Diseased Corneas Research Articles

Greffes lamellaires de cornée. Retour vers le futur

Transplantation of all of the corneal layers (penetrating keratoplasty) is the most common corneal grafting method, regardless of the indication. The surgical procedure is relatively simple but penetrating keratoplasty has several potential complications: --Unnecessary replacement of healthy recipient endothelium in diseases affecting only the anterior layers carries a risk of immune rejection. --Replacement of the anterior part of the cornea in diseases affecting only the posterior layers delays visual recovery Lamellar keratoplasty consists of replacing only the diseased corneal layers. Although it was the first procedure to be described, lamellar keratoplasty became less popular because it is technically difficult and often gives poor visual results due to imperfect lamellar dissection. Recent surgical advances have overcome these difficulties, and lamellar keratoplasty may well become the first-choice treatment for most stromal and endothelial diseases.

Comparison of corneal endothelial cell loss during phacoemulsification using continuous anterior chamber infusion versus those using ophthalmic viscosurgical device: Randomized controlled trial

AimWe conducted this study to evaluate and compare corneal endothelial cell loss between phacoemulsification with continuous anterior chamber infusion using anterior chamber maintainer (ACM) and phacoemulsification using ophthalmic viscosurgical device (OVD).Materials and MethodsThis was a prospective, randomized controlled trial. Fifty eyes of 47 patients of senile cataract undergoing phacoemulsification were included. Patients were randomly allocated into two groups of 25 eyes each. Cataract surgery was performed by phacoemulsification with anterior chamber (AC) continuous infusion with balanced salt solution (BSS) plus and ACM without OVD in Group A, and in Group B, phacoemulsification was performed using OVD with BSS plus. Corneal endothelial cell count and pachymetry were performed preoperatively and postoperatively on day 1, day 7, and day 30.ResultsThe mean increase in pachymetry was 4.86%, 2.94%, and 1.94%, (Group A) and 5.95%, 3.94%, and 0.51%, (Group B) on first, seventh, and 30th postoperative day respectively. The difference between the percentage increase in pachymetry between the two groups was not significant at day 1 (P = 0.441), day 7 (P = 0.298), and day 30 (P = 0.174) postoperatively. The density of endothelial cells decreased postoperatively (day 30) by 7.38% (Group A) and 7.47% (Group B) without any significant statistical difference (P = 0.983) between two groups.ConclusionUse of ACM for continuous AC infusion and omission of OVD during phacoemulsification did not cause significant difference in corneal swelling or endothelial cell loss in the immediate postoperative period up to one month.

Lentes de contato terapêuticas e as vantagens dos materiais de alto Dk

Therapeutic contact lenses are useful in a variety of ocular surface diseases. Their main indications are: to relieve the pain; protect ocular surface; promote corneal healing and epithelial regeneration; seal a leaking corneal wound and deliver ophthalmic drugs on the ocular surface. There are several kinds of lens designs and materials, and their choice is dependent on the specific disease to be treated, the duration of treatment and the physiologic needs of the diseased cornea. Bullous keratopathy, recurrent epithelial erosion syndrome, dry eye and postoperative epithelial defects are amongst their indications. Therapeutic contact lenses should not be indicated in the presence of active infectious keratitis or when the patient is not compliant. Corneal neovascularization, giant papillary conjunctivitis and infectious keratitis are serious complications, which can be prevented by correctly fitting and maintaining the therapeutic contact lenses. Silicon-hydrogel therapeutic contact lenses, due to their higher oxygen permeability, allow extended wear schedules, decreasing the need for frequent lens replacement.

