Donor Corneas For Transplantation Research Articles

Advancements in Hydrogels for Corneal Healing and Tissue Engineering.

Hydrogels have garnered significant attention for their versatile applications across various fields, including biomedical engineering. This review delves into the fundamentals of hydrogels, exploring their definition, properties, and classification. Hydrogels, as three-dimensional networks of crosslinked polymers, possess tunable properties such as biocompatibility, mechanical strength, and hydrophilicity, making them ideal for medical applications. Uniquely, this article offers original insights into the application of hydrogels specifically for corneal tissue engineering, bridging a gap in current research. The review further examines the anatomical and functional complexities of the cornea, highlighting the challenges associated with corneal pathologies and the current reliance on donor corneas for transplantation. Considering the global shortage of donor corneas, this review discusses the potential of hydrogel-based materials in corneal tissue engineering. Emphasis is placed on the synthesis processes, including physical and chemical crosslinking, and the integration of bioactive molecules. Stimuli-responsive hydrogels, which react to environmental triggers, are identified as promising tools for drug delivery and tissue repair. Additionally, clinical applications of hydrogels in corneal pathologies are explored, showcasing their efficacy in various trials. Finally, the review addresses the challenges of regulatory approval and the need for further research to fully realize the potential of hydrogels in corneal tissue engineering, offering a promising outlook for future developments in this field.

Suppressing Pro-Apoptotic Proteins by siRNA in Corneal Endothelial Cells Protects against Cell Death.

Corneal endothelial cells (CE) are critical for the cornea's transparency. For severe corneal damage, corneal tissue transplantation is the most promising option for restoring vision. However, CE apoptotic cell death occurs during the storage of donor corneas for transplantation. This study used small interfering (si)RNA-mediated silencing of pro-apoptotic proteins as a novel strategy to protect CE against apoptosis. Therefore, the pro-apoptotic proteins Bax and Bak were silenced in the human corneal endothelial cell line (HCEC-12) by transfection with Accell™siRNA without any adverse effects on cell viability. When apoptosis was induced, e.g., etoposide, the caspase-3 activity and Annexin V-FITC/PI assay indicated a significantly reduced apoptosis rate in Bax+Bak-siRNA transfected HCECs compared to control (w/o siRNA). TUNEL assay in HCECs exposed also significantly lower cell death in Bax+Bak-siRNA (7.5%) compared to control (w/o siRNA: 32.8%). In ex vivo donor corneas, a significant reduction of TUNEL-positive CEs in Bax+Bak-siRNA corneas (8.1%) was detectable compared to control-treated corneas (w/o siRNA: 27.9%). In this study, we demonstrated that suppressing pro-apoptotic siRNA leads to inhibiting CE apoptosis. Gene therapy with siRNA may open a new translational approach for corneal tissue treatment in the eye bank before transplantation, leading to graft protection and prolonged graft survival.

Optimization and evaluation of oxygen-plasma-modified, aligned, poly (Є-caprolactone) and silk fibroin nanofibrous scaffold for corneal stromal regeneration

The shortage of human donor corneas for transplantation necessitates the exploration of tissue engineering approaches to develop corneal substitutes. However, these substitutes must possess the necessary strength, transparency, and ability to regulate cell behaviour before they can be used in patients. In this study, we investigated the effectiveness of an oxygen plasma surface-modified poly-ε-caprolactone (PCL) combined with silk fibroin (SF) nanofibrous scaffold for corneal stromal regeneration. To fabricate the electrospun scaffolds, PCL and SF blends were used on a rotating mandrel. The optimization of the blend aimed to replicate the structural and functional properties of the human cornea, focusing on nanofibre alignment, mechanical characteristics, and in vitro cytocompatibility with human corneal stromal keratocytes. Surface modification of the scaffold resulted in improved transparency and enhanced cell interaction. Based on the evaluation, a composite nanofibrous scaffold with a 1:1 blend of PCL and SF was selected for a more comprehensive analysis. The biological response of keratocytes to the scaffold was assessed through cellular adhesion, proliferation, cytoskeletal organization, gene expression, and immunocytochemical staining. The scaffold facilitated the adhesion of corneal stromal cells, supporting cell proliferation, maintaining normal cytoskeletal organization, and promoting increased expression of genes associated with healthy corneal stromal keratocytes. These findings highlight the potential of a surface-modified PCL/SF blend (1:1) as a promising scaffolding material for corneal stromal regeneration. The developed scaffold not only demonstrated favourable biological interactions with corneal stromal cells but also exhibited characteristics aligned with the requirements for successful corneal tissue engineering. Further research and refinement of these constructs could lead to significant advancements in addressing the shortage of corneas for transplantation, ultimately improving the treatment outcomes for patients in need.

