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Intraocular epithelial ingrowth is a rare but serious complication of eye surgery or trauma. The diagnosis is challenging but can be aided by techniques such as anterior optical coherence tomography (AS-OCT). This study aimed to compare clinical and histopathological diagnostic data to evaluate the accuracy of AS-OCT. This retrospective analysis focused on patients presenting with suspected epithelial invasion between 2017 and 2022. Clinical data and histopathological findings were analyzed for diagnostic confirmation. Immunohistochemistry using the corneal-specific marker cytokeratin 3 and the conjunctival-specific marker cytokeratin 13 helped to identify the type of invading epithelial cells. Statistical analysis was used to evaluate the effectiveness of AS-OCT for diagnosis. This study included 51 patients who underwent keratoplasty or enucleation. Sixteen cases (31%) of epithelial ingrowth were histologically confirmed. The most common type was the cystic form (82%). Enucleation was required in 25% of patients, mainly due to diffuse epithelial invasion. Immunohistochemistry revealed a conjunctival origin in 82% of the infiltrated cells. AS-OCT showed a sensitivity of 78% and a specificity of 26% in detecting hyperreflective structures associated with epithelial invasion. This study highlights the diverse manifestations of epithelial ingrowth and the need for improved diagnostic techniques, with AS-OCT showing promising results but requiring further validation to avoid misdiagnosis.

Background: Acute keratoconus (acute KC), which affects approximately 1.6–2.8% of keratoconus (KC) patients, is a pathological condition of the cornea characterized by stromal edema due to entry of aqueous humor through a tear in Descemet’s membrane. Methods: We present a novel combination of surgical procedures that allows swifter visual recovery in a consecutive, retrospective case series. The new surgical procedure for acute KC consists of a combination of Muraine corneal sutures to smooth the corneal curvature and Excimer laser-assisted penetrating keratoplasty and was performed in six acute KC patients from 2019 to 2022 at the Department of Ophthalmology, University Hospital of Martin-Luther-University Halle-Wittenberg (UMH), Germany. We monitored data on preoperative status, operative details, intraoperative and postoperative complications and visual outcomes were analyzed. Results: The mean age was 41.5 ± 13.5 years (3 OD, 3 OS). Neurodermatitis was present in 3 patients (50%). All patients received significant visual benefits from the procedure. Preoperative BCVA was hand motion (logMAR 3.0) in all patients; postoperatively, BCVA improved significantly logMAR 0.03 ± 0.09 [range: 0.2–0.4; p < 0.001, FUP 20+/−10 months). Visual acuity remained stable throughout the roughly biannual follow-ups. One patient developed endothelial graft rejection after 2 years. During the last examination, all eyes had clear grafts and stable curvatures, K1 and K2 being 42.43 ± 4.17 D and 44.95 ± 4.07 D, respectively, and mean corneal astigmatism was 2.61 ± 1.74 D. The thinnest corneal thickness was 519 ± 31 µm. A graft size of 8.0 × 8.1 mm was the most beneficial. Conclusions: in patients with acute KC and hydrops, a penetrating keratoplasty with Muraine corneal sutures is successful in terms of graft clarity and visual outcome. Combining the procedures allows quicker visual recovery. Patients with a history of neurodermatitis should have preoperative and postoperative dermatologic treatment and close follow-up for possible complications.

This study investigates the possible toxic effects of the preoperative antiseptic substances povidone iodine (PVI) and polyhexanide (PHMB; Serasept® 2) on wound healing in ophthalmology. To assess this impact, human telomerase-immortalized corneal epithelial (hTCEpi) cells and human telomerase-immortalized conjunctival epithelial (hCjE) cells were exposed to 1% and 5% PVI or 0.04% PHMB for different periods to evaluate the cytotoxicity of these two antiseptics. Furthermore, the toxicity of these antiseptics was investigated in a human tissue-specific corneal epithelial construct and porcine eye culture model. The results reveal the high cytotoxicity of PVI and PHMB in the hTCEpi and hCjE in monolayer cell culture models, independent of the incubation time and concentration of these substances. However, after hTCEpi cell differentiation into a tissue-specific corneal epithelial construct, contact with these antiseptics for the relevant preoperative time did not alter cPARP1 or Ki67 expression. Furthermore, the wound-healing process in the porcine cornea was not significantly influenced after incubation with these antiseptics. In summary, corneal and conjunctival epithelial cell lines are very sensitive to PVI and PHMB, whereas no significant alterations were found in intact tissue-specific corneal epithelial constructs or porcine corneas. Therefore, we could not identify PVI and PHMB as reasons for postoperative eye irritation.