Menicon Z Rigid Gas Permeable Lenses for Keratoconus and Irregular Corneas: A Retrospective Case Series

The tisilfocon A (Menicon Z) rigid gas permeable (RGP) contact lens polymer has the highest oxygen permeability (Dk) value of any RGP lens material. To our knowledge there is no published literature documenting the success of this material in fitting contact lenses to highly irregular and diseased corneas. A retrospective medical record review of 40 consecutive patients (64 eyes) who had worn lenses made in the Menicon Z RGP material was performed. All patients had some form of corneal pathology and were fitted in a single tertiary care hospital-based contact lens service. Menicon Z contact lenses were fit to 33 eyes of 20 patients with keratoconus (KC) and 31 eyes of 23 patients with irregular corneas. The irregular cornea group comprised 28 eyes of 21 patients after corneal surgery, 1 eye with ocular cicatricial pemphigoid and 2 eyes of 1 patient with chronic staphylococcal blepharo-keratoconjuncitivitis. Treatment goals included acceptable corneal physiology, comfort, and visual acuity. In the KC group, there was an 82% (27/33 eyes) success rate as measured by the treatment goals. Mean duration of wear for successful patients was 2.5 years (0.5-5 years). Only 12% (4/33 eyes) failed directly due to inability of the Menicon Z material to perform adequately while another material was acceptable. In the irregular surface group (surgical and ocular surface disease), there was a 74% (23/31 eyes) success rate. Mean duration of wear for successful patients in this group is 2.79 years (3 months-7 years). Only 3% (1/31 eyes) failed directly due to inability of the Menicon Z material to perform adequately while another material was acceptable. The tisilfocon A hyper Dk lens material maintains clinically acceptable corneal health and clinically acceptable visual acuity for patients with KC and irregular corneal surfaces. In this series of patients with complicated corneal disease, the most failures were due to design issues and discomfort not related to the Menicon Z material. The Menicon Z material was inadequate in only 8% of eyes, whereas another RGP material was successful. Most patients with irregular corneas in a tertiary care center can be successfully fit with the tisilfocon A RGP material for visual rehabilitation.

Deep anterior lamellar keratoplasty

AbstractDespite the long term risk of endothelial rejection, full‐thickness corneal transplantation is still considered the gold standard surgery for replacing a diseased cornea. Deep anterior lamellar keratoplasty (DALK) is not a new procedure, but in the past, its use has been of limited popularity owing to the challenging, lengthy and tedious surgery involved. With the advent of newer surgical techniques and instruments for performing lamellar corneal surgery, DALK has gained in popularity. This presentation discusses the techniques for performing the procedure as well as the clinical outcomes, especially focusing on Big bubble DALK. Results will be compared with the outcomes of penetrating keratoplasty (PK). Results suggest that DALK is the logical choice in the surgical management of corneal stromal disease, in presence of a functional endothelium.

Adipose-Derived Stem Cells Are a Source for Cell Therapy of the Corneal Stroma

Most corneal diseases affect corneal stroma and include immune or infectious diseases, ecstatic disorders, traumatic scars, and corneal dystrophies. Cell-based therapy is a promising therapeutic approach to overcome the current disadvantages of corneal transplantation. We intended to search for a cell source to repopulate and regenerate corneal stroma. We investigated the ability of human processed lipoaspirate derived (PLA) cells to regenerate corneal stroma in experimental animals. In the first set of experiments, we tested the biosafety and immunogenicity of human PLA stem cells transplanted into the corneal stroma of rabbits. No immune response was elicited even though we used immune-competent animals. PLA cells survived up to 10 weeks post-transplant, maintained their shape, and remained intermingled in the stroma without disrupting its histological pattern. Interestingly, transparency was preserved even 10 weeks after the transplant, when PLA cells formed a discontinuous layer in the stroma. In the second set of experiments, regeneration of the corneal stroma by PLA cells was assessed, creating a niche by partial ablation of the stroma. After 12 weeks, human cells were disposed following a multilayered pattern and differentiated into functional keratocytes, as assessed by the expression of aldehyde-3-dehydrogenase and cornea-specific proteoglycan keratocan. Based on our results, we believe that adipose-derived adult stem cells can be a cell source for stromal regeneration and repopulation in diseased corneas. The low health impact of the surgical procedure performed to obtain the PLA cells provides this cell source with an additional beneficial feature for its possible future autologous use in human patients.

