Preliminary study of rabbits as an animal model of mammalian eye transplantation and literature review / Estudo preliminar de coelhos como modelo animal para transplante de olho em mamíferos e revisão da literatura

ABSTRACT Purpose: To describe an innovative animal model of eye transplantation used in rabbits. Methods: six Dutch-belted male rabbits were submitted to lateral orbitotomy in the right eye, wide retrobulbar anatomy exposure, dissection of the structures, identification and distal section of the optic nerve followed by anastomosis either by vicryl (group 1) or fibrin glue (group 2). Electroretinography recording was performed before the section of the optic nerve and every 30 seconds after, to monitor the function of retina. Left eye was used as control group. Results: After optic nerve resection and anastomosis, stable ERG amplitude of the right eye was lost after 302 seconds in group 1 and after 296 seconds on group 2. Left eye kept longer stable ERG amplitude curves. Conclusions: The animal model of whole eye transplantation was effective in describing a novel technique to be used in rabbits, with success of the anatomic procedure. Further studies will clarify the best anastomosis methods and maintenance of function of the receptor organ. Translational relevance: this animal model of whole eye transplantation provides a novel perspective for blind patients and the research models, since we describe a novel mammal animal model. This model can be used as basis of a human model of whole eye transplantation in future studies.

RESUMO Objetivo: Descrever uma técnica cirúrgica inovadora para transplante de olho em um modelo animal em coelhos. Métodos: Seis coelhos machos com Dutch Belted foram submetidos à orbitotomia lateral do olho direito, com ampla exposição da anatomia retrobulbar, dissecção do cone muscular, exposição e secção distal do nervo óptico seguida de anastomose por vicryl (Grupo 1) ou cola de fibrina (Grupo 2). O registro da eletrorretinografia foi realizado antes da secção do nervo óptico e a cada 30 segundos após, para monitorar a função da retina. O olho esquerdo foi usado como grupo controle. Resultados: Após a ressecção do nervo óptico, a estabilidade da amplitude da eletrorretinografia foi perdida no olho direito após 302 segundos no Grupo 1 e após 296 segundos no Grupo 2. O olho esquerdo manteve eletrorretinografia estável por períodos mais longos. Conclusão: O modelo animal de transplante total de olho foi eficaz em descrever uma nova técnica cirúrgica para ser utilizada em laboratório com coelhos, com sucesso do procedimento anatômico. Novos estudos esclarecerão os melhores métodos de anastomose e manutenção da função do órgão receptor.

Meibomian Gland Dysfunction in Ocular Graft vs. Host Disease: A Need for Pre-Clinical Models and Deeper Insights

Despite decades of experience with hematopoietic stem cell transplantation, we are still faced with the delicate equipoise of achieving stable ocular health post-transplantation. This is because ocular graft-versus-host disease (oGvHD) following hematopoietic stem cell transplantation frequently occurs (≥50%) among transplant patients. To date, our understanding of the pathophysiology of oGvHD especially the involvement of the meibomian gland is still limited as a result of a lack of suitable preclinical models among other. Herein, the current state of the etiology and, pathophysiology of oGvHD based on existing pre-clinical models are reviewed. The need for additional pre-clinical models and knowledge about the involvement of the meibomian glands in oGvHD are emphasized.

Development of a New Large-Animal Model for Composite Face and Whole-Eye Transplantation: A Novel Application for Anatomical Mapping Using Indocyanine Green and Liquid Latex.

BACKGROUND: Recent advances in the field of vascularized composite allotransplantation have revolutionized reconstructive surgery and demonstrated opportunity for restoration of orbital content and perhaps vision. The development of an animal model that would facilitate study of surgical technique, nerve regeneration, vision restoration, and immunosuppression is needed. The aim of this study is to describe three different large-animal cadaveric models suitable for composite face and whole-eye transplantation. In addition, the authors introduce a novel colored liquid latex and indocyanine green mixture for vasculature identification and visualization. METHODS: The authors studied three different flap designs using cadaveric Yorkshire swine and modified them for face and whole-eye transplant harvest. Flaps harvested included a chimeric flap, a monobloc flap, and a bipedicled free flap. The authors injected selected vasculature with colored latex to better delineate vascular anatomy supplying the orbit and face. Indocyanine green was added to the latex solution to allow for visualization of the vascular supply using near-infrared imaging. RESULTS: Colored latex and indocyanine green injections were successfully visualized in all cadaveric dissections. All three modified flap designs showed a well-defined and consistent vascular network within each face and whole-eye transplantation flap using laser-assisted near-infrared imaging. CONCLUSIONS: The authors present the feasibility of composite face and whole-eye transplantation models using a novel mixture of liquid latex and indocyanine green in a porcine cadaver. Further study in large animals is needed to appraise the surgical feasibility of this procedure and potential for clinically relevant outcomes, including vision restoration. The authors believe this study establishes a foundation for translation into live animal models for optic nerve regeneration.

