanical Properties of an Acellular Porcine Corneal Stroma.

    Science.gov (United States)

    Li, Qing; Wang, Hongmei; Dai, Zhenye; Cao, Yichen; Jin, Chuanyu

    2017-11-01

    To construct an acellular porcine corneal stroma (aPCS) as a human corneal stroma alternative and to further explore its biomechanical properties. A combination of DNA-RNA enzymes and ultrasound technology was used to strip the native porcine corneal cells. The microstructure of aPCS was observed by H&E staining, DAPI staining, and α-Gal tests. The mechanical properties were detected by a tension machine. Cytotoxicity of aPCS was measured by the MTT assay. The subcutaneous embedding experiment in rats was also used to detect immunity and degradation. The aPCS was transplanted into the rabbit cornea by lamellar keratoplasty, general observations were made at 3 days, 1 week, 1 month, and 3 months after implantation, respectively. The microstructure and mechanical properties of aPCS were not damaged during the decellularization process. The aPCS extracts had no significant cytotoxicity on human corneal stroma cells. Moreover, the subcutaneous embedding experiment in rats demonstrated that aPCS could not be degraded and induced no immune reaction in and around the transplanted discs. More important is that the aPCS reconstructed normal corneal stroma and maintained corneal transparency and thickness, with almost no neovascularization and inflammation at 3 months after surgery. The aPCS prepared in this study had good biocompatibility, safety, and low antigenicity, which has great potential for corneal disease treatment.

  19. Minimizing the risk of disease transmission during corneal tissue processing.

    Science.gov (United States)

    Lindquist, Thomas D; Miller, Thomas D; Elsen, Jennifer L; Lignoski, Paul J

    2009-06-01

    Corneal transplantation is undergoing significant change because the dysfunctional portion of the cornea may now be selectively transplanted. After recovery of corneoscleral tissue, further processing of such tissue as in microkeratome preparation of endothelial keratoplasty lenticules is defined as "open-container processing" by the Eye Bank Association of America. Airborne bacterial contamination during preparation of corneal tissue is a potential source of postoperative infection. This review addresses ways to minimize the risk of disease transmission as corneal tissue is processed for lamellar keratoplasty, endothelial keratoplasty, or femtosecond laser-assisted penetrating keratoplasty and to minimize risk to eye bank personnel or physicians preparing the tissue. Secondly, quality assurance measures are described that qualify the environment in which corneal tissue is being processed. We propose that the environment in which corneal tissue is being processed must be able to demonstrate acceptable levels of airborne microbial contamination annually as measured by settle plates to estimate airborne bacterial sedimentation. It is recommended that any environment where corneal tissue is prepared should meet the minimum standard of a conventional operating room which is <25 colony-forming unit per 90-mm settle plate per 1-hour exposure.

  20. Biomechanical relationships between the corneal endothelium and Descemet's membrane.

    Science.gov (United States)

    Ali, Maryam; Raghunathan, VijayKrishna; Li, Jennifer Y; Murphy, Christopher J; Thomasy, Sara M

    2016-11-01

    The posterior face of the cornea consists of the corneal endothelium, a monolayer of cuboidal cells that secrete and attach to Descemet's membrane, an exaggerated basement membrane. Dysfunction of the endothelium compromises the barrier and pump functions of this layer that maintain corneal deturgesence. A large number of corneal endothelial dystrophies feature irregularities in Descemet's membrane, suggesting that cells create and respond to the biophysical signals offered by their underlying matrix. This review provides an overview of the bidirectional relationship between Descemet's membrane and the corneal endothelium. Several experimental methods have characterized a richly topographic and compliant biophysical microenvironment presented by the posterior surface of Descemet's membrane, as well as the ultrastructure and composition of the membrane as it builds during a lifetime. We highlight the signaling pathways involved in the mechanotransduction of biophysical cues that influence cell behavior. We present the specific example of Fuchs' corneal endothelial dystrophy as a condition in which a dysregulated Descemet's membrane may influence the progression of disease. Finally, we discuss some disease models and regenerative strategies that may facilitate improved treatments for corneal dystrophies. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Comparison of 2 Different Methods of Transepithelial Corneal Collagen Cross-Linking: Analysis of Corneal Histology and Hysteresis.

    Science.gov (United States)

    Park, Young Min; Kim, Ho Yoon; Lee, Jong Soo

    2017-07-01

    To compare the effect of 2 different methods of transepithelial corneal collagen cross-linking (CXL) on corneal histology and hysteresis. The right eyes of New Zealand white rabbits (n = 50) were treated using one of the 2 transepithelial CXL methods. The eyes were then divided into 2 groups: group 1, transepithelial CXL using continuous ultraviolet A (UVA); group 2, accelerated transepithelial CXL using pulsed UVA. Both groups showed a postoperative increase in central corneal thickness. In both groups, an increase in corneal hysteresis was found after CXL, but the changes were not significant when compared with the baseline value. The corneal resistance factor and corneal-compensated intraocular pressure also remained unchanged in both groups after treatment. In both groups, the CXL effect reached the anterior-mid part of the corneal stroma. In group 1, CXL mostly affected the anterior-mid part stroma, whereas group 2 showed stromal keratocyte loss, and an acellular zone, in the deep stroma. More signs of keratocyte apoptosis were observed in group 2 than in group 1. Accelerated transepithelial CXL using pulsed UVA showed considerably deeper effect in the stroma with keratocyte loss than transepithelial CXL using continuous UVA.

  2. Refractive improvements and safety with topography-guided corneal crosslinking for keratoconus: 1-year results.

    Science.gov (United States)

    Nordström, Maria; Schiller, Maria; Fredriksson, Anneli; Behndig, Anders

    2017-07-01

    To assess the refractive improvements and the corneal endothelial safety of an individualised topography-guided regimen for corneal crosslinking in progressive keratoconus. An open-label prospective randomised clinical trial was performed at the Department of Clinical Sciences, Ophthalmology, Umeå University Hospital, Umeå, Sweden. Thirty-seven patients (50 eyes) with progressive keratoconus planned for corneal crosslinking were included. The patients were randomised to topography-guided crosslinking (photorefractive intrastromal crosslinking (PiXL); n=25) or uniform 9 mm crosslinking (corneal collagen crosslinking (CXL); n=25). Visual acuity, refraction, keratometry (K1, K2 and Kmax) and corneal endothelial morphometry were assessed preoperatively and at 1, 3, 6 and 12 months postoperatively. The PiXL treatment involved an asymmetrical treatment zone centred on the area of maximum corneal steepness with treatment energies ranging from 7.2 to 15.0 J/cm(2); the CXL treatment was a uniform 9 mm 5.4 J/cm(2) pulsed crosslinking. The main outcome measures were changes in refractive errors and corneal endothelial cell density. The spherical refractive errors decreased (pkeratoconus with decreased spherical refractive errors and improved visual acuity, without damage to the corneal endothelium. NCT02514200, Results. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  3. One donor cornea for 3 recipients: a new concept for corneal transplantation surgery.

    Science.gov (United States)

    Vajpayee, Rasik B; Sharma, Namrata; Jhanji, Vishal; Titiyal, Jeewan S; Tandon, Radhika

    2007-04-01

    To describe the use of a single donor corneal tissue in 3 patients with corneal pathologic conditions. A donor corneal tissue was divided into 3 parts using a microkeratome and a trephine. The anterior lamellar disc was transplanted into a patient with macular corneal dystrophy using the automated lamellar therapeutic keratoplasty technique. The posterior lamellar disc was transplanted into a patient with pseudophakic bullous keratopathy using the Descemet stripping automated endothelial keratoplasty technique. The peripheral corneoscleral rim was used for limbal stem cell transplantation in a child with limbal stem cell deficiency. All surgical procedures were performed successfully. At 3 months, the best-corrected visual acuities achieved following automated lamellar therapeutic keratoplasty, Descemet stripping automated endothelial keratoplasty, and limbal stem cell transplantation were 20/60, 20/40, and 20/200, respectively. The advent of customized component corneal transplantation techniques may allow the use of 1 donor cornea to treat multiple patients.

  4. Comparison of Endothelial Cell Loss by Specular Microscopy ...

    African Journals Online (AJOL)

    Aim: To compare the endothelial cell loss between phacoemulsification and manual small‑incision cataract surgery (SICS). Endothelial cell loss was also compared in phacoemulsification group by temporal clear corneal incision (CCI) and by superior scleral incision (SI) technique. Materials and Methods: A total of 200 ...

  5. [Treatment of corneal ulcers with platelet rich plasma].

    Science.gov (United States)

    Acosta, L; Castro, M; Fernandez, M; Oliveres, E; Gomez-Demmel, E; Tartara, L

    2014-02-01

    To assess the efficacy of platelet rich plasma (PRP) in the treatment of extensive corneal ulcers in albino rabbits. New Zealand rabbits, divided in 3 groups, were used for the study. Corneal ulcers of 10mm diameter were made. Rabbits blood was extracted for the preparation of the PRP of the corresponding group. The blood was processed by differential centrifugation. The first group, named control, was treated with sterile saline every 8h. The second group, named gel, was treated with deproteinized extract gel beef fat every 8h, and the third group, named PRP received one PRP drop on the first and third day of monitoring. The rabbits were monitored, by taking photographs, each day for the 7 days that the study lasted. A better outcome was observed in the group with deproteinized extract gel beef fat (GE group), and the PRP group (PL group), in comparison with the control group (CO group) (P<.05). The PRP showed to be just as effective as the commercial product (Solcoseryl®), for the regeneration of the extensive and deep corneal ulcers. Besides, it stands out as a no surgical procedure is required, and there is easy access, low cost and reduced doses. Copyright © 2013 Sociedad Española de Oftalmología. Published by Elsevier Espana. All rights reserved.

  6. Customised component corneal transplantation: a blessing for three patients.

    Science.gov (United States)

    Sati, Alok; Shankar, Sandeep; Jha, Ashok; Gurunadh, Velamakanni Satyanarayana

    2014-08-21

    The existing acute shortage of good quality donor corneas in a developing country like India, prompted us to attempt customised component corneal transplantation. Using this surgical strategy, one corneoscleral button was used for three recipients. Anterior and posterior lamellar discs were used for anterior lamellar keratoplasty and Descemet's stripping endothelial keratoplasty in patients with superficial corneal scar and pseudophakic bullous keratopathy, respectively. From the remnant peripheral corneoscleral rim, a patch graft was taken and used for a case of perforated corneal ulcer. Postoperatively, the two earlier mentioned cases achieved visual acuities of 20/30 and 20/60, respectively, whereas the latter mentioned patient with the patch graft achieved good tectonic stability. This case report highlights the optimal utilisation of a corneoscleral button by customising it for three recipients. Moreover, a patch graft has been introduced in the armamentarium of customised component corneal transplantation for the first time. 2014 BMJ Publishing Group Ltd.

  7. Hypocellular scar formation or aberrant fibrosis induced by an intrastromal corneal ring: a case report

    Directory of Open Access Journals (Sweden)

    Ramkumar Hema L

    2011-08-01

    Full Text Available Abstract Introduction Intrastromal corneal rings or segments are approved for the treatment of myopia and astigmatism associated with keratoconus. We describe a clinicopathological case of intrastromal corneal rings. For the first time, the molecular pathological findings of intrastromal corneal rings in the cornea are illustrated. Case presentation A 47-year-old African-American man with a history of keratoconus and failure in using a Rigid Gas Permeable contact lens received an intrastromal corneal ring implant in his left eye. Due to complications, penetrating keratoplasty was performed. The intrastromal corneal ring channels were surrounded by a dense acellular (channel haze and/or hypocellular (acidophilic densification collagen scar and slightly edematous keratocytes. Mild macrophage infiltration was found near the inner aspect of the intrastromal corneal rings. Molecular analyses of the microdissected cells surrounding the intrastromal corneal ring channels and central corneal stroma revealed 10 times lower relative expression of IP-10/CXCL10 mRNA and two times higher CCL5 mRNA in the cells surrounding the intrastromal corneal ring, as compared to the central corneal stroma. IP-10/CXCL10 is a fibrotic and angiostatic chemokine produced by macrophages, endothelial cells and fibroblasts. Conclusion An intrastromal corneal ring implant can induce hypocellular scar formation and mild inflammation, which may result from aberrant release of fibrosis-related chemokines.

  8. Vanishing corneal vessels

    Science.gov (United States)

    Nicholson, Luke; Chana, Rupinder

    2013-01-01

    We wish to highlight the importance of acknowledging the accompanying effects of topical phenylephrine drops on the eye other than its intended mydriasis. We reported a case of a 92-year-old woman with a corneal graft who was noted to have superficial corneal vascularisation which was not documented previously. After the instillation of topical tropicamide 1% and phenylephrine 2.5%, for funduscopy, the corneal vascularisation was not visible. When reassessed on another visit, tropicamide had no effect on the vessels and only phenylephrine did. We wish to highlight that when reviewing patients in cornea clinics, instilling phenylephrine prior to being seen may mask important corneal vascularisation. PMID:24121816

  9. Effect of DATS on experimental corneal neovascularization induced by corneal suture in rats cornea

    Directory of Open Access Journals (Sweden)

    Xiao-Jun Zhou

    2017-11-01

    Full Text Available AIM: To study the effect of diallyl trisulfide(DATSon experimental corneal neovascularization(CNVin rats induced by corneal suture and detect the expression of vascular endothelial growth factor(VEGFand p-AKT in rats cornea. METHODS: The rat model of corneal neovascularization(CNVwas induced by corneal suture. Rats were randomly divided into Group A: physiological saline control group containing DMSO(10 rats; Group B: 25μmol/L DATS treatment group(10 rats; Group C: 50μmol/L DATS treatment group(10 rats; Group D: 100μmol/L DATS treatment group(10 rats; Group E: 200μmol/L DATS treatment group(10 rats. The occurrence and development of CNV were observed by slit-lamp microscope at 7d after suture, and the area of CNV were calculated.Two weeks later, HE staining was used to observe the pathological organization form of each cornea, and RT-PCR and Western blot were used to detect the expression of VEGF mRNA and protein expression of VEGF and p-AKT between each groups. RESULTS: The blood vessel area of Group C, D and E was compared with that of Group A, the difference was statistically significant(PPPCONCLUSION: DATS can inhibit corneal neovascularization of the rats induced by suture. Its mechanism may be associated with suppression of VEGF secretion, down-regulation of VEGF and inactivation of p-AKT.

  10. Ipsilateral rotational autokeratoplasty for the management of corneal opacities.

    Science.gov (United States)

    Verma, N; Melengas, S; Garap, J A

    1999-02-01

    Penetrating keratoplasty is the logical solution for the management of corneal opacities. In situations such as in Papua New Guinea, where donor corneal tissue is scarce and corneal opacities are plenty, an alternative procedure for the management of corneal opacities in the form of ipsilateral rotational autokeratoplasty was considered. In the present prospective study, ipsilateral rotational autokeratoplasty was performed in 17 eyes over a 2 year period in a general hospital. The patient's cornea was trephined eccentrically and the corneal opacity was dialed out of the visual axis and was replaced by clear peripheral cornea. Most opacities were leucomata (76.4%). The average size of the opacity was 5.1 mm and the corneal button size was 7 mm.A final visual acuity of 6/18 or better was obtained in 64.7% of cases (at 12 months). No significant postoperative complications were encountered. No complex formula was needed to calculate the size of the button and, by simply adding 3 mm to the pupillary diameter in standard illumination, one could make an estimation of the graft diameter. Rotational autokeratoplasty has a definite role in places where donor corneal tissue is scarce, in patients in whom long-term steroids are a risk or in situations where follow up of patients is difficult. Rejection is a theoretical impossibility, but late endothelial failure could occur, requiring regrafting. Rotational autokeratoplasty should be seriously considered as an alternative to conventional penetrating keratoplasty.

  11. Cataract surgery after Descemet stripping endothelial keratoplasty

    Directory of Open Access Journals (Sweden)

    Sunita Chaurasia

    2012-01-01

    Full Text Available Management of endothelial dysfunction in phakic patients is sometimes a dilemma for corneal surgeons. Phakic patients with visually significant cataract and endothelial dysfunction are preferably managed by performing combined cataract surgery with endothelial keratoplasty. However, combined surgery may be deferred in eyes with early incipient cataract, younger age and where anterior chamber is poorly visualized. As cataract formation may be accelerated after endothelial keratoplasty, these eyes may need cataract surgery subsequently. Surgical intervention in eyes with endothelial keratoplasty is of concern as this may affect the graft adversely and threaten graft survival. In this report, we describe the intraoperative surgical details and postoperative clinical course of a patient who underwent phacoemulsification with intraocular lens implantation after Descemet stripping automated endothelial keratoplasty (DSAEK.

  12. Stability of vision, topography, and endothelial cell density from 1 year to 2 years after deep lamellar endothelial keratoplasty surgery.

    Science.gov (United States)

    Ousley, Paula J; Terry, Mark A

    2005-01-01

    To evaluate whether the visual, topographic, and endothelial cell count results observed 1 year after deep lamellar endothelial keratoplasty (DLEK) surgery remain stable up to 2 years after surgery. Prospective, noncomparative, interventional case series. Twenty eyes of 20 patients with corneal edema from Fuchs' endothelial dystrophy. Deep lamellar endothelial keratoplasty endothelial replacement surgery, with a 9.0-mm or 9.5-mm scleral access incision and a specialized intrastromal trephine, was performed. Snellen visual acuities, corneal topography, and endothelial cell counts were prospectively measured preoperatively and 1 year and 2 years after DLEK. Uncorrected and best spectacle-corrected visual acuity (BSCVA), refractive and topographic astigmatism, mean corneal curvature, topographic regularity and symmetry, and endothelial cell density. At 1 year postoperatively, BSCVA averaged 20/50 (range, 20/25-20/200), spherical equivalents (SE) averaged -0.194+/-1.521 diopters (D), manifest refraction (MR) astigmatism averaged 2.04+/-1.05 D (range, 0.0-4.0 D), topographic astigmatism averaged 2.3+/-1.1 D, mean corneal curvature was 43.2+/-1.8 D, the surface regularity index (SRI) averaged 1.16+/-0.41, and the surface asymmetry index (SAI) averaged 1.05+/-1.09. At 2 years postoperatively, BSCVA averaged 20/48 (range, 20/25-20/200), SE averaged -0.369+/-1.267 D, MR astigmatism averaged 1.76+/-0.66 D (range, 0.75-3.0 D), topographic astigmatism averaged 2.4+/-1.1 D, mean corneal curvature was 43.6+/-1.8 D, the SRI averaged 1.13+/-0.44, and the SAI averaged 0.76+/-0.59. There was no significant change in visual or topographic parameters between 1 year and 2 years postoperatively (P>0.05). Endothelial cell counts averaged 2335+/-468 cells/mm(2) at 1 year and 2151+/-457 cells/mm(2) at 2 years postoperatively (P = 0.041). Deep lamellar endothelial keratoplasty provides stable refractions, corneal topography, and endothelial cell densities as long as 2 years after surgery

  13. Corneal biomechanical properties from two-dimensional corneal flap extensiometry: application to UV-riboflavin cross-linking.

    OpenAIRE

    Kling, Sabine; Ginis, H.; Marcos, Susana

    2012-01-01

    Corneal biomechanical properties are usually measured by strip extensiometry or inflation methods. We developed a two-dimensional (2D) flap extensiometry technique, combining the advantages of both methods, and applied it to measure the effect of UV-Riboflavin cross-linking (CXL). Corneal flaps (13 pig/8 rabbit) from the de-epithelialized anterior stroma (96 μm) were mounted on a custom chamber, consisting of a BK7 lens, a reflective retina, and two reservoirs (filled with Riboflavin and sili...

