Vector analysis of astigmatism before and after LASIK: a comparison of two different platforms for treatment of high astigmatism.

PURPOSE: To compare the outcomes of astigmatic laser in-situ keratomileusis (LASIK) procedures between two different platforms using J0 and J45 vector analysis. METHODS: Patients were divided into four groups, depending on the type of astigmatism and laser platform on which they were treated. Astigmatism was between 2 and 7 diopters (D). One hundred and thirty-five patients with myopic astigmatism (246 eyes) and 102 patients with mixed astigmatism (172 eyes) underwent unremarkable LASIK correction on Wavelight Allegretto Eye-Q 400Hz and Schwind Amaris 750S laser platform. The preoperative and postoperative sphere, negative cylinder [C] and axis (ø) of manifest refractions were subjected to vector analysis by calculations of the standard J0 (cos [4π(ø-90)/360]xC/2) and J45 (sin[4π(ø-90)/360]xC/2). RESULTS: Reporting the key results, we found J0 significantly reduced after LASIK in both groups (p < 0.001) but not J45. There was no significant association between individual pairs of pre and postoperative J0 & J45 values. There was no significant difference between the outcomes of the two platforms. CONCLUSIONS: Wavelight Allegretto 400Hz and Schwind Amaris 750S showed excellent results for treating patients with astigmatism, regardless whether it is mixed or myopic astigmatism. The J45 did not reduce significantly possibly because of the low number of eyes with oblique astigmatism. There was no genuine difference post-operatively between groups treated on two different laser platforms according to the vector analyses.

Three-year corneal graft survival rate in high-risk cases treated with subconjunctival and topical bevacizumab.

PURPOSE: To evaluate the effect of combined subconjunctival and topical bevacizumab treatment on corneal graft survival rate in high-risk eyes. METHODS: Prospective, consecutive, interventional case series. Fifty eyes of 50 high-risk patients scheduled for penetrating keratoplasty (PK) were included in the study; two Stevens-Johnson syndromes (SJS), five corneal combustions due to chemical burn, seven post-traumatic vascularised leucomas, 11 post-infectious vascularised leucomas, 19 rejected grafts and six corneal ulcers. Additional surgeries such as autologous limbal stem cell and/or amniotic membrane transplantation were performed together with PK in ten cases. All eyes received subconjunctival injection of 0.5 ml bevacizumab (25 mg/ml) after PK. Eyes with more than two quadrants of neovascularisation (NV) received bevacizumab drops (25 mg/ml) postoperatively for up to 12 weeks. Donor grafts were followed up for best-corrected visual acuity, graft clarity, change in NV, endothelial cell density loss (ECD), and adverse events. Mean follow-up was 36.5 months (range 32-61). RESULTS: Best-corrected visual acuity increase was statistically significant in 82 % (41/50) of eyes 3 years after PK (paired t-test, p = 0.02). Thirty-five (70 %) high-risk grafts remained clear throughout the 3-year follow-up period. Decrease of corneal NV was observed in 84 % (42/50) of eyes treated with bevacizumab. ECD changed from preoperative 2,864 ± 301 down to 1,905 ± 187 cells/mm(2) at 3 postoperative years. A non-healing epithelial defect was recorded in one patient with SJS after 12 weeks of topical bevacizumab. CONCLUSION: Combined subconjunctival and topical bevacizumab treatment may improve corneal graft survival rate in the majority of high-risk cases.

Synthetic vs natural scaffolds for human limbal stem cells.