Demonstration of technical feasibility and viability of whole eye transplantation in a rodent model.

INTRODUCTION: Whole eye transplantation (WET) holds promise for vision restoration in devastating/disabling visual loss (congenital or traumatic) not amenable to surgical or neuroprosthetic treatment options. The eye includes multiple tissues with distinct embryonic lineage and differential antigenicity. Anatomically and immunologically, the eye is unique due to its avascular (cornea) and highly vascular (retina) components. Our goal was to establish technical feasibility, demonstrate graft viability, and evaluate histologic changes in ocular tissues/adnexae in a novel experimental model of WET that included globe, adnexal, optic nerve (ON), and periorbital soft tissues. METHODS: Outbred Sprague-Dawley rats (n = 5) received heterotopic vascularized WET from donors. Each WET included the entire globe, adnexa, ON, and periorbital soft tissues supplied by the common carotid artery and external jugular vein. Viability and perfusion were confirmed by clinical examination, angiography and magnetic resonance imaging (MRI). Globe, adnexal, and periorbital tissues were analyzed for histopathologic changes, and the ON was examined for neuro-regeneration at study endpoint (30 days) or Banff Grade 3 rejection in the periorbital skin (whichever was earlier). RESULTS: Gross examination confirmed transplant viability and corneal transparency. Average operative duration was 64.0 ±â€¯5.8 min. Average ischemia time was 26.0 ±â€¯4.2 min. MRI revealed loss of globe volume by 36.0 ±â€¯2.8% after transplantation. Histopathology of globe and adnexal tissues showed unique and differential patterns of inflammatory cell infiltration. The ON revealed a neurodegeneration pattern. CONCLUSION: The present study is the first in the literature to establish an experimental model of WET. This model holds significant potential in investigating mechanistic pathways, monitoring strategies or developing management approaches involving ocular viability, immune rejection, and ON regeneration after WET.

Self-organization and progenitor targeting generate stable patterns in planarian regeneration.

During animal regeneration, cells must organize into discrete and functional systems. We show that self-organization, along with patterning cues, govern progenitor behavior in planarian regeneration. Surgical paradigms allowed the manipulation of planarian eye regeneration in predictable locations and numbers, generating alternative stable neuroanatomical states for wild-type animals with multiple functional ectopic eyes. We used animals with multiple ectopic eyes and eye transplantation to demonstrate that broad progenitor specification, combined with self-organization, allows anatomy maintenance during regeneration. We propose a model for regenerative progenitors involving (i) migratory targeting cues, (ii) self-organization into existing or regenerating eyes, and (iii) a broad zone, associated with coarse progenitor specification, in which eyes can be targeted by progenitors. These three properties help explain how tissues can be organized during regeneration.

A New Composite Eyeball-Periorbital Transplantation Model in Humans: An Anatomical Study in Preparation for Eyeball Transplantation.

BACKGROUND: Vascularized composite allotransplantation offers a new hope for restoration of orbital content and perhaps vision. The aim of this study was to introduce a new composite eyeball-periorbital transplantation model in fresh cadavers in preparation for composite eyeball allotransplantation in humans. METHODS: The composite eyeball-periorbital transplantation flap borders included the inferior border, outlined by the infraorbital rim; the medial border, created by the nasal dorsum; the lateral border, created by the lateral orbital rim; and the superior border, created by the superior part of the eyebrow. The pedicle of the flap included the facial artery, superficial temporal artery, and external jugular vein. The skin and subcutaneous tissues of the periorbital region were dissected and the bony tissue was reached. A coronal incision was performed and the frontal lobe of the brain was reached by means of frontal osteotomy. Ophthalmic and oculomotor nerves were also included in the flap. After a "box osteotomy" around the orbit, the dissection was completed. Methylene blue and indocyanine green injection (SPY Elite System) was performed to show the integrity of the vascular territories after facial flap harvest. RESULTS: Adequate venous return was observed within the flap after methylene blue dye injection. Laser-assisted indocyanine green angiography identified a well-defined vascular network within the entire composite eyeball-periorbital transplantation flap. CONCLUSIONS: For the first time, a novel composite eyeball-periorbital transplantation model in human cadavers was introduced. Good perfusion of the flap confirmed the feasibility of composite eyeball-periorbital transplantation in the clinical setting. Although harvesting of the flap is challenging, it introduces a new option for reconstruction of the periorbital region including the eyeball.