  14. Tissue and cellular biomechanics during corneal wound injury and repair.

    Science.gov (United States)

    Raghunathan, Vijay Krishna; Thomasy, Sara M; Strøm, Peter; Yañez-Soto, Bernardo; Garland, Shaun P; Sermeno, Jasmyne; Reilly, Christopher M; Murphy, Christopher J

    2017-08-01

    Corneal wound healing is an enormously complex process that requires the simultaneous cellular integration of multiple soluble biochemical cues, as well as cellular responses to the intrinsic chemistry and biophysical attributes associated with the matrix of the wound space. Here, we document how the biomechanics of the corneal stroma are altered through the course of wound repair following keratoablative procedures in rabbits. Further we documented the influence that substrate stiffness has on stromal cell mechanics. Following corneal epithelial debridement, New Zealand white rabbits underwent phototherapeutic keratectomy (PTK) on the right eye (OD). Wound healing was monitored using advanced imaging modalities. Rabbits were euthanized and corneas were harvested at various time points following PTK. Tissues were characterized for biomechanics with atomic force microscopy and with histology to assess inflammation and fibrosis. Factor analysis was performed to determine any discernable patterns in wound healing parameters. The matrix associated with the wound space was stiffest at 7days post PTK. The greatest number of inflammatory cells were observed 3days after wounding. The highest number of myofibroblasts and the greatest degree of fibrosis occurred 21days after wounding. While all clinical parameters returned to normal values 400days after wounding, the elastic modulus remained greater than pre-surgical values. Factor analysis demonstrated dynamic remodeling of stroma occurs between days 10 and 42 during corneal stromal wound repair. Elastic modulus of the anterior corneal stroma is dramatically altered following PTK and its changes coincide initially with the development of edema and inflammation, and later with formation of stromal haze and population of the wound space with myofibroblasts. Factor analysis demonstrates strongest correlation between elastic modulus, myofibroblasts, fibrosis and stromal haze thickness, and between edema and central corneal

  15. Characterization of Corneal Indentation Hysteresis.

    Science.gov (United States)

    Ko, Match W L; Dongming Wei; Leung, Christopher K S

    2015-01-01

    Corneal indentation is adapted for the design and development of a characterization method for corneal hysteresis behavior - Corneal Indentation Hysteresis (CIH). Fourteen porcine eyes were tested using the corneal indentation method. The CIH measured in enucleated porcine eyes showed indentation rate and intraocular pressure (IOP) dependences. The CIH increased with indentation rate at lower IOP ( 25 mmHg). The CIH was linear proportional to the IOP within an individual eye. The CIH was positively correlated with the IOP, corneal in-plane tensile stress and corneal tangent modulus (E). A new method based on corneal indentation for the measurement of Corneal Indentation Hysteresis in vivo is developed. To our knowledge, this is the first study to introduce the corneal indentation hysteresis and correlate the corneal indentation hysteresis and corneal tangent modulus.

  16. Constructing an in vitro cornea from cultures of the three specific corneal cell types.

    Science.gov (United States)

    Schneider, A I; Maier-Reif, K; Graeve, T

    1999-10-01

    This paper presents a reliable method for establishing pure cultures of the three types of corneal cells. This is believed to be the first time, corneal cells have been cultured from fetal pig corneas. Cell growth studies were performed in different media. Subcultures of the three corneal cell types were passaged until the 30th generation without their showing signs of senescence. For engineering an in vitro cornea, corneal epithelial cells were cultured over corneal stromal cells in an artificial biomatrix of collagen with an underlying layer of corneal endothelial cells. The morphology, histology, and differentiation of the in vitro cornea were investigated to determine the degree of comparability to the cornea in vivo. The in vitro construct displayed signs of transition to an organotypic phenotype of which the most prominent was the formation of two basement membranes.

  17. Elastic modulus and collagen organization of the rabbit cornea: epithelium to endothelium.

    Science.gov (United States)

    Thomasy, Sara M; Raghunathan, Vijay Krishna; Winkler, Moritz; Reilly, Christopher M; Sadeli, Adeline R; Russell, Paul; Jester, James V; Murphy, Christopher J

    2014-02-01

    The rabbit is commonly used to evaluate new corneal prosthetics and study corneal wound healing. Knowledge of the stiffness of the rabbit cornea would better inform the design and fabrication of keratoprosthetics and substrates with relevant mechanical properties for in vitro investigations of corneal cellular behavior. This study determined the elastic modulus of the rabbit corneal epithelium, anterior basement membrane (ABM), anterior and posterior stroma, Descemet's membrane (DM) and endothelium using atomic force microscopy (AFM). In addition, three-dimensional collagen fiber organization of the rabbit cornea was determined using nonlinear optical high-resolution macroscopy. The elastic modulus as determined by AFM for each corneal layer was: epithelium, 0.57 ± 0.29 kPa (mean ± SD); ABM, 4.5 ± 1.2 kPa, anterior stroma, 1.1 ± 0.6 kPa; posterior stroma, 0.38 ± 0.22 kPa; DM, 11.7 ± 7.4 kPa; and endothelium, 4.1 ± 1.7 kPa. The biophysical properties, including the elastic modulus, are unique for each layer of the rabbit cornea and are dramatically softer in comparison to the corresponding regions of the human cornea. Collagen fiber organization is also dramatically different between the two species, with markedly less intertwining observed in the rabbit vs. human cornea. Given that the substratum stiffness considerably alters the corneal cell behavior, keratoprosthetics that incorporate mechanical properties simulating the native human cornea may not elicit optimal cellular performance in rabbit corneas that have dramatically different elastic moduli. These data should allow for the design of substrates that better mimic the biomechanical properties of the corneal cellular environment. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  18. corneal pyogenic granuloma

    African Journals Online (AJOL)

    GB

    2012-09-14

    Sep 14, 2012 ... Figure 3: A child with pyogenic cornea granuloma intra operatively. After excision of the mass, central corneal stromal defect developed. The patient was followed up for more than a month with topical antibiotic and cycloplegic. Subsequently, the defect healed and leucoma corneal opacity (figure.

  19. VRP09 Reduction of Corneal Scarring Following Blast and Burn Injuries to Cornea Using siRNAs Targeting TGFb and CTGF

    Science.gov (United States)

    2012-10-01

    compare the effectiveness of siRNAs oligonucleotides and AAV -vectored siRNAs in rabbit models of blast and burn corneal injuries for reduction of...next phase of developing an effective antiscarring drug therapy for corneal blast and burn injuries. Starting with in vitro results and by using...particles that we generated with a commercially available kit of biodegradable polymers were tested in vitro using fresh rabbit globes that were

  20. Femtosecond laser lenticule transplantation in rabbit cornea: experimental study.

    Science.gov (United States)

    Liu, Huiying; Zhu, Wenqing; Jiang, Alice C; Sprecher, Alicia J; Zhou, Xingtao

    2012-12-01

    To evaluate the feasibility of femtosecond laser-induced lenticule transplantation in the rabbit cornea and to observe the relative histologic characteristics of corneal tissue and nerve repair after transplantation. Eight healthy, purebred, New Zealand white rabbits underwent femtosecond laser small-incision lenticule extraction (SMILE) surgery in the right eye. Lenticules were inserted into a femtosecond laser-created corneal stromal pocket in the left eye, which was defined as femtosecond laser corneal lenticule transplantation. Postoperative observation and examination were completed to evaluate the surgery. In the early postoperative period, inflammation of the cornea was noted, tissue around the lenticule was edematous, and cells were activated. Tissue edema remained at postoperative day 10. By 1 month, edema had resolved, activated cells gradually became quiescent, and nerve fiber regeneration was observed. By 3 months, the lenticule integrated into the recipient cornea, extracellular matrix gradually cleared, and thicker nerve fibers were noted. By 6 months postoperative, morphology and distribution of the corneal stromal fibers were close to normal, and the number of nerve fibers was reduced. Femtosecond laser corneal lenticule transplantation in rabbits is feasible, as the lenticule was shown to thrive and integrate with the recipient stroma. Nerve regeneration begins after 1 month. Copyright 2012, SLACK Incorporated.

  1. The molecular basis of corneal transparency.

    Science.gov (United States)

    Hassell, John R; Birk, David E

    2010-09-01

    The cornea consists primarily of three layers: an outer layer containing an epithelium, a middle stromal layer consisting of a collagen-rich extracellular matrix (ECM) interspersed with keratocytes and an inner layer of endothelial cells. The stroma consists of dense, regularly packed collagen fibrils arranged as orthogonal layers or lamellae. The corneal stroma is unique in having a homogeneous distribution of small diameter 25-30 nm fibrils that are regularly packed within lamellae and this arrangement minimizes light scattering permitting transparency. The ECM of the corneal stroma consists primarily of collagen type I with lesser amounts of collagen type V and four proteoglycans: three with keratan sulfate chains; lumican, keratocan, osteoglycin and one with a chondroitin sulfate chain; decorin. It is the core proteins of these proteoglycans and collagen type V that regulate the growth of collagen fibrils. The overall size of the proteoglycans are small enough to fit in the spaces between the collagen fibrils and regulate their spacing. The stroma is formed during development by neural crest cells that migrate into the space between the corneal epithelium and corneal endothelium and become keratoblasts. The keratoblasts proliferate and synthesize high levels of hyaluronan to form an embryonic corneal stroma ECM. The keratoblasts differentiate into keratocytes which synthesize high levels of collagens and keratan sulfate proteoglycans that replace the hyaluronan/water-rich ECM with the densely packed collagen fibril-type ECM seen in transparent adult corneas. When an incisional wound through the epithelium into stroma occurs the keratocytes become hypercellular myofibroblasts. These can later become wound fibroblasts, which provides continued transparency or become myofibroblasts that produce a disorganized ECM resulting in corneal opacity. The growth factors IGF-I/II are likely responsible for the formation of the well organized ECM associated with transparency

  2. Corneal biomechanics - a review.

    Science.gov (United States)

    Kling, Sabine; Hafezi, Farhad

    2017-05-01

    In recent years, the interest in corneal biomechanics has strongly increased. The material properties of the cornea determine its shape and therefore play an important role in corneal ectasia and related pathologies. This review addresses the molecular origin of biomechanical properties, models for their description, methods for their characterisation, techniques for their modification, and computational simulation approaches. Recent research has focused on developing non-contact techniques to measure the biomechanical properties in vivo, on determining structural and molecular abnormalities in pathological corneas, on developing and optimising techniques to reinforce the corneal tissue and on the computational simulation of surgical interventions. A better understanding of corneal biomechanics will help to improve current refractive surgeries, allow an earlier diagnosis of ectatic disorders and a better quantification of treatments aiming at reinforcing the corneal tissue. © 2017 The Authors Ophthalmic & Physiological Optics © 2017 The College of Optometrists.

  3. Adipose-derived mesenchymal stem cell administration does not improve corneal graft survival outcome.

    Directory of Open Access Journals (Sweden)

    Sherezade Fuentes-Julián

    Full Text Available The effect of local and systemic injections of mesenchymal stem cells derived from adipose tissue (AD-MSC into rabbit models of corneal allograft rejection with either normal-risk or high-risk vascularized corneal beds was investigated. The models we present in this study are more similar to human corneal transplants than previously reported murine models. Our aim was to prevent transplant rejection and increase the length of graft survival. In the normal-risk transplant model, in contrast to our expectations, the injection of AD-MSC into the graft junction during surgery resulted in the induction of increased signs of inflammation such as corneal edema with increased thickness, and a higher level of infiltration of leukocytes. This process led to a lower survival of the graft compared with the sham-treated corneal transplants. In the high-risk transplant model, in which immune ocular privilege was undermined by the induction of neovascularization prior to graft surgery, we found the use of systemic rabbit AD-MSCs prior to surgery, during surgery, and at various time points after surgery resulted in a shorter survival of the graft compared with the non-treated corneal grafts. Based on our results, local or systemic treatment with AD-MSCs to prevent corneal rejection in rabbit corneal models at normal or high risk of rejection does not increase survival but rather can increase inflammation and neovascularization and break the innate ocular immune privilege. This result can be partially explained by the immunomarkers, lack of immunosuppressive ability and immunophenotypical secretion molecules characterization of AD-MSC used in this study. Parameters including the risk of rejection, the inflammatory/vascularization environment, the cell source, the time of injection, the immunosuppression, the number of cells, and the mode of delivery must be established before translating the possible benefits of the use of MSCs in corneal transplants to clinical

  4. Triple procedure: cataract extraction, intraocular lens implantation, and corneal graft.

    Science.gov (United States)

    Oie, Yoshinori; Nishida, Kohji

    2017-01-01

    Corneal triple procedures consist of combined keratoplasty surgery and cataract surgery. Currently, anterior lamellar keratoplasty, endothelial keratoplasty, and conventional penetrating keratoplasty, are very often simultaneously performed with cataract surgery. The present article reviews the recent advances and current developments in corneal triple procedures. Many new findings have been reported. Core vitrectomy before host trephination and retroillumination during continuous curvilinear capsulorhexis has enabled us to perform penetrating keratoplasty triple procedures more safely. Type-1 big-bubble, baring Dua's layer, facilitates well tolerated anterior lamellar triple keratoplasty without Descemet membrane perforation. Descemet membrane endothelial keratoplasty triple procedure can provide excellent expected postoperative refraction. New techniques, including manual medium incision cataract surgery or glued intrascleral fixation of the intraocular lens with pupilloplasty, can be used for endothelial keratoplasty triple procedure, and frequent topical steroid administration can contribute to the reduction of postoperative cystoid macular edema. Recent technical advancements could contribute to safer and more effective corneal triple procedures. Triple surgery combined with novel procedures, including regenerative medicine and bioengineered corneal transplantation, will be performed in the near future.

  5. Corneal melting after collagen cross-linking for keratoconus: a case report

    Directory of Open Access Journals (Sweden)

    Zissimopoulos Athanassios

    2011-04-01

    Full Text Available Abstract Introduction Corneal collagen cross-linking is a rather new technique that uses riboflavin and ultraviolet A light for collagen fiber stabilization in keratoconus corneas. Other than reversible side effects, the preliminary results of corneal collagen cross-linking studies suggest that it is a rather safe technique. In this report, we demonstrate a case of corneal melting after corneal collagen cross-linking for keratoconus corneas associated with an acute inflammatory response. Case presentation A 23-year-old Caucasian man with keratoconus cornea stage 1 to 2 underwent uneventful corneal collagen cross-linking treatment according to the Dresden protocol. The next day the patient had intense photophobia, watering and redness of the eye, and his visual acuity was limited to counting fingers. Slit lamp biomicroscopy revealed severe corneal haze accompanied by non-specific endothelial precipitates following an acute inflammatory response. Mild inflammation could be detected in the anterior chamber. Moreover, the re-epithelialization process could barely be detected. His corneal state gradually deteriorated, resulting in descemetocele and finally perforation. Conclusion In this report, we present a case of a patient with corneal melting after standard corneal collagen cross-linking treatment for keratoconus corneas following an acute inflammatory response. Despite modifying postoperative treatment, elaboration of all apparent associated causes by the treating physicians and undergoing extensive laboratory testing, the patient developed descemetocele, which led to perforation. Our report suggests that further research is necessary regarding the safety of corneal collagen cross-linking in keratoconus corneas.

  6. Biocompatibility of poly(ethylene glycol) and poly(acrylic acid) interpenetrating network hydrogel by intrastromal implantation in rabbit cornea.

    Science.gov (United States)

    Zheng, Luo Luo; Vanchinathan, Vijay; Dalal, Roopa; Noolandi, Jaan; Waters, Dale J; Hartmann, Laura; Cochran, Jennifer R; Frank, Curtis W; Yu, Charles Q; Ta, Christopher N

    2015-10-01

    We evaluated the biocompatibility of a poly(ethylene glycol) and poly(acrylic acid) (PEG/PAA) interpenetrating network hydrogel designed for artificial cornea in a rabbit model. PEG/PAA hydrogel measuring 6 mm in diameter was implanted in the corneal stroma of twelve rabbits. Stromal flaps were created with a microkeratome. Randomly, six rabbits were assigned to bear the implant for 2 months, two rabbits for 6 months, two rabbits for 9 months, one rabbit for 12 months, and one rabbit for 16 months. Rabbits were evaluated monthly. After the assigned period, eyes were enucleated, and corneas were processed for histology and immunohistochemistry. There were clear corneas in three of six rabbits that had implantation of hydrogel for 2 months. In the six rabbits with implant for 6 months or longer, the corneas remained clear in four. There was a high rate of epithelial defect and corneal thinning in these six rabbits. One planned 9-month rabbit developed extrusion of implant at 4 months. The cornea remained clear in the 16-month rabbit but histology revealed epithelial in-growth. Intrastromal implantation of PEG/PAA resulted in a high rate of long-term complications. © 2015 Wiley Periodicals, Inc.

  7. Effect of lactated Ringer's solution and compound electrolyte solution on the corneal endothelium in phacoemulsification

    Directory of Open Access Journals (Sweden)

    Yang Xia

    2017-11-01

    Full Text Available AIM: To compare the effect of compound electrolyte solution and lactated Ringer's solution on corneal function in cataract phacoemulsification, and to provide scientific basis for clinical selection of appropriate perfusion fluid. METHODS: The patients with senile cataract were randomly divided into control group with lactated Ringer's solution as anterior chamber perfusion and experimental group with compound electrolyte as anterior chamber perfusion. Surgical removal of cataract and phacoemulsification with intraocular lens implantation were taken. The corneal endothelial cell density, central corneal thickness, hexagonal cell ratio and endothelial cell coefficient of variation were measured at preoperative and postoperative points. RESULTS: Totally 60 patients successfully completed all follow-ups, the experimental group of 30 cases, the control group of 30 cases. The density of corneal endothelial cells in experimental group was significantly higher than those in the lactated Ringer's solution group at 1 and 3d after operations(P=0.030, 0.046. The coefficient of variation of corneal endothelial cells in lactated Ringer's solution group was higher than that in compound electrolyte group at 1 and 14d after operation(P=0.025, 0.014. The visual acuity of the compound electrolyte group was better than that of the lactated Ringer's solution on the first day after operation(P=0.04. CONCLUSION: In the phacoemulsification of senile cataract, the compound electrolyte perfusion has better histocompatibility, which can maintain the stability of corneal endothelial cell structure and reduce corneal endothelial cell injury. The compound electrolyte perfusion solution is more suitable for senile cataract phacoemulsification surgery.

  8. Stimulation of the corneal blinking reflex by ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Tobias, C.; Luce, J.; Yanni, N.; Brustad, T.; Lyman, J.; Kimura, S.

    1961-05-04

    Accelerated alpha particles from the Berkeley heavy-ion linear accelerator were used in a series of experiments designed to elucidate the conditions by which radiation can stimulate or modify nerve action in mammals. Single millisecond pulses in excess of 40,000 radsor pulse trains of less than 1 sec duration elicited the corneal blinking reflex when delivered to the cornea of unanesthetized rabbits. The lowest threshold dose was observed when the Bragg ionization peak was placed at 140 {mu} depth.

