Reproducibility of Endothelial Assessment during Corneal Organ Culture: Comparison of a Computer-Assisted Analyzer with Manual Methods

Deb‐Joardar, Nilanjana; Thuret, Gilles; Gavet, Yann; Acquart, Sophie; Garraud, Olivier; Egelhoffer, Harald; Gain, Philippe

doi:10.1167/iovs.06-1043

Cited by 32 publications

(9 citation statements)

References 18 publications

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“…In the current study, the precision estimated as the ratio between the residual standard deviation and the average estimated level (Table 1, 5th column) for polymegethism and pleomorphism was consistent with findings in other studies where inter‐operator and inter‐system variances were examined (de Sanctis et al. 2006; Deb‐Joardar et al. 2007a,b).…”

Section: Discussionsupporting

confidence: 90%

“…A common model for analysing the random error in endothelial morphometry is to divide the random error into inter‐system variation, inter‐operator variation and measurement error. Inter‐system variation, which involves inter‐operator variation, has been analysed in several recent studies (Deb‐Joardar et al. 2007a,b; Hirneiss et al.…”

Section: Discussionmentioning

confidence: 99%

“…Inter‐operator and inter‐system studies typically result in higher variations (de Sanctis et al. 2006; Deb‐Joardar et al. 2007a,b) associated with variabilities among systems and operators.…”

Section: Discussionmentioning

confidence: 99%

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Simulation of specular microscopy images of corneal endothelium, a tool for control of measurement errors

Bucht

Söderberg

Manneberg

2011

Acta Ophthalmologica

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Purpose:We aimed at developing simulation software capable of producing images of corneal endothelium close to identical to images captured by clinical specular microscopy with defined morphometrical characteristics. It was further planned to demonstrate the usefulness of the simulator by analysing measurement errors associated with a trained operator using a commercially available semi-automatic algorithm for analysis of simulated images.Methods: Software was developed that allows creation of unique images of the corneal endothelium expressing morphology close to identical with that seen in images of corneal specular microscopy. Several hundred unique images of the corneal endothelium were generated with randomization, spanning a physiological range of endothelial cell density. As an example of the usefulness of the simulator for analysis of measurement errors in corneal specular microscopy, a total of 12 of all the images generated were randomly selected such that the endothelial cell density expressed was evenly distributed over the physiological range of endothelial cell density. The images were transferred to a personal computer. The imagenet-640 software was used to analyse endothelial cell size variation, percentage of hexagonal endothelial cells, and endothelial cell density. Results:The simulator developed allows randomized generation of corneal specular microscopy images with a preset expected average and variation of cell structure. Calculated morphometric information of each cell is stored in the simulator. The image quality can secondarily be varied with a toolbox of filters to approximate a large spectrum of clinically captured images. As an example of the use of the simulator, measurement errors associated with one trained operator using the imagenet-640 software, and focusing on endothelial cell density, were examined. The functional dependence between morphometric information estimated with the imagenet-640 software algorithm and real morphometric information as provided by the simulator was analysed with regression. It was demonstrated that that the estimations of endothelial cell size variation was associated with a scaling error and that the random error was strongly dependent on the operator. Conclusion:The newly developed simulator for randomized generation of morphometrically defined corneal specular microscopy images for the first time makes it possible to estimate a spatial scaling error of an available semi-automatic algorithm and to determine the random measurement error of important morphometric estimates in a defined reference sample of images. It is anticipated that the simulator will be a valuable tool for the generation of a large set of morphometrically well-characterized corneal specular microscopy images that can be used for calibration among research centres, for minimization of random errors and for measurement of quality control. Simulated images will be useful for the development of fully automatic analysis of corneal endothelial cell morphometry.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

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Simulation of specular microscopy images of corneal endothelium, a tool for control of measurement errors

Bucht

Söderberg

Manneberg

2011

Acta Ophthalmologica

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“…Irrespective of apparent EC density (ECD), the best three images in terms of overall image quality were chosen. ECD and morphometry (coefficient of variation (CV) of cell area and hexagonal cell percentage) were determined using a Sambacornea endothelial analyzer (Sambatechnologies, Meylan, France) [25,26,27,28,29] on at least 300 cells. The entire corneal endothelium stained with trypan blue was also photographed with a digital camera (Canon EOS 350D, Tokyo, Japan).…”