Histological, immunohistochemical and clinical considerations on amniotic membrane transplant for ocular surface reconstruction.

Amniotic membrane (AM) transplantation has been used successfully worldwide in ophthalmology plastic surgery for over 100 years. This review presents the histological and the immunohistochemical features of AM compared to those of the conjunctiva and discusses the techniques of processing and preservation, its mechanism of action in ocular reconstruction, its clinical ophthalmic indications, but also advantages and limitations of grafting with this biomaterial.

Whole-eye transplantation: a look into the past and vision for the future.

Blindness afflicts ~39 million people worldwide. Retinal ganglion cells are unable to regenerate, making this condition irreversible in many cases. Whole-eye transplantation (WET) provides the opportunity to replace diseased retinal ganglion cells, as well as the entire optical system and surrounding facial tissue, if necessary. Recent success in face transplantation demonstrates that this may be a promising treatment for what has been to this time an incurable condition. An animal model for WET must be established to further enhance our knowledge of nerve regeneration, immunosuppression, and technical aspects of surgery. A systematic review of the literature was performed to evaluate studies describing animal models for WET. Only articles in which the eye was completely enucleated and reimplanted were included. Study methods and results were compared. In the majority of published literature, WET can result in recovery of vision in cold-blooded vertebrates. There are a few instances in which mammalian WET models demonstrate survival of the transplanted tissue following neurovascular anastomosis and the ability to maintain brief electroretinogram activity in the new host. In this study we review in cold-blooded vertebrates and mammalian animal models for WET and discuss prospects for future research for translation to human eye transplantation.

Total Human Eye Allotransplantation: Developing Surgical Protocols for Donor and Recipient Procedures.

BACKGROUND: Vascularized composite allotransplantation of the eye is an appealing, novel method for reconstruction of the nonfunctioning eye. The authors' group has established the first orthotopic model for eye transplantation in the rat. With advancements in immunomodulation strategies together with new therapies in neuroregeneration, parallel development of human surgical protocols is vital for ensuring momentum toward eye transplantation in actual patients. METHODS: Cadaveric donor tissue harvest (n = 8) was performed with orbital exenteration, combined open craniotomy, and endonasal approach to ligate the ophthalmic artery with a cuff of paraclival internal carotid artery, for transection of the optic nerve at the optic chiasm and transection of cranial nerves III to VI and the superior ophthalmic vein at the cavernous sinus. Candidate recipient vessels (superficial temporal/internal maxillary/facial artery and superficial temporal/facial vein) were exposed. Vein grafts were required for all anastomoses. Donor tissue was secured in recipient orbits followed by sequential venous and arterial anastomoses and nerve coaptation. Pedicle lengths and calibers were measured. All steps were timed, photographed, video recorded, and critically analyzed after each operative session. RESULTS: The technical feasibility of cadaveric donor procurement and transplantation to cadaveric recipient was established. Mean measurements included optic nerve length (39 mm) and caliber (5 mm), donor artery length (33 mm) and caliber (3 mm), and superior ophthalmic vein length (15 mm) and caliber (0.5 mm). Recipient superficial temporal, internal maxillary artery, and facial artery calibers were 0.8, 2, and 2 mm, respectively; and superior temporal and facial vein calibers were 0.8 and 2.5 mm, respectively. CONCLUSION: This surgical protocol serves as a benchmark for optimization of technique, large-animal model development, and ultimately potentiating the possibility of vision restoration transplantation surgery. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, V.

Clinical Considerations for Vascularized Composite Allotransplantation of the Eye.

Vascularized composite allotransplantation represents a potential shift in approaches to reconstruction of complex defects resulting from congenital differences as well as trauma and other acquired pathology. Given the highly specialized function of the eye and its unique anatomical components, vascularized composite allotransplantation of the eye is an appealing method for restoration, replacement, and reconstruction of the nonfunctioning eye. Herein, we describe conventional treatments for eye restoration and their shortcomings as well as recent research and events that have brought eye transplantation closer to a potential clinical reality. In this article, we outline some potential considerations in patient selection, donor facial tissue procurement, eye tissue implantation, surgical procedure, and potential for functional outcomes.