Background: Conjunctival melanoma (CM) is associated with a high rate of local recurrence and poor survival rate. Novel therapeutic options are needed to reduce recurrence rate. The objective of the study was to demonstrate the improved effectiveness of electrochemotherapy (ECT) on CM using repetitive application. Methods: Tumor spheroids of three CM cell lines (CRMM1, CRMM2, CM2005.1) were treated repetitively with ECT using the chemotherapeutic agent bleomycin on days 3, 5, and 7 of culture. Application of bleomycin alone and electroporation alone served as controls. The cytotoxic effect was analyzed on day 10 compared to untreated control using an independent t-test. The spheroid outgrowth rate was measured. Result: CM tumor spheroid size (median value: 78%, SD: 32%) and viability (median value: 11%, SD: 11%) were dramatically reduced after repetitive ECT treatment (p-value < 0.001). Decreased proliferation capacity (down to 8%) and an increase of apoptotic cells were observed. In most repetitive ECT-treated spheroids, no viable or proliferating cells were detected. Only 33–40% of repetitive ECT-treated spheroids exhibited single outgrowing cells with a delay of time up to 38 days. Conclusion: Repetitive ECT application effectively induces cytotoxic effects in CM spheroids by inducing apoptosis, inhibiting proliferation and decreasing the percentage of surviving tumor cells. Thus, repetitive ECT results in improved antitumor effectiveness in CM and could be an alternative therapy option.

Limbal stem cell deficiency (LSCD) severely impairs vision and can lead to blindness. LSCD causes include chemical burns, infections, multiple previous operations and congenital malformations. Allogeneic limbal transplantation is a procedure for treating LSCD where prepared limbal tissue is attached using a double running suture during allogeneic penetrating keratoplasty (PKP). A total of 22 patients underwent ALT surgery between February 2019 and June 2022 at the University Hospital Halle (Saale). Regular follow-up was performed postoperatively every three months and included visual acuity testing, pressure measurement, slit lamp microscopic examination, fundoscopy, corneal topography and anterior segment optical coherence tomography (AS-OCT). The mean patient age was 69.5 years, and the mean follow-up was 19 months. All included patients had LSCD and multiple previous surgeries. Patient LSCD etiology was 59% infectious and 41% traumatic. ALTs integrated into corneal surfaces in all patients, demonstrated on AS-OCT. Since most patients initially received allogeneic limbal transplants, none of the operated eyes had surgical complications. Overall, visual acuity improved postoperatively from an initial 2.06 to 1.44 logarithm of the minimum angle of resolution (logMAR). Allogeneic limbal transplantation can be used to treat LSCD and its integration into the surrounding corneal tissue can be observed on AS-OCT.