Semiautomatic assessment of endothelial density and morphology in organ-cultured corneas - potential predictors for transplantation suitability and clinical outcome?

The quality of the endothelial cell layer is a major criterion for the approval of organ-cultured human donor-corneas for transplantation. We wanted to compare the predictive capacities of initial endothelial density and endothelium cell morphology for the approval of donor corneas for transplantation and for the clinical outcome after transplantation. The endothelial density and endothelium morphology in organ culture were examined by semiautomatic assessment of 1031 donor corneas. We performed a statistical analysis for correlations of donor-data and cultivation parameters regarding their predictive capacities for the final approval of donor corneas for transplantation and the clinical outcome of 202 transplanted patients. Corneal endothelium cell density proved to be the only parameter with a certain predictive capacity with regard to the final decision, whether donor corneas are suitable for transplantation - however, the correlation was low (area under the curve [AUC] = 0.655). Endothelial cell morphology lacked any predictive power (AUC = 0.597). The clinical outcome regarding visual acuity seemed to be largely independent from both corneal endothelial cell density and morphology. Sub-analyses on transplanted patients stratified for their diagnoses vindicated these findings. Higher endothelial density (above a cut-off level of 2000 cells/mm2), as well as better endothelial morphology do not seem to be critical for transplant-corneal functionality in organ culture and up to 2years after transplantation. Comparable long-term studies on graft survival are recommended to determine, whether the present endothelial density cut-off levels might be too stringent.

In Vitro Profiling of the Extracellular Matrix and Integrins Expressed by Human Corneal Endothelial Cells Cultured on Silk Fibroin-Based Matrices.

Developing a scaffold for culturing human corneal endothelial (HCE) cells is crucial as an alternative cell therapeutic approach to bridge the growing gap between the demand and availability of healthy donor corneas for transplantation. Silk films are promising substrates for the culture of these cells; however, their tensile strength is several-fold greater than the native basement membrane which can possibly influence the dynamics of cell-matrix interaction and the extracellular matrix (ECM) secreted by the cells in long-term culture. In our current study, we assessed the secretion of ECM and the expression of integrins by the HCE cells on Philosamia ricini (PR) and Antheraea assamensis (AA) silk films and fibronectin-collagen (FNC)-coated plastic dishes to understand the cell-ECM interaction in long-term culture. The expression of ECM proteins (collagens 1, 4, 8, and 12, laminin, and fibronectin) on silk was comparable to that on the native tissue. The thicknesses of collagen 8 and laminin at 30 days on both PR (4.78 ± 0.55 and 5.53 ± 0.51 μm, respectively) and AA (4.66 ± 0.72 and 5.71 ± 0.61 μm, respectively) were comparable with those of the native tissue (4.4 ± 0.63 and 5.28 ± 0.72 μm, respectively). The integrin expression by the cells on the silk films was also comparable to that on the native tissue, except for α3 whose fluorescence intensity was significantly higher on PR (p ≤ 0.01) and AA (p ≤ 0.001), compared to that on the native tissue. This study shows that the higher tensile strength of the silk films does not alter the ECM secretion or cell phenotype in long-term culture, confirming the suitability of using this material for engineering the HCE cells for transplantation.