Estimation of the potential eye and tissue donor pool in an Australian emergency department.

OBJECTIVE: EDs have long been considered a potential source of eye and tissue donors, but no specific evidence to support this was identified in the Australian setting. The present study aims to bridge that knowledge gap, by analysing medical and social histories of those who have died over a 5 year period so as to determine donation eligibility in this population. METHODS: A retrospective audit was undertaken of all patients who died within the Royal Melbourne Hospital ED between 2010 and 2014. ED records, pharmacy records and electronic medical histories were audited for the presence of eye and tissue donation exclusion criteria and the distribution of these criteria within the target population. RESULTS: Over the 5 year period, of 326 deaths that occurred in the ED, one third was suitable for eye donation (n = 106) and one in seven (n = 45) for tissue donation. Of the age appropriate patients, five criteria were identified that excluded up to 85% of the population not eligible to donate. These were: haematological malignancies, neurodegenerative conditions, non-haematological malignancies, chronic renal failure and eye disease. CONCLUSION: The present study has identified a large pool of potential eye and tissue donors; a pool mostly unrecognised by emergency clinicians. An extensive list of exclusion criteria restricts donor potential. However, the present study has identified that only five fundamentally limit donation in the ED population. Utilisation of this knowledge will allow for the development of clinical triggers that will improve identification, and increase realisation, of potential donors.

Ethical Considerations of Whole-Eye Transplantation.

Whole eye transplantation (WET) remains experimental. Long presumed impossible, recent scientific advances regarding WET suggest that it may become a clinical reality. However, the ethical implications of WET as an experimental therapeutic strategy remain largely unexplored. This article evaluates the ethical considerations surrounding WET as an emerging experimental treatment for vision loss. A thorough review of published literature pertaining to WET was performed; ethical issues were identified during review of the articles.

Allogenic iPSC-derived RPE cell transplants induce immune response in pigs: a pilot study.

Stem cell strategies focused on replacement of RPE cells for the treatment of geographic atrophy are under intense investigation. Although the eye has long been considered immune privileged, there is limited information about the immune response to transplanted cells in the subretinal space of large animals. The purpose of this study was to evaluate the survival of allogenic induced pluripotent stem cell-derived RPE cells (iPSC-RPE) delivered to the subretinal space of the pig as well as determine whether these cells induce an immune response in non-diseased eyes. GFP positive iPSC-RPE, generated from outbred domestic swine, were injected into the subretinal space of vitrectomized miniature swine. Control eyes received vehicle only. GFP positive iPSC-RPE cells were identified in the subretinal space 3 weeks after injection in 5 of 6 eyes. Accompanying GFP-negative cells positive for IgG, CD45 and macrophage markers were also identified in close proximity to the injected iPSC-RPE cells. All subretinal cells were negative for GFAP as well as cell cycle markers. We found that subretinal injection of allogenic iPSC-RPE cells into wild-type mini-pigs can induce the innate immune response. These findings suggest that immunologically matched or autologous donor cells should be considered for clinical RPE cell replacement.

Dendritic cell physiology and function in the eye.

The eye and the brain are immunologically privileged sites, a property previously attributed to the lack of a lymphatic circulation. However, recent tracking studies confirm that these organs have good communication through classical site-specific lymph nodes, as well as direct connection through the blood circulation with the spleen. In addition, like all tissues, they contain resident myeloid cell populations that play important roles in tissue homeostasis and the response to foreign antigens. Most of the macrophage and dendritic cell (DC) populations in the eye are restricted to the supporting connective tissues, including the cornea, while the neural tissue (the retina) contains almost no DCs, occasional macrophages (perivascularly distributed), and a specialized myeloid cell type, the microglial cell. Resident microglial cells are normally programmed for immunological tolerance. The privileged status of the eye, however, is relative, as it is susceptible to immune-mediated inflammatory disease, both infectious and autoimmune. Intraocular inflammation (uveitis and uveoretinitis) and corneal graft rejection constitute two of the more common inflammatory conditions affecting the eye leading to considerable morbidity (blindness). As corneal graft rejection occurs almost exclusively by indirect allorecognition, host DCs play a major role in this process and are likely to be modified in their behavior by the ocular microenvironment. Ocular surface disease, including allergy and atopy, also comprise a significant group of immune-mediated eye disorders in which DCs participate, while infectious disease such as herpes simplex keratitis is thought to be initiated via corneal DCs. Intriguingly, some more common conditions previously thought to be degenerative (e.g. age-related macular degeneration) may have an autoimmune component in which ocular DCs and macrophages are critically involved. Recently, the possibility of harnessing the tolerizing potential of DCs has been applied to experimental models of autoimmune uveoretinitis with good effect. This approach has considerable potential for use in translational clinical therapy to prevent sight-threatening disease caused by ocular inflammation.