Background: To investigate whether vaccination against SARS-CoV-2 is associated with the onset of retinal vascular occlusive disease (RVOD). Methods: In this multicentre study, data from patients with central and branch retinal vein occlusion (CRVO and BRVO), central and branch retinal artery occlusion (CRAO and BRAO), and anterior ischaemic optic neuropathy (AION) were retrospectively collected during a 2-month index period (1 June–31 July 2021) according to a defined protocol. The relation to any previous vaccination was documented for the consecutive case series. Numbers of RVOD and COVID-19 vaccination were investigated in a case-by-case analysis. A case–control study using age- and sex-matched controls from the general population (study participants from the Gutenberg Health Study) and an adjusted conditional logistic regression analysis was conducted. Results: Four hundred and twenty-one subjects presenting during the index period (61 days) were enrolled: one hundred and twenty-one patients with CRVO, seventy-five with BRVO, fifty-six with CRAO, sixty-five with BRAO, and one hundred and four with AION. Three hundred and thirty-two (78.9%) patients had been vaccinated before the onset of RVOD. The vaccines given were BNT162b2/BioNTech/Pfizer (n = 221), followed by ChadOx1/AstraZeneca (n = 57), mRNA-1273/Moderna (n = 21), and Ad26.COV2.S/Johnson & Johnson (n = 11; unknown n = 22). Our case–control analysis integrating population-based data from the GHS yielded no evidence of an increased risk after COVID-19 vaccination (OR = 0.93; 95% CI: 0.60–1.45, p = 0.75) in connection with a vaccination within a 4-week window. Conclusions: To date, there has been no evidence of any association between SARS-CoV-2 vaccination and a higher RVOD risk.

In the tissue donation field, to prevent pathogen transmission, all donors are screened by postmortem swabs for SARS-CoV-2 using qRT–PCR. Corneas from donors who tested positive for SARS-CoV-2 were subjected to further investigations. Corneal transplants and culture medium from positive donors were cultured under appropriate safety conditions for further analyses. Cornea tissue samples, including sclera/limbus/cornea, and culture media were taken at different time points for testing for SARS-CoV-2 using qRT–PCR, immunohistochemistry (IHC) and subgenomic RNA (sgRNA) analysis. Between January and May 2021, in four donors with initial negative premortem rapid tests, SARS-CoV-2 was detected post-mortem using qRT–PCR. In these cases, SARS-CoV-2 was observed at the beginning of cultivation in both tissue and culture medium using qRT–PCR and IHC. The virus was mainly localized in the limbus epithelial cells, with a stable detection level. Premortem rapid tests are potentially insufficient to exclude SARS-CoV-2 infection in corneal donors. While, for SARS-CoV-2, the risk of infection via transplants is considered low, a residual risk remains for presymptomatic new infections. However, our investigations provide the first indications that, with organ cultures, the risk of virus transmission is minimized due to the longer minimum culture period.

The purpose of our research was the development of Amphotericin B-loaded in situ gelling nanofibers for the treatment of keratomycosis. Different formulation strategies were applied to increase the drug load of the sparingly water-soluble Amphotericin B in electrospun Gellan Gum/Pullulan fibers. These include bile salt addition, encapsulation in poly(lactic-co-glycolic acid) (PLGA) nanoparticles and formation of a polymeric Amphotericin B polyelectrolyte complex. The Amphotericin B polyelectrolyte complex (AmpB-Eu L) performed best and was very effective against the fungal strain Issatchenkia orientalis in vitro. The complex was characterized in detail by attenuated total reflection infrared spectroscopy, X-ray powder diffraction, and differential scanning calorimetry. A heat induced stress test was carried out to ensure the stability of the polyelectrolyte complex. To gain information about the cellular tolerance of the developed polyelectrolyte complex a new, innovative multilayered-stratified human cornea cell model was used for determination of the cellular toxicity in vitro. For a safe therapy, the applied ophthalmic drug delivery system has to be sterile. Sterilization by electron irradiation caused not degradation of pure Amphotericin B and also for the bile salt complex. Furthermore, the developed Amphotericin B polyelectrolyte complex was not degraded by the irradiation process. In conclusion, a new polyelectrolyte Amphotericin B complex has been found which retains the antifungal activity of the drug with sufficient stability against irradiation-sterilization induced drug degradation. Furthermore, in comparison with the conventional used eye drop formulation, the new AmpB-complex loaded nanofibers were less toxic to cornea cells in vitro. Electrospinning of the Amphotericin B polyelectrolyte complex with Gellan Gum/ Pullulan leads to the formation of nanofibers with in situ gelling properties, which is a new and promising option for the treatment of keratomycosis.