AIM: To investigate the impact of synthetic electrospun polyurethane (PU) and polycaprolactone (PCL) nanoscaffolds, before and after hydrolytic surface modification, on viability and differentiation of cultured human eye epithelial cells, in comparison with natural scaffolds: fibrin and human amniotic membrane. METHODS: Human placenta was taken at elective cesarean delivery. Fibrin scaffolds were prepared from commercial fibrin glue kits. Nanoscaffolds were fabricated by electrospinning. Limbal cells were isolated from surpluses of human cadaveric cornea and seeded on feeder 3T3 cells. The scaffolds used for viability testing and immunofluorescence analysis were amniotic membrane, fibrin, PU, and PCL nanoscaffolds, with or without prior NaOH treatment. RESULTS: Scanning electron microscope photographs of all tested scaffolds showed good colony spreading of seeded limbal cells. There was a significant difference in viability performance between cells with highest viability cultured on tissue culture plastic and cells cultured on all other scaffolds. On the other hand, electrospun PU, PCL, and electrospun PCL treated with NaOH had more than 80% of limbal cells positive for stem cell marker p63 compared to only 27%of p63 positive cells on fibrin. CONCLUSION: Natural scaffolds, fibrin and amniotic membrane, showed better cell viability than electrospun scaffolds. On the contrary, high percentages of p63 positive cells obtained on these scaffolds still makes them good candidates for efficient delivery systems for therapeutic purposes.

How to convert the obliquely crossed to non-crossed astigmatism? A simple method using vector analysis.

The authors discussed about the problem of special form in astigmatism classification. This special type of astigmatism is the form of obliquely crossed astigmatism. In which the meridians, major and minor, are not right angles. In this astigmatism is not possible to prescribing for cylindrical (toric) spectacle lens. Authors describe the Thompson formula for oblique crossed cylinder and observe that this formula is to complicate for calculation new cylinder power. In this reason, the authors create the new formula and simple procedure for this calculation. This simple formula based on vector analysis and read: DM3 = DM2 x cos2 beta.

Curvature analyses of the corneal front and back surface.

In 25 eyes, age range 60-80 years, in purpose of preoperative treatment prior to cataract surgery, measurements of the radius of curvature as well as main meridians of the anterior and posterior corneal surfaces were performed. Average value of the curvature of the anterior corneal surface (R1) was R1 = 7.84 mm, while posterior radius (R2) was 6.4 mm. Main meridian of the anterior corneal surface was in 70% in plus value (+), while postrerior main meridian was always in minus value. It is very important to highlight that main corneal meridians R1 and R2 always cross outside the 90 degrees angle. That is why the combination of the calculation based on the two main meridians superposition (overrefraction) as a form of crossed cylinders has to be calculated (Astigmatismus directus seu obliquus decusatus) using special formula. One of that kind of formulas is Thompson's. Authors want to emphasise the importance of the optical calculation of the posterior corneal surface in the refractive surgery.

Stereoacuity and Multifocal Intraocular Lenses - a Systematic Review.

Background: Among various visual functions, stereoacuity, or the ability to perceive depth, is the most sophisticated binocular function. Many publications discuss the influence of retinal image formation by multifocal intraocular lenses on glare and contrast sensitivity, but only a few present results of testing binocular vision in patients with multifocal intraocular lenses. Objective: This article is designed to review the results of testing binocular vision in patients with multifocal intraocular lenses implanted in cataract surgery. Methods: This article was performed based on a literature review and Internet search through scientific databases such as PubMed, Scopus, Web of Science, and Google Scholar. Results: Some reports found that patients implanted with the monofocal lens, when measured with a near addition, presented statistically significant better stereoacuity scores than those implanted with any of the multifocal intraocular lens types. When the TNO test was used for measurement, statistically significant better stereoacuity was disclosed with the refractive multifocal intraocular lens than with the diffractive-based multifocal intraocular lens design. Stereoacuity scores, even within the same types of lenses, were significantly better with the Titmus test than with the TNO test. Conclusion: Stereoacuity is not affected by multifocality-induced retinal blur as it is by other causes of image degradation such as small residual refractive error very early opacification of ocular media or dry eye. Multifocal intraocular lenses do not cause more functional aniseikonia than would be expected with a monofocal intraocular lens. Since stereoacuity is compromised with unilateral multifocal intraocular lens implantation bilateral implantation should be attempted.

The Reliability of PCL/Anti-VEGF Electrospun Scaffolds to Support Limbal Stem Cells for Corneal Repair.