  9. Indications for Corneal Transplantation at a Tertiary Referral Center in Tehran

    Directory of Open Access Journals (Sweden)

    Mohammad Zare

    2010-01-01

    Full Text Available Purpose: To report the indications and techniques of corneal transplantation at a tertiary referral center in Tehran over a 3-year period. Methods: Records of patients who had undergone any kind of corneal transplantation at Labbafinejad Medical Center, Tehran, Iran from March 2004 to March 2007 were reviewed to determine the indications and types of corneal transplantation. Results: During this period, 776 eyes of 756 patients (including 504 male subjects with mean age of 41.3±21.3 years underwent corneal transplantation. The most common indication was keratoconus (n=317, 40.8% followed by bullous keratopathy (n=90, 11.6%, non-herpetic corneal scars (n=62, 8.0%, infectious corneal ulcers (n=61, 7.9%, previously failed grafts (n=61, 7.9%, endothelial and stromal corneal dystrophies (n=28, 3.6%, and trachoma keratopathy (n=26, 3.3%. Other indications including Terrien′s marginal degeneration, post-LASIK keratectasia, trauma, chemical burns, and peripheral ulcerative keratitis constituted the rest of cases. Techniques of corneal transplantation included penetrating keratoplasty (n=607, 78.2%, deep anterior lamellar keratoplasty (n=108, 13.9%, conventional lamellar keratoplasty (n=44, 5.7%, automated lamellar therapeutic keratoplasty (n=8, 1.0%, and Descemet stripping endothelial keratoplasty (n=6, 0.8% in descending order. The remaining cases were endothelial keratoplasty and sclerokeratoplasty. Conclusion: In this study, keratoconus was the most common indication for penetrating keratoplasty which was the most prevalent technique of corneal transplantation. However, deep anterior lamellar keratoplasty is emerging as a growing alternative for corneal pathologies not involving the endothelium.

  10. Proangiogenic Function of T Cells in Corneal Transplantation.

    Science.gov (United States)

    Di Zazzo, Antonio; Tahvildari, Maryam; Subbarayal, Brinda; Yin, Jia; Dohlman, Thomas H; Inomata, Takenori; Mashaghi, Alireza; Chauhan, Sunil K; Dana, Reza

    2017-04-01

    Corneal neovascularization increases the risk of T cell-mediated allograft rejection. Here, we investigate whether T cells promote angiogenesis in transplantation. Conventional effector T cells were collected from draining lymph nodes of allogeneic or syngeneic corneal transplanted BALB/c mice. T cells were either cocultured with vascular endothelial cells (VECs) to assess VEC proliferation or used in a mixed lymphocyte reaction assay. Messenger RNA (mRNA) expression of vascular endothelial growth factor (VEGF)-A, -C, and VEGF receptor 2 (VEGF-R2) in VECs was assessed by real-time PCR. VEGF-A protein expression was determined by enzyme-linked immunosorbent assay. Flow cytometry was used to analyze VEGF-R2 expression in corneal CD31 cells, and VEGF-A and IFNγ expression in corneal CD4 T cells. Allogeneic T cells from high-risk (HR) grafted mice induced more VEC proliferation than those from syngeneic transplant recipients (P = 0.03). Vascular endothelial growth factor-A mRNA and protein expression were higher in T cells from draining lymph nodes (P = 0.03 and P = 0.04, respectively) and cornea (protein; P = 0.04) of HR compared with low-risk (LR) grafted hosts. Vascular endothelial growth factor-A, VEGF-C, and VEGF-R2 mRNA expression were increased in VECs when cocultured with T cells from HR transplants compared with LR transplants and naive mice. In addition, IFNγ blockade in T cell/VEC coculture increased VEC proliferation and VEGF-A protein expression, whereas blocking VEGF-A significantly reduced VEC proliferation (P = 0.04). Allogeneic T cells from corneal transplant hosts promote VEC proliferation, probably via VEGF-A signaling, whereas IFNγ shows an antiangiogenic effect. Our data suggest that T cells are critical mediators of angiogenesis in transplantation.

  11. A novel method in preparation of acellularporcine corneal stroma tissue for lamellar keratoplasty.

    Science.gov (United States)

    Shao, Yi; Tang, Jing; Zhou, Yueping; Qu, Yangluowa; He, Hui; Liu, Qiuping; Tan, Gang; Li, Wei; Liu, Zuguo

    2015-01-01

    Our objective was to develop a novel lamellar cornealbiomaterial for corneal reconstruction.Theporcine acellular corneal stroma discs (ACSDs) were prepared from de-epithelized fresh porcine corneas (DFPCs) by incubation with 100% fresh human serum and additional electrophoresis at 4°C. Such manipulation removed theanterior corneal stromal cells without residual of DNA content and α-Galantigen. Human serum decellularizing activity on porcineanterior corneal stroma cells is through apoptosis, and associated with the presence of α-Gal epitopes in anterior stroma. ACSDs displayed similar optical, biomechanical properties and ultrastructure to DFPCs, and showed good histocompatibility in rabbit corneal stromal pockets and anterior chamber. Rabbit corneallamellar keratoplasty (LKP) using ACSDs showed no rejection and high transparency of cornea at 2 months after surgery. In vivo confocal laser scanning microscopy and immunostaining analysis showed complete re-epithelization and stromal cell in growth of ACSDs without inflammatory cell infiltration, new blood vessel ingrowth and excessive wound healing. In conclusion, this novel decellularization method may be valuable for preparation of xenogenic corneal tissue for clinical application, ACSDs resulted from this method may be served as a matrix equivalent for LKP in corneal xenotransplantation.

  12. A novel method in preparation of acellularporcine corneal stroma tissue for lamellar keratoplasty

    Science.gov (United States)

    Shao, Yi; Tang, Jing; Zhou, Yueping; Qu, Yangluowa; He, Hui; Liu, Qiuping; Tan, Gang; Li, Wei; Liu, Zuguo

    2015-01-01

    Our objective was to develop a novel lamellar cornealbiomaterial for corneal reconstruction.Theporcine acellular corneal stroma discs (ACSDs) were prepared from de-epithelized fresh porcine corneas (DFPCs) by incubation with 100% fresh human serum and additional electrophoresis at 4°C. Such manipulation removed theanterior corneal stromal cells without residual of DNA content and α-Galantigen. Human serum decellularizing activity on porcineanterior corneal stroma cells is through apoptosis, and associated with the presence of α-Gal epitopes in anterior stroma. ACSDs displayed similar optical, biomechanical properties and ultrastructure to DFPCs, and showed good histocompatibility in rabbit corneal stromal pockets and anterior chamber. Rabbit corneallamellar keratoplasty (LKP) using ACSDs showed no rejection and high transparency of cornea at 2 months after surgery. In vivo confocal laser scanning microscopy and immunostaining analysis showed complete re-epithelization and stromal cell in growth of ACSDs without inflammatory cell infiltration, new blood vessel ingrowth and excessive wound healing. In conclusion, this novel decellularization method may be valuable for preparation of xenogenic corneal tissue for clinical application, ACSDs resulted from this method may be served as a matrix equivalent for LKP in corneal xenotransplantation. PMID:26885261

  13. Effects of aberrant Pax6 gene dosage on mouse corneal pathophysiology and corneal epithelial homeostasis.

    Directory of Open Access Journals (Sweden)

    Richard L Mort

    /- genotypes have only relatively minor effects on LESC clone numbers but cause more severe corneal endothelial and stromal defects. This should prompt further investigations of the pathophysiology underlying human aniridia and ARK.

  14. Bilateral reversible corneal edema associated with amantadine use.

    Science.gov (United States)

    Esquenazi, Salomon

    2009-12-01

    In this article, we report a case of bilateral severe reversible corneal edema caused by amantadine therapy. A 39-year-old women was referred to us for evaluation of bilateral corneal edema. Her past medical history was significant for multiple sclerosis, anorexia, and seizures. She developed painless progressive bilateral loss of vision for the past 6 months. She was evaluated by several ophthalmologists elsewhere who felt that the patient's visual loss was secondary to a nutritional deficiency as opposed to related to multiple sclerosis. She was started on vitamin B-12 medication without improvement in her symptoms. She was then evaluated by neuro-ophthalmology. The examination revealed severe bilateral corneal edema and was referred to our corneal service for further evaluation of her corneal condition. Our examination revealed best corrected visual acuity of 20/400 bilaterally. Corneal thickness was 940 microm in the right eye and 802 microm in the left. Color vision was intact. Conjunctivas were white bilaterally. Cornea evaluation revealed diffuse stromal edema and Descemet's folds and microcystic subepithelial edema with to guttae noted. Anterior chambers were deep and quiet. A specular microscopy revealed significant pleomorphism and polymegathism with an endothelial cell count of 1,504 cells in the right eye and 1,596 in the left eye. Review of the patient's medical information revealed therapy with amantadine 2 months prior to the appearance of the patient's symptoms as a means to control the patient's tremors. The patient experienced rapid resolution of the corneal edema within the next 2 months after discontinuation of the agent with recovery of best corrected visual acuity of 20/40 in the right eye and 20/30 in the left. In cases of unexplained corneal edema and in the absence of any identifiable ocular cause, a review of toxic effects of systemic medications should be performed. Early diagnosis may prevent irreversible endothelial damage. Amantadine

  15. Corneal Cell Morphology in Keratoconus: A Confocal Microscopic Observation

    Science.gov (United States)

    Ghosh, Somnath; Mutalib, Haliza Abdul; Kaur, Sharanjeet; Ghoshal, Rituparna; Retnasabapathy, Shamala

    2017-01-01

    Purpose To evaluate corneal cell morphology in patients with keratoconus using an in vivo slit scanning confocal microscope. Methods A cross-sectional study was conducted to evaluate the corneal cell morphology of 47 keratoconus patients and 32 healthy eyes without any ocular disease. New keratoconus patients with different disease severities and without any other ocular co-morbidity were recruited from the ophthalmology department of a public hospital in Malaysia from June 2013 to May 2014. Corneal cell morphology was evaluated using an in vivo slit-scanning confocal microscope. Qualitative and quantitative data were analysed using a grading scale and the Nidek Advanced Visual Information System software, respectively. Results The corneal cell morphology of patients with keratoconus was significantly different from that of healthy eyes except in endothelial cell density (P = 0.072). In the keratoconus group, increased level of stromal haze, alterations such as the elongation of keratocyte nuclei and clustering of cells at the anterior stroma, and dark bands in the posterior stroma were observed with increased severity of the disease. The mean anterior and posterior stromal keratocyte densities and cell areas among the different stages of keratoconus were significantly different (P 0.05) among the three stages of keratoconus. Conclusion Confocal microscopy observation showed significant changes in corneal cell morphology in keratoconic cornea from normal healthy cornea. Analysis also showed significant changes in different severities of keratoconus. Understanding the corneal cell morphology changes in keratoconus may help in the long-term monitoring and management of keratoconus. PMID:28894403

  16. BMP7 gene transfer via gold nanoparticles into stroma inhibits corneal fibrosis in vivo.

    Directory of Open Access Journals (Sweden)

    Ashish Tandon

    Full Text Available This study examined the effects of BMP7 gene transfer on corneal wound healing and fibrosis inhibition in vivo using a rabbit model. Corneal haze in rabbits was produced with the excimer laser performing -9 diopters photorefractive keratectomy. BMP7 gene was introduced into rabbit keratocytes by polyethylimine-conjugated gold nanoparticles (PEI2-GNPs transfection solution single 5-minute topical application on the eye. Corneal haze and ocular health in live animals was gauged with stereo- and slit-lamp biomicroscopy. The levels of fibrosis [α-smooth muscle actin (αSMA, F-actin and fibronectin], immune reaction (CD11b and F4/80, keratocyte apoptosis (TUNEL, calcification (alizarin red, vonKossa and osteocalcin, and delivered-BMP7 gene expression in corneal tissues were quantified with immunofluorescence, western blotting and/or real-time PCR. Human corneal fibroblasts (HCF and in vitro experiments were used to characterize the molecular mechanism mediating BMP7's anti-fibrosis effects. PEI2-GNPs showed substantial BMP7 gene delivery into rabbit keratocytes in vivo (2×10(4 gene copies/ug DNA. Localized BMP7 gene therapy showed a significant corneal haze decrease (1.68±0.31 compared to 3.2±0.43 in control corneas; p88%; p<0.0001, and immunoblotting of BMP7-transefected HCFs grown in the presence of TGFβ demonstrated significantly enhanced pSmad-1/5/8 (95%; p<0.001 and Smad6 (53%, p<0.001, and decreased αSMA (78%; p<0.001 protein levels. These results suggest that localized BMP7 gene delivery in rabbit cornea modulates wound healing and inhibits fibrosis in vivo by counter balancing TGFβ1-mediated profibrotic Smad signaling.

  17. Initial results of small incision deep lamellar endothelial keratoplasty (DLEK).

    Science.gov (United States)

    Fogla, Rajesh; Padmanabhan, Prema

    2006-02-01

    To evaluate the results of replacing the posterior stroma and endothelium, using small incision deep lamellar endothelial keratoplasty (DLEK) surgical technique, in patients with corneal endothelial dysfunction. Noncomparative case series. Fifteen eyes of 15 patients (six males and nine females) with endothelial dysfunction were included in this study. Through a 5-mm scleral incision, a deep lamellar pocket was created across the cornea, followed by excision of an 8.0-mm disk of posterior lamellar corneal tissue. Same size lamellar donor disk was prepared and placed in position without the need of suture fixation. Best spectacle-corrected visual acuity (BSCVA), refraction, endothelial cell density, corneal topography, and corneal thickness were analyzed. Average BSCVA preoperative was 20/200 (range 20/40 to hand movements (HM)), improving to 20/50 (range 20/20 to 20/120) at a mean follow-up of 7.2 months. Average refractive astigmatism at last follow-up was 1.46+/-1.21 diopters (range, 0 to 4 diopters). Preoperative average donor endothelial cell density was 2047+/-311 cells/mm2, and that at last follow-up was 1732+/-514 cells/mm2. Preoperative average pachymetry was 801.4+/-211.3 microm, improving to 553+/-90.4 microm at last follow-up. Initial results with small incision DLEK procedure indicate that it is a safe procedure that provides healthy donor endothelial cell count and function postoperatively, with encouraging visual results.

  18. Diagnosis and Management of Iridocorneal Endothelial Syndrome

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    Marta Sacchetti

    2015-01-01

    Full Text Available The iridocorneal endothelial (ICE syndrome is a rare ocular disorder that includes a group of conditions characterized by structural and proliferative abnormalities of the corneal endothelium, the anterior chamber angle, and the iris. Common clinical features include corneal edema, secondary glaucoma, iris atrophy, and pupillary anomalies, ranging from distortion to polycoria. The main subtypes of this syndrome are the progressive iris atrophy, the Cogan-Reese syndrome, and the Chandler syndrome. ICE syndrome is usually diagnosed in women in the adult age. Clinical history and complete eye examination including tonometry and gonioscopy are necessary to reach a diagnosis. Imaging techniques, such as in vivo confocal microscopy and ultrasound biomicroscopy, are used to confirm the diagnosis by revealing the presence of “ICE-cells” on the corneal endothelium and the structural changes of the anterior chamber angle. An early diagnosis is helpful to better manage the most challenging complications such as secondary glaucoma and corneal edema. Treatment of ICE-related glaucoma often requires glaucoma filtering surgery with antifibrotic agents and the use of glaucoma drainage implants should be considered early in the management of these patients. Visual impairment and pain associated with corneal edema can be successfully managed with endothelial keratoplasty.

  19. Corneal decompensation following filtering surgery with the Ex-PRESS® mini glaucoma shunt device

    Directory of Open Access Journals (Sweden)

    Tojo N

    2015-03-01

    Full Text Available Naoki Tojo, Atsushi Hayashi, Akio Miyakoshi Department of Ophthalmology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan Purpose: To report a case of corneal decompensation due to the Ex-PRESS® mini glaucoma shunt device (Ex-PRESS.Patient and methods: A 75-year-old man had pseudoexfoliation glaucoma in his right eye. He underwent filtration surgery with Ex-PRESS. His intraocular pressure was 7 mmHg after 9 months.Results: We observed partial decompensation of the corneal endothelium adjacent to the filtering bleb. Specular microscopy revealed a marked decrease in the endothelial cell density at the center of the cornea.Conclusion: Anterior segment optical coherence tomography is very useful for evaluating corneal edema and the position of Ex-PRESS. It is important to follow up with an examination of the corneal endothelial cells. Keywords: Ex-PRESS, bullous keratopathy, trabeculectomy, complication, cornea 

  20. Comparative Evaluation of Corneal Endothelium in Patients with Diabetes Undergoing Phacoemulsification.

    Science.gov (United States)

    Sahu, Pramod K; Das, Gopal K; Agrawal, Sumeet; Kumar, Sabitabh

    2017-01-01

    The purpose of this study is to determine if phacoemulsification with posterior chamber foldable intraocular lens implantation has a greater impact on the corneal endothelium of type 2 diabetic patients as compared to nondiabetic controls. This study compared the corneal endothelial status in sixty patients with diabetes with good glycemic control and sixty nondiabetic controls before and after (1 week, 1 month, 2 month, and 3 month) uneventful phacoemulsification. Central corneal endothelial cell density, percentage hexagonality, and coefficient of variation were measured using a noncontact specular microscope. Central corneal thickness was taken as a surrogate marker for endothelium functional status. Data were age and sex matched. Patients with diabetes showed significantly higher loss in endothelial cell count as compared to nondiabetic controls. Furthermore, the patients with diabetes showed a slower recovery trend in the endothelial healing response as evidenced by lower change in the coefficient of variation. Significant correlation was found between energy used and change in endothelial count as well as coefficient of variation in nondiabetics only. In spite of good glycemic control, patients with diabetes have significantly more endothelial damage in comparison to nondiabetic controls with similar nuclear grading and phaco energy used. This warrants a more careful use of phaco energy in patient with diabetes.

  1. Corneal transplant - slideshow

    Science.gov (United States)

    ... ency/presentations/100082.htm Corneal transplant - series—Normal anatomy To use the sharing features on this page, ... Bethesda, MD 20894 U.S. Department of Health and Human Services National Institutes of Health Page last updated: ...

  2. A human corneal equivalent constructed from SV40-immortalised corneal cell lines.

    Science.gov (United States)

    Zorn-Kruppa, Michaela; Tykhonova, Svitlana; Belge, Gazanfer; Bednarz, Jürgen; Diehl, Horst A; Engelke, Maria

    2005-02-01

    Within the last decade, extensive research in the field of tissue and organ engineering has focused on the development of in vitro models of the cornea. The use of organotypic, three-dimensional corneal equivalents has several advantages over simple monolayer cultures. The aim of this study was to develop a corneal equivalent model composed of the same cell types as in the natural human tissue, but by using immortalised cell lines to ensure reproducibility and to minimise product variation. We report our success in the establishment of an SV40-immortalised human corneal keratocyte cell line (designated HCK). A collagen matrix, built up with these cells, displayed the morphological characteristics of the human stromal tissue and served as a biomatrix for the immortalised human corneal epithelial and endothelial cells. Histological cross-sections of the whole-cornea equivalents resemble human corneas in tissue structure. This organotypic in vitro model may serve as a research tool for the ophthalmic science community, as well as a model system for testing for eye irritancy and drug efficacy.

  3. A Simple Way to Clear the Media for Vitrectomy in Eyes with Corneal Edema

    Directory of Open Access Journals (Sweden)

    Touka Banaee

    2012-01-01

    Full Text Available Herein we introduce a simple approach for clearing an edematous cornea during vitreoretinal surgery in eyes with decompensated corneal endothelium, allowing the surgeon to postpone penetrating keratoplasty. This technique was performed in 3 eyes by filling the anterior chambers with air or silicone oil, and sufficiently cleared the media for completion of vitrectomy. This simple technique enables completion of the vitrectomy without a temporary keratoprosthesis and penetrating keratoplasty in eyes with corneal edema due to endothelial decompensation.

  4. Non-invasive assessment of the donor corneal endothelium using ocular redox fluorometry.

    OpenAIRE

    Shimazaki, J; Laing, R A; Tsubota, K; Kenyon, K R

    1996-01-01

    AIMS: To investigate the usefulness of ocular redox fluorometry for evaluating donor corneal endothelial viability. METHODS: Corneas from 42 recipients of penetrating keratoplasty and four donor corneas were examined by ocular redox fluorometry. Autofluorescence from reduced pyridine nucleotides (PN) and oxidised flavoproteins (Fp) of the human corneal endothelium were measured non-invasively, and the PN/Fp ratio was used as a tissue metabolic indicator. Specular microscopy and electron micro...

