bluuebunny/crossref_metadata_2025_split huggingface dataset:
Myopia macular atrophy in the two‐continent population‐based study
Authors: Panda‐Jonas Songhomitra , Mukharram Bikbov , Gyulli M. , Wang Yaxing , Xu Jie , Jonas Rahul , Jonas Jost 
Published: January 2025

Aims/Purpose: To examine myopic macular atrophy (myopic macular degeneration (MMD) stage‐4) and myopic patchy atrophies (MMD‐stage‐3) in three ethnically different cohorts recruited in a population‐based manner.Methods: The population‐based Ural Eye and Medical Study (UEMS) and the Beijing Eye Study (BES) included individuals aged 40+ years, and the Ural Very Old Study (UVOS) examined individuals aged 85+ years.Results: Among 5794 UEMS participants, 19 eyes had MMD‐stage‐4, with 17 (89%) eyes showing a foveal BMD; two eyes could not fully be explored. All 19 eyes showed localized SRPs. Among 21 eyes with MMD‐stage‐3, BMD and SRP prevalence was 9/21 (44%) and 7/21 (33%), respectively. Among 930 UVOS participants, 17 eyes had MMD‐stage‐4, with 16 (94%) eyes showing foveal BMDs and SRPs; one eye could not clearly be assessed. Among 18 eyes with MMD‐stage‐3, BMD and SRP prevalence was 3/18 (17%) and 2/18 (11%), respectively. Among 3468 BES participants, 8 eyes had MMD‐stage‐4, with all eyes showing foveal BMDs and SRPs. Among 14 eyes with MMD‐stage‐3, BMD and SRP prevalence was 10/14 (71%) and 7/21 (33%), respectively. In all three study cohorts, BMD and SRP prevalence was higher (P < 0.001) in MMD‐stage‐4 than MMD‐stage‐3. In MMD‐stage‐3, 12 (55%) of 22 eyes with BMDs showed SRPs, while 12 (86%) of 14 eyes with SRP showed BMDs. SRP prevalence correlated with BMD prevalence (OR:78.3;95%CI:16.1,382).Conclusions: Independent of the ethnic background, all assessable eyes with myopic macular atrophy showed foveal BMDs associated with SRPs, while patchy atrophies could be differentiated into eyes with BMDs and SRPs and eyes without BMDs and without SRPs. The results suggest that SRP as sequel of myopic choroidal neovascularization is a complication of BMDs, and that patchy atrophies may develop due to a primary BMD with secondary RPE defect, or due to an expansion defect in the RPE without concomitant underlying BMD.
Gene editing in ipscs derived from patients affected by inherited retinal dystrophies exhibits rescue of the disease‐associated phenotype
Authors: Siles Laura , Ruiz‐Nogales Sheila , Méndez‐Vendrell Pilar , Pomares Esther 
Published: January 2025

Aims/Purpose: Inherited retinal dystrophies (IRD) are rare disorders triggering blindness and other ocular affectations that currently have no effective treatment. Gene editing allows the precise and permanent correction of pathogenic variants in the DNA of the patient itself. By using two different gene editing technologies, we aimed to repair pluripotent stem cell lines (iPSCs) derived from IRD patients.Methods: We used CRISPR/Cas9 and TALEN methodologies for the correction of mutations in iPSCs from seven patients affected by IRD like Retinitis Pigmentosa or Stargardt disease. Mutant and corrected isogenic cell clones were differentiated into retinal cell models (photoreceptors, RPE cells and retinal organoids) to analyze the potential rescue of the disease‐associated phenotype.Results: We efficiently corrected all the iPS cell lines and without detected off‐target effects. In the majority of cases, at least 50% of the clones exhibit correction without undesired genomic aberrations. The best gene editing results were in the small insertion/deletion or homozygous mutations, reaching the 80% of efficiency. In addition, the design of the sgRNA guides decisively influence gene editing outcomes, and, in case of heterozygous carriers, the specific recognition of the mutant allele only. Also, we achieved successful gene editing by using the patient's own wild‐type allele and not the exogenous ssODN template. Finally, corrected clones differentiated into several retinal cell models exhibit the rescue of the phenotype associated to the disease.Conclusions: Gene editing represents a promising tool for the treatment of IRD. TALENs were less effective than CRISPR/Cas9, but they could be useful in cases where CRISPR not due to genomic concerns. Altogether, these data significantly contribute and encourage the study and application of gene editing as a therapeutic strategy for IRD, and suggest an unexpected application of CRISPR‐mediated correction of pathogenic variants in ssODN‐free assays.
