bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cryopreservation of corneal endothelial cells in vitro, ex vivo, and on a tissue engineered endothelial graft
Authors: Sagnial Tomy , Coban Melis , Aouimeur Inès , Coulomb Louise , Philippe Gain , He Zhiguo 
Published: January 2025

Aims/Purpose: In response to the global shortage of corneas, alternatives such as cell injection therapy and tissue‐engineered endothelial keratoplasty (TEEK) have emerged, both relying on the mass production of primary cultured corneal endothelial cells (CECs). Cryopreservation of corneas (the primary cell culture source), cultured CECs (in vitro), and TEEK grafts (final products) is crucial to facilitate their industrialization and clinical application. While corneal cryopreservation has long been a challenge, advancements in cryoprotectants warrant a renewed investigation. This study aims to assess various cryopreservation methods for CECs n the three different states.Methods: Ten cryopreservation media and varying cooling rates (‐2°C/min, ‐1°C/min, or ‐0.5°C/min) were first tested on cultured CECs. After thawing, cell survival rates were assessed using Trypan blue staining and an automated cell counter. Subsequently, the best conditions identified for cultured CECs were applied to native CECs adhered to their Descemet's membrane and CECs on TEEKs. Post‐thawing viability was assessed using triple labeling with Hoechst, Ethidium, and Calcein‐AM (Pipparelli et al. IOVS 2011).Results: A gradual cooling rate of ‐1°C per minute and the cryopreservation medium CryoStor CS10 provided the best conditions for in vitro CECs, ensuring an average viability of 91 ± 1% post‐cryopreservation. However, applying the same conditions to native CECs on corneas or CECs on TEEK grafts resulted in a viability of less than 80%. Upon thawing, CECs tended to detach easily from the DM or bio‐engineered grafts, leading to significant areas without cells.Conclusions: Cryopreservation is effective for in vitro cultured CECs but remains very challenging for CECs attached to the DM or TEEKs. The next step will be to address cell attachment issues during cryopreservation.References
Aurélien Pipparelli, Gilles Thuret, David Toubeau, Zhiguo He, Simone Piselli, Sabine Lefèvre, Philippe Gain, Marc Muraine; Pan‐Corneal Endothelial Viability Assessment: Application to Endothelial Grafts Predissected by Eye Banks. Invest. Ophthalmol. Vis. Sci. 2011;52(8):6018‐6025. https://doi.org/10.1167/iovs.10-6641.
Adipose‐derived mesenchymal stem cells (admscs) and retinal pigment epithelial cells (RPE‐1) interactions in stress environments via tunneling nanotubes
Authors: Gozel Merve , Senkoylu Karya , Kesim Cem , Hasanreisoğlu Murat 
Published: January 2025

Aims/Purpose: The study aims to elucidate the potential role of TNT‐mediated intercellular communication between the AdMSCs and RPE‐1 cells in reducing stress‐induced damage.Methods: Oxidative stress and serum starvation were used as stress to create TNTs between the AdMSC and RPE‐1 cells. Immunofluorescence and scanning electron microscopy were used to show that TNT formation. Following stress conditions, the cells' viability was examined by CTG assay, and the levels of reactive oxygen species (ROS) were measured by H2DCFH‐DA probes. Microscopy showed that JC‐1 labelled mitochondrial exchange occurs between cells via TNTs. To prevent TNTs, a transmembrane culture system was used and evaluated how to effect on cell viability and changes of ROS levels.Results: We examined how TNTs support mitochondrial transport and intercellular communication between AdMSCs and RPE‐1 during stress. We discovered that TNT‐mediated mitochondrial translocation from AdMSCs to RPE‐1 contributes to both increased cell survival and a decrease in ROS levels. We showed that AdMSCs and RPE‐1 had a direct connection that was essential for stress recovery. After stress‐induced damage, co‐culture improved vitality and maintained retinal function. Because mechanical restriction of TNT synthesis reduced cell viability and raised ROS levels, it is clear that TNTs are essential for cellular defence.Conclusions: Overall, our study provides valuable insights into the cellular mechanisms underlying stress response and underscores the therapeutic promise of AdMSC derived TNTs in preserving retinal health.
Integrating single‐cell and spatial transcriptomics delineates spatial gene expression patterns and molecular signatures of various tissues in mouse eyes
Authors: Ouyang Shuai 
Published: January 2025