  5. Graft Biomechanics Following Three Corneal Transplantation Techniques.

    Science.gov (United States)

    Feizi, Sepehr; Montahai, Talieh; Moein, Hamidreza

    2015-01-01

    To compare corneal biomechanical properties following three different transplantation techniques, including Descemet stripping automated endothelial keratoplasty (DSAEK), deep anterior lamellar keratoplasty (DALK) and penetrating keratoplasty (PK) in comparison to normal eyes. This cross-sectional comparative study included 118 eyes: 17 eyes of 17 patients received DSAEK, 23 eyes of 21 patients underwent DALK using Anwar's big bubble technique, and 45 eyes of 36 patients had PK; 33 right eyes of 33 normal subjects served as the control group. Using the ocular response analyzer (ORA, Reichert Ophthalmic Instruments, Buffalo, New York, USA), corneal hysteresis (CH) and corneal resistance factor (CRF) were measured and compared among the study groups at least 3 months after all sutures were removed. Mean patient age was 26.9 ± 5.0 years in the control group, 28.8 ± 4.2 in the PK group, 27.2 ± 6.5 in the DALK group, and 62.5 ± 16.8 in the DSAEK group (P biomechanical parameters comparable to normal subjects (9.84 ± 1.59 and 9.89 ± 1.73 mmHg, respectively). Graft biomechanical parameters after DSAEK are lower than those following PK and DALK. After PK and DALK in keratoconic eyes, these metrics are increased to normal values. These differences may have implications for interpreting intraocular pressure or planning graft refractive surgery after keratoplasty.

  6. In vivo imaging of the effect of LPS on arterial endothelial cells: molecular imaging of heat shock protein 60 expression

    NARCIS (Netherlands)

    Wick, M.C.; Mayerl, Christina; Backovic, Aleksandar; Zee, R. van der; Jaschke, Werner; Dietrich, Hermann; Wick, G.

    2008-01-01

    Bacterial endotoxins are known as stress factors for endothelial cells. In 20 normocholesterolemic New Zealand White (NZW) rabbits, endothelial stress was induced by intravenous (i.v.) injection of lipopolysaccharide (LPS), while eight NZW rabbits were sham-treated or served as untreated

  7. Confocal microscopy and electrophysiological study of single patient corneal endothelium cell cultures

    Science.gov (United States)

    Tatini, Francesca; Rossi, Francesca; Coppi, Elisabetta; Magni, Giada; Fusco, Irene; Menabuoni, Luca; Pedata, Felicita; Pugliese, Anna Maria; Pini, Roberto

    2016-04-01

    The characterization of the ion channels in corneal endothelial cells and the elucidation of their involvement in corneal pathologies would lead to the identification of new molecular target for pharmacological treatments and to the clarification of corneal physiology. The corneal endothelium is an amitotic cell monolayer with a major role in preserving corneal transparency and in regulating the water and solute flux across the posterior surface of the cornea. Although endothelial cells are non-excitable, they express a range of ion channels, such as voltage-dependent Na+ channels and K+ channels, L-type Ca2 channels and many others. Interestingly, purinergic receptors have been linked to a variety of conditions within the eye but their presence in the endothelium and their role in its pathophysiology is still uncertain. In this study, we were able to extract endothelial cells from single human corneas, thus obtaining primary cultures that represent the peculiarity of each donor. Corneas were from tissues not suitable for transplant in patients. We characterized the endothelial cells by confocal microscopy, both within the intact cornea and in the primary endothelial cells cultures. We also studied the functional role of the purinergic system (adenosine, ATP and their receptors) by means of electrophysiological recordings. The experiments were performed by patch clamp recordings and confocal time-lapse microscopy and our results indicate that the application of purinergic compounds modulates the amplitude of outward currents in the isolated endothelial cells. These findings may lead to the proposal of new therapies for endothelium-related corneal diseases.

  8. Changing Indications and Surgical Techniques for Corneal Transplantation Between 2004 and 2009 at a Tertiary Referral Center

    Science.gov (United States)

    Zare, Mohammad; Javadi, Mohammad A.; Einollahi, Bahram; Karimian, Farid; Rafie, Ali R. B.; Feizi, Sepehr; Azimzadeh, Ahmad

    2012-01-01

    Purpose: The aim of this study is to report the indications, techniques, and clinical outcomes of corneal transplantation and investigate any changing trends in surgical techniques over a 6 year period. Materials and Methods: Records of patients who had undergone any kind of corneal transplantation at Labbafinejad Medical Center, Tehran, Iran, from January 2004 to December 2009 were reviewed to determine the indications and types of corneal transplantation. Postoperative best-corrected visual acuity, refractive error, graft clarity, and complications were reported. Results: During this period, 1859 eyes of 1624 patients with a mean age of 41.3 ± 21.3 years underwent corneal transplantation. The most common indication was keratoconus (38.4%) followed by aphakic/pseudophakic bullous keratopathy (11.7%), previous failed grafts (10.6%), infectious corneal ulcers (10.1%), non-herpetic corneal scars (7.6%), trachoma keratopathy (4.7%), stromal corneal dystrophies (4.6%), post-herpetic corneal scar (3.7%), Fuchs’ endothelial dystrophy (0.8%), and congenital hereditary endothelial dystrophy (0.4%). Techniques of corneal transplantation included penetrating keratoplasty (PKP; 70.9%), deep anterior lamellar keratoplasty (DALK; 20.1%), conventional lamellar keratoplasty (LKP; 4.4%), and Descemet's stripping automated endothelial keratoplasty (DSAEK; 2.3%). Over the study period, there was a significant increase in the relative frequency of infectious corneal ulcers, failed grafts, and trachoma keratopathy. Additionally, a significant reduction was observed in PKP and LKP procedures, and volume of DALK and DSAEK increased significantly. At final follow-up, 69.0% of grafts were clear in the PKP group. This figure was 82.6%, 82.7%, and 97.6% in the DALK, LKP, and DSAEK groups, respectively. Conclusion: Keratoconus was the most common indication and PKP was the most prevalent technique used for corneal transplantation. However, significant changes in the indications and

  9. Treatment of corneal chemical alkali burns with a crosslinked thiolated hyaluronic acid film.

    Science.gov (United States)

    Griffith, Gina L; Wirostko, Barbara; Lee, Hee-Kyoung; Cornell, Lauren E; McDaniel, Jennifer S; Zamora, David O; Johnson, Anthony J

    2018-02-08

    The study objective was to test the utilization of a crosslinked, thiolated hyaluronic acid (CMHA-S) film for treating corneal chemical burns. Burns 5.5mm in diameter were created on 10 anesthetized, male New Zealand white rabbits by placing a 1N NaOH soaked circular filter paper onto the cornea for 30s. Wounds were immediately rinsed with balanced salt solution (BSS). CMHA-S films were placed in the left inferior fornix of five injured and five uninjured animals. Five animals received no treatment. At 0h, 48h, 96h, and on day 14 post chemical burn creation, eyes were evaluated by white light imaging, fluorescein staining, and optical coherence tomography (OCT). Corneal histology was performed using H&E and Masson's Trichrome stains. Image analysis indicated biocompatible CMHA-S treatment resulted in significant decreases in the areas of corneal opacity at 48h, 96h, and on day 14 postoperatively. A significant increase in re-epithelialization was seen 14days post injury. CMHA-S treated corneas showed significantly less edema than untreated burns. No pathological differences were observed in corneal histological samples as a result of CMHA-S treatment. CMHA-S films facilitate re-epithelialization and decrease the area of corneal opacity in our corneal alkali burn rabbit model. Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  10. Distrofia corneal de Schnyder

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    Michel Guerra Almaguer

    Full Text Available La principal entidad hereditaria con depósitos de lípidos en el estroma corneal es la distrofia cristalina central, conocida como distrofia de Schnyder, quien la describió en Suiza en 1927. Se caracteriza por depósitos blanco-amarillentos en el estroma corneal central y superficial. Se presenta un paciente de 28 años, del sexo masculino y piel negra, con antecedente de salud anterior. Acudió a consulta y refirió una disminución de la visión y cambio de coloración progresiva de ambos ojos, de años de evolución. En la exploración oftalmológica de ambos ojos se apreciaron lesiones blanquecinas anulares a nivel del estroma corneal, con ligera turbidez corneal central. Los estudios refractivos realizados constataron un astigmatismo hipermetrópico simple. El resto del examen oftalmológico fue negativo. Para el diagnóstico de certeza se empleó el microscopio confocal. Se concluye que el caso presenta una distrofia corneal estromal de tipo cristalina, de Schnyder.

  11. Corneal Graft Rejection: Incidence and Risk Factors

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    Alireza Baradaran-Rafii

    2008-12-01

    Full Text Available

    PURPOSE: To determine the incidence and risk factors of late corneal graft rejection after penetrating keratoplasty (PKP. METHODS: Records of all patients who had undergone PKP from 2002 to 2004 without immunosuppressive therapy other than systemic steroids and with at least one year of follow up were reviewed. The role of possible risk factors such as demographic factors, other host factors, donor factors, indications for PKP as well as type of rejection were evaluated. RESULTS: During the study period, 295 PKPs were performed on 286 patients (176 male, 110 female. Mean age at the time of keratoplasty was 38±20 (range, 40 days to 90 years and mean follow up period was 20±10 (range 12-43 months. Graft rejection occurred in 94 eyes (31.8% at an average of 7.3±6 months (range, 20 days to 39 months after PKP. The most common type of rejection was endothelial (20.7%. Corneal vascularization, regrafting, anterior synechiae, irritating sutures, active inflammation, additional anterior segment procedures, history of trauma, uncontrolled glaucoma, prior graft rejection, recurrence of herpetic infection and eccentric grafting increased the rate of rejection. Patient age, donor size and bilateral transplantation had no significant influence on graft rejection. CONCLUSION: Significant risk factors for corneal graft rejection include

  12. Corneal endothelium and central corneal thickness changes in type 2 diabetes mellitus

    Directory of Open Access Journals (Sweden)

    El-Agamy A

    2017-03-01

    Full Text Available Amira El-Agamy,1,2,* Shams Alsubaie3,* 1Department of Optometry and Vision Sciences, College of Applied Medical Sciences, King Saud University, Riyadh, Saudi Arabia; 2Mansoura Ophthalmic Center, Faculty of Medicine, Mansoura University, Mansoura, Egypt; 3Department of Optometry and Vision Sciences, Sulaiman Al Habib Hospital, Riyadh, Saudi Arabia *These authors contributed equally to this work Purpose: This study was conducted to compare the corneal endothelial cell density (ECD, morphological features, and central corneal thickness (CCT in type 2 diabetes mellitus (DM with age-matched, nondiabetic control subjects using EM-3000 Specular Microscope. Study design: This was a prospective, hospital-based, nonrandomized, case–control, observational, and quantitative study. Subjects and methods: The study included 57 patients (57 eyes with type 2 DM and 45 control (nondiabetic subjects (45 eyes. The corneal endothelial structure and CCT were examined in all eyes by noncontact specular microscopy using EM-3000 Specular Microscope. The endothelial structure was studied for ECD, coefficient of variation of cell area (CV, and percentage of hexagonal cells. Results: The study included 36 eyes without diabetic retinopathy (DR, 14 eyes with nonproliferative DR, and 7 eyes with proliferative DR. There were 26 eyes with a duration of ≤10 years and 31 eyes with a duration of >10 years. Also, there were 24 eyes with HbA1c ≤7.5% and 33 eyes with HbA1c >7.5%. ECD was significantly lower in the diabetic cornea than in control group (P=0.014. CV was higher in diabetic cornea (P=0.008. The diabetic cornea group had lower percentage of hexagonal cells than the control group, but the difference was not statistically significant (P=0.603. Also, diabetic cornea was thicker than control group, but not statistically significant (P=0.301. Conclusion: This study documented that type 2 DM causes a significant reduction of ECD and increased CV (polymegathism. Also

  13. [Corneal wound healing after penetrating keratoplasty with EGF application. Experimental studies].

    Science.gov (United States)

    Szaflik, J; Fryczkowski, A W; Liberek, I; Czubak, M; Brix, M; Broniek, G; Fryczkowski, P

    1999-01-01

    To check post penetrating keratoplasty (PK) corneal wound healing characteristics after epidermal growth factor (EGF) application and to compare it with controls. The PK was performed in the group of 72 young, healthy New Zealand rabbits (36 females and 36 males). Slit-lamp examination, tonometry and corneal topography by Tomey's corneal modeling system (TMS-1) were carried out before and after surgery. The PK was performed in both eyes. Half of animals were used as a bilateral donor for the other half, with a rule: right eye cornea to the right eye and left eye cornea to the left eye. As a result, after completed surgery 36 rabbits had bilateral grafts. The animals were divided into 3 equal groups (12 in each). Two drops of the human recombined EGF dissolved in the saline solution with concentration varied from 500 to 1500 ng in each drop were applied to the right eye according to schedule. The left eye was used as a control and did not receive EGF. Time of observation varied from 24 hours to 6 months. The tensinometry and the histopathologic study--light and electron microscopy were performed to determine corneal scarring. The wound healing pattern after PK was characteristic and constant in each group. The corneal wound healing significantly accelerated in the EGF treated group of rabbits compared with the controls (p times/day, after two weeks of application we noted increase of the wound strength up to 600 folds, comparing with controls. Well-organized scar was histologically seen on the 21st post-surgery day. The post-operative corneal astigmatism was less expressed in the eyes treated with EGF comparing to controls. These preliminary results of our experimental study indicated accelerated effect on the corneal wound healing after PK with topical, low dose hrEGF application. Clinical observation of utilization of similar low doses of the hrEGF after PK--is in progress.

  14. Clinical effect of femtosecond laser assisted penetrating corneal transplantation operation

    Directory of Open Access Journals (Sweden)

    Hong-Jian Zhou

    2014-10-01

    Full Text Available AIM: To observe the clinical effect of femtosecond laser assisted penetrating keratoplasty. METHODS: Twenty-four cases(24 eyeswith corneal lesions were performed with femtosecond laser assisted penetrating keratoplasty. Preoperative and postoperative endothelial cell density and visual quality were compared.RESULTS: One week after operation, corneal grafts were clear in 21 eyes(87.5%, mild cloudy in 3 eyes(12.5%; visual acuity ≥0.5 in 18 eyes(75.0%, 0.2~0.4 in 6 eyes(25.0%. After 3mo the mean corneal astigmatism was 2.16±0.21D(range 2.25~3.09D. Compared to conventional penetrating keratoplasty which mean corneal astigmatism was average 3.67±0.38D after operation, there was significant difference between two groups(PPCONCLUSION: Femtosecond laser assisted penetrating corneal transplantation operation can improve patient's visual quality. And compared to traditional penetrating keratoplasty astigmatism decreased significantly, incision can be made in individual shape more precisely and neatly.

  15. Deficiency of Cholesteryl Ester Transfer Protein Protects Against Atherosclerosis in Rabbits.

    Science.gov (United States)

    Zhang, Jifeng; Niimi, Manabu; Yang, Dongshan; Liang, Jingyan; Xu, Jie; Kimura, Tokuhide; Mathew, Anna V; Guo, Yanhong; Fan, Yanbo; Zhu, Tianqing; Song, Jun; Ackermann, Rose; Koike, Yui; Schwendeman, Anna; Lai, Liangxue; Pennathur, Subramaniam; Garcia-Barrio, Minerva; Fan, Jianglin; Chen, Y Eugene

    2017-06-01

    CETP (cholesteryl ester transfer protein) plays an important role in lipoprotein metabolism; however, whether inhibition of CETP activity can prevent cardiovascular disease remains controversial. We generated CETP knockout (KO) rabbits by zinc finger nuclease gene editing and compared their susceptibility to cholesterol diet-induced atherosclerosis to that of wild-type (WT) rabbits. On a chow diet, KO rabbits showed higher plasma levels of high-density lipoprotein (HDL) cholesterol than WT controls, and HDL particles of KO rabbits were essentially rich in apolipoprotein AI and apolipoprotein E contents. When challenged with a cholesterol-rich diet for 18 weeks, KO rabbits not only had higher HDL cholesterol levels but also lower total cholesterol levels than WT rabbits. Analysis of plasma lipoproteins revealed that reduced plasma total cholesterol in KO rabbits was attributable to decreased apolipoprotein B-containing particles, while HDLs remained higher than that in WT rabbits. Both aortic and coronary atherosclerosis was significantly reduced in KO rabbits compared with WT rabbits. Apolipoprotein B-depleted plasma isolated from CETP KO rabbits showed significantly higher capacity for cholesterol efflux from macrophages than that from WT rabbits. Furthermore, HDLs isolated from CETP KO rabbits suppressed tumor necrosis factor-α-induced vascular cell adhesion molecule 1 and E-selectin expression in cultured endothelial cells. These results provide evidence that genetic ablation of CETP activity protects against cholesterol diet-induced atherosclerosis in rabbits. © 2017 American Heart Association, Inc.

  16. Airbag induced corneal ectasia.

    Science.gov (United States)

    Mearza, Ali A; Koufaki, Fedra N; Aslanides, Ioannis M

    2008-02-01

    To report a case of airbag induced corneal ectasia. Case report. A patient 3 years post-LASIK developed bilateral corneal ectasia worse in the right eye following airbag deployment in a road traffic accident. At last follow up, best corrected vision was 20/40 with -4.00/-4.00 x 25 in the right eye and 20/25 with -1.25/-0.50 x 135 in the left eye. This is a rare presentation of trauma induced ectasia in a patient post-LASIK. It is possible that reduction in biomechanical integrity of the cornea from prior refractive surgery contributed to this presentation.

  17. Challenges in pediatric endothelial keratoplasty

    Directory of Open Access Journals (Sweden)

    Vikas Mittal

    2014-01-01

    Full Text Available We performed endothelial keratoplasty (EK in three eyes of two siblings (2.5 years, male and 3.5 years, female with congenital hereditary endothelial dystrophy (CHED and report the intraoperative and postoperative difficulties. Repeated iris prolapse, apprehension of crystalline lens touch due to positive vitreous pressure, and need for frequent air injections to attach the graft were intraoperative challenges in all three eyes. These were addressed by use of Sheet′s glide instead of Busin′s glide during graft insertion and suturing of main and side ports before air injection. One eye had graft dislocation on second postoperative day due to eye rubbing by the child. Graft was repositioned with air and a venting incision was created. Postoperative examination required repeated general anesthesia. Corneal edema resolved completely in all three eyes. Present case series highlights the possible intraoperative and postoperative challenges and their solutions in pediatric EK for CHED.

  18. Modeling of corneal and retinal pharmacokinetics after periocular drug administration.

    Science.gov (United States)

    Amrite, Aniruddha C; Edelhauser, Henry F; Kompella, Uday B

    2008-01-01

    To develop pharmacokinetics models to describe the disposition of small lipophilic molecules in the cornea and retina after periocular (subconjunctival or posterior subconjunctival) administration. Compartmental pharmacokinetics analysis was performed on the corneal and retinal data obtained after periocular administration of 3 mg of celecoxib (a selective COX-2 inhibitor) to Brown Norway (BN) rats. Berkeley Madonna, a differential and difference equation-based modeling software, was used for the pharmacokinetics modeling. The data were fit to different compartment models with first-order input and disposition, and the best fit was selected on the basis of coefficient of regression and Akaike information criteria (AIC). The models were validated by using the celecoxib data from a prior study in Sprague-Dawley (SD) rats. The corneal model was also fit to the corneal data for prednisolone at a dose of 2.61 mg in albino rabbits, and the model was validated at two other doses of prednisolone (0.261 and 26.1 mg) in these rabbits. Model simulations were performed with the finalized model to understand the effect of formulation on corneal and retinal pharmacokinetics after periocular administration. Celecoxib kinetics in the BN rat cornea can be described by a two-compartment (periocular space and cornea, with a dissolution step for periocular formulation) model, with parallel elimination from the cornea and the periocular space. The inclusion of a distribution compartment or a dissolution step for celecoxib suspension did not lead to an overall improvement in the corneal data fit compared with the two-compartment model. The more important parameter for enhanced fit and explaining the apparent lack of an increase phase in the corneal levels is the inclusion of the initial leak-back of the dose from the periocular space into the precorneal area. The predicted celecoxib concentrations from this model also showed very good correlation (r = 0.99) with the observed values in

  19. Endothelial cell loss and visual outcome of deep anterior lamellar keratoplasty versus penetrating keratoplasty: a randomized multicenter clinical trial

    NARCIS (Netherlands)

    Cheng, Y.Y.; Visser, N.; Schouten, J.S.A.G.; Wijdh, R.J.; Pels, E.; Cleynenbreugel, H. van; Eggink, C.A.; Zaal, M.J.; Rijneveld, W.J.; Nuijts, R.M.