Corneal Imaging with Anterior Segment Optical Coherence Tomography for Lamellar Keratoplasty Procedures

To assess and describe the uses of anterior segment optical coherence tomography (OCT) in the evaluation of the cornea before and after lamellar corneal transplantation procedures. Prospective, noncomparative, observational case series. Seven eyes of seven patients undergoing anterior and posterior lamellar corneal transplantation procedures at the Singapore National Eye Centre were included in the study. High-resolution anterior segment OCT scans of the cornea and anterior segment were performed both before and after lamellar transplantation procedures on the cornea with the Visante anterior segment OCT system (Visante OCT; Carl Zeiss Meditec, Inc, Dublin, California, USA), and the imaging findings were correlated with the clinical picture. Measurements of lamella thickness were performed with the software provided. Anterior segment OCT images were able to provide valuable information on donor apposition, Descemet membrane detachment after deep anterior lamellar keratoplasty (DALK), posterior lamellar dislocation, primary graft failure, and anterior chamber crowding with consequent chamber angle encroachment and pupillary block after Descemet stripping automated endothelial keratoplasty (DSAEK). Anterior segment OCT is a valuable imaging tool for assessing the feasibility of lamellar transplantation surgery in the diseased cornea and in the management of surgical complications after such procedures.

A Novel Population of Repair Cells Identified in the Stroma of the Human Cornea

The transmembrane protein CD133 is expressed on somatic stem cells of various adult human tissues. To investigate whether human corneal stroma also contains CD133-expressing cells and to analyze their functional features, stromal cells were isolated by collagenase digestion, immunophenotyped, and transferred to different culture systems to determine their stem cell properties as well as their differentiation potentials. For comparison, the embryonic keratocyte cell line EK1.Br, the dermal stromal cell line NHDF, and stromal cells of diseased corneas were studied. On average, 5.3% of the normal stromal cells expressed the stem cell marker CD133 and 3.6% co-expressed CD34. Expression of CD133 but not CD34 was also demonstrated for EK1.Br cells, whereas NHDF cells were negative for both markers. Further analysis of CD133(+) normal corneal cells revealed that a significant proportion displayed a monocytic phenotype with co-expression of CD45 and CD14. In diseased corneas, up to 26.8% of the stromal cells showed expression of CD133, and virtually all CD133(+) cells co-expressed CD14 but not CD45. Moreover, using a standard clonogenic assay, normal stromal cells had the capacity to form colonies of the macrophage lineage. These colonies could be further differentiated into lumican-expressing keratocytes. Our data suggest that the human corneal stroma harbors CD133(+) monocytic progenitor cells, which possess the potential to differentiate into the fibrocytic lineage. Thus, CD133(+) /CD45(+) /CD14(+) cells might represent stromal repair cells that differentiate into keratocytes via a CD133(+)/CD45()/CD14(+) intermediate stage. The findings from our study may shed new light on regenerative processes of the human corneal stroma.

Laminins in normal, keratoconus, bullous keratopathy and scarred human corneas

The laminin composition (LMalpha1-alpha5, beta1-beta3, gamma1 and gamma2 chains) of normal corneas and corneal buttons from keratoconus, bullous keratopathy (BKP), Fuchs' dystrophy + BKP, Fuchs' dystrophy without BKP and scar after deep lamellar keratoplasty (DLKP) was investigated with immunohistochemistry. The epithelial basement membranes (BMs) of both normal and diseased corneas contained LMalpha3, alpha5, beta1, beta3, gamma1 and gamma2 chains. The epithelial BM morphology was altered in the different diseases. Scarring was associated with irregular BM and ectopic stromal localization of different laminin chains. The Descemet's membrane (DM) contained LMalpha5, beta1 and gamma1 chains in all cases and additionally LMbeta3 and gamma2 chains in the majority of keratoconus corneas. The interface in the DLKP cornea had patches of LMalpha3, alpha4, alpha5, beta1 and beta2 chains, and an extra BM-like structure under the Bowman's membrane. These results suggest that laminin chains participate in the process of corneal scarring and in the pathogenesis of some corneal diseases. The novel finding of LMalpha3, beta3 and gamma2 in the DM of keratoconus buttons indicates that this membrane is also involved in the disease and that some cases of keratoconus may have a congenital origin, without normal downregulation of the LMbeta3 chain.