Aims/Purpose: The eye is a highly organized structure composed of well‐defined anatomical area with coordinated but distinct‐functional roles. This study aimed to elucidate spatial gene expression patterns and molecular signatures in different ocular tissues.Methods: We performed spatial transcriptomics sequencing on normal mouse eye samples and integrated single‐cell transcriptomics and spatial transcriptomics for comprehensive bioinformatics analysis of eye. Findings regarding the discovery of several ligand‐receptor pairs and gene expression patterns were validated by in situ sequencing and immunofluorescence experiments.Results: Firstly, we revealed the spatial gene expression patterns of ocular tissues and identified core spatial regulons of various tissues within the eye. Based on spatial transcriptomics analysis, we uncovered that the central retina prominently mediates synaptic signaling and ion channel transmission functions while the peripheral retina has significant visual reception and oxidative phosphorylation functions. In addition, this undertaking characterized critical neurotransmitters and spatial ligand‐receptor pairs action modes of distinct cell types within the retina, which might contribute to the maintenance of retinal electrical signal transmission. Furthermore, we simulated the spatial differentiation trajectories of lens and corneal cells, and delineated the spatial location dependent changes in gene expression profiles. Subsequently, we identified several novel spatial region‐specific regulators in lens and cell type‐specific regulators in cornea. Finally, we unveiled an ocular spatial organizational communication signaling system that dependent on candidate long‐distance inter‐tissue signaling molecule communication.Conclusions: This undertaking is likely to aid forthcoming studies on the mechanisms of eye development, differentiation, regeneration and disease occurrence. Our dataset also serves as a scientific resource for atlas, but will also be useful in screening for molecular targets of eye lesions occurring in specific spatial locations, which will greatly improve our understanding of the mechanisms underlying the maintenance of ocular homeostasis in health and disease.
A low dose bimatoprost 0.01% & timolol 0.1% eye gel compared to bimatoprost 0.03% & timolol 0.5% eye drops: A phase III randomized study
Authors: Stalmans Ingeborg , Cordeiro M. Francesca 
Published: January 2025

Aims/Purpose: To compare the efficacy and safety of preservative‐free (PF) fixed combination, bimatoprost 0.01%/timolol 0.1% eye gel (BTFC eye gel) with bimatoprost 0.03%/timolol 0.5% eye drops (BTFC eye drops) in patients with open‐angle glaucoma (OAG) or ocular hypertension (OHT).Methods: Phase III, international, multicentre, randomised, 2 parallel groups, investigator‐masked, 3‐month treatment duration study. Patients with OHT or OAG, insufficiently controlled with a first‐line monotherapy, and requiring a dual therapy, were randomized to receive either PF BTFC eye gel (bimatoprost 0.01%/timolol 0.1%) or PF BTFC eye drops (bimatoprost 0.03%/timolol 0.5%), once daily for 12 weeks. The primary efficacy endpoint was the change from baseline (Day 1) to Week 12 in IOP at 08:00, in the study eye. Noninferiority was based on a predetermined criteria (upper 95% confidence interval margin of 1.5 mmHg at Week 12 at 08:00). Further efficacy and safety endpoints were assessed as secondary outcomes.Results: The mean change from baseline to Week 12 in IOP at 08:00 was −10.96 ± 3.43 mmHg for BTFC eye gel and −11.14 ± 3.56 mmHg for BTFC eye drops. Noninferiority of T4030 was demonstrated, based on the predetermined criteria. IOP‐lowering efficacy was also similar between treatment groups at Week 6 (08:00; 10:00; 16:00) and Week 12 (10:00; 16:00). The safety profile was similar between T4030 and Ganfort; however, more patients withdrew from the study due to a treatment‐related adverse event (AE) with BTFC eye drops (8 patients [21 AEs]), compared to BTFC eye gel (no patients). Global tolerance was considered as satisfactory/very satisfactory by the investigator for more patients in the T4030 (96.4%) versus Ganfort (91.6%) at Week 6, with a statistically significant between group difference (p = 0.020).Conclusions: A low dose PF bimatoprost 0.01%/timolol 0.1% eye gel can be considered as a safe and effective tool in the therapeutic management of glaucoma and OHT.
Differential profile of mirnas in aqueous humour: A new potential epigenetic mechanism for pseudoexfoliation glaucoma
Authors: Villanueva Carolina Garcia , Andrés‐Blasco Irene , Martucci A. , Lopez J. F. Ramos , Moreno Vicente Zanon , Montero M. Gonzalez , Nucci C. , Durán Maria Dolores Pinazo , Feijoo Julian Garcia 
Published: January 2025