Crosslinker-free collagen gelation for corneal regeneration

Development of an artificial cornea can potentially fulfil the demand of donor corneas for transplantation as the number of donors is far less than needed to treat corneal blindness. Collagen-based artificial corneas stand out as a regenerative option, having promising clinical outcomes. Collagen crosslinked with chemical crosslinkers which modify the parent functional groups of collagen. However, crosslinkers are usually cytotoxic, so crosslinkers need to be removed from implants completely before application in humans. In addition, crosslinked products are mechanically weak and susceptible to enzymatic degradation. We developed a crosslinker free supramolecular gelation strategy using pyrene conjugated dipeptide amphiphile (PyKC) consisting of lysine and cysteine; in which collagen molecules are intertwined inside the PyKC network without any functional group modification of the collagen. The newly developed collagen implants (Coll-PyKC) are optically transparent and can effectively block UV light, are mechanically and enzymatically stable, and can be sutured. The Coll-PyKC implants support the growth and function of all corneal cells, trigger anti-inflammatory differentiation while suppressing the pro-inflammatory differentiation of human monocytes. Coll-PyKC implants can restrict human adenovirus propagation. Therefore, this crosslinker-free strategy can be used for the repair, healing, and regeneration of the cornea, and potentially other damaged organs of the body.

Regenerative Corneal Prosthesis as an Alternative to Human Donor Cornea for Transplantation to Treat Blindness

Regenerative Corneal Prosthesis as an Alternative to Human Donor Cornea for Transplantation to Treat Blindness

Integration and remodelling of a collagen anterior lamellar keratoplasty graft in an animal model – A preliminary report

There is an international shortage of donor corneas for transplantation to treat the 1.5–2.0 million new cases of blindness secondary to corneal disease. Research has therefore been directed towards the development of artificial corneas using alternative materials such as collagen. The biocompatibility of an acellular collagen-based scaffold for anterior lamellar keratoplasty was investigated in vivo in a rabbit model. This scaffold has previously shown promise as a corneal substitute in vitro. Slit-lamp and Optical Coherence Tomography examinations were carried out at 2 weeks, 1, 2, 3, and 6 months post-operatively. Graft-host integration was investigated using immunohistochemistry of the cornea at 6 months. Results showed that the graft was biocompatible, supported corneal re-epithelialisation, and showed no signs of rejection. Migration of stromal cells into areas of the graft was observed, however this was accompanied by extensive graft digestion. Whilst the scaffold was biocompatible, further modifications to the material or supplementation with matrix metalloproteinase inhibitors are required to bring us closer to a stable and fully integrated corneal substitute.

Engineering a Corneal Stromal Equivalent Using a Novel Multilayered Fabrication Assembly Technique.

To overcome the serious shortage of donor corneas for transplantation, alternatives based on tissue engineering need to be developed. Decellularized corneas are one potential alternative, but their densely packed collagen architecture inhibits recellularization in vitro. Therefore, a new rapid method of recellularizing these constructs to ensure high cellularity throughout the collagen scaffold is needed. In this study, we developed a novel method for fabricating corneal constructs by using decellularized porcine corneal sheets assembled using a bottom-up approach by layering multiple sheets between cell-laden collagen I hydrogel.Corneal lenticules were cut from porcine corneas by cryosectioning, then decellularized with detergents and air-dried for storage as sheets. Human corneal stromal cells were encapsulated in collagen I hydrogel and cast between the dried sheets. Constructs were cultured in serum-free medium supplemented with ascorbic acid and insulin for 2 weeks. Epithelial cells were then seeded on the surface and cultured for an additional week. Transparency, cell viability, and phenotype were analyzed by qPCR, histology, and immunofluorescence. Constructs without epithelial cells were sutured onto an ex vivo porcine cornea and cultured for 1 week.Lenticules were successfully decellularized, achieving dsDNA values of 13 ± 1.2 ng/mg dry tissue, and were more resistant to degradation than the collagen I hydrogels. Constructs maintained high cell viability with a keratocyte-like phenotype with upregulation of keratocan, decorin, lumican, collagen I, ALDH3A1, and CD34 and the corneal epithelial cells stratified with a cobblestone morphology. The construct was amenable to surgical handling and no tearing occurred during suturing. After 7 days ex vivo, constructs were covered by a neoepithelium from the host porcine cells and integration into the host stroma was observed.This study describes a novel approach toward fabricating anterior corneal substitutes in a simple and rapid manner, obtaining mature and suturable constructs using only tissue-derived materials.Impact statementNew strategies are needed to face the important worldwide shortage of donor tissues for corneal transplantation. This study describes a novel approach based on decellularized sheets of porcine cornea interspaced with cell-laden collagen I hydrogels. These constructs matured into a transplantable tissue.