Intérêt des greffes de membranes amniotiques dans les pathologies oculaires de surface

Amniotic membrane transplantation is now a widely adopted technique in the field of eye surface diseases. Depending on the indication, the amniotic membrane can be used as either a graft or a patch. When used as a graft, the amniotic membrane serves as a substrate for regrowth of deficient epithelium; the aim is to integrate this membrane. The basal membrane reinforces the adhesion and differentiation of the corneal epithelial cells, facilitates their migration, and prevents their apoptosis. When used as a patch, the amniotic membrane is sutured epithelium-down so as to maximize the concentration of biological factors delivered by this membrane: the membrane covers the diseased cornea and acts as a biological bandage and analgesic. The best indications for amniotic membrane grafts are acute chemical burns and trophic corneal ulcers refractory to all medical treatment. When these ulcers are perforated or in the early stages of perforation, it is best to use multiple layers of amniotic membrane, restoring the thickness of the cornea. In cases of confirmed limbal deficiency, amniotic membrane grafts may be a useful complement to the necessary limbal stem cell grafts. In the future, amniotic membranes will provide an indispensable support for the expansion of cultured stem cells. Amniotic membrane grafts may also be used to reconstruct the conjunctiva following the exeresis of symblepharons or conjunctival tumors. However, the use of this technique is currently limited to diseases with little inflammation and no extensive fibrosis.

Aldehyde dehydrogenase (ALDH) 3A1 expression by the human keratocyte and its repair phenotypes

Transparency is essential for normal corneal function. Recent studies suggest that corneal cells express high levels of so-called corneal crystallins, such as aldehyde dehydrogenase (ALDH) and transketolase (TKT) that contribute to maintaining cellular transparency. Stromal injury leads to the appearance of repair phenotype keratocytes, the corneal fibroblast and myofibroblast. Previous studies on keratocytes from species such as bovine and rabbit indicate that the transformation from the normal to repair phenotype is accompanied by a loss of corneal crystallin expression, which may be associated with loss of cellular transparency. Here we investigated if a similar loss occurs with human keratocyte repair phenotypes. Human corneal epithelial cells were collected by scraping and keratocytes were isolated by collagenase digestion from cadaveric corneas. The cells were either processed immediately (freshly isolated keratocytes) or were cultured in the presence of 10% fetal bovine serum or transforming growth factor-β to induce transformation to the corneal fibroblast and myofibroblast phenotypes, respectively. RT-PCR, western blotting and immunolabeling were used to detect mRNA and protein expression of ALDH isozymes and TKT. ALDH enzyme activity was also quantitated and immunolabeling was performed to determine the expression of ALDH3A1 in human corneal tissue sections from normal and diseased corneas. Human corneal keratocytes isolated from three donors expressed ALDH1A1 and ALDH3A1 mRNA, and one donor also expressed ALDH2 and TKT. Corneal epithelial cells expressed ALDH1A1, ALDH2, ALDH3A1 and TKT. Compared to normal keratocytes, corneal fibroblast expression of ALDH3A1 mRNA was reduced by 27% ( n = 5). ALDH3A1 protein expression as detected by western blotting was markedly reduced in passage zero fibroblasts and undetectable in higher passages ( n = 3). TKT protein expression was reduced in fibroblasts compared to keratocytes ( n = 2). ALDH3A1 enzyme activity was not detectable in corneal fibroblasts ( n = 6) but was readily detected in corneal epithelial cells (0.29 ± 0.1 U/mg protein, n = 4) and keratocytes (0.05 ± 0.009 U/mg protein, n = 7). ALDH3A1 expression was also reduced in corneal fibroblasts and myofibroblasts as determined by immunolabeling of the cells in culture ( n = 3) and in diseased corneal tissues in situ ( n = 2). We conclude that expression of the crystallin ALDH3A1 is decreased in repair phenotype human keratocytes, compared to normal human keratocytes. Extrapolating from studies of bovine and rabbit, the reduced expression of ALDH3A1 may contribute to the loss of corneal transparency experienced by human patients after injury and refractive surgeries.