Aims/Purpose: Pseudoexfoliation glaucoma (PEG), is the most prevalent of the secondary glaucomas, characterized by the accumulation of fibrillar extracellular material in the anterior eye chamber. The miRNAs are regulators of gene expression, as epigenetic effectors. Our aim is to identify candidate miRNAs that may be involved in PEG physiopathology.Methods: A collaborative, longitudinal, prospective, case‐control study was set up in 120 participants, of both sexes, that were classified as: PEG patients (n = 60) and healthy individuals (CG n = 60). Sociodemographic and opththalmologic data were recorded. Aqueous humor was collected at the onset of glaucoma surgery, labelled and stored at. ‐80° until processing. Total ARN was obtained, and expression and quantitation of the miRNAs ‐122‐5p, ‐146a‐5p and ‐320a‐3p, were assayed by real time quantitative polimerase chain reaction (RT qPCR) in both study groups. Databases DIANA TOOLS, miRDB, TargetScanHuman 7.2 were used to connect the target genes with its biological functions. Data were processed by IBM SPSS 28.0 statistical program.Results: Mean age was 73 ± 11 years. Gender distribution was 32 % men/68 % women. Mean intraocular pressure was 17 mmHg in the right eye (RE) and 19 mmHg in the left eye (LE). Mean retinal nerve fiber layer thickness was 73 μm in RE and 71 μm in LE. MiRNAs ‐122‐5p (p = 0.02),146a‐5p (p = 0.02) and ‐320a‐3p (p = 0.04) showed a significantly higher expression in the aqueous humor of PEG versus CG.Conclusions: miRNAs ‐122‐5p, ‐146a‐5p and ‐320a‐3p are involved in PEG pathogenesis, epigenetically affecting the expression of target genes and protein translation, by means of their biological functions, in relation to extracellular matrix, metalloproteinases, glutamate, and the signaling pathways MAPK and RhoA/ROCK. We may suggest to investigate new biological therapies on the basis of the above miRNAs in PEG patients.
Identification of diagnostic and prognostic biomarkers in dry eye disease in primary open‐angle glaucoma: A cross‐sectional case‐control study
Authors: Sáez Sara Mora , Andrés‐Blasco Irene , Cerdà‐Ibáñez Marta , Peris‐Martínez Cristina , Durán Maria Dolores Pinazo 
Published: January 2025