Section: Methodsmentioning

confidence: 99%

Poloxamines for Deswelling of Organ-Cultured Corneas

et al. 2012

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Background: Poloxamines are amphiphilic tetrofunctional block copolymers composed of four polyoxyethylene-polyoxypropylene arms joined to a central ethylene diamine bridge. Their safe profile allows diverse pharmaceutical and biomedical applications. Aim: To assess their use for corneal deswelling using a porcine model of organ culture (OC). Methods: Five poloxamines (T90R4, T904, T908, T1107 and T1307) were dissolved in a standard commercial OC medium (control) to reach 350 mosm kg^–1. In vitro cytotoxicity was tested using MTT assay on human corneal epithelial and endothelial cell (EC) lines and on primary human corneal fibroblasts. Paired porcine corneas stored in OC for 3 days were assigned for 48 h to a poloxamine medium or to a standard deswelling medium containing 5% dextran T500. Corneal EC density, morphometry, mortality, stromal thickness and transparency were evaluated before and after deswelling. Post-deswelling, EC viability/mortality was determined using a fluorescent live/dead assay. Results: Besides similar corneal thickness reduction and transparency improvement, T908, T1107 and T1307 decreased EC loss (5.4 ± 1.7% vs. 9.9 ± 2.6% in controls (p < 0.001)) and mortality, improved EC morphometry and reduced endothelial lesions compared to dextran. Conclusion: On this porcine model, poloxamines T908, T1107 and T1307 appear as good candidates to replace dextran for the deswelling. Experiments on human corneas are now necessary to confirm their efficiency and safety profile in OC.

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“…In this study, every cornea was procured by in situ excision with single-use sterile instrument, without any contact with retina or optic nerve, which are be considered as specific risk factors to possible iatrogenic spread of sporadic and variant prion disease [35,36]. We did risk analysis before the use of those corneas to develop our active storage machine to make evolve eyebanking, with assessment of their baseline intrinsic quality (endothelial cell density [37][38][39][40], transparency [41], presence of scar or not, presence of previous refractive surgery or not) and safety (no infection [42][43][44]), following tests used in daily routine in eyebanks. Furthermore, with these common medical contraindications people may wish to give but feel excluded from donation: donation for research can be a way to give them back this opportunity.…”

Section: Plos Onementioning

confidence: 99%

Corneal donation for research versus for transplantation: A-year prospective study of acceptance rates in a French University Hospital

et al. 2020

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Fresh corneal donation is essential for basic and preclinical research, but more unknown to public and the medical teams than donation for transplantation: it may raise concerns. We prospectively compared the acceptance rates and the characteristics of targeted corneal donation for research versus donation for transplantation during one year. The Agence de la Biomé decine authorized us to procure fresh corneas targeted for research, only from the donors with medical contraindications for transplantation, in order not to increase grafts shortage. Three nurses from the hospital coordination team of Saint-Etienne University Hospital, obtained consent for research and transplantation in parallel, screening all intra-hospital deaths cases, following standard protocol to check no refusal from families, despite the French opt-out system. They contacted 127 families for research and 244 for transplantation, in 71% of cases by telephone. Consent was obtained in 62% of cases for research and 54% for transplantation (P = 0.135). The main contraindication for transplantation was the cognitive disorders (66%) followed by the blood cancers (8%). This new specific activity, providing new source of fresh corneas for research immediately usable without any eyebank storage steps, didn't reduce the number of corneas procured for transplantation versus previous years (P = 0.998). Donors in the research group were 10 years older (P<0.001) without difference regarding endothelial cell quality (P = 0.071), allowing maximal clinical relevance for protocols using these fresh human scientific corneas provided by targeted donation.

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Reproducibility of Endothelial Assessment during Corneal Organ Culture: Comparison of a Computer-Assisted Analyzer with Manual Methods

Cited by 32 publications

References 18 publications

Simulation of specular microscopy images of corneal endothelium, a tool for control of measurement errors

Simulation of specular microscopy images of corneal endothelium, a tool for control of measurement errors

Poloxamines for Deswelling of Organ-Cultured Corneas

Corneal donation for research versus for transplantation: A-year prospective study of acceptance rates in a French University Hospital

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