Genetically modified mesenchymal stem cells with cxcr4 and tsg‐6 as future cell therapy for the ocular surface: in vitro study of the cell migration and the regenerative capacity in an inflammation model
Authors: Figueroa‐Haro Jessica Nataly , de la Parte Esther González , Marceñido Beatriz , Vázquez Carmen García , de la Rosa Sara Galindo , Paniagua Marina López , Calonge Margarita , López‐Miguel Alberto , Miguel Teresa Nieto , de la Mata Sampedro Ana 
Published: January 2025

Aims/Purpose: Regeneration of the corneal epithelium, mediated by limbal epithelial stem cells, is essential for the maintenance of corneal transparency. Loss or dysfunction of these cells or their niche, known as limbal stem cell deficiency (LSCD), causes corneal inflammation, ulceration, neovascularization, chronic pain, and conjunctivalization, leading to corneal blindness. Mesenchymal stem cells (MSCs) are a safe and effective treatment for LSCD, however, there is controversy regarding their migration potential. The aim of this work is to enhance the migration and regenerative capacity of adipose tissue‐derived MSCs (AT‐MSCs) by overexpressing the TNF‐stimulated gene 6 (TSG‐6) and the chemokine receptor CXCR4, while maintaining their phenotype.Methods: AT‐MSCs were transduced with TSG‐6 and CXCR4 using lentiviral vectors. The immunophenotype of the transduced AT‐MSCs was analyzed evaluating the expression of CD14, CD19, CD34, CD45, HLA‐DR, CD73, CD90 and CD105. Moreover, the migration capacity and the regenerative potential of the modified AT‐MSCs were analyzed using a Transwell cell migration assay in the presence or absence of the pro‐inflammatory cytokine TNFα, and a wound healing assay in human corneal epithelial cells (HCE).Results: TSG‐6 and CXCR4‐modified AT‐MSCs expressed the MSCs surface markers, maintaining the immunophenotype. Both cells, CXCR4 or TSG‐6‐modified, showed significantly higher migration than control AT‐MSCs in standar conditions. Under inflammatory conditions, a significant increase in the migratory capacity of TSG‐6‐modified AT‐MSCs was observed compared to the control and the CXCR4‐modified ones. The HCE wound healing was faster in the presence of TSG‐6‐modified AT‐MSCs.Conclusions: TSG‐6 and CXCR4‐modified AT‐MSCs maintain the MSCs immunophenotype and improve their migration capacity under standar conditions. Transduction of AT‐MSCs with TSG‐6 enhances both migratory and regenerative capacity of AT‐MSCs under inflammatory conditions.