Aims/Purpose: To integrate clinical and molecular data that allow us to identify diagnostic and prognostic biomarkers of dry eye disease (DED) in primary open‐angle glaucoma (POAG).Methods: A cross‐sectional case‐control study consisting of 182 participants of both sexes, was done. Volunteers were classified into 4 groups: healthy controls (CG; n = 50), DED group (n = 42), POAG group (PG; n = 44), and DED+POAG (n = 46). All participants underwent a complete and systematic medical history, filled out the questionnaires, and were subjected to ophthalmologic examination. Reflex tear samples were collected and subsequently processed using enzyme‐linked immunosorbent assay (ELISA) to quantify the levels of interleukins (IL) ‐1β and ‐18. Statistical analysis was performed with the SPSS 28.0 program.Results: Mean age of the study participants was 60+15 years, with a total of 56 males (31%) and 126 females (69%). The use of preservative‐containing eye drops was observed in 13% of the POAG patients, and 21% of the DED+POAG group. Clinical data [Schirmer test, tear break‐up time (TBUT), tear meniscus height (TMH) measured by optical coherence tomography (OCT)], and the tear expression of IL‐1β and IL‐18 were significantly different between groups. According to the ocular surface disease index (OSDI) questionnaire, dry eyes were more severe (OSDI=42) and the IL‐1β and IL‐18 tear levels were significantly higher in 37% of the DED+POAG patients.Conclusions: We suggest that assessment of TMH, OSDI, and the tear expression of IL‐1β and IL‐18 can be potential biomarkers for identifying patients at risk of DED in the POAG subgroup undergoing treatment with antihypertensive eye drops.
Neurovascular coupling response: A follow‐up study
Authors: Jordão João , Rodrigues Sérgio , Domingues Joana , Serranho Pedro , Guimarães Pedro , Bernardes Rui 
Published: January 2025

Aims/Purpose: This work aims to assess the consistency of the neurovascular coupling (NVC) response of a healthy subject group at two visits 5 months apart.Methods: Fundus images (4x4 degrees) of the macular region (3 baseline acquisitions and 3 acquisitions each preceded by flicker stimulation) of 13 healthy adults (one eye each, age (m+/‐sd): 33.08+/‐11.4 yrs.) were acquired with the rtx1 AO camera (ImagineEyes, Orsay, France). The rtx1 was configured for a 20 second, 15Hz, and 4 μW.cm^(‐2) flicker stimulation over a 29x20 degrees area of the imaged eye fundus. Acquisitions for each subject were repeated 159+/‐17 (m+/‐sd) days later.The lumen of retinal arterioles within the macular region was determined using custom‐developed software for the automatic segmentation of lumen‐wall interfaces. An experienced technician visually assessed all segmentations to reject subpar results. Lumen diameters were calculated from the distance between the lumen‐wall interfaces as measured perpendicular to the vessel axis along a vessel segment. NVC responses were calculated per eye as the relative difference (%) of the lumen diameter after stimulation to the respective baseline.Results: A total of 22 out of 156 images (14%) were rejected and excluded from further analysis. NVC responses ranged from 0.21% to 8.28% (3.36+/‐2.75 % (m+/‐sd)) for the first visit and from ‐0.24% to 9.13% (3.03+/‐2.54 %) for the follow‐up. Response differences between time points ranged from ‐6.15% (8.28% to 2.13%) to 4.21% (0.98% to 5.19%), with an average (sd) of 0.34(3.03) % for the group. A paired sample Wilcoxon test between time points is not statistically significant (p = 0.650), supporting that the responses are consistent between time points.Conclusions: The obtained results show a consistent NVC response for a 5‐month interval.
Orthokeratology and myopia control: A comprehensive meta‐analysis of axial growth in children and adolescents
Authors: Queirós António , Mota‐Silva Inês , Pereira‐da‐Mota Ana Filipa 
Published: January 2025