Since only few reported studies propose anti-vascular endothelial growth factor (anti-VEGF) delivery through electrospun scaffolds, this study greatly contributes to the potential prevention of patient's vision loss, as it explores electrospun polycaprolactone (PCL) coated with anti-VEGF for the blockage of abnormal cornea vascularization. In terms of physicochemical properties, the biological component increased the PCL scaffold fiber diameter (by ~24%) and pore area (by ~82%), while ut slightly reduced its total porosity as the anti-VEGF solution filled the voids of the microfibrous structure. The addition of the anti-VEGF increased the scaffold stiffness almost three-fold at both strains of 5 and 10%, as well as its biodegradation rate (~36% after 60 days) with a sustained release profile after Day 4 of phosphate buffered saline incubation. In terms of scaffold application function, the PCL/Anti-VEGF scaffold proved to be more favorable for the adhesion of cultured limbal stem cells (LSCs); this was confirmed by the SEM images, where the cells showed flat and elongated conformations. Further support of the LSC growth and proliferation was confirmed by the identified p63 and CK3 markers after cell staining. These results demonstrate the advantageous effect of the surface-adsorbed anti-VEGF to stop vision loss and help damaged corneal tissue repair.

Resolution of pseudophakic cystoid macular edema with combination therapy of topical corticosteroids and nonsteroidal anti-inflammatory drugs.

A 69 years old women underwent uneventful cataract surgery of her left eye with phacoemulsification and posterior chamber intraocular lens implantation in topical anesthesia. Patient was postoperatively treated with combination of antibiotic and steroid in decreasing dosages during five weeks: one drop five times a day the first week, three times a day second to forth week and one time a day the fifth week. In each checkup, performed first postoperative day, 7 days, 5 weeks and 12 weeks after the operation, visual acuity with and without correction, tonometry, corneal transparency, biomicroscopy of posterior pole and measure of macular thickness by optical coherence tomography (OCT) were performed. At first day follow-up visit, the patient's visual acuity was 20/25 but 6 weeks after the operation, the patient's vision had worsened to 20/60 after a slow steroid tapper. At that time OCT showed foveal thickening and cystic changes specific for cystoid macular edema (CME). Combination of corticosteroid and non-steroidal anti-inflammatory drug four times daily was included in therapy. The dose was tapered off over the ensuing 8 weeks. The total treatment duration was 12 weeks. At the patient's 2-month follow-up visit, vision has improved to 20/20 and the fovea appeared flat. OCT showed complete resolution of foveal thickening and cystic changes. Combination of corticosteroid and NSAID is effective and safe therapy for treating pseudophakic CME. Patient showed significant improvement in visual acuity and retinal thickness at 2 months post treatment.

1-year follow-up study of endothelial cell density loss after penetrating keratoplasty.

High endothelial cell density (ECD) is essential for the corneal graft clarity. We evaluated ECD loss in 120 eyes that underwent penetrating keratoplasty (PK) in Eye Clinic Svjetlost in a one year follow up period. Patients were divided into 3 groups of high (N = 35), intermediate (N = 31) and low risk (N = 54) for graft failure. Postoperative central endothelial density, coefficient of variation in cell area (polymegathism), percentage of hexagonal cells (pleomorphism) in comparison to preoperative donor cell measurements were determined in the following postoperative time-points of 1, 2, 3, 6, 9 and 12 months. There were no significant differences in the preoperative ECD values, storage time, donor age or surgical procedures between groups. Throughout all time points intermediate group had the greatest statistically significant ECD loss as compared to high and low risk groups. There were no significant differences between high and low risk group. After 12 month post PK, intermediate risk group had 28.38% ECD loss as compared to 24.07% in high and 23.03% ECD loss in low risk group. Coefficient of variation in cell area (CV) was for high risk group 0.34, intermediate 0.40 and low risk 0.31 which was not significantly different between groups. Percentage of plemorphism in high risk was 54%, intermediate 58% and in low risk 48% which was significantly different as compared to other two groups. Our study showed that corneal pathology is among others, very important prognostic factor for ECD after PK. However, longer follow up period is needed.

Anti-VEGF in treatment of diabetic macular edema.