    2011-01-01

    OBJECTIVE: To compare endothelial cell (EC) loss, visual and refractive outcomes, and complications after deep anterior lamellar keratoplasty (DALK) and penetrating keratoplasty (PK). DESIGN: Randomized, multicenter clinical trial. PARTICIPANTS: Fifty-six eyes of 56 patients with a corneal stromal

  20. Endothelial Cell Loss and Visual Outcome of Deep Anterior Lamellar Keratoplasty versus Penetrating Keratoplasty : A Randomized Multicenter Clinical Trial

    NARCIS (Netherlands)

    Cheng, Yanny Y. Y.; Visser, Nienke; Schouten, Jan S.; Wijdh, Robert-Jan; Pels, Elisabeth; van Cleynenbreugel, Hugo; Eggink, Catharina A.; Zaal, Michel J. W.; Rijneveld, Wilhelmina J.; Nuijts, Rudy M. M. A.

    Objective: To compare endothelial cell (EC) loss, visual and refractive outcomes, and complications after deep anterior lamellar keratoplasty (DALK) and penetrating keratoplasty (PK). Design: Randomized, multicenter clinical trial. Participants: Fifty-six eyes of 56 patients with a corneal stromal

  1. Surgical alternatives to penetrating keratoplasty II: endothelial keratoplasty.

    Science.gov (United States)

    Goins, Kenneth M

    2008-06-01

    Penetrating keratoplasty (PK) became the standard of care for optical and tectonic rehabilitation of corneal blindness and visual impairment in the second half of the twentieth century. Posterior corneal disorders or endotheliopathies are the reason for one-third to one-half of all corneal transplants today in the US. Any procedure that replaces the endothelium ideally should accomplish the following results: (1) a smooth surface topography without significant change in astigmatism, (2) a highly predictable corneal power, (3) a healthy donor endothelium that resolves all edema, (4) a tectonically stable globe, safe from injury and infection, and (5) an optically pure cornea. Although PK consistently can achieve results 3 and 5 above, the other goals of stable topography, predictable corneal power and tectonic stability, have remained elusive despite our best efforts at ingenious suturing and trephination techniques. Endothelial keratoplasty (EK) is a new surgical procedure designed to replace diseased corneal endothelium with healthy donor endothelium through either a lamellar corneal flap approach or through limbal scleral incision, leaving the surface of the recipient cornea untouched by surface corneal sutures. This manuscript evaluates the impact and future of EK in ophthalmology.

  2. Corneal grafting and aggressive medication for corneal defects in graft-versus-host disease following bone marrow transplantation.

    Science.gov (United States)

    Tarnawska, D; Wylegała, E

    2007-12-01

    To evaluate the clinical outcomes of corneal grafting for severe dry eye complications in bone marrow transplant recipients. Eleven eyes of nine patients with severe corneal complications of chronic graft-versus-host disease were treated from 2000 to 2005. The subjects underwent penetrating keratoplasty (n=9 eyes; seven for perforation and two for leucoma) or deep anterior lamellar keratoplasty (n=2 eyes) for deep postinflammatory stromal scarring without endothelial abnormalities. Patients were observed for graft survival, visual acuity, and postoperative complications. During the follow-up period (mean=21.6 months), nine grafts (82%) remained clear or almost entirely clear and gained more than two logarithmic units of best-corrected visual acuity. Two regrafts were necessary as the primary grafts became cloudy after 9 and 11 months. Persistent epithelial defects occurred in seven eyes (64%), cataract in six (55%), ocular hypertension in five (45%), corneal calcareous degeneration in two (18%), loss of clarity in two (18%), and sterile ulceration in one (9%). Corneal grafting was effective for restoring corneal clarity and improving visual function in this series of patients. Although the complication ratio was high and some patients required regrafting, there was a clinical improvement in the majority of patients.

  3. Uso de viscoelásticos na facoemulsificação em cães portadores de catarata: efeitos sobre a pressão intraocular, a morfologia das células endoteliais e a espessura corneana Use of viscoelastic substances for the phacoemulsification in dogs with cataract: effects on the intraocular pressure, morphology of endothelial cells, and corneal thickness

    Directory of Open Access Journals (Sweden)

    J.L.V. Chiurciu

    2010-06-01

    Full Text Available Avaliaram-se as células endoteliais, a espessura corneana e a pressão intraocular (PIO de cães portadores de catarata madura, empregando-se viscoelástico à base de hialuronato de sódio 3% e sulfato de condroitina 4% e hidroxipropilmetilcelulose 2%, utilizando-se 20 cães, distribuídos entre os dois grupos dos viscoelásticos. A técnica cirúrgica adotada foi a da facoemulsificação bimanual. As avaliações tonométricas foram efetuadas antes e após o ato cirúrgico, aos 1, 7, 14, 21, 28 e 60 dias de pós-operatório, e a microscopia especular, antes e após 7, 28 e 60 dias. Não houve diferença estatística entre os grupos quanto à PIO, com exceção aos 14 dias, em que se observou maior PIO com o uso de hialuronato de sódio 3% e sulfato de condroitina 4%. Não houve diferença entre os grupos quanto aos parâmetros relacionados ao endotélio, com diminuição discreta da densidade celular endotelial e aumento da área celular com a utilização de hidroxipropilmetilcelulose 2%. A utilização de ambos os dispositivos viscoelásticos analisados é recomendada para o procedimento de facoemulsificação em cães.The endothelial cells, the corneal thickness, and the intraocular pressure (IOP were evaluated in dogs with cataract, using viscoelastic substances based on 3% sodium hyaluronate and 4% chondroitin sulfate and comparing them with 2% hydroxypropylmethylcellulose. Twenty dogs were distributed in two groups of ten, each using one viscoelastic material. The surgical technique was bimanual phacoemulsification. The tonometric evaluations were made before and at one, seven, 14, 21, 28, and 60 days after the surgery and the specular microscopy before and after seven, 28, and 60 days. No statistical difference between groups was found according to IOP, except at 14 days, which was significantly higher with the use of 3% sodium hyaluronate and 4% chondroitin sulfate. There was no statistical difference between the groups considering

  4. Alternatives to allograft corneal transplantation.

    Science.gov (United States)

    Jhanji, Vishal; Sharma, Namrata; Agarwal, Tushar; Vajpayee, Rasik B

    2010-07-01

    Corneal transplantation is the most commonly performed solid organ transplantation in the world. Despite a glorious history of more than a 100 years, the success of conventional corneal transplantation surgery is marred by problems like graft rejection,graft infection and associated glaucoma due to long-term use of topical corticosteroids.In addition there is a dearth of donor corneal tissue in some parts of the world which subsequently adds on to the existing burden on the eye banks every year. We propose alternatives to the conventional corneal transplantation surgery for the management of corneal scarring. The potential use of alternatives to allograft corneal transplantation surgery has been described by corneal surgeons around the world. These techniques consist of nonsurgical interventions like contact lens fitting. Surgical alternatives include excimer laser phototherapeutic keratectomy, optical iridectomy, rotational autokeratoplasty and contralateral autokeratoplasty. Although these techniques are not practiced routinely, however, their appropriate utilization would clearly help the corneal surgeons to get rid of certain problems associated with allograft corneal transplantation. Careful selection of patients can yield encouraging results with the use of these alternative techniques. Visual outcomes may not be as good as after a routine keratoplasty; nevertheless, this setback is outweighed by advantages such as absence of corneal graft rejection. We also believe that the use of these techniques would at least partially resolve the issue of scarcity of donor corneal tissue in the developing world.

  5. Corneal Biomechanical Response Following Collagen Cross-Linking With Rose Bengal-Green Light and Riboflavin-UVA.

    Science.gov (United States)

    Bekesi, Nandor; Kochevar, Irene E; Marcos, Susana

    2016-03-01

    To compare the biomechanical corneal response of two different corneal cross-linking (CXL) treatments, rose bengal-green light (RGX) and riboflavin-UVA (UVX), using noninvasive imaging. A total of 12 enucleated rabbit eyes were treated with RGX and 12 with UVX. Corneal dynamic deformation to an air puff was measured by high speed Scheimpflug imaging (Corvis ST) before and after treatment. The spatial and temporal deformation profiles were evaluated at constant intraocular pressure of 15 mm Hg, and several deformation parameters were estimated. The deformation profiles were modeled numerically using finite element analysis, and the hyperelastic corneal material parameters were obtained by inverse modeling technique. The corneal deformation amplitude decreased significantly after both CXL methods. The material parameters obtained from inverse modeling were consistent with corneal stiffening after both RGX and UVX. Within the treated corneal volume, we found that the elasticity decreased by a factor of 11 after RGX and by a factor of 6.25 after UVX. The deformation of UVX-treated corneas was smaller than the RGX-treated corneas. However, the reconstructed corneal mechanical parameters reveal that RGX produced in fact larger stiffening of the treated region (100-μm depth) than UVX (137-μm depth). Rose bengal-green light stiffens the cornea effectively, with shorter treatment times and shallower treated areas. Dynamic air puff deformation imaging coupled with mechanical simulations is a useful tool to characterize corneal biomechanical properties, assess different treatments, and possibly help optimize the treatment protocols.

  6. Riboflavin-UV--a crosslinking for fixation of biosynthetic corneal collagen implants.

    Science.gov (United States)

    Wand, Kerstin; Neuhann, Raphael; Ullmann, Andrea; Plank, Katharina; Baumann, Michael; Ritter, Roland; Griffith, May; Lohmann, Chris P; Kobuch, Karin

    2015-05-01

    To evaluate riboflavin-UV-A crosslinking as an alternative suture-free fixation method for biosynthetic corneal collagen implants. A range of cell-free corneal implants consisting of recombinant human collagen type III were examined. In vitro, the implants were crosslinked with different riboflavin solutions and irradiations. Ex vivo, the biosynthetic corneal implants were placed on the anterior cornea of porcine and rabbit eyes after performing deep anterior lamellar keratoplasty with a trephine, femtosecond laser, or excimer laser. UV-A crosslinking was performed with isotonic or hypotonic riboflavin at either standard or rapid procedure. The corneas were excised, fixed in PFA 4%, and embedded in paraffin. Crosslinking effects on the implants and the adhesion between implant and corneal bed were evaluated by slit-lamp biomicroscopy, optical coherence tomography (OCT) images, and histologically. After the crosslinking procedure, the implants showed different degrees of thinning. The accuracy of cutting the corneal bed was highest with the excimer laser. Good adhesion of the implant in the corneal bed could be demonstrated in OCT images. This was more accurate in porcine eyes than in rabbit eyes. Histologically, crosslinks between implant and corneal stroma were demonstrated. There was no difference between standard and rapid crosslinking procedures. Riboflavin-UV-A crosslinking as a fixation method for biosynthetic corneal collagen implants was demonstrated to be promising. It can reduce suture-related complications such as haze formation and surface irregularity. Stability of the implants, especially shrinkage after riboflavin-UV-A crosslinking, needs to be further evaluated. Biostability, integration, and long-term outcome are further evaluated in in vivo animal experiments.

  7. Spontaneous Corneal Hydrops in a Patient with a Corneal Ulcer

    Directory of Open Access Journals (Sweden)

    Hatim Batawi

    2016-01-01

    Full Text Available Purpose: We report the case of a 77-year-old man with no history of keratoconus or other ectatic disorders who presented with corneal hydrops in the setting of a corneal ulcer. The risk factors, pathogenesis and treatment options of corneal hydrops are discussed. Method: This is an observational case report study. Results: A 77-year-old man presented with a 1-day history of severe pain, redness, mucous discharge and photophobia in the right eye. A slit-lamp examination of the right eye showed an area of focal corneal edema and protrusion. Within the area of edema and protrusion, there was an infiltrate with an overlying epithelial defect consistent with an infectious corneal ulcer. The Seidel test showed no leakage, so a clinical diagnosis of corneal hydrops associated with nonperforated corneal ulcer was made. With appropriate antibiotic treatment, the corneal ulcer and hydrops both resolved over a 1-month period. Conclusion: Corneal hydrops can occur in the setting of corneal infections.

  8. Massive corneal edema treated with corneal cross-linking.

    Science.gov (United States)

    Laborante, A; Buzzonetti, L; Longo, C

    2012-01-01

    Massive corneal edema disrupts the fine architecture of corneal stroma that guarantees its transparency, causing opacities that seriously impair clear vision and are usually solved by corneal transplant. Corneal cross-linking, a treatment developed to halt keratoconus progression, results in a loss of water and a compaction of corneal stroma. It might therefore be useful to improve the pathologic edematous condition of some corneas, ameliorating visual acuity and allowing more time for a surgical procedure of keratoplasty. Six patients with visual impairing corneal edemas further to lens phacoemulsification, penetrating keratoplasty, or post-infective neovascularization were treated with corneal cross-linking alone, or in combination with amniotic membrane apposition with or without anti-angiogenic therapy. All patients partly resolved the edematous condition, improving both corneal transparency and visual acuity. Corneal cross-linking appears to be a useful method to treat massive corneal edemas, so that keratoplasty can be at least delayed, and need not to be an emergency treatment in these cases.

  9. Study of light scattering using C-Quant® in patients with Fuchs' endothelial dystrophy: A pilot study.

    Science.gov (United States)

    Castaño-Martín, B; Gros-Otero, J; Martínez, J; Teus, M

    2017-11-01

    The purpose of this study was to determine the light scattering in patients with Fuchs' endothelial dystrophy without clinically significant corneal oedema, and evaluate its relationship with endothelial cell count, corneal thickness, and corneal biomechanical parameters. The values of light scattering were measured by C-Quant® (Oculus Optikgeräte GmbH, Germany) in 32 eyes of 17 patients diagnosed with Fuchs' endothelial dystrophy without clinically significant corneal oedema. Corneal biomechanical properties were determined using ORA (ocular response) and Corvis ST® (tonometry). A light scattering value outside the normal range was observed in 93.8% of eyes studied. No statistically significant association (P>.05) was found between the values of the measured light scattering by C-Quant® and endothelial count, pachymetry, or corneal biomechanical properties. In this study, changes were found in the values of light scattering values of patients with corneal Fuchs' endothelial dystrophy. This change does not appear to correlate significantly with disease severity. Copyright © 2017 Sociedad Española de Oftalmología. Publicado por Elsevier España, S.L.U. All rights reserved.

  10. Corneal biomechanical properties from air-puff corneal deformation imaging

    Science.gov (United States)

    Marcos, Susana; Kling, Sabine; Bekesi, Nandor; Dorronsoro, Carlos

    2014-02-01

    The combination of air-puff systems with real-time corneal imaging (i.e. Optical Coherence Tomography (OCT), or Scheimpflug) is a promising approach to assess the dynamic biomechanical properties of the corneal tissue in vivo. In this study we present an experimental system which, together with finite element modeling, allows measurements of corneal biomechanical properties from corneal deformation imaging, both ex vivo and in vivo. A spectral OCT instrument combined with an air puff from a non-contact tonometer in a non-collinear configuration was used to image the corneal deformation over full corneal cross-sections, as well as to obtain high speed measurements of the temporal deformation of the corneal apex. Quantitative analysis allows direct extraction of several deformation parameters, such as apex indentation across time, maximal indentation depth, temporal symmetry and peak distance at maximal deformation. The potential of the technique is demonstrated and compared to air-puff imaging with Scheimpflug. Measurements ex vivo were performed on 14 freshly enucleated porcine eyes and five human donor eyes. Measurements in vivo were performed on nine human eyes. Corneal deformation was studied as a function of Intraocular Pressure (IOP, 15-45 mmHg), dehydration, changes in corneal rigidity (produced by UV corneal cross-linking, CXL), and different boundary conditions (sclera, ocular muscles). Geometrical deformation parameters were used as input for inverse finite element simulation to retrieve the corneal dynamic elastic and viscoelastic parameters. Temporal and spatial deformation profiles were very sensitive to the IOP. CXL produced a significant reduction of the cornea indentation (1.41x), and a change in the temporal symmetry of the corneal deformation profile (1.65x), indicating a change in the viscoelastic properties with treatment. Combining air-puff with dynamic imaging and finite element modeling allows characterizing the corneal biomechanics in-vivo.

  11. Adherens junction proteins are expressed in collagen corneal equivalents produced in vitro with human cells.

    Science.gov (United States)

    Giasson, Claude J; Deschambeault, Alexandre; Carrier, Patrick; Germain, Lucie

    2014-01-01

    To test whether adherens junction proteins are present in the epithelium and the endothelium of corneal equivalents. Corneal cell types were harvested from human eyes and grown separately. Stromal equivalents were constructed by seeding fibroblasts into a collagen gel on which epithelial and endothelial cells were added on each side. Alternatively, bovine endothelial cells were used. At maturity, sections of stromal equivalents were processed for Masson's trichrome or indirect immunofluorescence using antibodies against pan-, N-, or E-cadherins or α- or β-catenins. Alternatively, stromal equivalents were dissected, to separate the proteins from the epithelium, endothelium, and stroma with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Western blots of the transferred proteins exposed to these primary antibodies were detected with chemiluminescence. Native corneas were processed similarly. Three or four layers of epithelial cells reminiscent of the native cornea (basal cuboidal and superficial flatter cells) lay over a stromal construct containing fibroblastic cells under which an endothelium is present. Western blots and indirect immunofluorescence revealed that, similarly to the native cornea, the epithelium reacted positively to antibodies against catenins (α and β) and E-cadherin. The endothelium of corneal constructs, whether of human or bovine origin, reacted mildly to catenins and N-cadherin. This collagen-based corneal equivalent simulated the native cornea. Cells from the epithelial and endothelial layers expressed adherens junction proteins, indicating the presence of cell-cell contacts and the existence of polarized morphology of these layers over corneal equivalents.

  12. [Relationship between corneal neovascularization and various relevant biological factors in surrounding cornea stroma of rats].

    Science.gov (United States)

    Wang, Ting; Shi, Wei-yun; Li, Su-xia; Liu, Ming-na

    2009-02-01

    To study the relationship between corneal neovascularization and various biological factors in corneal stroma of rats. It was an experimental study. Corneal neovascularization was induced by alkali burn in 40 rats. Transforming growth factor-beta1 (TGF-beta1), alpha-smooth muscle actin (alpha-SMA) and fibroblast activation protein (FAP) in the stroma surrounding corneal neovascularization were detected by immunohistochemical studies on day 1, 3 and 7 after chemical burn. Platelet-endothelial cell adhesion molecule-1 (CD31) was used to identify the vascular endothelial cells. RT-PCR was used to identify FAP in the cornea 3 and 7 days after chemical burn. Picrosirius staining and polarization microscopy were used to detect changes of collagen types I and III in the cornea. After alkali burn, TGF-beta1 was first expressed in the cornea stroma. Then, some stroma cells expressed both alpha-SMA and FAP. The FAP(+) keratocytes were found surrounding the CD31(+) endothelium of angiogenesis. RT-PCR study showed that FAP mRNA was only present in neovascularized cornea and not in normal cornea. Polarization microscopy revealed that the collagen types I and III were rearranged in neovascularized cornea. Various biological factors in corneal stroma are changed when the cornea shows neovascularization. FAP(+) keratocytes are present in the stroma, and the appearance of these cells parallels the growth of vascular endothelial cells. Collagen types I and III are rearranged during the process of angiogenesis.