Aims/Purpose: The aim of this study was to evaluate the longitudinal effect on axial eye length in myopic children and adolescents treated with orthokeratology (OK) lenses compared to spectacles (GL), through a systematic review and meta‐analysisMethods: We conducted a systematic search of PubMed and Google Scholar databases for randomized controlled trials and cohort studies investigated the effects of longitudinal axial length difference, using the keyword “Axial Length” AND “orthokeratology” AND “spectacles”. From the initial 91 studies, we excluded 13 review studies, 10 meta‐analysis studies, 7 systematic reviews and 37 studies that did not meet the inclusion criteria. Of the 24 selected studies, detailed information was available at 24, 18, 12, 6, 3 and 1 month, with 10, 8, 19, 17, 2 and 3 studies respectively, allowing for the comparison between 2172 eyes treated with orthokeratology and 2095 eyes with glasses.Results: The analysis of the 24 studies (which included 59 comparisons from 1 month to 24 months) showed no statistically significant differences in initial axial length [AL(GL) = 24.39 ± 0.34 mm ; AL(OK) = 24.44 ± 0.38 mm] and initial refractive error [M(GL) = ‐2.59 ± 0.70 D ; M(OK) = ‐2.53 ± 0.44 D] between the two treatment groups (p > 0.565, independent samples t‐test) and for the evolution time intervals (p > 0.591, ANOVA). This analysis encompassed 4267 eyes with a mean age of 10.23 ± 1.21 years. Forest plot provided in an overall effect size of myopic defocus AL=‐0.16 com IC 95% ‐0.19 a ‐0.14, Z=12.71 p < 0.001. A random effects model was used considering the heterogeneity observed (I2=97%; p < 0.001, df=58). There was a greater myopia control effect at 24 months (AL=‐0.26, 95% IC ‐0.29 to ‐0.22, Z=13.71 p < 0.001; I2=0%; p = 0.95) than to treatments of less than 3 months (AL=‐0.04, 95% IC ‐0.06 to ‐0.01, Z=2.60 p = <0.009; I2=86%; p < 0.001).Conclusions: While the treatment at 24 months shows an homogeneous behaviour among studies, shorter follow‐up times renders a larger heterogeneity and for that reason, longer follow‐up periods should be preferred to estimate a more solid effect size. Orthokeratology treatment has been shown to be effective in myopia control progression for short or longer longitudinal treatments in children and young people with spectacles comparison.
Retinal image quality with myopia control lenses, in the synthetic accommodative wavefront model
Authors: García Maria Mechó , Fernandes Paulo , Faria‐Ribeiro Miguel Ángel 
Published: January 2025

Aims/Purpose: Current myopia control contact lenses (CLs) are based on multifocality designs that affect decrease retinal image quality (RIQ). This study aimed to evaluate the changes in RIQ and accommodative response (AR) with two myopia control CLs.Methods: 100 synthetic eyes were generated using anc accommodative wavefront model to simulate the changes in RIQ with accommodation. The synthetic eyes were “fitted”, using direct wavefront summation, with two myopia control CLs: the MiSight with +2.00D treatment zones and the Mylo with an EDOF design. Peak image quality was calculated for the naked eye and with CLs, in the relaxed state and a target vergence of 40 cm, with an entrance pupil diameter of 4.28 mm. Additionally, the AR that maximizes RIQ was calculated.Results: The mean RIQ [ranged from 0 to 1] for the relaxed eye was 0.42 ± 0.10 without a CLs. It halved to 0.26 ± 0.05 with the Misight and reduced by threefold to 0.15 ± 0.04 with the Mylo. For 40cm the RIQ was 0.34 ± 0.09 without a CL, 0.24 ± 0.07 with the Misight and 0.19 ± 0.03 with the Mylo.For target vergences with demands close the effective add power of the CL, the accommodative RIQ through‐focus produced a second peak prior to the maximum peak, correspondent to the focusing power of the treatment zones1, yielding a RIQ of 0.08 ± 0.03 and 0.12 ± 0.04 for the Misight and Mylo, respectively. The AR without CL was 2.33 ± 0.20 D, 1.75 ± 0.22 D and 1.75 ± 0.22 D with the Misight and the Mylo, respectively.Conclusions: Both myopia control CLs reduced the RIQ in the relaxed and accommodated states. For 40 cm, both CLs induce a secondary peak in the RIQ through‐focus. With Mylo, the secondary peak appeared less than 0.1D from the maximum peak, possibly due to its EDOF design. The AR was reduced with both CLs 0.58 D (lag) compared to the naked eye condition. These results indicate that, both lenses impose significant limitations RIQ and AR.References
Faria‐Ribeiro M, Amorim‐de‐Sousa A, González‐Méijome JM. Predicted accommodative response from image quality in young eyes fitted with different dual‐focus designs. Ophthalmic Physiol Opt. 2018;38(3):309‐16.
Silver bismuth sulfide quantum dot‐based bioelectronics for light‐mediated neuronal stimulation
Authors: Kaleli Humeyra Nur , Balamur Rıdvan , Kaya Tarık Safa , Önal Asım , Pehlivan Çiğdem , Sahin Afsun , Nizamoğlu Sedat , Hasanreisoğlu Murat 
Published: January 2025