Diabetic macular edema is the leading cause of moderate visual deterioration in patients with diabetic retinopathy. Ranibizumab) blocks vascular endothelial growth factor (VEGF) induced hyperpermeability of blood vessels. In this prospective case series we investigated the efficacy and safety of anti-VEGF treatment in reduction of central retinal thickness (CRT) and improvement in visual acuity (VA) in patients with diabetic macular edema (DME). 9 patients were followed up for 6 months and treated monthly with intravitreal ranibizumab. VA and CRT were measured at each visit. Treatment was discontinued as the peak improvement of either parameter was reached and reinstituted in case of deterioration/recurrence of edema. Study endpoints included: VA using ETDRS chart, CRT and number of injections at 6 months. Mean VA from all 9 patients increased by 0.3 lines of logMAR (p < 0.05 compared to baseline), and CRT decreased from 515 +/- 123 microm to 310 +/- 110 microm. The improvement of VA after ranibizumab injection was in correlation with a decrease in CRT. Mean of 4 injections were needed to control the disease during the follow-up period. Ranibizumab treatment was effective in VA and reducing CRT. Several injections were needed to control the disease. Regular OCT examinations and retreatment are advised in order to maintain initially reached VA.

First results of Intracor procedure in Croatia.

This study reports early outcomes of a cohort of presbyopic patients treated with Intracor. The study took place from December 2010 to May 2011 and was conducted in University Eye Hospital "Svjetlost", Zagreb, Croatia. 95 eyes were enrolled in this prospective clinical trial (49 patients with non dominant eye and 23 with bilateral treatment). All patients gave informed consent prior to enrollment. Follow up consisted of uncorrected and corrected distant and near visual acuity, record of topographic changes, visual disturbances and patient satisfaction at 1 week, 1 and 3 months after the surgery. In this study Intracor procedure presented as both safe and effective with all eyes gaining several lines of uncorrected near visual acuity (UNVA), and achieving good uncorrected distant visual acuity(UDVA) as well. UDVA was affected by a mild myopic shift, which was effective in reducing mild preexisting hyperopia in some patients but led to a mild myopic outcome in previously emmetropic patients. Statistically significant improvement in UDVA and UNVA was observed in all time points. At 3 months of postoperative follow up all patients gained several lines of UNVA with monocular UNVA Jaeger system 1.67 +/- 0.28. UDVA showed slight improvement over time and initial myopic shift showed tendency of slight decrease with all patients achieving 1.0. Overall patients satisfaction was very high (98%) with only a few (3 patients, 5 eyes) reporting mild halo and glare at 3 months postop.Intracor procedure has proven its short-term safety and efficacy in treating presbyopia. However, longer follow up period is needed.

The expression of interleukin-1 alpha, TNF and VEGF in corneal cells of patients with bullous keratopathy.

Bullous keratopathy (BK) is a chronic corneal edema with or without subepithelial bullae as a result of a loss of the endothelial cells. 15 patients with BK after cataract surgery with intraocular lens implantation, due to Fuchs dystrophy (n = 3) or corneal endothelial trauma (n = 12) were included in the study. All patients were treated by amniotic membrane transplantation (AMT). Corneal epithelial cells in patients suffering from BK secreted 3.91 +/- 3.09 pg/mL of IL-1 alpha, 4446 +/- 16.8 pg/mL of TNF and 81.43 +/- 37.81 pg/mL of VEGF-I. Levels of all 3 investigated cytokines were significantly higher as compared to controls (p < 0.005). Amniotic membranes that were used to treat investigated patients contained 638.98 +/- 613.98 pg/mL of IL-1ra, 0.026 +/- 0.009 pg/mL of sTNF and 81.39 +/- 21.01 pg/mL of VEGF-R. Beneficial clinical effect of the AMT in treating BK could be explained by its natural production of pro-inflammatory cytokine antagonists such as IL-ra, sTNF antagonist and VEGF-R.

Transplantation of amniotic membrane in corneal ulcers and persistant epithelial defects.

Amniotic membrane transplantation (AMT) leads to reduction of inflammatory symptoms and causes faster epithelisation in corneal ulcers and persistant epithelial defect. 21 patients with corneal ulcer (n = 18) or non-healing epithelial defect (n = 3) unresponsive to conventional treatment were included in the study. All patients were treated by AMT. Corneal epithelial cells in patients suffering from corneal ulcer secreted 3.51 +/- 1.79 of IL-1alpha, 64.27 +/- 31.53 pg/mL of TNFalpha and 209.07 +/- 201.82 pg/mL of VEGF. Levels of all 3 investigated cytokines were significantly higher as compared to controls (p < 0.005). Amniotic membranes that were used contained 775.69 +/- 613.98 pg/mL of IL-1alpha, 0.036 +/- 0.033 pg/mL of sTNF and 175.01 +/- 166.63 pg/mL of VEGF-R. Supporting effect of the AMT could be explained by the fact that AM secretes its natural antinflammatory antagonists IL-1ra, sTNF and VEGF-R.