In Vitro Efficacy of Lufenuron against Filamentous Fungi and Blood Concentrations after PO Administration in Horses

Lufenuron is a benzoylphenyl urea-derived insecticide that has been recently introduced as a novel treatment for fungal infections in horses. The purposes of this study were to determine (1) the in vitro efficacy of lufenuron against Aspergillus and Fusarium spp. and (2) the ability of lufenuron to reach efficacious blood concentrations after PO administration in horses. Fungal colonies isolated from diseased equine corneas were tested against lufenuron solutions up to 700 μg/mL. Twenty-one adult horses received 1 of 3 PO lufenuron treatment regimens: 5 mg/kg body weight (BW) q24h for 3 days, 20 mg/kg BW q24h for 3 days, or 60 mg/kg BW q24h for 1 day. Blood samples were collected up to 96 hours after drug administration and analyzed by high-performance liquid chromatography. Statistical analyses of lufenuron blood concentrations were performed by analysis of variance and Fischer's Least Significant Difference test, with statistical significance set at P < .05. Lufenuron showed no effect on the in vitro growth of Aspergillus or Fusarium spp. Lufenuron was detected in the blood of all but 1 horse and showed no adverse effects. The maximum blood lufenuron concentration (83.5 ± 58.7 μg/L) was lower than the concentrations proven to be ineffective in vitro in this study. Further therapeutic use of lufenuron as an antifungal agent in horses should be based on proven efficacy against specific strains of clinically relevant fungi with pharmacokinetic data demonstrating sufficient lufenuron concentrations in target tissues.

A Dataset of Human Cornea Proteins Identified by Peptide Mass Fingerprinting and Tandem Mass Spectrometry

Diseases of the cornea are extremely common and cause severe visual impairment worldwide. To explore the basic molecular mechanisms involved in corneal health and disease, the present study characterizes the proteome of the normal human cornea. All proteins were extracted from the central 7-mm region of 12 normal human donor corneas containing all layers: epithelium, Bowman's layer, stroma, Descemet's membrane, and endothelium. Proteins were fractionated and identified using two different procedures: (i) two-dimensional gel electrophoresis and protein identification by MALDI-MS and (ii) strong cation exchange or one-dimensional SDS gel electrophoresis followed by LC-MS/MS. All together, 141 distinct proteins were identified of which 99 had not previously been identified in any mammalian corneas by direct protein identification methods. The characterized proteins are involved in many processes including antiangiogenesis, antimicrobial defense, protection from and transport of heme and iron, tissue protection against UV radiation and oxidative stress, cell metabolism, and maintenance of intracellular and extracellular structures and stability. This proteome study of the healthy human cornea provides a basis for further analysis of corneal diseases and the design of bioengineered corneas.

Partial enrichment of a population of human limbal epithelial cells with putative stem cell properties based on collagen type IV adhesiveness

The concept that corneal epithelium stem cells reside in limbus has been recognized for more than a decade, but isolation of these stem cells has not been accomplished. This study was an initial attempt to isolate a population of human limbal epithelial cells enriched for certain putative stem cell properties based on their phenotype. Epithelial cells harvested from fresh human limbal rings and their primary cultures were allowed to adhere to collagen IV-coated dishes for 20 min and 2 hr, sequentially. The rapidly adherent cells (RAC), slowly adherent cells and non-adherent cells were evaluated for certain stem cell properties: (a) BrdU-label retention, (b) expression of basal cell (integrin β1, p63, ABCG2) and differentiation (involucrin, keratin 12) markers, and (c) colony forming efficiency (CFE) and growth capacity on a 3T3 fibroblast feeder layer. Among unfractionated cells and the three selected populations, the RAC, accounting for about 10% of whole population, were enriched 5-fold in BrdU label-retaining cells, displayed the highest number of integrin β1 and p63 positive and involucrin negative cells, expressed high levels of ΔNp63 and ABCG2 mRNA, and lacked involucrin and K12 expression, and possessed the greatest CFE and growth capacity. These findings demonstrated for the first time that human limbal epithelial cells with stem cell properties can be partially enriched by their adhesiveness to collagen IV. The RAC population enriched for certain putative stem cell properties may prove useful in the future for transplantation to diseased and damaged corneas with limbal stem cell deficiency.