Current Trends and Future Perspective of Mesenchymal Stem Cells and Exosomes in Corneal Diseases.

The corneal functions (transparency, refractivity and mechanical strength) deteriorate in many corneal diseases but can be restored after corneal transplantation (penetrating and lamellar keratoplasties). However, the global shortage of transplantable donor corneas remains significant and patients are subject to life-long risk of immune response and graft rejection. Various studies have shown the differentiation of multipotent mesenchymal stem cells (MSCs) into various corneal cell types. With the unique properties of immunomodulation, anti-angiogenesis and anti-inflammation, they offer the advantages in corneal reconstruction. These effects are widely mediated by MSC differentiation and paracrine signaling via exosomes. Besides the cell-free nature of exosomes in circumventing the problems of cell-fate control and tumorigenesis, the vesicle content can be genetically modified for optimal therapeutic affinity. The pharmacology and toxicology, xeno-free processing with sustained delivery, scale-up production in compliant to Good Manufacturing Practice regulations, and cost-effectiveness are the current foci of research. Routes of administration via injection, topical and/or engineered bioscaffolds are also explored for its applicability in treating corneal diseases.

Decellularization and recellularization of cornea: Progress towards a donor alternative.

The global shortage of donor corneas for transplantation has led to corneal bioengineering being investigated as a method to generate transplantable tissues. Decellularized corneas are among the most promising materials for engineering corneal tissue since they replicate the complex structure and composition of real corneas. Decellularization is a process that aims to remove cells from organs or tissues resulting in a cell-free scaffold consisting of the tissues extracellular matrix. Here different decellularization techniques are described, including physical, chemical and biological methods. Analytical techniques to confirm decellularization efficiency are also discussed. Different cell sources for the recellularization of the three layers of the cornea, recellularization methods used in the literature and techniques used to assess the outcome of the implantation of such scaffolds are examined. Studies involving the application of decellularized corneas in animal models and human clinical studies are discussed. Finally, challenges for this technology are explored involving scalability, automatization and regulatory affairs.

Novel Use of Telemedicine for Corneal Tissue Evaluation in Eye Banking: Establishing a Standardized Approach for the Remote Evaluation of Donor Corneas for Transplantation.

To determine the feasibility of using telemedicine consultations in the evaluation of recovered donor corneas for transplant suitability. This study aims to establish and test the minimum imaging requirements for telemedical consultations of corneal tissue by remote eye bank medical directors. Digital images from the slit lamp, optical coherence tomography, and/or specular microscope were assembled into telemedical consults and emailed to 4 eye bank medical directors (M.A.T., J.W., C.S.S., N.K.R.). Feedback on the minimum image requirements for each corneal finding was collected. After establishing a standardized imaging and presentation protocol, test cases were presented to the medical directors to examine the validity of these remote consults. To establish a benchmark for the study's parameters, one medical director (J.W.) examined each case in person after his initial remote review. Examiners were masked to each other's responses. Minimum image requirements for determination of corneal findings were defined and were specific to each anatomic layer of the cornea (epithelial, stromal, or endothelial). Using a defined set of digital images for a set of common corneal findings, the rate of agreement between remote evaluators, eye bank staff, and the in-person evaluator was 100% (11 of 11 examples). For ambiguous test cases, remote evaluators agreed on 80% of the cases (4 of 5). Results from this pilot study suggest that telemedical review of corneal tissue using high-quality digital images may be adequate for accurate identification of specific corneal findings commonly encountered by eye banks.

A presentation of culture-positive corneal donors and the effect on clinical outcomes.