Distinct regulatory cascades govern extraocular and pharyngeal arch muscle progenitor cell fates.

Genetic regulatory networks governing skeletal myogenesis in the body are well understood, yet their hierarchical relationships in the head remain unresolved. We show that either Myf5 or Mrf4 is necessary for initiating extraocular myogenesis. Whereas Mrf4 is dispensable for pharyngeal muscle progenitor fate, Tbx1 and Myf5 act synergistically for governing myogenesis in this location. As in the body, Myod acts epistatically to the initiating cascades in the head. Thus, complementary pathways, governed by Pax3 for body, and Tbx1 for pharyngeal muscles, but absent for extraocular muscles, activate the core myogenic network. These diverse muscle progenitors maintain their respective embryonic regulatory signatures in the adult. However, these signatures are not sufficient to ensure the specific muscle phenotypes, since the expected differentiated phenotype is not manifested when satellite cells are engrafted heterotopically. These findings identify novel genetic networks that may provide insights into myopathies which often affect only subsets of muscles.

Impediments to eye transplantation: ocular viability following optic-nerve transection or enucleation.

Maintenance of ocular viability is one of the major impediments to successful whole-eye transplantation. This review provides a comprehensive understanding of the current literature to help guide future studies in order to overcome this hurdle. A systematic multistage review of published literature was performed. Three specific questions were addressed: (1) Is recovery of visual function following eye transplantation greater in cold-blooded vertebrates when compared with mammals? (2) Is outer retina function following enucleation and reperfusion improved compared with enucleation alone? (3) Following optic-nerve transection, is there a correlation between retinal ganglion cell (RGC) survival and either time after transection or proximity of the transection to the globe? In a majority of the studies performed in the literature, recovery of visual function can occur after whole-eye transplantation in cold-blooded vertebrates. Following enucleation (and reperfusion), outer retinal function is maintained from 4 to 9 h. RGC survival following optic-nerve transection is inversely related to both the time since transection and the proximity of transection to the globe. Lastly, neurotrophins can increase RGC survival following optic-nerve transection. This review of the literature suggests that the use of a donor eye is feasible for whole-eye transplantation.

Enxertos homólogos de esclera e dura-máter em olhos de coelhos: análise histopatológica comparativa / Homologous scleral and dura mater grafts in rabbit eyesa comparative histopathological analysis

OBJETIVO: Avaliar a reação tecidual à implantação de enxertos homólogos de esclera e dura-máter. MÉTODOS: Foram realizadas cirurgias experimentais em 41 coelhos albinos da raça Nova Zelândia, sendo que dois coelhos foram selecionados, ao acaso, para serem doadores dos enxertos a serem utilizados no experimento. Trinta coelhos foram divididos em dois grupos: grupo D e grupo E, sendo submetidos à implantação dos tecidos homólogos e nove animais constituíram o grupo controle (grupo DE), submetidos à cirurgia sem enxertia. Os olhos foram enucleados e amostras foram colhidas em duas, seis e 12 semanas após o experimento, constituindo os subgrupos I, II e III. RESULTADOS: Foi realizada análise histopatológica qualitativa, além de estudo semiquantitativo comparativo da vascularização e infiltrado inflamatório na esclera e dura-máter transplantadas. Procedeu-se, também, a histomorfometria das medidas do lado dos enxertos com sistema analisador de imagens. Não houve diferença estatisticamente significante comparando-se a vascularização, infiltrado inflamatório e medidas do lado, entre os subgrupos I de esclera em relação à dura-máter, assim como nos subgrupos II. Não foram comparados os subgrupos III, pois foram observados somente fragmentos dos enxertos de dura-máter. CONCLUSÕES: O enxerto de dura-máter mostrou intensa reabsorção e progressiva substituição por tecido conjuntivo frouxo. O enxerto de esclera mostrou discreta reabsorção na periferia com formação de membrana fibrosa mais evidente, integrando este à esclera do hospedeiro.