In Vitro Efficacy of Lufenuron against Filamentous Fungi and Blood Concentrations after PO Administration in Horses

Lufenuron is a benzoylphenyl urea-derived insecticide that has been recently introduced as a novel treatment for fungal infections in horses. The purposes of this study were to determine (1) the in vitro efficacy of lufenuron against Aspergillus and Fusarium spp. and (2) the ability of lufenuron to reach efficacious blood concentrations after PO administration in horses. Fungal colonies isolated from diseased equine corneas were tested against lufenuron solutions up to 700 microg/mL. Twenty-one adult horses received 1 of 3 PO lufenuron treatment regimens: 5 mg/kg body weight (BW) q24h for 3 days, 20 mg/kg BW q24h for 3 days, or 60 mg/ kg BW q24h for 1 day. Blood samples were collected up to 96 hours after drug administration and analyzed by high-performance liquid chromatography. Statistical analyses of lufenuron blood concentrations were performed by analysis of variance and Fischer's Least Significant Difference test, with statistical significance set at P < .05. Lufenuron showed no effect on the in vitro growth of Aspergillus or Fusarium spp. Lufenuron was detected in the blood of all but 1 horse and showed no adverse effects. The maximum blood lufenuron concentration (83.5 +/- 58.7 microg/L) was lower than the concentrations proven to be ineffective in vitro in this study. Further therapeutic use of lufenuron as an antifungal agent in horses should be based on proven efficacy against specific strains of clinically relevant fungi with pharmacokinetic data demonstrating sufficient lufenuron concentrations in target tissues.

A novel flow cytometry based assay for quantification of corneal angiogenesis in the mouse model of herpetic stromal keratitis

In this study a novel flow cytometry based quantitative assay for measuring corneal angiogenesis is demonstrated. Corneas of Balb/c mice were lightly scarified and infected with 5×10 5, 5×10 4 and 5×10 3 pfu Herpes simplex virus 1 (HSV-1) RE virus. Development of corneal angiogenesis was studied on day 15 post infection by direct visualisation of infected cornea with a slit-lamp biomicroscope. Endothelial cells constituting the newly developed blood vessels in cornea were stained with murine anti CD31 antibody on frozen corneal sections and corneal whole mounts at day 15 post infection. Total number of endothelial cells was quantified at day 15 post infection by flow cytometry. Mice infected with different doses of HSV-1 RE developed severe to mild corneal angiogenesis at day 15 post infection. Endothelial cells constituting the newly formed blood vessels expressed CD31 at day 15 post infection. Flow cytometry revealed that, the number of CD31 positive cells isolated from diseased corneas increased with the increase in Neovascularisation index. The flow cytometry analysis used in this present study is a useful, accurate and cost effective method for quantifying corneal angiogenesis.