Donor-to-host transmission of infectious agents is a rare but well-recognised complication of corneal transplantation and may carry a grave visual prognosis. In this case series, we describe the clinical features and risk factors of using culture-positive donor corneas for transplantation. Retrospective chart review of a series of patients who underwent either penetrating keratoplasty (PK) or Descemet's stripping automated endothelial keratoplasty (DSAEK) with positive microbiology cultivation during routine assessment of donor corneal tissue obtained at the time of surgery. Donor and recipient characteristics, tissue preparation and surgical parameters, clinical signs and outcomes were registered. Eleven patients who received culture-positive corneal grafts were identified: six with Candida, three with Gram-positive bacteria and two with Gram-negative bacteria. Three patients developed clinical keratitis after routine DSAEK using corneas contaminated with Candida species. The median death-to-preservation time (DPT) of these three donor corneas was 18.08 (range 18.08 to 20.90) h, while in the remaining eight donors, it was 12.27 (range 9.32 to 20.47) h. Despite the initiation of antifungal treatment, all three cases required explantation of the graft and a subsequent re-DSAEK. The use of donor corneas that are culture-positive for Candida carries a risk for developing postoperative keratitis and the risk may be higher in DSAEK. Unlike the cold storage technique employed for donor corneas described in this case series, organ culture technique requires microbiological screening and supplementation of an antifungal agent which may reduce the risk of donor-to-host transmission of fungal infection.

Cost Minimization Analysis of Precut Cornea Grafts in Descemet Stripping Automated Endothelial Keratoplasty.

Descemet stripping automated endothelial keratoplasty (DSAEK) is the most common corneal transplant procedure. A key step in the procedure is preparing the donor cornea for transplantation. This can be accomplished via 1 of 3 alternatives: surgeon cuts the cornea on the day of surgery, the cornea is precut ahead of time in an offsite facility by a trained technician, or a precut cornea is purchased from an eye bank. Currently, there is little evidence on the costs and effectiveness of these 3 strategies to allow healthcare providers decide upon the preferred method to prepare grafts.The aim of this study was to compare the costs and relative effectiveness of each strategy.The Singapore National Eye Centre and Singapore Eye Bank performed both precut cornea and surgeon-cut cornea transplant services between 2009 and 2013.This study included 110 subjects who received precut cornea and 140 who received surgeon-cut cornea. Clinical outcomes and surgical duration were compared across the strategies using the propensity score matching. The cost of each strategy was estimated using the microcosting and consisted of facility costs and procedural costs including surgical duration. One-way sensitivity analysis and threshold analysis were performed.The cost for DSAEK was highest for the surgeon-cut approach ($13,965 per procedure), followed by purchasing precut corneas ($12,659) and then setting up precutting ($12,421). The higher procedural cost of the surgeon-cut approach was largely due to the longer duration of the procedure (surgeon-cut = 72.54 minutes, precut = 59.45 minutes, P < 0.001) and the higher surgeon fees. There was no evidence of differences in clinical outcomes between grafts that were precut or surgeon-cut. Threshold analysis demonstrated that if the number of cases was below 31 a year, the strategy that yielded the lowest cost was purchasing precut cornea from eye bank. If there were more than 290 cases annually, the cheapest option would be to setup precutting facility.Our findings suggest that it is more efficient for centers that are performing a large number of cornea transplants (more than 290 cases) to set up their own facility to conduct precutting.

Enhanced regeneration of corneal tissue via a bioengineered collagen construct implanted by a nondisruptive surgical technique.