  13. Preparation and characterization of a novel tobramycin-containing antibacterial collagen film for corneal tissue engineering.

    Science.gov (United States)

    Liu, Yang; Ren, Li; Long, Kai; Wang, Lin; Wang, Yingjun

    2014-01-01

    Corneal disease is a major cause of blindness and keratoplasty is an effective treatment method. However, clinical treatment is limited due to a severe shortage of high-quality allogeneic corneal tissues and the bacterial infection after corneal transplantation. In this study, we develop a novel artificial and antibacterial collagen film (called Col-Tob) for corneal repair. In the Col-Tob film, the tobramycin, which is an aminoglycoside antibiotic to treat various types of bacterial infections, was cross-linked by 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide and N-hydroxysuccinimide onto the collagen. Physical properties, antibacterial property and biocompatibility of the films were characterized. The results indicate that the film is basically transparent and has appropriate mechanical properties. Cell experiments show that human corneal epithelial cells could adhere to and proliferate well on the film. Most importantly, the film exhibits excellent antibacterial effect in vitro. Lamellar keratoplasty shows that the Col-Tob film can be sutured in rabbit eyes and are epithelialized completely in 15 ± 5 days, and their transparency is restored quickly in the first month. Corneal rejection reaction, neovascularization and keratoconus are not observed within 3 months. This film, which can be prepared in large quantities and at low cost,should have potential application in corneal repair. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  14. [An experimental research of recombinant human epidermal growth factor on corneal wound healing].

    Science.gov (United States)

    Zheng, R; Jin, X; Yang, B; Li, B; Li, L; Xu, Z; Zhu, H

    1998-05-01

    To investigate the effects of recombinant human epidermal growth factor (rhEGF) eye drops on corneal wound healing. Twenty-four white rabbits were randomly divided into 4 groups, 6 rabbits 12 eyes each. Anterior keratectomy of 8 mm in diameter and 1/3 cornea in thickness was performed on each eye. Each of the following concentrations of rhEGF: 1, 10, 100 microg/ml eye drops or normal saline (control) was applied four times daily for a week respectively for one group. The wound area was determined by computer imaging analysis. The mean epithelial healing rate of rhEGF 1, 10, 100 microg/ml groups was 9.31, 9.96, 9.31 mm(2)/day respectively, significantly greater than 8.11 mm(2)/day of the control group. The action of rhEGF of 10 microg/ml was somewhat better than that of 1 or 100 microg/ml, and no significant difference was noticed among the three rhEGF groups. Moderate inflammation and corneal neovascularization were induced in the rhEGF 100 microg/ml treated group. rhEGF 1 - 10 microg/ml can accelerate corneal wound healing in the rabbit with no adverse side-effects. It may be used to treat serious corneal trauma and ulcer clinically.

  15. Corneal biomechanical properties in 3 corneal transplantation techniques with a dynamic Scheimpflug analyzer.

    Science.gov (United States)

    Maeda, Naoyuki; Ueki, Ryotaro; Fuchihata, Mutsumi; Fujimoto, Hisataka; Koh, Shizuka; Nishida, Kohji

    2014-11-01

    To compare the corneal biomechanical properties of eyes that have undergone penetrating keratoplasty (PK), deep anterior lamellar keratoplasty (DALK), and Descemet stripping automated endothelial keratoplasty (DSAEK). This case-control study comprised 20 post-PK eyes, 14 post-DALK eyes, 15 post-DSAEK eyes, and 50 normal control eyes. A dynamic Scheimpflug analyzer (the Corvis ST) was used to evaluate the corneal biomechanical properties including deformation amplitude (DA) and radius at the highest concavity (R hc). In post-PK eyes, the mean DA was 1.20 ± 0.13 mm, which was significantly higher than those of the control eyes (1.07 ± 0.09) and the post-DSAEK eyes (1.08 ± 0.12). The DA (1.18 ± 0.18) in the post-DALK eyes was significantly higher than in the control eyes. The R hc in the post-PK (6.34 ± 0.37 mm), -DALK (6.04 ± 1.22), and -DSAEK (6.44 ± 0.58) eyes was significantly smaller than in the control eyes (7.57 ± 0.78). The dynamic Scheimpflug analyzer provides a method to obtain new biomechanical information on the cornea such as the DA and R hc, and these parameters differed among eyes that had undergone 3 different types of corneal surgery. Abnormalities in these parameters after the different corneal transplantation techniques may indicate larger deviations in the stress-strain reaction of the cornea and more uncertainty in the intraocular pressure measurements than in normal eyes.

  16. Chondroitin Sulfate-Based Biocompatible Crosslinker Restores Corneal Mechanics and Collagen Alignment.

    Science.gov (United States)

    Wang, Xiaokun; Majumdar, Shoumyo; Ma, Garret; Sohn, Jeeyeon; Yiu, Samuel C; Stark, Walter; Al-Qarni, Awad; Edward, Deepak P; Elisseeff, Jennifer H

    2017-08-01

    To evaluate the crosslinking effect of functionalized chondroitin sulfate (CS) in an ex vivo rabbit cornea model. Chondroitin sulfate molecules were chemically modified with the N-hydroxysuccinimide (NHS) group. Enucleated rabbit eyes were crosslinked with 2, 5, or 10 mg/mL CS-NHS solution for 30 or 60 minutes. The CS-NHS penetration, corneal swelling ratio, Young's modulus, and ultrastructure of the crosslinked corneas were characterized. In addition, rabbit corneas were further treated with a collagenase-chondroitinase solution to create an ex vivo keratoconus (KC)-like model. The KC model corneas were crosslinked with a standard riboflavin-ultraviolet (UV) method or alternatively with CS-NHS. Corneal mechanics, ultrastructure, and keratocyte gene expression were evaluated after UV and CS-NHS crosslinking. CS-NHS effectively penetrated into the corneal stroma within 60 minutes of treatment initiation. CS-NHS crosslinking reduced the swelling ratio by 35%, increased Young's modulus by 20%, and increased collagen fibril diameter and density. CS-NHS crosslinking improved corneal mechanics of KC model corneas to levels comparable to those with UV crosslinking. Moreover, CS-NHS crosslinking demonstrated significant downregulation of proinflammatory gene expression of keratocytes, indicating a potential protective effect imparted by CS-NHS during crosslinking. Our results demonstrated that CS-NHS can reinforce normal and KC model corneal mechanics, and restore collagen density and alignment in KC model corneas without causing extensive keratocyte apoptosis and proinflammatory gene upregulation. Therefore, CS-NHS crosslinking can potentially provide an effective, safe, and biocompatible means of corneal reinforcement.

  17. Computational Model for Corneal Transplantation

    Science.gov (United States)

    Cabrera, Delia

    2003-10-01

    We evaluated the refractive consequences of corneal transplants using a biomechanical model with homogeneous and inhomogeneous Young's modulus distributions within the cornea, taking into account ablation of some stromal tissue. A FEM model was used to simulate corneal transplants in diseased cornea. The diseased cornea was modeled as an axisymmetric structure taking into account a nonlinearly elastic, isotropic formulation. The model simulating the penetrating keratoplasty procedure gives more change in the postoperative corneal curvature when compared to the models simulating the anterior and posterior lamellar graft procedures. When a lenticle shaped tissue was ablated in the graft during the anterior and posterior keratoplasty, the models provided an additional correction of about -3.85 and -4.45 diopters, respectively. Despite the controversy around the corneal thinning disorders treatment with volume removal procedures, results indicate that significant changes in corneal refractive power could be introduced by a corneal transplantation combined with myopic laser ablation.

  18. The Association of Delayed Corneal Surface and Visual Recovery After Corneal Transplantation With Longer Donor Storage Time-A Prospective Analysis.

    Science.gov (United States)

    Lam, Douglas K T; Chan, Tommy C Y; Wong, Janice S C; Chow, Galen Y; Wong, Victoria W Y

    2014-01-01

    This study aimed to assess the influence of donor corneal storage time on endothelial cell count (ECC), corneal epithelial recovery, and visual rehabilitation after corneal transplantation in the first postoperative year. A collaborative prospective study involving a local eye bank and a tertiary ophthalmic unit was conducted. Donor cornea buttons were stored in Optisol-GS (Chiron Ophthalmics Inc, Irvine, Calif) storage media for a maximum of 14 days before transplantation. Before corneal distribution, the eye bank collected information on death-to-harvesting time, death-to-surgery time, donor central corneal thickness, and donor ECC at various time points. Subjects who underwent penetrating keratoplasty and endothelial keratoplasty were recruited and monitored for 1 year. Postoperative epithelial healing, visual acuity, ECC, and hospital stay were evaluated. Thirty-one eyes of 31 patients completed the study. There was a significant positive correlation between donor storage time and epithelial healing (Spearman ρ = 0.39, P = 0.031). Faster epithelial healing was significantly correlated with posttransplantation visual improvements at months 1, 3, and 6 and shorter hospital stay (Spearman ρ = 0.74, P time and ECC loss preoperatively and posttransplantation. The duration of graft storage in Optisol-GS storage media up to 14 days had no significant effects on long-term visual acuity and ECC postoperatively. Shorter storage time had significant correlation with earlier epithelial healing and faster visual rehabilitation.

  19. SUITABILITY OF CORNEAL TISSUE FOR TRANSPLANTATION PROCURED FROM HANGING CASES

    Directory of Open Access Journals (Sweden)

    Rekha Gyanchand

    2017-06-01

    Full Text Available BACKGROUND Requirement of donor cornea is essential to target the corneal blind. The best method to procure such corneas is from any major hospitals, which has a mortuary facility. The eye donation with hanging as the cause of death is very common in a mortuary setup. Some factors that are concerning regarding corneas procured from death due to hanging is the prolonged exposure of the cornea at the time of death, the exact time of death is not known, most of the cadavers are refrigerated for investigations as these arrive at the mortuary usually at night. Due to these reasons, the corneal surgeons are hesitant to use corneas procured from death due to hanging for corneal transplantation. Analysing these corneas would contribute to a great extent to the donor cornea pool in providing sight to the corneal blind, especially as majority are young individuals who commit suicide by hanging. In this study, the donor corneas were analysed with regards to corneal epithelial defect, endothelial cell morphology and utilisation of these corneas for transplantation. The aim of the HCRP study is to analyse the effect of death due to hanging on donor cornea. 1. Corneal epithelial status. 2. Corneal endothelial cell morphology. 3. Utilisation of corneas for transplantation. MATERIALS AND METHODS Donor corneas from 22 donors who died due to hanging were procured from hospital mortuary. All the 44 corneas were transplanted. Various parameters like demography, death to enucleation time, cadaver preservation in cold storage, endothelial cell density and utilisation of cornea for transplantation were noted. Design- Retrospective study. Statistical Analysis- Descriptive statistics, Pearson and Spearman correlation and Chi-square test were used to test the hypotheses. RESULTS Out of the 44 corneas analysed, 75% of the donors were refrigerated as a part of medicolegal investigations protocol. The average DTP time was 12 hours in refrigerated group and 5 hours in non

  20. Human corneal epithelial subpopulations

    DEFF Research Database (Denmark)

    Søndergaard, Chris Bath

    2013-01-01

    subpopulations in human corneal epithelium using a combination of laser capture microdissection and RNA sequencing for global transcriptomic profiling. We compared dissociation cultures, using either expansion on γ-irradiated NIH/3T3 feeder cells in serum-rich medium or expansion directly on plastic in serum......-free EpiLife medium, using a range of physiologically relevant oxygen concentrations (2%, 5%, 10%, 15% and 20%). Using immunocytochemistry and advanced fluorescence microscopy, cells were characterized regarding growth, cell cycle distribution, colony-forming efficiency (CFE), phenotypes...... was not dependent on the system used for propagation (Bath et al. 2013a). Laser capture microdissection was used to isolate cellular subpopulations in situ from the spatially defined differentiation pathway in human corneal epithelium according to an optimized protocol for maintenance of expression profiles...

  1. Biomechanics of Corneal Ring Implants

    OpenAIRE

    Daxer, Albert

    2015-01-01

    Purpose: To evaluate the biomechanics of corneal ring implants by providing a related mathematical theory and biomechanical model for the treatment of myopia and keratoconus. Methods: The spherical dome model considers the inhomogeneity of the tunica of the eye, dimensions of the cornea, lamellar structure of the corneal stroma, and asphericity of the cornea. It is used in this study for calculating a strengthening factor sf for the characterization of different ring-shaped corneal implant de...

  2. [Comparative evaluation of different corneal cross-linking techniques with respect to biomechanical stability of the cornea].

    Science.gov (United States)

    Pashtaev, N P; Pozdeeva, N A; Sinitsyn, M V; Zotov, V V; Gagloev, B V

    2016-01-01

    To perform a comparative analysis of the following cross-linking techniques: standard cross-linking (SCXL), local transepithelial femto cross-linking (LTF) and transepithelial intrastromal femto cross-linking with MyoRing implantation (TIF+MyoRing) in experimental animals and patients with progressive stage II--III keratoconus (KC), paying particular attention to the changes in biomechanical stability of the cornea. The experimental series was performed on 20 eyes of 10 rabbits. The animals were divided into 4 groups of 5. Group 1 served as the control, group 2 consisted of animals that underwent SCXL, group 3 -- of those after LTF, and group 4 -- of those after TIF+MyoRing. Femto step was performed using the IntraLase FS 60 kHz femtosecond laser, cross-linking - with the Evolution machine. The follow-up period was 1 month. Clinical study enrolled 48 patients (45 eyes) with stage II-III KC. Depending on the surgical technique all the patients were divided into 3 groups. In group I (14 eyes, 9 patients), TIF+MyoRing was performed, in group II (16 eyes, 20 patients) --SCXL, in group III (15 eyes, 19 patients) -- LTF. The follow-up period was 6 months. The experiment showed a significant increase in rupture resistance of the cornea in all three groups. Six months after surgery, patients from group I demonstrated a 1.0±0.22 mmHg higher corneal resistance factor (CRF) and a 1.0±0.12 mmHg higher corneal hysteresis (CH). In group II, the said parameters increased by 0.8±0.10 mmHg and 0.6±0.16 mmHg, respectively; in group III -- by 0.8±0.25 mmHg and 0.6±0.26 mmHg, respectively. In neither group the density of endothelial cells has reduced significantly over the follow-up period. C-ONCLUSION: All three methods (LTF, TIF and SCXL) equally increase the biomechanical strength of the cornea and are safe. In patients with progressive stage II-III KC, TIF+MyoRing surgery has yielded an even higher improvement of biomechanical parameters in a 6-month follow-up period, which

  3. Corneal biomechanics: a review

    OpenAIRE

    Piñero Llorens, David Pablo; Alcón, Natividad

    2014-01-01

    Biomechanics is often defined as ‘mechanics applied to biology’. Due to the variety and complexity of the behaviour of biological structures and materials, biomechanics is better defined as the development, extension and application of mechanics for a better understanding of physiology and physiopathology and consequently for a better diagnosis and treatment of disease and injury. Different methods for the characterisation of corneal biomechanics are reviewed in detail, including those that a...

  4. Corneal surface reconstruction - a short review

    Directory of Open Access Journals (Sweden)

    Madhavan H N

    2009-01-01

    . Cells or tissues can be embedded in a liquid Mebiol gel solution at lower than 20°C and cultured three dimensionally in a hydrogel state at 37°C. The sol-gel transition temperature can be controlled by altering chemical composition of thermo-reversible gelation polymer (TGP. Mebiol Gel prevents the growth of the fibroblasts and it is not toxic to cells. In our earlier study, continuous culture cell lines 18 and we demonstrated that Mebiol Gel supported the growth of corneal limbal epithelial cells and these cells expressed both limbal and corneal phenotype, suggesting that limbal epithelial cells cultivated in Mebiol Gel would be a promising material for corneal surface tissue engineering. With this in view, we developed a novel method of cultivation of human limbal cells in a synthetic scaffold made of a thermo reversible polymer, that allows the limbal epithelial cells to survive, proliferate and differentiate into corneal epithelial cells and demonstrated the effectiveness of this polymer for corneal tissue engineering using an experimentally induced limbal deficiency in a rabbit model. In this report, the detailed procedure of the animal model and the results are discussed. We evaluated the efficacy of autologous expanded corneal epithelial cell transplants derived from harvested limbal biopsy cultured on a thermo-reversible polymer (Mebiol Gel for the management of unilateral limbal stem cell disease (LSCD.Corneal limbal biopsies from 12 rabbits were cultured on Mebiol Gel at 37°C. Cells were harvested from the dishes after 3 weeks by reducing temperature to 4°C. Autologous transplantation was undertaken to reconstruct the experimentally induced limbal stem cell deficiency in the rabbit eyes. The corneas of both eyes of all rabbits were harvested later for histological and RT-PCR studies. Reparative surgery was a total success in 7 (58.3%; score, 8-10, partial success in 2 (16.7%; score, 6-7 and failure in 3 (25%; score, <5. Histological and RT-PCR study

  5. Femtosecond laser-assisted sutureless anterior lamellar keratoplasty for superficial corneal opacities.

    Science.gov (United States)

    Jabbarvand, Mahmoud; Hashemian, Hesam; Khodaparast, Mehdi; Ghadimi, Hadi; Khalilipour, Elias

    2014-11-01

    To evaluate the visual and refractive outcomes, endothelial cell count (ECC), ocular surface changes, corneal aberrations, and biomechanical profile changes after femtosecond laser-assisted anterior lamellar keratoplasty surgery for superficial corneal scars. Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran. Prospective case series. Patients with superficial corneal scars had femtosecond laser-assisted anterior lamellar keratoplasty. Visual and refractive results, ECC, ocular surface changes, corneal aberrations, and biomechanical profiles were assessed preoperatively and for 1 year postoperatively. Nineteen eyes (19 patients) were evaluated. A significant decline occurred in refractive astigmatism and corneal astigmatism after 1 year. There was a nonsignificant reduction in corneal hysteresis and the corneal resistance factor from preoperatively to 1 year postoperatively. The corneal-compensated intraocular pressure (IOP) and Goldmann-correlated IOP increased during the follow-up; the increase was not significant. A statistically insignificant reduction in the root mean square for trefoil and spherical aberrations occurred between 1 month and 1 year postoperatively (P=.1 and P=.4, respectively). The decreases in primary coma and total higher-order aberrations approached significance (P=.08 and P=.07, respectively). There were no significant changes in the central corneal thickness, ECC, or ocular surface parameters. No intraoperative complications occurred. Femtosecond laser-assisted anterior lamellar keratoplasty was an efficient and safe procedure for improving the quality of vision in patients with anterior corneal pathology, and the results remained stable during the 1-year follow-up. No author has a financial or proprietary interest in any material or method mentioned. Copyright © 2014 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

  6. Decellularization of porcine corneas and repopulation with human corneal cells for tissue-engineered xenografts.

    Science.gov (United States)

    Yoeruek, Efdal; Bayyoud, Tarek; Maurus, Christine; Hofmann, Johanna; Spitzer, Martin S; Bartz-Schmidt, Karl-Ulrich; Szurman, Peter

    2012-03-01

    To evaluate the potential use of decellularized porcine corneas (DPCs) as a carrier matrix for cultivating human corneal cells in tissue engineering. Corneal cells were isolated from human corneoscleral rims. Porcine corneas were decellularized using hypotonic tris buffer, ethylene diamine tetra-acetic acid (EDTA, 0.1%), aprotinin (10 KIU/ml) and 0.3% sodium dodecyl sulphate. Haematoxylin-eosin (HE) and 4,6-diamidino-2-phenylindole (DAPI) staining were performed to confirm removal of the corneal cells. Quantitative analysis was performed to determine levels of desoxyribonucleic acid (DNA) using DNA Purification Kit (Fermentas, St. Leon-Rot, Germany). Alcian blue staining was carried out to analyse the structure of the extracellular matrix (ECM). Corneal stromal cells were injected into the DPCs; limbal corneal epithelial cells and corneal endothelial cells were seeded onto the anterior and posterior surfaces of the DPCs, respectively. Evaluation was undertaken at days 14 and 30. The phenotypical properties of the cultivated corneal cells were investigated using Immunolocalization of type I collagen, keratocan, lumican, cytokeratin 3 (AE5) and type VIII collagen. Haematoxylin-eosin and DAPI staining showed efficient elimination of porcine corneal cells, whereas alcian blue confirmed gross preservation of the ECM. The quantitative analysis of the DNA content showed a significant reduction (mean before decellularization: 75.45 ± 13.71 ng/mg; mean after decellularization: 9.87 ± 2.04 ng/mg, p types of corneal cells were efficiently cultured and expanded on the DPCs. Decellularized porcine corneas might serve as a potential scaffold for tissue engineering of the cornea, possibly providing xenogenic substrate for corneal transplantation. © 2011 The Authors. Acta Ophthalmologica © 2011 Acta Ophthalmologica Scandinavica Foundation.