Selective laser trabeculoplasty in the treatment of pseudoexfoliation glaucoma in patients allergic to all anti-glaucoma drops.

Secondary chronic open-angle glaucoma associated with pseudoexfoliation (PEX) syndrome accounts for approximately 25% of all glaucomas and represents the most common identifiable cause of glaucoma overall. Selective laser trabeculoplasty (SLT) is effective in reducing intraocular pressure (IOP) in glaucomatous patients and has the advantage of preserving surrounding structures. We report here SLT treatment of a 82 year old female with a secondary developed open-angle pseudoexfoliation glaucoma allergic to all anti glaucoma eye drops especially those which contain bensalconium chloridum as preservative. Since patient was allergic also to methyl-cellulose, we performed SLT with water as a mediator. Patient had PEX syndrome for 10 years, immature cataracts on both eyes, and best corrected visual acuity (BCVA) 0.7 on the right and 0.2 on the left eye. We have monitored intraocular pressure (IOP), the changes in the visual field and optic nerve. Preoperative IOP was 28 mmHg on the right and 30 mmHg on the left eye. The follow up period was 24 months with time points for measured parameters every 3 months. After 18 months IOP remained in the normal values (average 17 mmHg) on the right eye, but on the left eye it increased up to 28 mmHg. SLT re-treatment was carried out on the left eye and the IOP stabilized again on the values between 16-18mmHg. There were no significant change in the visual field and optic nerve configuration before and after SLT (C/D value for right eye: 0.3-0.4; C/D left eye: 0.5). Based on this case report, SLT seems to be very effective treatment for maintaining regular IOP in patient with PEX who is allergic to all types of medications.

Amniotic membrane transplantation in the treatment of persistent epithelial defect on the corneal graft.

It has been shown that amniotic membrane transplantation (AMT) improves healing of the epithelium defects as it serves as a basement membrane for endothelial cells growth, prevents inflammatory cell infiltration and reduces apoptosis in keratocytes. Having in mind the healing properties of AM we investigated the efficacy of AMT in persistent epithelial defect (PED) on the corneal graft. 80 corneal grafts were prospectively followed up for presence of PED 10 months after surgery. PED was detected in 12 cases (15%) having surgery for: rejected graft (n = 4), keratoconus (n = 3), keratoconus following PK on a second eye (n = 3), corneal perforation (n = 1) and Stevens-Johnson keratopathy (n = 1). Epithelial defect (ED) developed 14 +/- 7 days after surgery in 10 cases and 1.5 month in other two. All patients were primarily conservatively treated with subconjuctival steroids and artificial tears for 10 days and systemic steroid therapy if needed after, until the period of 2 weeks. 4 patients were healed. Since ED was unresponsive to all previous treatments for more than 2 weeks, one layer of AM was placed on the corneal lesion in 5 patients, and in 3 cases of deep PED several layers of AM were placed. Healing of the defect was obtained in 7/8 (87.5%) eyes. In 1 patient second AM transplantation was necessary. Mean epithelization time was 2 weeks (range 1-3 weeks) in monolayer and 3 weeks (range 2-4 weeks) for multilayer cases. 5 out of 8 patients retained the same best corrected visual acuity (BCVA) while 3/8 patients improved their vision more than 2 lines. Preoperative corneal thickness of 255 +/- 40 mm increased to 455 +/- 90 mm. AM transplantation facilitates healing of corneal epithelium. PED on the corneal graft unresponsive to conventional treatment can be effectively cured when covered with one or more amniotic membrane layers.

Penetrating keratoplasty and Verisyse iris-claw lens--is it safe for corneal graft?