Surgical detachment of irido‐ and lenticulocorneal adhesions in peters anomaly
Authors: Kotola Elsa‐Leea , Turunen Joni A. , Lindahl Päivi , Krootila Kari , Majander Anna 
Published: January 2025

Aims/Purpose: To report anatomical and visual acuity outcome of surgical detachment of iridocorneal or lenticulocorneal adhesions in pediatric patients with Peters anomaly.Methods: An observational study of pediatric patients with Peters anomaly treated with surgical detachment of iridocorneal or lenticulocorneal adhesions between 2011 and 2024 at the Helsinki University Hospital. Ocular and extraocular diagnoses, age, sex, best corrected visual acuity (BCVA), intraocular pressure, ocular surgeries, surgical complications, family history, genetics and anterior segment optical coherence tomography data were collected from the medical records.Results: Fourteen eyes from nine patients underwent the surgery. The median age at the time of surgery was 14 months (range 5‐56) and the median follow‐up period was 156 months (range 6‐166). Of the treated eyes, 12 had iridocorneal adhesions, while two eyes also had lenticulocorneal adhesions. No surgical complications were reported. Anterior segment anatomy and corneal clarity were stable or improved during the follow‐up. Among the fourteen eyes, three eyes achieved a BCVA better than 0.3 (Snellen decimal), while none had a BCVA worse than 0.05. BCVA was not available for three treated eyes due to the patients' young age. The following additional ocular conditions were recorded: microphthalmia, cataract, glaucoma, retinal degeneration, nystagmus, corectopia and strabismus. Four eyes underwent further ocular surgeries, such as cataract surgery with intraocular lens implantation, anterior vitrectomy, pupilloplasty, and temporal iridectomy. Five patients had systemic findings. Pathogenic variants in the PITX3 and RARB genes were detected in three patients and one patient, respectively.Conclusions: This study demonstrates that surgical detachment of irido‐ and even lenticulocorneal adhesions in Peters anomaly is safe and results stable anatomical and visual acuity outcome over the period of critical visual development.
Extraction and quantification of human tear proteins sampled by schirmer test strips. A validated and standardized method
Authors: Porru Emanuele , Ciavarella Carmen , Comito Rossana , Astolfi Gloria , Violante Francesco , Versura Piera 
Published: January 2025

Aims/Purpose: The identification of biomarkers in tears help in diagnosis and management of ocular diseases, and beyond [1]. We aimed to develop and validate a method for extracting and quantifying proteins from human tears collected using Schirmer test strips, which are typically discarded after clinical use.Methods: Tears from 18 healthy volunteers were collected by aspiration with a micropipette and by paper strips after use for Schirmer test (5 minutes at closed eyes). Aspirated tears were centrifuged and stored at ‐80°C until processing. Tears from strips were eluted using a developed extraction buffer (ABC 100mM, Chaps X‐100 0,25%), and both samples were analyzed using the Agilent 2100 Bioanalyzer. The conversion of the wetted portion (mm) into volume tears (μL) was accurately assessed. The method's accuracy, precision, and sensitivity were estimated, and it was further validated using mass spectrometry.Results: The elution buffer enabled high recovery values for proteins such as albumin, lysozyme, zinc‐alpha2‐glycoprotein (ZAG), lipocalin A, transferrin, and lactoferrin, as compared to the results from the aspirated tears. The method demonstrated high accuracy (bias%<10%), precision (CV%<10%), and sensitivity, with a matrix effect of less than 5% for the studied proteins. The recovery rates were greater than 90% for lysozyme and albumin, and greater than 80% for ZAG, lipocalin, transferrin, and lactoferrin. Validation using mass spectrometry confirmed the reliability of protein identifications and quantifications.Conclusions: The developed protocol is simple, cost‐effective, and achieves significant protein recovery, making it suitable for routine clinical analyses in the clinical setting. The method's validation through mass spectrometry highlights its robustness and applicability for identifying and quantifying tear proteins.References
Rentka, Aniko, et al. "Evaluation of commonly used tear sampling methods and their relevance in subsequent biochemical analysis." Annals of clinical biochemistry 54.5 (2017): 521‐529.