  7. Airbag-induced corneal flap.

    Science.gov (United States)

    Liyanage, Sidath E; Mearza, Ali A

    2009-02-01

    To describe a case of airbag-induced corneal flap in a previously normal cornea. Case report. A 27-year-old woman presented with complete loss of vision in her left eye following a road traffic accident which involved airbag deployment. There was no previous ocular history. Examination revealed a large corneal flap of 6mm in diameter, extending to the depth of anterior stroma. This was accompanied by a traumatic optic neuropathy. One month follow-up revealed complete reattachment of the corneal flap. This is the first reported case of a corneal flap induced by airbag deployment in a cornea with previously normal architecture.

  8. Evaluation of Corneal Endothelium in Children and Adolescents with Type 1 Diabetes Mellitus

    OpenAIRE

    Urban, Beata; Raczyńska, Dorota; Bakunowicz-Łazarczyk, Alina; Raczyńska, Krystyna; Krętowska, Małgorzata

    2013-01-01

    Purpose. To evaluate the systemic and local factors that contribute to the damage of endothelial cells in diabetic patients and to compare the endothelial structure of the cornea in diabetic and nondiabetic patients. Materials and Methods. The endothelial cell density (ECD) and central corneal thickness (CCT) were investigated in 123 eyes of type 1 diabetic patients and in 124 eyes of nondiabetic patients. The mean diabetic patients age was 15.34 ± 3.06 years versus 14.58 ± 2.01 years in the ...

  9. Mononuclear cells in the corneal response to endotoxin

    Energy Technology Data Exchange (ETDEWEB)

    Howes, E.L.; Cruse, V.K.; Kwok, M.T.

    1982-04-01

    A severe keratitis can be produced after the direct injection of bacterial endotoxin, or lipopolysaccharide (LPS), in rabbits. Corneal inflammation can progress to scarring and vascularization within a 2 to 3 week period. Pretreatment with systemic adrenal corticosteroids (triamcinolone) prevents this response. Limbal cellular and vascular events were studied during the first 20 hr after injection of LPS in treated and nontreated rabbits. Perivascular limbal inflammatory cells were counted and limbal vascular permeability was assessed by extravasation of 131I-albumin and 125I-fibrinogen in the cornea. Corticosteroids decreased but did not prevent the early protein extravasation and profoundly altered the inflammatory cell population around blood vessels at the limbus. Mononuclear cells, particularly mononuclear phagocytes, were sharply reduced. It is proposed that these cell types play an important role in the perpetuation and amplification of the inflammatory response in this reaction.

  10. Comparative analysis of corneal morphological changes after transversal and torsional phacoemulsification through 2.2 mm corneal incision

    Directory of Open Access Journals (Sweden)

    Assaf A

    2013-01-01

    Full Text Available Ahmed Assaf, Maged Maher RoshdyOphthalmology Department, Ain Shams University, Cairo, EgyptPurpose: This paper compares and evaluates the corneal morphological changes occurring after cataract surgery through a 2.2 mm corneal incision. We use two platforms for comparison and evaluation, transversal and torsional phacoemulsification.Patients and methods: This study includes 139 consecutive cataractous eyes (nuclear color 2–4, according to the Lens Opacities Classification System III [LOCSIII] of 82 patients undergoing cataract surgery through a 2.2 mm corneal incision. Two different phacoemulsification platforms were used and assigned randomly: we used the WhiteStar Signature® system with the Ellips™ FX transversal continuous ultrasound (US mode for group I (mean age: 65.33 ± 6.97 years, and we used the Infiniti® system with the OZil® Intelligent Phaco (IP torsional US mode for group II (mean age: 64.02 ± 7.55 years. The corneal endothelium and pachymetry were evaluated preoperatively and at 1 month postoperatively. Incision size changes were also evaluated.Results: All surgeries were uneventful. Before intraocular lens implantation, the mean incision size was 2.24 ± 0.06 mm in both groups (P = 0.75. In terms of corneal endothelial cell density, neither preoperative (I vs II: 2304.1 ± 122.5 cell/mm2 vs 2315.6 ± 83.1 cell/mm2, P = 0.80 nor postoperative (I vs II: 2264.1 ± 124.3 cell/mm2 vs 2270.3 ± 89.9 cell/mm2, P = 0.98 differences between the groups were statistically significant. The mean endothelial cell density loss was 1.7% ± 1.6% and 2.0% ± 1.4% in groups I and II, respectively. Furthermore, no significant differences between groups I and II were found preoperatively (P = 0.40 and postoperatively (P = 0.68 in central pachymetry. With surgery, the mean increase in central pachymetry was 28.1 ± 23.6 µm and 24.0 ± 24.0 µm in groups I and II, respectively (P = 0.1.Conclusion: Ellips™ FX transversal and OZil® IP torsional

  11. Changes in Corneal Topography and Biomechanical Properties after Collagen Cross Linking for Keratoconus: 1-Year Results

    Science.gov (United States)

    Sedaghat, Mohammadreza; Bagheri, Mansooreh; Ghavami, Shahri; Bamdad, Shahram

    2015-01-01

    Purpose: To evaluate changes in corneal topography and biomechanical properties after collagen cross-linking (CXL) for progressive keratoconus. Patients and Methods: Collagen cross-linking was performed on 97 eyes. We assessed uncorrected visual acuity (UCVA) and best corrected visual acuity (BCVA). Corneal topography indices were evaluated using placido disc topography, scanning slit anterior topography (Orbscan II), and rotating Scheimpflug topography (Pentacam). Specular microscopy and corneal biomechanics were evaluated. Results: A 1-year-follow-up results revealed that UCVA improved from 0.31 to 0.45 and BCVA changed from 0.78 to 0.84 (P corneal thickness decreased from 458.11 to 444.46 μm. Corneal volume decreased from 56.66 to 55.97 mm3 (P Corneal hysteresis changed from 8.18 to 8.36 (P = 0.552) and corneal resistance factor increased from 6.98 to 7.21 (P = 0.202), so these changes were not significant. Conclusion: Visual acuity and K values improved after CXL. In spite of the nonsignificant increase in endothelial cell count and increase in the CV, CLX seems to be a safe treatment for keratoconus. Further studies with larger sample sizes and longer follow-up periods are recommended. PMID:25949080

  12. Measurement of surgically induced corneal deformations using three-dimensional confocal microscopy.

    Science.gov (United States)

    Petroll, W M; Roy, P; Chuong, C J; Hall, B; Cavanagh, H D; Jester, J V

    1996-03-01

    The goal of this study was to develop and apply a new set of experimental techniques for measuring the local deformations induced by partial-thickness corneal incisions in situ. Eight adult cat eyes were enucleated and cannulated, with corneal viability maintained as close to in vivo conditions as possible and intraocular pressure (IOP) carefully controlled. Experimental measurements were made pre/post radial keratotomy (RK) surgery in situ at IOPs of 15, 30, and 45 mm Hg. Incision depth and cross-sectional profiles were measured at the midpoint of selected incisions using three-dimensional (3-D) tandem scanning confocal microscopy (TSCM); central corneal curvature was estimated using a commercial corneal topographical analysis system, and corneal thickness was assessed by both 3-D TSCM and ultrasonic pachymetry. Corneas were then processed for light microscopy and incision depth was measured histologically. Finite element models were developed for comparison with the experimental measurements. There was no significant change in central corneal thickness (-5.3 +/- 3.9%, n = 8) over the course of the experiments, demonstrating that normal endothelial cell function and normal stromal hydration was maintained. The in situ TSCM incision depth measurements were significantly correlated with the histological measurements (slope = 0.95, R = 0.854, p mechanical behavior of the cornea after refractive surgery. These data should provide the foundation for future studies into the relationships between local tissue mechanics and corneal wound healing.

  13. In Vitro Corneal Tomography of Donor Cornea Using Anterior Segment OCT.

    Science.gov (United States)

    Janunts, Edgar; Langenbucher, Achim; Seitz, Berthold

    2016-05-01

    The aim of this study was to establish a tomographic screening method for revealing potential pathologies in corneal donors before keratoplasty so they may be excluded as candidates for corneal transplantation. Donor corneal tomographies were measured in a viewing chamber filled with preservation medium and with the use of a clinical optical coherence tomography (OCT) device. Custom-written software was developed to extract corneal surfaces from the raw data, which were analyzed in the central and peripheral regions. An adaptive nonlinear edge-enhancement algorithm was used to observe scars within the corneal volume. The thickness distribution map was analyzed to detect keratoconus and corneas with extreme topographic irregularities. Measurements were repeated 5 times to assess reproducibility. Eight corneas were investigated: 6 randomly selected intact donors, unsuitable for implantation because of low endothelial cell densities, and 2 keratoconus corneas, excised from patients during corneal transplantation. A major thickness abnormality was detected in one of the intact donor corneas, so it was excluded from further analysis. The keratoconus corneas were clearly evident in optical coherence tomography cross-sectional images, and similarly, they could easily be identified by analyzing the thickness map. Overall, the measurements were reliable and had a Cronbach's alpha coefficient greater than 0.8. Donor corneal examination using sterile viewing chambers was found to be suitable as a pre-keratoplasty advanced screening routine. A proof of concept was demonstrated, which could identify both irregular corneas and those affected by keratoconus.

  14. The Rabbit Stream Cipher

    DEFF Research Database (Denmark)

    Boesgaard, Martin; Vesterager, Mette; Zenner, Erik

    2008-01-01

    The stream cipher Rabbit was first presented at FSE 2003, and no attacks against it have been published until now. With a measured encryption/decryption speed of 3.7 clock cycles per byte on a Pentium III processor, Rabbit does also provide very high performance. This paper gives a concise...... description of the Rabbit design and some of the cryptanalytic results available....

  15. Incisor adjustment in rabbits.

    Science.gov (United States)

    Brown, Cyndi

    2009-06-01

    Rabbit incisor teeth are open-rooted and, in healthy animals, grow continuously. Incisor adjustments are often necessary to maintain the health and well-being of rabbits with incisor malocclusion. This column will describe some techniques used to manage incisor malocclusion in the rabbit.

  16. Evaluation of the corneal endothelium in patients with diabetes mellitus type I and II.

    Science.gov (United States)

    Módis, László; Szalai, Eszter; Kertész, Katalin; Kemény-Beke, Adám; Kettesy, Beáta; Berta, András

    2010-12-01

    The aim of the present study is to determine corneal physiology and endothelial morphology after proper image analysis technique in type I and II diabetic patients. The HbA1c level and the grade of retinopathy were also recorded and correlated with the endothelial parameters. 41 eyes of 21 patients with type I and 59 eyes of 30 patients with type II diabetes mellitus (mean age was 40.97 ± 15.46 and 64.36 ± 10.47 years) were examined and compared to age-matched controls. Endothelial cell density (ECD), mean cell area, coefficient of variation of cell area, central corneal thickness, intraocular pressure, and grade of retinopathy were recorded. There was a statistically significant decreased endothelial cell density in type I disease (2428 ± 219 cell/mm2) in comparison with healthy subjects (2495 ± 191 cell/mm2, P=0.02). The diabetic corneas were thicker than normal (P=0.001). The HbA1c level was inversely correlated with the ECD (r=-0.60; Pevaluated values. The present study disclosed the alteration of the corneal endothelial morphology in type I diabetes mellitus as compared to normal subjects. The results indicated that type I diabetic corneas are more susceptible to environmental changes than type II corneas.

  17. Correlations between corneal hysteresis, intraocular pressure, and corneal central pachymetry.

    Science.gov (United States)

    Touboul, David; Roberts, Cynthia; Kérautret, Julien; Garra, Caroline; Maurice-Tison, Sylvie; Saubusse, Elodie; Colin, Joseph

    2008-04-01

    To analyze the correlation between corneal hysteresis (CH) measured with the Ocular Response Analyzer (ORA, Reichert) and ultrasonic corneal central thickness (CCT US) and intraocular pressure measured with Goldmann applanation tonometry (IOP GA). Bordeaux 2 University, Ophthalmology Department, Bordeaux, France. This study comprised 498 eyes of 258 patients. Corneal hysteresis, corneal resistance factor (CRF), and IOP corneal-compensated (IOPcc) were provided by the ORA device; CCT US and IOP GA were also measured in each eye. The study population was divided into 5 groups: normal (n = 122), glaucoma (n = 159), keratoconus (n = 88), laser in situ keratomileusis (LASIK) (n = 78), and photorefractive keratectomy (n = 39). The Pearson correlation was used for statistical analysis. Corneal hysteresis was not strongly correlated with IOP or CCT US. The mean CH in the LASIK (8.87 mm Hg) and keratoconus (8.34 mm Hg) groups was lower than in the glaucoma (9.48 mm Hg) and normal (10.26 mm Hg) groups. The lower the CH, the lower its correlation with IOPcc and IOP GA. A CH higher than the CRF was significantly associated with the keratoconus and post-LASIK groups. Corneal hysteresis, a new corneal parameter, had a moderate dependence on IOP and CCT US. Weaker corneas could be screened with ORA parameters, and low CH could be considered a risk factor for underestimation of IOP. The CCT US should continue to be considered a useful parameter.

  18. Changes in corneal hysteresis after clear corneal cataract surgery.

    Science.gov (United States)

    Hager, Annette; Loge, Kristina; Füllhas, Marc-Oliver; Schroeder, Bernd; Grossherr, Martin; Wiegand, Wolfgang

    2007-09-01

    To assess the changes in corneal hysteresis (CH) as measured by the Ocular Response Analyzer (ORA; Reichert Ophthalmic Instruments, Buffalo, New York, USA) to describe the influence of clear corneal cataract surgery on corneal viscoelastic properties and intraocular pressure (IOP) measured by noncontact tonometry (NCT) and Goldmann applanation tonometry (GAT). Retrospective, interventional, comparative study. One hundred and one eyes of 101 consecutive patients who underwent routine clear corneal cataract surgery were evaluated. CH, NCT, and central corneal thickness (CCT) were measured by ORA before surgery and at postoperative day 1. A control group of 48 pseudophakic eyes (surgery >3 months previously) was included. CCT increased from 556.82 +/- 32.5 microm before surgery to 580.26 +/- 45.5 microm after surgery (P corneal cataract surgery, CH is diminished, whereas CCT is increased significantly. Postoperative corneal edema leads to a change of corneal viscoelastic properties, resulting in a lower damping capacity of the cornea. It is supposed that GAT and NCT measurements are significantly different because of postoperative changes in viscoelastic properties of the cornea.

  19. In-vivo corneal pulsation in relation to in-vivo intraocular pressure and corneal biomechanics assessed in-vitro. An animal pilot study.

    Science.gov (United States)

    Rogala, Maja M; Danielewska, Monika E; Antończyk, Agnieszka; Kiełbowicz, Zdzisław; Rogowska, Marta E; Kozuń, Marta; Detyna, Jerzy; Iskander, D Robert

    2017-09-01

    The aim was to ascertain whether the characteristics of the corneal pulse (CP) measured in-vivo in a rabbit eye change after short-term artificial increase of intraocular pressure (IOP) and whether they correlate with corneal biomechanics assessed in-vitro. Eight New Zealand white rabbits were included in this study and were anesthetized. In-vivo experiments included simultaneous measurements of the CP signal, registered with a non-contact method, IOP, intra-arterial blood pressure, and blood pulse (BPL), at the baseline and short-term elevated IOP. Afterwards, thickness of post-mortem corneas was determined and then uniaxial tensile tests were conducted leading to estimates of their Young's modulus (E). At the baseline IOP, backward stepwise regression analyses were performed in which successively the ocular biomechanical, biometric and cardiovascular predictors were separately taken into account. Results of the analysis revealed that the 3rd CP harmonic can be statistically significantly predicted by E and central corneal thickness (Models: R2 = 0.662, p biomechanics in-vitro was confirmed. In particular, spectral analysis revealed that higher amplitude and power of the 3rd CP harmonic indicates higher corneal stiffness, while the 1st CP harmonic correlates positively with the corresponding harmonic of the BPL signal. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. In situ ultrahigh-resolution optical coherence tomography characterization of eye bank corneal tissue processed for lamellar keratoplasty.

    Science.gov (United States)

    Brown, Jamin S; Wang, Danling; Li, Xiaoli; Baluyot, Florence; Iliakis, Bernie; Lindquist, Thomas D; Shirakawa, Rika; Shen, Tueng T; Li, Xingde

    2008-08-01

    To use optical coherence tomography (OCT) as a noninvasive tool to perform in situ characterization of eye bank corneal tissue processed for lamellar keratoplasty. A custom-built ultrahigh-resolution OCT (UHR-OCT) was used to characterize donor corneal tissue that had been processed for lamellar keratoplasty. Twenty-seven donor corneas were analyzed. Four donor corneas were used as controls, whereas the rest were processed into donor corneal buttons for lamellar transplantation by using hand dissection, a microkeratome, or a femtosecond laser. UHR-OCT was also used to noninvasively characterize and monitor the viable corneal tissue immersed in storage medium over 3 weeks. The UHR-OCT captured high-resolution images of the donor corneal tissue in situ. This noninvasive technique showed the changes in donor corneal tissue morphology with time while in storage medium. The characteristics of the lamellar corneal tissue with each processing modality were clearly visible by UHR-OCT. The in situ characterization of the femtosecond laser-cut corneal tissue was noted to have more interface debris than shown by routine histology. The effects of the femtosecond laser microcavitation bubbles on the corneal tissue were well visualized at the edges of the lamellar flap while in storage medium. The results of our feasibility study show that UHR-OCT can provide superb, in situ microstructural characterization of eye bank corneal tissue noninvasively. The UHR-OCT interface findings and corneal endothelial disc thickness uniformity analysis are valuable information that may be used to optimize the modalities and parameters for lamellar tissue processing. The UHR-OCT is a powerful approach that will allow us to further evaluate the tissue response to different processing techniques for posterior lamellar keratoplasty. It may also provide information that can be used to correlate with postoperative clinical outcomes. UHR-OCT has the potential to become a routine part of tissue

  1. Intrastromal delivery of bevacizumab using microneedles to treat corneal neovascularization.

    Science.gov (United States)

    Kim, Yoo C; Grossniklaus, Hans E; Edelhauser, Henry F; Prausnitz, Mark R

    2014-09-11

    This study tested the hypothesis that highly targeted intrastromal delivery of bevacizumab using coated microneedles allows dramatic dose sparing compared with subconjunctival and topical delivery for treatment of corneal neovascularization. Stainless steel microneedles 400 μm in length were coated with bevacizumab. A silk suture was placed in the cornea approximately 1 mm from the limbus to induce corneal neovascularization in the eyes of New Zealand white rabbits that were divided into different groups: untreated, microneedle delivery, topical eye drop, and subconjunctival injection of bevacizumab. All drug treatments were initiated 4 days after suture placement and area of neovascularization was measured daily by digital photography for 18 days. Eyes treated once with 4.4 μg bevacizumab using microneedles reduced neovascularization compared with untreated eyes by 44% (day 18). Eyes treated once with 2500 μg bevacizumab using subconjunctival injection gave similar results to microneedle-treated eyes. Eyes treated once with 4.4 μg subconjunctival bevacizumab showed no significant effect compared with untreated eyes. Eyes treated with 52,500 μg bevacizumab by eye drops three times per day for 14 days reduced the neovascularization area compared with untreated eyes by 6% (day 18), which was significantly less effective than the single microneedle treatment. Visual exam and histological analysis showed no observable effect of microneedle treatment on corneal transparency or microanatomical structure. This study shows that microneedles can target drug delivery to corneal stroma in a minimally invasive way and demonstrates effective suppression of corneal neovascularization after suture-induced injury using a much lower dose compared with conventional methods. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.