The loss of the corneal endothelial cells, especially in a first postoperative year, has been observed in patients who underwent penetrating keratoplasty (PK). The implantation of new generation of "iris claw" phakic IOL (Verisyse) in refractive cases has been shown to cause clinicaly insignificant endothelial cell loss. In our prospective case series we investigated the endothelial cell loss and clinical outcome in patients that either underwent PK and implantation of PCIOL or PK and implantation of Verisyse IOL. In the first group of 9 patients scheduled for PK, implantation of Verisyse was performed due to the absence of the posterior capsule support. 2 of these patients had angle supported ACIOL, 4 patients were aphakic and 3 had posttraumatic cataract with ruptured posterior capsule. The second group of 12 patients had standard "triple" procedure (PK + ECCE + PCIOL). BCVA of both groups of patients prior the operation was hand movement in 12 patients, light perception in 7 patients and 0.05 in 3 patients. The preoperative endothelial cell count of the donor grafts obtained from the eye bank was 2800 cells/mm2 on average. The follow up was 6-10 months. Six months after the operation all "Verysise" patients maintained transparent graft. Postoperative visual acuity improvement was recorded in 18 out of 21 eyes (85.7%). Best spectacle corrected visual acuity of > or = 0.3 was achived in 55.5% in the Verisyse group and in 50.0% of "triple procedure" group. The endothelial cell count and morphology were estimated on the specular microscope on a monthly basis. Mean endothelial cells loss in patients with PK and Verysise was 40 +/- 8% and in patients with "triple" procedure was 42 +/- 12% at 10 postoperative months. There was no significant difference in the endothelial cell loss and clinical outcome between the group of patients who had PK and Verysise as compared to those with implanted PCIOL.

Comparison of different presbyopia treatments: refractive lens exchange with multifocal intraocular lens implantation versus LASIK monovision.

There are several methods for presbyopia treatment. Refractive lens exchange (RLE) followed by multifocal intraocular lens (MFIOL) implantation enables high rate of spectacle independence but have some visual disturbances. Laser in Situ Keratomileusis (LASIK) monovision gives patient ability to have good distant vision with dominant eye and good near vision with nondominant eye. In this prospective randomized study we wanted to compare clinical outcomes in patients who underwent either of the mentioned procedures. The first group included 50 patients (N = 100 eyes) who underwent RLE with MFIOL implantation and the second group included 50 patients (N = 100 eyes) who underwent LASIK monovision as presbyopia treatment. Uncorrected distant, near and intermediate visual acuity, patient's subjective satisfaction and visual disturbances were measured. Follow up was 6 months. Patients in RLE group had better near uncorrected visual acuity (UCVA) and patients in LASIK monovision group had better distant and intermediate vision. Patients in RLE group reported visual disturbances (halo, glare). Patients satisfaction and spectacle independence was high in both groups. Refractive lens exchange with multifocal intraocular lens implantation and LASIK monovision are effective methods for presbyopia treatment. LASIK monovision with -0.50D - 1.25D of residual dioptry at nondominant eye in patients under 50 years enables good vision at all distances without affecting stereovision. Patient selection and preoperative counseling are key to success.

Trichophyton Spp. fungal keratitis in 22 years old female contact lenses wearer.

Fungal keratitis represents one of the most difficult forms of microbial keratitis to diagnose and treat successfully. It is difficult to obtain correct diagnosis and topical antifungal preparations. Fungi can cause severe stromal necrosis and enter the anterior chamber by penetrating an intact Descemet membrane. The most common pathogens are filamentous fungi (Aspergillus and Fusarium spp.) and Candida albicans. The incidence of Trichophyton spp. keratitis is 5%. A 22 years old female contact lenses wearer after keratitis developed corneal melting syndrome, spontaneous perforation of the cornea and complicated cataract of the left eye. Conjunctival swab was sterile as well as first sample of corneal tissue and sample from the anterior chamber. Urgent therapeutic perforating keratoplasty (PK), was performed together with extracapsular cataract extraction and the implantation of the intraocular lens in the posterior chamber. The patient was treated with ciprofloxacin and diflucan (systemic therapy); with dexamethason and atropin (subconjunctivaly) and chlorhexidine, brolene, levofloxacin, polimyxin B, and dexamethason/neomycin (topically). Microbiology evaluation was performed once again following excisional biopsy of the intracameral portion of the lesion. The presence of Trichophyton spp. was finally confirmed. Itraconazole and garamycin were included in the systemic therapy. Corneal graft was clear for 17 days but decompensated 28 days after the PK. After two weeks microorganisms invaded the vitreous and caused endophthalmitis. Despite urgent pars plana vitrectomy patient developed endophthalmitis, lost light sensation and developed phthysis. Evisceration and the implantation of silicon prosthesis was done. Perforating keratoplasty is a method of choice in treating severe infectious keratitis unresponsive to conservative treatment but without the eradication of microorganisms it cannot restore the vision or save the eye. Trichophyton spp. may cause a severe disease of the anterior and posterior part of the eye which may finish with the lost of vision/eye. Prompt diagnosis and treatment of Trichophyton spp. keratitis are essential for a good visual outcome.