Severe shortage of donor corneas for transplantation, particularly in developing countries, has prompted the advancement of bioengineered tissue alternatives. Bioengineered corneas that can withstand transplantation while maintaining transparency and compatibility with host cells, and that are additionally amenable to standardized low-cost mass production are sought. In this study, a bioengineered porcine construct (BPC) was developed to function as a biodegradable scaffold to promote corneal stromal regeneration by host cells. Using high-purity medical-grade type I collagen, high 18% collagen content and optimized EDC-NHS cross-linker ratio, BPCs were fabricated into hydrogel corneal implants with over 90% transparency and four-fold increase in strength and stiffness compared with previous versions. Remarkably, optical transparency was achieved despite the absence of collagen fibril organization at the nanoscale. In vitro testing indicated that BPC supported confluent human epithelial and stromal-derived mesenchymal stem cell populations. With a novel femtosecond laser-assisted corneal surgical model in rabbits, cell-free BPCs were implanted in vivo in the corneal stroma of 10 rabbits over an 8-week period. In vivo, transparency of implanted corneas was maintained throughout the postoperative period, while healing occurred rapidly without inflammation and without the use of postoperative steroids. BPC implants had a 100% retention rate at 8 weeks, when host stromal cells began to migrate into implants. Direct histochemical evidence of stromal tissue regeneration was observed by means of migrated host cells producing new collagen from within the implants. This study indicates that a cost-effective BPC extracellular matrix equivalent can incorporate cells passively to initiate regenerative healing of the corneal stroma, and is compatible with human stem or organ-specific cells for future therapeutic applications as a stromal replacement for treating blinding disorders of the cornea.

Silk fibroin film as an efficient carrier for corneal endothelial cells regeneration

Tissue engineered corneas are a promising alternative to the limited donor corneas for transplantation. Here, we fabricated a transparent silk fibroin film with varied sericin content — low sericin silk fibroin film (LSS) and high sericin silk fibroin film (HSS) — to evaluate its ability to serve as an efficient carrier for corneal endothelial cells. Both LSS and HSS were prepared from Bombyx mori containing different percentages of sericin −6.25% and 9.83%, respectively. Tissue culture polystyrene was used as a reference substratum. In vitro studies, such as a morphological evaluation, initial attachment, expression of mRNAs and proteins (ZO-1 and Na+/K+-ATPase), were performed using rabbit primary corneal endothelial cells as models on silk fibroin films. Results showed that the corneal endothelial cells grown on as-fabricated LSS and HSS were well-maintained, indicating no effect of the difference in sericin content in the films. However, the HSS displayed better initial cell attachment due to the adhesive properties of sericin. These results demonstrated that rabbit corneal endothelial cells grew well on a silk substratum although the percentage of sericin showed no significant difference.

Awareness regarding eye donation among stakeholders in Srikakulam district in South India

BackgroundThere is a huge need for the availability of transplantable donor corneas worldwide to reduce the burden of corneal blindness due to corneal opacity. Voluntary eye donation depends on the awareness levels of various stakeholders in the community. This study aimed to assess the awareness level regarding eye donation among various stakeholders in Srikakulam district in the state of Andhra Pradesh, India.Methods355 subjects were selected from the district using multi stage random sampling. A pre tested semi structured questionnaire was used to collect information regarding each individual’s awareness, knowledge, and perception regarding eye donation. Each response was scored individually and a total score was calculated. Univariate and multivariate regression analysis was used to determine the factors associated with willingness towards eye donation and increased awareness levels.ResultsOf the 355 subjects interviewed, 192 (54%) were male and 163 (46%) were female. The mean age of the stakeholders was 35.9 years (SD ±16.1) and all the study subjects were literate. Ninety-three percent of subjects were aware of the concept of eye donation. Knowledge levels were similar among the teaching community and persons engaged in social service, but lower among students (p < 0.05). Among the stakeholders, there was considerable ambiguity regarding whether persons currently wearing spectacles or suffering from a chronic illnesses could donate their eyes. Older age group (p < 0.001), female gender (p < 0.001) and education (p < 0.001) were associated with increased knowledge levels. 82% of the subjects were willing to donate their eyes and this was unaffected by gender or geographical location (rural vs urban).ConclusionsAwareness levels and willingness to donate eyes are high among the stakeholders in Srikakulam district in India. The services of stakeholders could be utilized, in conjunction with other community based eye donation counselors, to promote awareness regarding eye donation among the general population.