Three-dimensional (3D) cell cultures represent small avascular tumors in vitro and simulate some of the biological characteristics of solid tumors, enhancing the evaluation of anticancer drug efficacy. Automated image analysis can be used for the assessment of tumor growth and documentation of changes in the size parameters of 3D tumor spheroids following anticancer treatments such as electrochemotherapy. The objective of this article is to assess the effect of various electroporation (EP) conditions (500–750 Volts/cm, 8–20 pulses, 100 µs pulse duration, 5 Hz repetition rate) combined with different bleomycin concentrations (1–2.5 ug/mL) on normal epithelial (HCjE-Gi) and conjunctival melanoma (CRMM1, CRMM2) 3D-cell cultures, through an automated image analysis and a comparison with standard histological assays. A reduction in tumor mass with loss of cell definition was observed after ECT (750 Volts/cm with eight pulses and 500 Volts/cm with 20 pulses) with bleomycin (1 μg/mL and 2.5 μg/mL) in the histological and immunohistochemical analyses of 3D CRMM1 and CRMM2 spheroids, whereas an increase in volume and a decrease in sphericity was documented in the automated image analysis and 3D visualization of both melanoma cell lines. For all other treatment conditions and for the HCjE-Gi cell line, no significant changes to their morphological features were observed. Image analysis with integrated software tools provides an accessible and comprehensive platform for the preliminary selection of homogenous spheroids and for the monitoring of drug efficacy, implementing the traditional screening methods.

Uveal melanoma (UM) is the most common primary intraocular tumor that arises from neoplastic melanocytes in the choroid, iris, and ciliary body. Electrochemotherapy (ECT) has been successfully established for the treatment of skin and soft tissue metastatic lesions, deep-seated tumors of the liver, bone metastases, and unresectable pancreas lesions. The aim of this study was to evaluate the effect of ECT in vitro in 3D spheroid culture systems in primary and metastatic UM cell lines. We also investigated the chick embryo chorioallantoic membrane (CAM) as an in vivo model system for the growth and treatment of UM tumors using ECT. The cytotoxic effect of ECT in 3D spheroids was analyzed seven days following treatment by assessment of the size and MTT [(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium reduction] assay. The cytotoxicity of ECT after intratumoral or intraarterial administration was evaluated histologically. In vitro and in vivo ECT caused a significant reduction in tumor size and viability compared to electroporation or chemotherapy in both sections of our study. The current results underline the effectiveness of ECT in the treatment of UM and prepare the way for further investigation of its potential application in UM.

AbstractPurpose??The purpose of the study was to investigate area and intensity of parapapillary autofluorescence in patients with ocular hypertension and controls.Patients and methods??Forty-two consecutive patients with ocular hypertension and 25 controls were investigated in a controlled prospective cross-sectional trial. All patients and controls were members of the Erlangen Glaucoma registry and received complete ophthalmologic examination and detailed standardized glaucoma examination, including 24-h IOP measurement, perimetrie (G1, three phases) and 15? stereographic slides of the optic disc. Additionally, patients and controls were investigated with VEP with blue on yellow pattern stimulation and confocal scanning laser ophthalmoscope (HRA II, Heidelberg Engineering, Dossenheim, Germany) to detect lipofuscin in the parapapillary atrophic zone. Eyes with retinal diseases or pathologic changes of retinal pigmentepithelium were excluded. Extension of parapapillary area with increased autofluorescence compared (qualitatively) to background autofluorescence around the optic disc and the level of autofluorescence (quantitatively) in the area of qualitatively increased autofluorescence was measured. Patients and controls did not differ significantly concerning age.Results??The area of parapapillary autofluorescence was significant larger in patients with ocular hypertension than in the control group (0.21?0.17?mm2versus 0.07?0.06?mm2;P<0.001, Mann-WhitneyU-test). An increased latency of blue-on-yellow pattern VEP was seen in patients with OHT with increasing parapapillary autofluorescence area (r=0.496;P<0.001, Spearman-Rho test).Conclusion??Parapapillary autofluorescence is increased in patients with ocular hypertension. Patients with large area of parapapillary autofluorescence had also an increased latency in blue on yellow pattern VEP. [ABSTRACT FROM AUTHOR]