Expression of HLA-G in human cornea, an immune-privileged tissue

Human leukocyte antigen (HLA)-G retains the capacity to modulate immune responses, favoring the establishment of tolerance in solid-tissue allotransplants. To better understand the mechanisms that promote corneal allograft survival, we investigated whether HLA-G was an immunoregulatory factor involved in corneal immunology. We therefore sought HLA-G expression in corneal tissues. Corneal transplantation consists in replacing the center of a diseased cornea with normal corneal tissue. Two corneal parts are not used in such surgery: diseased central corneal tissue and peripheral normal cornea. For this study, we used healthy corneas obtained from deceased donors and diseased corneas obtained from patients with pseudophakic bullous keratopathy or keratoconus who had undergone corneal transplantation. Immunohistochemical analysis carried out on the cryopreserved corneas showed a positive immunohistochemical staining with anti–HLA-G, anti–HLA-A, -B, and -C, and anti–HLA class I monoclonal antibodies. Staining was obtained for keratocytes, epithelial cells, and endothelial cells from both healthy and pathologic human corneas, revealing the presence of HLA class I proteins, including HLA-G. HLA-G transcripts were detected in normal cornea by reverse transcriptase–polymerase chain reaction with a classical pattern of alternative splicing. The detection of HLA-G protein in adult corneas leads to the conclusion that this protein may contribute to the maintenance of the privileged immune status of cornea.

A novel microscope system for time-lapse observation of corneal cells in a living mouse

A novel microscope system is presented for observation of corneal cells in a living mouse. It enables tracking of individual cells in all layers of the cornea at various times, thus allowing the generation of time-lapse recordings. The system consists of three major components: an upright fluorescence microscope for visualization of corneal cells, a mouse-holding unit for immobilization of the animal and the eye, and a set of gimbals which permit observation of a wide area of corneal surface without refocusing. The same cells could be observed at different times with the help of fiducial marks in the cornea, allowing their changes in position to be determined under natural and experimental conditions. This technique should prove useful in investigation of the cell movement in normal and diseased corneas, including the study of wound healing after an injury or surgery.

Bioengineered corneas: how close are we?

Bioengineered corneas are substitutes for human donor tissue that are designed to replace part or the full thickness of damaged or diseased corneas. They range from prosthetic devices that solely address replacement of the cornea's function to tissue-engineered hydrogels that allow some regeneration of the host tissue. In addition, there are also bioengineered lenticules that may be implanted into the cornea to improve vision by altering the refractive properties of the eye, an alternative procedure to refractive surgery. In recent years, there have been significant developments in many areas of bioengineered corneas, such as the clinical trials of an artificial cornea designed as a prosthesis, the development of completely natural corneal replacements, and the development of biosynthetic matrices that permit host tissue regeneration. For correction of refractive errors, a synthetic corneal onlay that allows stable overgrowth of epithelium appears to be promising.

Ex vivo expansion of corneal limbal epithelial/stem cells for corneal surface reconstruction.

The management of severe ocular surface disease due to limbal stem cell deficiency has changed dramatically. The concept of limbal stem cells, as the source of corneal epithelium revolutionised the therapeutic approach of ocular surface reconstruction. Deficiency of limbal stem cells results in blinding ocular surface diseases. Grafting viable limbal tissue, from either fellow healthy eye or a donor eye, with the resident stem cell population may replenish limbal stem cells and can restore the corneal surface to normality. Transplanting the limbal tissue can be achieved through a variety of procedures that include cadaveric keratolimbal allograft (KLAL), live or living related conjunctival limbal allograft (Ir-CLAL) and limbal autograft. Advances in tissue engineering techniques have offered a viable alternative to overcome the limitation of limbal tissue available for transplantation. Epithelial stem cells harvested from a small limbal biopsy can be expanded in vitro on a suitable carrier and then transplanted to the diseased cornea to successfully restore the corneal surface. This article is a chronological review of the important steps that brought ex vivo expanded stem cell transplantation in ocular, particularly corneal surface reconstruction. The MEDLINE data base was searched for the years 1966-2002, using key words cornea, cell culture, ex-vivo expansion, limbus, stem cell, ocular surface and transplantation. Several articles that were not found by MEDLINE search were taken from references from other articles. Inclusion or exclusion of article was based on the relevance to the subject. Corneal epithelial reconstruction with ex vivo expanded limbal cells is a potential tool in ocular surface reconstruction, although the technique is currently investigational. Strategies to achieve conjunctival epithelial restoration and tear film replenishment will allow ophthalmic surgeons to truly reconstruct the ocular surface.