Engineering multicellular organoid for corneal niche modelling and regenerative therapies
Authors: Gulzar Ayesha , Goktan Isilay , Yalcin Esra , Sahin Afsun , Kizilel Seda 
Published: January 2025

Purpose: The corneal niche is composed of various cell types, including limbal stem cells (LSCs), corneal epithelial cells (CECs), and neuronal cells. LSCs are pivotal for corneal epithelial regeneration, CECs maintain the corneal transparency, and neuronal cells contribute to sensory and trophic functions. Despite advances in corneal research, the interplay among these cell types within the corneal niche remains insufficiently understood. This study aims to engineer organoids combining LSCs, CECs, and neuronal cells to create a robust model for studying the corneal niche and to explore their transplantation potential.Methods: LSCs, CECs, and SHSY‐5Y neuronal cells were separately seeded in their growth mediums. After reaching confluency, cells were trypsinized and co‐cultured to form multicellular organoids using the hanging drop technique [1]. Various parameters including cell number, cell‐cell ratio and incubation time were optimized for organoid formation. The organoids were characterized for their cellular composition, structural integrity, and viability using immunohistochemical staining and confocal microscopy. Functional assays, including cell proliferation and differentiation markers, were performed to assess their regenerative potential.Results: To determine the optimal conditions for organoid formation, we experimented with different ratios of LSCs, CECs, and SHSY‐5Y cells, specifically 1:1:1, 2:2:1, 4:2:1, and 8:2:1. The results showed that the engineered organoids formed under all four ratios exhibited a well‐organized structure with high viability for a week. Functional assays including Ki67 and CD44 revealed active proliferation and differentiation, indicating the regenerative potential of the spheroids.Conclusions: This study underscores the potential use of tri‐coculture organoid as a promising therapeutic strategy for limbal stem cells deficiency and a valuable tool for investigating the complex cellular interactions within the corneal environment.References
M. Birgul Akolpoglu, Yasemin Inceoglu, Seda Kizilel, An all‐aqueous approach for physical immobilization of PEG‐lipid microgels on organoid surfaces, Colloids and Surfaces B: Biointerfaces, Volume 186, 2020, 110708, ISSN 0927‐7765, https://doi.org/10.1016/j.colsurfb.2019.110708.
Morphometric histological description of fuchs endothelial dystrophy guttae
Authors: Jonas Rahul , Howaldt Antonia , Matthaei Mario , Schlereth Simona , Bachmann Björn , Cursiefen Claus 
Published: January 2025

Aims: Morphometric histological description of Fuchs corneal endothelial dystrophy guttae as well as anterior banded and posterior layer of Descemet's membraneMethods: After routine DMEK procedures, 47 consecutive explants were taken for histopathological examination as part of clinical routine from 38 patients. 6μm thick sections were cut and stained with PAS and HE. The morphometric analysis was done using a Leica DM‐2500 microscope. Guttae were counted in a line measuring 400μm randomly selected in the middle of the tissue. The largest guttae with the best cross section was used to measure height and width at the base, the middle and the top. The anterior banded layer (ABL) and the posterior (banded and non‐banded as one) layer (PL) were measured below that gutta. The endothelial cells closest to that gutta were measured to get the height and width. Measurements were done using Discus Software (Ingenieursbüro Hilgen). Statistical analysis was done using linear regression on SPSS (Version 29.0 IBM).Results: The mean amount of guttae in the 400μm section was 11.76 ± 4.8. The mean thickness of ABL was 2.92 ± 0.96μm and of PL 8.59 ± 2.7μm. The guttae measured in height 8.68 ± 2.99μm, in width (base) 14.55 ± 4.22μm, in width (middle) 13.13 ± 4.54μm and in width (top) 14.72 ± 4.69μm. The endothelial cells measured in height 3.28 ± 0.98μm and in width 8.02 ± 2.8μm. The width (middle) of the guttae correlated with the width (base) (p < 0.001), width (top) (p < 0.001) and the width of the endothelial cells (p = 0.041). The PL correlated with the height of the guttae (p = 0.028), which also correlated with the height of the endothelial cells (p = 0.044). The width (top) correlated, additionally to the width (middle), with the width of the endothelial cells (p = 0.031). The PL did not correlate with the age of the patients. Most of the correlations kept their statistical significance after Bonferroni correction.Conclusions: The thicker the PL was, the higher were the guttae (r2 = 0,14). The wider the width (base) was, the wider the width(top) (r2 = 0,209) and width (middle) (r2 =0,455) was. Considering the observed variability in corneal guttae shape and dimensions even at the same corneal location at, there might be differences in the pathophysiological processes warranting further analysis.