  2. Thiopental Sodium Anesthesia Following Premedication of Rabbits with Vitamin C

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    Maryam Asfari

    2010-06-01

    Full Text Available Rabbits are widely used as laboratory animals for experimental surgery. Anesthesia of rabbits may present complications unless the method is easy to apply and safe to use. In present study, effects of different dosages of vitamin C on thiopental sodium induced anesthesia in 25 male New Zealand white rabbits were studied. In the animals that had not received vitamin C treatment before thiopental sodium induced general anesthesia, return mean time of front limb pedal, corneal and ear pinch reflexes were 6.40 ± 1.67, 6.60 ± 2.96 and 8.00 ± 2.58 minutes, respectively. Pre-treatment of rabbits with 30 and 240 mg kg-1 (IV of vitamin C followed by thiopental sodium 20 mg kg-1 (IV resulted in significant (P < 0.05 increase in front limb pedal reflex return mean time to 13.00 ± 2.24 and ear pinch to 11.60 ± 4.16 minutes, respectively. There was also significant (P < 0.05 decrease in the heart rate following induction of anesthesia in the animals pre-treated with 30 and 90 mg kg-1 (IV vitamin C and no change in the animals pre-treated with 240 mg kg-1 (IV vitamin C. Serum analysis indicated a significant (P < 0.05 increase in blood glucose. These results suggest that premedication of rabbits with vitamin C despite potentiating of thiopental sodium anesthesia in rabbits is not dose dependent.

  3. Intrastromal Injection of China Painting Ink in Corneas of Male Rabbits: Clinical and Histological Study

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    Alahmady Hamad Alsmman Hassan

    2016-01-01

    Full Text Available Background. Many patients with corneal opacity or complicated cataract in blind eye ask for cosmoses. In this study we tried to investigate the staining of corneas of male rabbits by Rotring China painting ink and to study the histological changes. Method. 10 eyes of 10 male Baladi Egyptian rabbits were injected (0.1 mL intrastromally in the cornea by the use of China painting ink (Rotring Tinta China through insulin syringe (27-gauge needle by single injection; clinical follow-up is for 6 months and lastly the rabbits were scarified and the stained eyes were enucleated for histological analysis. Results. Clinically the stain was stable in color and distribution in corneas with no major complications. Histological results of the stained rabbit corneas showed blackish pigmentation in the corneal stroma without any inflammatory cellular infiltration. Some fibroblast cells had pigment granules in their cytoplasm in the adjacent layers. Conclusion. Corneal staining by China painting ink is effective and safe in staining of male rabbits cornea; however further study in human corneas with longer follow-up period is advisable.

  4. Role of corneal collagen fibrils in corneal disorders and related pathological conditions

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    Hong-Yan Zhou

    2017-05-01

    Full Text Available The cornea is a soft tissue located at the front of the eye with the principal function of transmitting and refracting light rays to precisely sense visual information. Corneal shape, refraction, and stromal stiffness are to a large part determined by corneal fibrils, the arrangements of which define the corneal cells and their functional behaviour. However, the modality and alignment of native corneal collagen lamellae are altered in various corneal pathological states such as infection, injury, keratoconus, corneal scar formation, and keratoprosthesis. Furthermore, corneal recuperation after corneal pathological change is dependent on the balance of corneal collagen degradation and contraction. A thorough understanding of the characteristics of corneal collagen is thus necessary to develop viable therapies using the outcome of strategies using engineered corneas. In this review, we discuss the composition and distribution of corneal collagens as well as their degradation and contraction, and address the current status of corneal tissue engineering and the progress of corneal cross-linking.

  5. Investigation of protective effects of sodium hyaluronate eyedrop against corneal epithelial disorders using an electrophysiological method.

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    Araki, Ryosuke; Nakashima, Mikiro; Teshima, Mugen; Owaki, Yuichi; Nakashima, Mihoko N; Uematsu, Masafumi; Kitaoka, Takashi; Nakamura, Tadahiro; Kitahara, Takashi; Sasaki, Hitoshi

    2012-06-01

    The purpose of this study was to investigate the protective effects of sodium hyaluronate eyedrop against corneal epithelial disorders caused by antiglaucomatous eyedrops using an electrophysiological method. Three kinds of antiglaucomatous eyedrops, including benzalkonium chloride (BAC) as an ophthalmic preservative, a BAC-free antiglaucomatous eyedrop, and a sodium hyaluronate eyedrop, were used in this study. Eyedrops were applied to excised rabbit corneas, and the electrophysiological property of the cornea was monitored using an Ussing chamber with a turnover system that mimics human tear turnover. With this system, changes in transepithelial electrical resistance (TER) in the corneal surface were recorded. The corneal TER after applying antiglaucomatous eyedrops tended to decrease concomitantly with increasing the concentration of the BAC included as a preservative. On the other hand, there was no significant change in the corneal TER for the initial 60 min after applying sodium hyaluronate eyedrop compared with those of the control. Moreover, the pretreatment with a sodium hyaluronate eyedrop reduced the extent of decrease in the corneal TER observed after application of antiglaucomatous eyedrops alone. Those results indicate that a sodium hyaluronate eyedrop has the potential to protect the corneal surface from antiglaucomatous eyedrops, including BAC as an ophthalmic preservative.

  6. [Refractive changes after Descemet membrane endothelial keratoplasty].

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    Röck, T; Bartz-Schmidt, K U; Röck, D; Yoeruek, E

    2014-01-01

    Penetrating keratoplasty has different refractive disadvantages in contrast to posterior lamellar keratoplasty. For example, a decentered corneal trephination and a tilted trephination or unevenly tightened corneal sutures can cause an uncontrolled high astigmatism as well as a refractive change. Also the postoperative refraction may change over time as a result of wound healing, suture loosening or suture removal. The aim of this retrospective study was to investigate a possible refractive change after Descemet membrane endothelial keratoplasty (DMEK). A total of 139 pseudophakic eyes from 125 patients with endothelial decompensation had undergone DMEK surgery at the Tübingen Eye Clinic. After a mean postoperative time of 13.15 ± 2.98 months after DMEK discreet mean changes in the spherical equivalent of + 0.37 ± 0.87 diopters and the cylinder to - 0.45 ± 0.57 diopters were observed. The mean central corneal thickness decreased from 670 ± 70 µm to 544 ± 55 µm. In conclusion after DMEK a discreet induced hyperopic refractive shift due to the reversal of stromal swelling was observed.

  7. Effects of Topical 1% Sodium Hyaluronate and Hydroxypropyl Methylcellulose in Treatment of Corneal Epithelial Defects.

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    Shahraki, Kourosh; Hosseini, Seyed-Rafi; Amini Fard, Atefeh; Shademan, Hashem; Shahraki, Kianoush; Salari, Amir Masood; Amini Fard, Mohammad-Naeim

    2016-01-01

    We aimed to compare the therapeutic effects of topical 1% sodium hyaluronate (Healon) or hydroxypropyl methylcellulose (HPMC) for the treatment of alkali-induced epithelial corneal defects. An alkali burn was produced in 30 corneas of 30 New Zealand White rabbits, using a 7.5-mm-diameter trephine. The rabbits were randomly divided into three groups. Four times a day, one group was treated with 1% sodium hyaluronate, one with HPMC, and one (the control group) with physiologic saline. During the treatment period, the size of the epithelial defect was observed every day, up to day 17, using a slit-lamp biomicroscope (with fluorescein). Sodium hyaluronate significantly accelerated the wound healing process compared with saline and increased the healing rate to an even greater extent compared with HPMC. Sodium hyaluronate, but not HPMC, is an effective wound-healing adjuvant for alkali-induced corneal epithelial defects.

  8. Ghrelin inhibits atherosclerotic plaque angiogenesis and promotes plaque stability in a rabbit atherosclerotic model.

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    Wang, Li; Chen, Qingwei; Ke, Dazhi; Li, Guiqiong

    2017-04-01

    Intraplaque angiogenesis associates with the instability of atherosclerotic plaques. In the present study, we investigated the effects of ghrelin on intraplaque angiogenesis and plaque instability in a rabbit model of atherosclerosis. The rabbits were randomly divided into three groups, namely, the control group, atherosclerotic model group, and ghrelin-treated group, with treatments lasting for 4 weeks. We found that the thickness ratio of the intima to media in rabbits of the ghrelin-treated group was significantly lower than that in rabbits of the atherosclerotic model group. The number of neovessels and the levels of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor 2 (VEGFR2) decreased dramatically in rabbits of the ghrelin-treated group compared to those of the atherosclerotic model group. Ghrelin significantly decreased the plaque content of macrophages, matrix metalloproteinase (MMP)-2, and MMP-9, in a rabbit model of atherosclerosis. In addition, the level of the pro-inflammatory factor monocyte chemoattractant protein (MCP)-1 was significantly lower in rabbits of the ghrelin-treated group than in rabbits of the atherosclerotic model group. In summary, ghrelin can inhibit intraplaque angiogenesis and promote plaque stability by down-regulating VEGF and VEGFR2 expression, inhibiting the plaque content of macrophages, and reducing MCP-1 expression at an advanced stage of atherosclerosis in rabbits. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Optimal use of donor corneal tissue: one cornea for two recipients.

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    Sharma, Namrata; Agarwal, Prakashchand; Titiyal, Jeewan S; Kumar, Chandrashekhar; Sinha, Rajesh; Vajpayee, Rasik B

    2011-10-01

    To evaluate the feasibility, safety, and efficacy of the use of 1 donor cornea for 2 recipients who required anterior lamellar and posterior lamellar keratoplasties, respectively. Twelve eyes with anterior corneal stromal pathology and 12 eyes with irreversible endothelial dysfunction were evaluated for transplant surgery at a tertiary eye care referral center. Twelve healthy donor corneas were split into 2 parts, that is, anterior lamellar button (350-μm-thick) and posterior lamellar button (150-μm-thick) using a microkeratome (Moria, Antony, France). The anterior lamellar button was used to perform automated lamellar therapeutic keratoplasty in 12 eyes, and the posterior lamellar button was used to undertake Descemet stripping automated endothelial keratoplasty in 12 eyes. The parameters evaluated were feasibility of the procedure, intraoperative and postoperative complications, if any, and visual outcome. It was possible to use 12 donor tissues for 24 patients as envisaged. No intraoperative or postoperative complications were observed. In the automated lamellar therapeutic keratoplasty group, 83.3% (10 of 12) of patients achieved a best-corrected visual acuity (BCVA) of >20/60, and in the Descemet stripping automated endothelial keratoplasty group 75% (9 of 12) of patients achieved a BCVA of >20/60. Our study demonstrates that 1 donor corneal tissue can be successfully used for 2 patients as a routine practice with appropriate and optimal case selection. Such techniques may help to reduce the magnitude of corneal blindness in developing countries where there are shortages of donor corneal tissue.

  10. Infiltrating inflammatory cell phenotypes and apoptosis in rejected human corneal allografts.

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    Larkin, D F; Alexander, R A; Cree, I A

    1997-01-01

    The aim of this study was to survey the histopathological and immunohistochemical features of rejected human corneal allografts. Following graft failure in each case due to rejection, paraffin-embedded specimens of 17 corneal transplants which had been replaced were examined by light microscopy and immunohistochemistry. Specimens were either first (n = 9), second (n = 4) or third (n = 4) grafts and were removed at varying intervals from 4 weeks following documented rejection. Those grafts which were removed earliest following onset of rejection had the most intense graft inflammatory infiltrates. Immunohistochemical staining showed a high proportion of graft stroma-infiltrating cells expressing leucocyte common antigen, and many of these cells also bore T cell or macrophage markers. Leucocyte-keratocyte apposition and regional loss of keratocytes were observed in all rejection specimens, but not in non-rejected control grafts. In situ end-labelling of DNA double-strand breaks and morphological features identified keratocyte apoptosis in 5 of 12 specimens examined for this phenomenon. Corneal endothelial cells were absent in 7 specimens and present in reduced numbers in the remaining 10 specimens. Endothelial cell monolayer attenuation and keratocyte loss are consistent findings in grafts removed subsequent to clinically observed endothelial rejection. Death of donor corneal cells is mediated, at least in part, by apoptosis. The stromal inflammatory infiltrate consists mainly of T lymphocytes and macrophages, which may be responsible for induction of keratocyte apoptosis.

  11. Efeitos da atorvastatina, fluvastatina, pravastatina e simvastatina sobre a função endotelial, a peroxidação lipídica e a aterosclerose aórtica em coelhos hipercolesterolêmicos Effects of atorvastatin, fluvastatin, pravastatin, and simvastatin on endothelial function, lipid peroxidation, and aortic atherosclerosis in hypercholesterolemic rabbits

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    Paulo Afonso Ribeiro Jorge

    2005-04-01

    wall of hypercholesterolemic rabbits after adjusting the dosages of those statins to reduce total serum cholesterol levels to similar values. METHODS: Male rabbits were divided into the following 6 groups of 10 animals (n=10: 1 GH (control - hypercholesterolemic animals; 2 GA - atorvastatin; 3 GF - fluvastatin; 4 GP - pravastatin; 5 GS - simvastatin; and 6 GN - normal. The animals were fed a standard food preparation enriched with 0.5% cholesterol and 2% coconut oil for 45 days. Fifteen days after beginning the experiment, atorvastatin, fluvastatin, pravastatin and simvastatin were administered for 15 days through gavage, and the dosages were adjusted to obtain similar cholesterol values in each group. At the end of the experiment, a blood sample was withdrawn for determining total cholesterol and separating the lipoproteins, and a segment of the thoracic aorta was removed to be used for studying endothelial function and lipid peroxidation, and for measuring aortic atherosclerosis in histological sections. RESULTS: The statins significantly reduced total serum cholesterol levels, LDL-cholesterol levels, and aortic atherosclerosis. The MDA content was also significantly reduced in native and oxidized LDL, as well as in the arterial wall. Endothelium-dependent relaxation was significantly greater in the treated group compared with that in the hypercholesterolemic group. CONCLUSION: The statins, at dosages adjusted, had a significant and similar effect in reducing lipid peroxidation in native and oxidized LDL-C and in arterial walls, in decreasing aortic atherosclerosis, and in reverting endothelial dysfunction.

  12. In Vivo Characteristics of Corneal Endothelium/Descemet Membrane Complex for the Diagnosis of Corneal Graft Rejection.

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    Abou Shousha, Mohamed; Yoo, Sonia H; Sayed, Mohamed S; Edelstein, Sean; Council, Matthew; Shah, Ravi S; Abernathy, Joshua; Schmitz, Zachary; Stuart, Patrick; Bentivegna, Rocio; Fernandez, Maria P; Smith, Christopher; Yin, Xiaotang; Harocopos, George J; Dubovy, Sander R; Feuer, William J; Wang, Jianhua; Perez, Victor L

    2017-06-01

    To evaluate the utility of endothelial/Descemet membrane complex (En/DM) characteristics in diagnosing corneal graft rejection. Diagnostic reliability study. One hundred thirty-nine eyes (96 corneal grafts post penetrating keratoplasty or Descemet stripping automated endothelial keratoplasty: 40 clear, 23 actively rejecting, 24 rejected, and 9 nonimmunologic failed grafts; along with 43 age-matched control eyes) were imaged using high-definition optical coherence tomography. Images were used to describe En/DM and measure central corneal thickness (CCT) and central En/DM thickness (DMT). En/DM rejection index (DRI) was computed to detect the relative En/DM thickening to the entire cornea. In actively rejecting grafts, DMT and DRI were significantly greater than controls and clear grafts (28, 17, and 17 μm and 1.5, 1 and 1, respectively; P < .001). Rejected grafts had the highest DMT and DRI compared to all groups (59 μm and 2.1; P < .001). DMT and DRI showed excellent accuracy, significantly better than that of CCT, in differentiating actively rejecting from clear grafts (100% and 96% sensitivity; 92.5% and 92.5% specificity), actively rejecting from rejected grafts (88% and 83% sensitivity; 91% and 83% specificity), and nonimmunologic failed from rejected grafts (100% and 100% sensitivity; 88% and 100% specificity). DMT correlated significantly with rejection severity (P < .001). In corneal grafts, in vivo relative thickening of the En/DM is diagnostic of graft rejection as measured by DMT and DRI. These indices have excellent accuracy, sensitivity, and specificity in detecting graft immunologic status, superior to CCT. DMT is a quantitative index that correlates accurately with the severity of rejection. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Sphingosine-1 phosphate prevents ethanol-induced corneal epithelial apoptosis

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    Pierre Fournie

    2012-01-01

    Full Text Available Background: Apoptosis is a programmed cell death in multicellular organisms, found in a wide variety of conditions, including inflammatory process, everywhere in the body, including the cornea and conjunctiva. Aim: To evaluate the effect of a new topical formulation of sphingosine-1 phosphate on preventing apoptosis of the corneal epithelium. Setting: Medical University. Materials and Methods: We tested several formulations suitable for topical application. Twenty-five rabbits were distributed among five groups. Group 1 comprised the controls. In Group 2, 20% ethanol was applied topically for 20 seconds; in Group 3, 50 μM topical sphingosine-1 phosphate was applied 2 hours prior to 20% ethanol application. In Group 4, 200 μM topical sphingosine-1 phosphate was applied 2 hours before the 20% ethanol application. In Group 5, only 200 μM topical sphingosine-1 phosphate was applied. Apoptosis was evaluated using the terminal deoxynucleotidyl transferase biotin-dUTP Nick End Labeling (TUNEL assay. Pairwise comparisons were performed using t-tests with Scheffe′s correction. Data were analyzed using STATA 9.0 statistical software. Results: A suspension of sphingosine-1 phosphate in the presence of Montanox 80 was stable and could be formulated without sonication. Epithelial apoptosis was detected only in Groups 2 and 3. Conclusion: Sphingosine-1 phosphate can prevent ethanol-induced apoptosis in the corneal epithelium of rabbits.

  14. Corneal stromal acupuncture combined with amniotic membrane transplantation for treating bullous keratopathy

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    Chao-Qing Wang

    2014-06-01

    endothelial cells reduced and form swelled more obviously compared with preoperatively.CONCLUSION: Corneal acupuncture combined with amniotic membrane transplantation can effectively control the symptoms of BK, prevent the recurrence of BK, and especially it is a simple, safe and practical way for patients with poor visual function.

  15. Deep lamellar endothelial keratoplasty visual acuity, astigmatism, and endothelial survival in a large prospective series.

    Science.gov (United States)

    Terry, Mark