PURPOSE: To evaluate tissue reaction to implantation of homologous scleral and dura mater grafts. METHODS: Experimental surgeries were performed on 41 albine New Zealand rabbits; two rabbits were selected at random to be graft donors. Thirty rabbits were divided into two groups: group D and group E, receiving grafts of homologous tissues. The remaining nine animals comprised the control group (DE) and were submitted to surgery but with no graft. The eyes were enucleated and samples collected two, six and 12 weeks after the experiment, comprising subgroups I, II and III. RESULTS: A qualitative histopathological analysis was performed together with a comparative semi-quantitative study on vascularization and inflammatory infiltrate in the transplanted sclera and dura mater. Histomorphometry was conducted based on measurements of the grafts with an image analyzing system. There was no statistically significant difference when comparing vascularization, inflammatory infiltrate and measurements between subgroup I in relation to sclera and dura mater, nor in subgroup II. Subgroup III was not compared, since only fragments of dura mater grafts were considered. CONCLUSIONS: The dura mater graft presented intense absorption and progressive replacement of loose connective tissue. And the scleral graft showed discreet absorption in the periphery with formation of a more evident fibrous membrane, integrating the graft into the host sclera.

[Homologous scleral and dura mater grafts in rabbit eyes: a comparative histopathological analysis]. / Enxertos homólogos de esclera e dura-máter em olhos de coelhos: análise histopatológica comparativa.

PURPOSE: To evaluate tissue reaction to implantation of homologous scleral and dura mater grafts. METHODS: Experimental surgeries were performed on 41 albine New Zealand rabbits; two rabbits were selected at random to be graft donors. Thirty rabbits were divided into two groups: group D and group E, receiving grafts of homologous tissues. The remaining nine animals comprised the control group (DE) and were submitted to surgery but with no graft. The eyes were enucleated and samples collected two, six and 12 weeks after the experiment, comprising subgroups I, II and III. RESULTS: A qualitative histopathological analysis was performed together with a comparative semi-quantitative study on vascularization and inflammatory infiltrate in the transplanted sclera and dura mater. Histomorphometry was conducted based on measurements of the grafts with an image analyzing system. There was no statistically significant difference when comparing vascularization, inflammatory infiltrate and measurements between subgroup I in relation to sclera and dura mater, nor in subgroup II. Subgroup III was not compared, since only fragments of dura mater grafts were considered. CONCLUSIONS: The dura mater graft presented intense absorption and progressive replacement of loose connective tissue. And the scleral graft showed discreet absorption in the periphery with formation of a more evident fibrous membrane, integrating the graft into the host sclera.

Retinoblastoma: from bench to bedside.

Retinoblastoma (Rb) is the most common primary ocular malignancy of children and is caused by a mutation in the gene RB1. Approximately 40% of cases are associated with one or more constitutional mutations, and are therefore heritable, whereas the other 60% are sporadic. Rb is exclusively found in young children. In some cases, Rb tumours metastasise to extraocular organs including bone, lung and brain. Although there is no effective treatment for metastatic disease, non-metastatic cases can be cured by removal of the eye(enucleation). Newer treatment strategies emphasise salvaging the affected eye whenever possible. Animal models of Rb have been developed with xenograft and transgenic techniques. Each model has both strengths and weaknesses for exploring the mechanisms of disease development and progression and the efficacy of new treatment strategies.

Detection of human immunodeficiency virus, hepatitis B virus, and hepatitis C virus in donor eyes using polymerase chain reaction.

Detection of human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV) in donor eyes was performed. DNAs were extracted from the uvea, and they were amplified using the polymerase chain reaction (PCR). Amplified viral DNAs were detected with liquid hybridisation and chemiluminescent assay in which no radioactive materials were used. This method was shown to have a sensitivity limit of fewer than 10 copies of HIV, making it much more sensitive than the current techniques employed in eye banks. The method was applied to 120 donor eyes, including four from donors seropositive for HBV. The HBV gene was detected in one case in which the donor's blood had not been tested for HBV. HIV and HCV genes were not detected in any of the samples. The assay could be an effective screening test for the detection of these viruses in eye bank eyes.