Quality control in corneal eye banking: impact of preservation time on graft endothelium
Authors: Cabanas João Castro , Cardoso Daniel Ferreira , Martins Pedro Moreira , Soares Ricardo Machado , Ribeiro Ana Clara , Arêde Carlos , Sequeira Joaquim 
Published: January 2025

Aims/Purpose: To evaluate the impact of preservation duration on the endothelial quality of corneal grafts and to identify the most effective parameters for quality control of these grafts.Methods: Retrospective analysis of specular microscopy data from corneal grafts collected by the Cornea and Amniotic Membrane Department of Unidade Local de Saúde Gaia e Espinho between 2021 and 2024. According to department protocol, corneal grafts were stored at 4°C in Optisol‐GS (Bausch and Lomb, Rochester, USA) solution, and specular microscopy was performed at 24 hours (D0), day 5 (D5), and day 10 (D10) post‐harvest. Only grafts with at least two specular microscopy exams (D0‐D5) before transplantation were considered. Data collected included donor age, endothelial cell density (ECD), hexagonality (HEX), and coefficient of variation (VC) at all time points. Data was compared across all time points using repeated‐measures ANOVA adjusted for confounders, with a significance level set at 0.05.Results: Of 93 corneal grafts, 58 met the inclusion criteria, with 12 having specular microscopy analysis at D10. The mean donor age was 61.7 ± 12.9 years. Mean ECD was 2846 ± 434 cells/mm2 at D0, 3025 ± 535 cells/mm2 at D5, and 2880 ± 440 cells/mm2 at D10. Mean HEX was 58.7 ± 11.7% at D0, 51.1 ± 10.5% at D5, and 49.6 ± 12.3% at D10. Mean VC was 34.0 ± 6.7% at D0, 37.9 ± 6.2% at D5, and 38.5 ± 8.7% at D10. Variations in HEX and VC between D0 and D5 were statistically significant (p&lt;0.001), while no significant difference was found between D5 and D10 (p = 0.915 and p = 0.274, respectively). These differences were independent of graft donor age (p>0.05).Conclusions: In this study, preservation time affects the endothelial quality as represented by the decrease of cell HEX and increased VC, especially in the first days of preservation (D0‐D5) while this effect is more attenuated between D5 and D10. Moreover, these two parameters might be more reliable markers of endothelial cell quality in corneal grafts during preservation compared to ECD as this measure is more prone to exam variation as seen in our results.