PurposeTo assess the degree of corneal diameter shrinkage induced by Q-switched mid-infrared laser corneal trephination for penetrating keratoplasty in an experimental model.MethodsCorneal trephination was performed in 80 enucleated porcine eyes fixed in a holder centered on an automated globe rotation device, by Q-switched (2.94 µm) Er:YAG laser along open masks. Four types of masks were used to protect the underlying corneal tissue: metal masks (donor and recipient) and ceramic masks (donor and recipient). Two spot diameters (0.65 mm and 0.96 mm) were combined with two energy settings (40 mJ/pulse and 50 mJ/pulse) for each of the masks used. Repetition rate was fixed at 5 Hz. Diameters of donor buttons/recipient beds (horizontal and vertical) were measured immediately after the trephination and compared to the given mask size.ResultsMinimum corneal shrinkage was found in the recipient metal mask group (mean ± SD=0.3±0.4%) with 50 mJ pulse energy and 0.65 mm spot diameter (in the horizontal diameter), while the maximum shrinkage (5.3±2.8%) was found in the donor metal mask group with 50 mJ pulse energy and 0.96 mm spot diameter. Corneal shrinkage was less pronounced in recipient beds than in donor buttons ( P<0.01). The differences in shrinkage between the use of ceramic and metal masks were insignificant ( P>0.05). Mean induced corneal diameter discrepancies between the donor button and the recipient bed (with metal and ceramic masks) were 2.5% and 2.5% in vertical diameter and 3.4% and 2.4% in horizontal diameter.ConclusionsThe Q-switched Er:YAG laser experimental corneal trephination for penetrating keratoplasty may induce minor degrees of corneal diameter shrinkage in donor buttons and recipient openings. Oversizing of donor masks by 0.25?0.35 mm (i.e. 3?4% of graft size) may be a valid option to avoid refractive consequences. [ABSTRACT FROM AUTHOR]

BackgroundMacrodiscs with physiologic macrocups and elevated intraocular pressure (IOP) are frequently suspicious for glaucoma. Patients with an elevated c/d ratio and an IOP of more than 21 mmHg are often treated for glaucoma. We investigated a possible relationship between macrodiscs, IOP and central corneal thickness.MethodsUsing the Erlangen glaucoma registry, 341 out of 1,096 consecutive patients with unremarkable and repeated visual field (Octopus 500, G1-3, MD<2.1) were selected. Most of these patients had been sent as glaucoma suspects. The following data were collected: corneal ultrasound pachymetry (Tomey, AL-2000), IOP profile, visual acuity and central corneal power. The morphometric analysis was performed by planimetry using the Littmann formula. Optic nerve heads with a disc area of more than 3.1 mm 2 were defined as macrodiscs.ResultsPatients were subdivided into the following groups: group A: healthy macrodiscs, IOP <22 mmHg (87 patients); group B: healthy macrodiscs, IOP >21 mmHg (66 patients); group C: normal-sized healthy discs, IOP <22 mmHg (93 patients); group D: normal-sized healthy discs, IOP >21 mmHg (95 patients). Mean corneal thickness was: group A: 575±36 µm, group B: 596±37 µm, group C: 557±31 µm and group D: 585±38 µm ( P=0.003). Mean central corneal power measured: group A: 41.7±1.3 D, group B: 42.4±1.3 D, group C: 43.0±1.2 D and group D: 42.8±1.2 D ( P<0.05). The maximum of IOP was: group A: 17.6±3.1, group B: 26.2±4.1, group C: 17.2±3.4 and group D: 29.7±5.5 mmHg. Optic disc size measurements were: group A: 3.91±0.66, group B: 3.65±0.6, group C: 2.56±0.30 mm 2 and group D: 2.39±0.39 µm, respectively.ConclusionsMacrodiscs with elevated IOP have a higher corneal curvature (mean: +0.7 D) and a thicker central cornea (mean: +21 µm) compared to macrodiscs without elevated IOP. An exact evaluation of optic disc morphology in combination with pachymetric and keratometric measurements in relation to IOP could avoid an overtreatment in patients with suspect primary open-angle glaucoma. [ABSTRACT FROM AUTHOR]