Aims/Purpose: Nanomaterial based bioelectronics have been effectively developed as optoelectronic bio‐interfaces for neural stimulation and can be fabricated from diverse substances to adapt to cellular environments. In this study, we employed silver bismuth sulfide (AgBiS2) quantum dot (QD)‐based neural interfaces stimulated by near‐infrared light.Methods: Primary neuron isolation and culture on AgBiS2 QD based photovoltaic device and indium tin oxide (ITO) control device was performed. Cell viability was assessed by CTG, MTT, Live/Dead assay and LDH leakage assays. Cellular stress with light stimulation was assessed by measuring intracellular reactive oxygen species. Cell specific biomarkers, NeuN, beta‐III Tubulin and F‐actin, were examined by immunofluorescence staining to demonstrate short‐ and long‐term morphological changes and neural network improvements. Number of neuron count and neurite length measurement were analyzed to compare groups. Under light stimulation (λ = 780 nm), neural dynamics and electrophysiological activity were examined through intracellular calcium flow and patch clamp.Results: Throughout the 14‐day culture period, neurons remained healthy and viable, preserving their characteristics and forming extensive networks with neurite outgrowth on both the device and ITO. Light stimulation was found to have no adverse effects on viability or intracellular stress levels. The device demonstrated successful photostimulation of neurons with calcium release and generating action potential under 780 nm light illumination which is validating its potential to be used as optoelectronic bio‐interfaces for neural applications.Conclusions: Our study holds high potential for the development of QD‐based retinal prosthesis, enabling near infrared light‐controlled activation in vision related diseases.Funding Information: This study was funded by the European Union (ERC, MESHOPTO, 101045289).References
Balamur R, Oh JT, Karatum O, Wang Y, Onal A, Kaleli HN, Pehlivan C, Şahin A, Hasanreisoglu M, Konstantatos G, Nizamoglu S. Capacitive and Efficient Near‐Infrared Stimulation of Neurons via an Ultrathin AgBiS2 Nanocrystal Layer. ACS Applied Materials & Interfaces. 2024 May 29
Evaluation of the conjunctival bacterial flora in the New Zealand white rabbit
Authors: Cejudo Raquel Maroto , Encalada Denisse Michelle Espinosa , Rodriguez Oscar Esparcia , Gómez‐Juarez Sango Mónica , Pretel Fernando Andrés , Valenciano Carlos Cava 
Published: January 2025

Aims/Purpose: We aimed to characterize the normal bacterial flora of conjunctiva and eye lids of the white New Zealand rabbit.Methods: Twelve rabbits (Twenty‐four eyes) were employed. Before sampling, an ophthalmological examination of the ocular surface was performed to exclude the presence of any lesions.Microbiological sampling by sterile swabs was performed on the superior and inferior conjunctival fornix. Samples were placed in eSwab 1 mL transport medium (eSwab® – Copan's Liquid Amies Elution Swab) and submitted for culture. They were cultured on blood agar, chocolate agar and sabouraud agar. Species identification was carried out by mass spectrophotometry (MALDI‐TOF, BioMèrieux®) and/or conventional techniques. They were seeded in triplicate and the quantification of the germs was expressed in terms of CFU/mL.Results: Seven different species were observed. The most frequent was Moraxella cuniculi (705 ± 1129 CFU/mL), followed by Corynebacterium spp (71.43 ± 267.3 CFU/mL) and coagulase negative Staphylococcus (5 ± 10.92 CFU/mL). Other less frequently species observed were Staphylococcus aureus, Streptococcus viridans, Tsukamurella spp and Bacillus spp.Statistical analysis showed a significant statistical difference between M. cuniculli and the other species observed. (Krussal Wallis p < 0.001).Conclusions: M. cuniculi was the species with the highest CFU/mL in the conjunctival flora of the rabbit.