In‐situ printing of gelma‐based hydrogels for cornea regeneration
Authors: Goodarzi Hamid , Om Boda , Griffith May , Boutopoulos Christos 
Published: January 2025

Aims/Purpose: Corneal perforation is a medical emergency that can lead to blindness. Treatment options encompass cyanoacrylate or fibrin glue, both linked to side effects, including cytotoxicity. Gelatin methacryloyl‐based biomaterials offer an alternative to these adhesives [1, 2]. Achieving optical clarity and smooth surface integrity is crucial for the restoration of vision in treated corneal wounds.Methods: Currently, the delivery of corneal biomaterials utilizes standard syringe systems, which lack the accuracy to reconstruct the cornea's shape. Therefore, the use of a laser bioprinting technology, developed in the Boutopoulos lab [3], for achieving precise in‐situ corneal wound repair was examined. We used a photocrosslinkable ink comprising Gelatin Methacryloyl (GelMa), hydroxyethyl acrylate (HEA), and Lithium phenyl‐2,4,6‐trimethylbenzoylphosphinate (LAP) as a photoinitiator.Results: Printability was optimized by generating nanoliter‐volume individual droplets using 230 μJ laser energy, a flow rate 20 microliter per minute, and a temperature of 37°C. Rheology, optical characterization, and bursting pressure measurements assessed its potential to seal corneal perforations. Results indicated a hydrogel storage modulus of 1.31 ± 0.31 KPa for printed LiQD cornea and a bursting pressure of 38 ± 6 mmHg when used to seal full thickness cornea perforation in cadaveric pig eyes. Optical clarity akin to the native cornea was observed (%light transmission: 93.12 ± 1.02 printed vs 92.61 ± 1.50). The OCT results showed that the LIST technique could potentially fill the corneal wounds and reconstruct the natural curvature of the wounded cornea.Conclusions: In conclusion, a precise printing system for delivering adhesive corneal regenerative biomaterials to wounds was developed and characterized.References1. Sharifi, S., et al., 2021. 6(11): p. 3947‐3961.2. Barroso, I.A., et al., 2022. 9(2).3. Ebrahimi Orimi, H., et al., Sci Rep, 2020. 10(1): p. 9730.
Do estrogens receptor agonists have a neuroprotective effect on mouse rgc after optic nerve axotomy?
Authors: Rodríguez‐Ramírez Kristy T. , Galindo‐Romero Caridad , Vidal‐Sanz Manuel , Agudo‐Barriuso Marta 
Published: January 2025

Aims/Purpose: Estrogens are known to have neuroprotective effects in the central nervous system1,2 (CNS) and only this steroid hormone has the capability of rescue neuronal cell death caused by an injury. Estrogen receptors (ERs) are expressed in both sexes and are distributed throughout the CNS, while the ER‐β is the most predominant in the brain.The aim of this study is:1. To compare the course of death of RGCs in female and male mice after optic nerve axotomy during the fast (up to 9 days) and slow (from 9 day onwards) phases of RGC loss.2. To analyse the expression of ER‐α and ER‐β in intact retinas.3. To study the neuroprotective effect of ER agonism in the two phases of RGC loss.Methods: Optic nerve crush (ONC) was performed on the left eye of pigmented male and female mice, and the course of retinal ganglion cell3 (RGC) loss analysed up to 21 days after the injury. In intact retinas of both sexes the number of RGCs expressing the estrogen receptors alpha (ER‐α) and beta (ER‐β) was studied. To test the neuroprotective potential of both ERs, ER‐β or ER‐α agonists1,2 were injected into the vitreous just after ONC and 9 days later, and the retinas analysed at 21 days.Results: The time course of axotomy‐induced RGC death differs between male and female mice. In males, axotomy‐induced RGC loss is significant after 3 days, whereas in females it is delayed until 5 days. In both sexes, ERs are expressed by more than 70% of RGCs. ER‐β agonism neuroprotects axotomised RGCs at 21 days in both sexes: RGC survival in ER‐β treated animals is 19% higher in males and 17% higher in females than in vehicle treated animals.Conclusions: The loss of RGCs following axotomy is significant at an earlier in males than in females. RGCs from both sexes express both ERs. Intravitreal administration of ER‐β agonist delays axotomy‐induced RGCs loss during the second, slow phase of death.References1. Chakrabarti M., et al. Estrogen receptor agonists for attenuation of neuroinflammation and neurodegeneration. Brain Res Bull. 2014; 109:22‐31.2. Das A., et al. Estrogen receptor agonists and estrogen attenuate TNF‐α‐induced apoptosis in VSC4.1 motoneurons. J Endocrinol. 2011; 208(2):171‐82.3. Galindo‐Romero C., et al. Axotomy‐induced retinal ganglion cell death in adult mice: quantitative and topographic time course analyses. Exp Eye Res. 2011; 92(5):377‐87.