In-body tissue-engineered collagenous connective tissue membranes (BIOSHEETs) for potential corneal stromal substitution

There is a severe shortage of donor cornea for transplantation in many countries. Collagenous connective tissue membranes, named BIOSHEETs, grown in vivo were successfully implanted in rabbit corneal stroma for in vivo evaluation of their suitability as a corneal stromal substitute to solve this global donor shortage. BIOSHEETs were prepared by embedding silicone moulds into dorsal subcutaneous pouches in rabbits for 1 month and stored in glycerol. After re-swelling in saline and trephining, disk-shaped BIOSHEETs (4 mm diameter) were allogeneically implanted into stromal pockets prepared in the right cornea of seven rabbits. Clinical tests for corneal thickness and transparency, and tissue analyses were performed. Because the BIOSHEETs (thickness, 131 ± 14 µm) obtained were opaque immediately after implantation, the transparency of the cornea decreased. The total thickness of the BIOSHEET-implanted cornea increased from 364 ± 21.0 µm to 726 ± 131 µm. After 4 weeks' implantation, the thickness of the cornea stabilized (493 ± 80 µm at 4 weeks and 447 ± 46 µm at 8 weeks). The transparency of the cornea increased progressively with time of implantation. The random orientation of collagen fibrils in the original BIOSHEETs tended to be homogeneous, similar to that of the native stroma. No inflammatory cells accumulated and fibroblast-like cells infiltrated the implant. The BIOSHEETs showed high biocompatibility with stromal tissues; however, further studies are needed to test its functional aspects. Although this research is only intended as a proof of concept, BIOSHEETs may be considered a feasible corneal stromal replacement, especially for treating visual impairment caused by stromal haze. Copyright © 2013 John Wiley & Sons, Ltd.

Are Polymegethism, Pleomorphism, and “Poor Swelling” Valid Discard Parameters in Immediate Postmortem Evaluation of Human Donor Corneal Endothelium?

To study the validity of endothelial polymegethism, pleomorphism, and "poor swelling" as tissue discard parameters in the immediate postmortem evaluation of human donor corneal endothelium. We retrospectively evaluated the quality of the endothelium at first and second evaluations for all processed corneas exhibiting moderate polymegethism, pleomorphism, or "poor swelling" in our eye bank over a 5-year period. Out of 2008 eyes qualifying for our study, 422 corneas (21%) showed polymegethism, pleomorphism, or poor swelling at the first tissue evaluation immediately after excision of the corneoscleral button. In 363 (86%) of these corneas, a normal endothelial mosaic was observed at the second tissue evaluation after 7 to 21 days of organ culture, whereas only 59 (14%) still showed persistent polymegethism, pleomorphism, or "poor swelling" at that time point. A recovery of normal endothelial cell mosaic and "normal swelling" at the second evaluation suggests that cellular contour parameters do not relate to tissue viability, but rather to a cellular stress reaction. If so, the validity of endothelial cellular contour morphology as an early parameter in assessing the suitability of a donor cornea for transplantation may be reconsidered.

Light microscopy of the corneal pathologies that we meet in our daily practice as an eye banker

AbstractPurpose Organ culture is the method of choice for the preservation of a human donor cornea for transplantation throughout Europe. This method is often closely connected with the assessment of the cornea using light microscopy. The corneal endothelium, its polymeghatism, pleomorphism, cornea guttata and other endothelial abnormalities as well as epithelial and stromal pathologies will be presented to show the usefulness of light microscopy in the evaluation of corneal pathologies.Methods Photographs from light microscopy (both phase contrast and bright field) taken before and after corneal storage in organ culture will be compared and discussed in relation to changes in endothelial morphology and the swelling of the intercellular spaces. The individual corneal layers (the corneal epithelium, the stroma and the endothelium) of pathological corneas will be compared to those of control corneas.Results Among corneal pathologies, cornea guttata, corneal dystrophies (Fuchs endothelial corneal dystrophy, posterior polymorphous corneal dystrophy) and crystalline keratopathy will be shown using pathological explants. Moreover, the corneal pathologies will be correlated with histological findings. Finally, bacterial and fungal contamination of the cornea during organ culture will be shown.Conclusion Light microscopy is an essential part of qualitative and quantitative corneal assessment, which allows corneas with a variety of pathologies, mostly endothelial in origin, to be excluded from grafting. This work was supported by the project PRVOUK‐P24LF13 of Charles University in Prague.