Poly(ε-caprolactone) Titanium Dioxide and Cefuroxime Antimicrobial Scaffolds for Cultivation of Human Limbal Stem Cells.

Limbal Stem Cell Deficiency (LSCD) is a very serious and painful disease that often results in impaired vision. Cultivation of limbal stem cells for clinical application is usually performed on carriers such as amniotic membrane or surgical fibrin gel. Transplantation of these grafts is associated with the risk of local postoperative infection that can destroy the graft and devoid therapeutic benefit. For this reason, electrospun scaffolds are good alternatives, as proven to mimic the natural cells surroundings, while their fabrication technique is versatile with regard to polymer functionalization and scaffolds architecture. This study considers the development of poly(ε-caprolactone) (PCL) immune-compatible and biodegradable electrospun scaffolds, comprising cefuroxime (CF) or titanium dioxide (TiO2) active components, that provide both bactericidal activity against eye infections and support of limbal stem cells growth in vitro. The PCL/CF scaffolds were prepared by blend electrospinning, while functionalization with the TiO2 particles was performed by ultrasonic post-processing treatment. The fabricated scaffolds were evaluated in regard to their physical structure, wetting ability, static and dynamic mechanical behaviour, antimicrobial efficiency and drug release, through scanning electron microscopy, water contact angle measurement, tensile testing and dynamic mechanical analysis, antimicrobial tests and UV-Vis spectroscopy, respectively. Human limbal stem cells, isolated from surgical remains of human cadaveric cornea, were cultured on the PCL/CF and PCL/TiO2 scaffolds and further identified through immunocytochemistry in terms of cell type thus were stained against p63 marker for limbal stem cells, a nuclear transcription factor and cytokeratin 3 (CK3), a corneal epithelial differentiation marker. The electrospun PCL/CF and PCL/TiO2 successfully supported the adhesion, proliferation and differentiation of the cultivated limbal cells and provided the antimicrobial effect against Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans.

Preparation and preservation of amniotic membrane.

The human amniotic membrane (AM) is the innermost layer of the placenta and consists of a single epithelial layer, a thick basement membrane and an avascular stroma. Due to the number of its properties, AM is increasingly used in the treatment of severe ocular surface diseases. The amniotic basement membrane facilitates migration and growth of epithelial cells, therefore promoting epithelialization. The avascular stroma of the AM reduces fibrovascular ingrowth and abnormal neovascularization. Amniotic epithelium produces anti-inflammatory and growth factors beneficial to the treatment of inflammatory corneal diseases. AM is prepared from a fresh placenta under sterile conditions, washed with balanced salt solution containing penicillin, streptomycin, neomycin and amphotericin B, placed in tissue culture and glycerol at a ratio of 1:1, and stored at -80 degrees C. A donor serological test for human immunodeficiency virus and hepatitis B and C viruses has to be all negative. After transplantation of the amniotic membrane (AMT) onto the eye surface, AM will be slowly absorbed within approximately 4-6 weeks. Depending on consumption, amniotic membranes are used up to 1 year after preparation, although many have recommended storage for an indefinite period. Since AM is not a completely transparent tissue, the patient's visual acuity may decrease after AMT; the patient should be aware of this temporary effect prior to surgery.