Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control

Journal: Advanced Healthcare Materials

Article Title: Enhanced Bioprinting of 3D Corneal Stroma Patches with Reliability, Assessing Product Consistency and Quality through Optimized Electron Beam Sterilization

doi: 10.1002/adhm.202403118

Figure Lengend Snippet: Fabrication process and experimental timeline of 3D corneal stroma patches. a) Schematic of the bioprinting process using pre‐sterilized and EB‐treated Co‐dECM hydrogels (2.0%) with keratocytes, showing the printing parameters ( v = 5.0 mm s −1 , 25G nozzle, 40 kPa pressure) to create a 1.5 cm × 1.5 cm stroma patch with ≈500 µm thickness, followed by thermal crosslinking at 37 °C for 30 min and its intended application as a corneal stroma substitute. b) Representative images of the 3D bioprinting process at 4 °C showing the sequential printing of three 3D cornea stroma and final single batch production, with ten batches manufactured in total. c) Photograph of a fabricated 3D corneal stroma patch maintained at 4 °C. Scale bar, 1 cm. (scale bar included). d) Timeline schematic showing the manufacturing of 10 batches at 4 °C on day 0, followed by sampling points on day 3 and day 17 ( n = 30 patches per timepoint) with patches maintained at 37 °C.

Article Snippet: The 3D corneal stroma patch was fabricated by encapsulating commercially available human keratocytes (ScienCell, Catalog No. 6520) at passage two or three, at a concentration of 5 × 106 cells mL −1 .

Techniques: Sampling

Journal: Advanced Healthcare Materials

Article Title: Enhanced Bioprinting of 3D Corneal Stroma Patches with Reliability, Assessing Product Consistency and Quality through Optimized Electron Beam Sterilization

doi: 10.1002/adhm.202403118

Figure Lengend Snippet: Cell viability analysis of keratocytes in 3D printed corneal stroma patches. a) Representative 3D confocal microscopy images showing LIVE/DEAD staining of keratocytes in Pre‐sterilized and EB (5 kGy) sterilized patches at days 3 and 17. Live cells are shown in green, and dead cells in red, with both top‐view (3D image) and cross‐sectional views (YZ axis) presented. Scale bars, 500 µm. b) Quantitative analysis of live keratocyte percentages in 3D corneal stroma patches at days 3 and 17, with statistical significance indicated by asterisks ( *p < 0.05, ****p < 0.0001 ). Error bars represent standard deviation. c) Keratocyte viability measurements over 17 days using CCK‐8 assay (OD 450 ), comparing pre‐sterilized and EB (5 kGy) sterilized patches. Error bars indicate standard deviation.

Article Snippet: The 3D corneal stroma patch was fabricated by encapsulating commercially available human keratocytes (ScienCell, Catalog No. 6520) at passage two or three, at a concentration of 5 × 106 cells mL −1 .

Techniques: Confocal Microscopy, Staining, Standard Deviation, CCK-8 Assay

Journal: Advanced Healthcare Materials

Article Title: Enhanced Bioprinting of 3D Corneal Stroma Patches with Reliability, Assessing Product Consistency and Quality through Optimized Electron Beam Sterilization

doi: 10.1002/adhm.202403118

Figure Lengend Snippet: Characterization of 3D‐bioprinted corneal stroma patches before and after electron beam (EB) sterilization. a) Transmittance spectra comparison between pre‐sterilized and EB sterilized samples at different temperatures. The graph shows transmittance (%) as a function of wavelength (nm) from 300–800 nm. Solid lines represent measurements at 37 °C for both pre‐sterilized (black) and EB (5 kGy) sterilized (blue) samples. Dotted lines indicate measurements at 4 °C for pre‐sterilized (black) and EB (5 kGy) sterilized (blue) samples. b) Schematic representation and fluorescence microscopy analysis of 3D corneal stroma patches on day 17. The left panels show illustrations of bioprinted patches with Lumican‐positive keratocytes. Center panels display Lumican (green) and DAPI (blue) staining showing keratocyte distribution and alignment. The right panels present color survey analysis and circular color mapping of the fiber orientation. Scale bars, 100 µm. c) Quantitative analysis of collagen fiber orientation distribution comparing pre‐sterilized (black line) and EB‐sterilized (5 kGy, blue line) patches. d) Scanning electron microscopy (SEM) images revealing the ultrastructural organization of collagen fibers in pre‐sterilized and EB (5 kGy) sterilized patches. Scale bars, 500 nm.

Article Snippet: The 3D corneal stroma patch was fabricated by encapsulating commercially available human keratocytes (ScienCell, Catalog No. 6520) at passage two or three, at a concentration of 5 × 106 cells mL −1 .

Techniques: Comparison, Fluorescence, Microscopy, Staining, Electron Microscopy

Journal: Frontiers in Veterinary Science

Article Title: Antimicrobial activity of cell-free supernatant derived from Ligilactobacillus animalis SWLA-1 in a novel ex vivo canine corneal infection model

doi: 10.3389/fvets.2024.1346313

Figure Lengend Snippet: Confirmation of cell viabilities of human keratocytes using the CCK-8 assay. Results were obtained 72 h post-treatment with CFS derived from L. animalis SWLA-1, as compared to mock-treated cells at the indicated dilutions. Data are presented as mean ± standard deviation for three independent experiments. Data were analyzed by one-way analysis of variance (n.s: non-significant, *** p <0.001).

Article Snippet: Human keratocytes (ScienCell, Catalog #6520) were obtained and cultured in a fibroblast medium (ScienCell, Catalog #2301).

Techniques: CCK-8 Assay, Derivative Assay, Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: Targeting HMGB1-NFκb Axis and miR-21 by Glycyrrhizin: Role in Amelioration of Corneal Injury in a Mouse Model of Alkali Burn

doi: 10.3389/fphar.2022.841267

Figure Lengend Snippet: GLY reduces miR-21 expression in keratocytes, and blockade of miR-21 preventes keratocytes differentiation to myofibroblasts. (A) qRT-PCR shows significantly increased miR-21 level in HK stimulated by TNFα or TGF-β1. (B) mRNA expression levels of miR-21 and HMGB1 in HK treated by TNFα with or without GLY. (C) The expression level of miR-21 in HK after miR-21 agomir transfection. (D–E) Western blot and qRT-PCR were used to examine α-SMA expression after miR-21 transfection. HK were transfected with miR-21 agomir, antagomir, and their negative controls (NC) before treated with TGF-β1. Western blotting shows significantly increased of α-SMA in HK overexpressed miR-21 and the quantified data shown in (E) mRNA expression levels of α-SMA in HK (F) Data were analyzed using one-way ANOVA followed by Bonferroni’s multiple comparison test ( n = 3/group). The results are expressed as the means ± SD of at least three independent experiments, and qRT-PCR was conducted in triplicate. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human keratocytes (HK, catalog #6502) and culture medium (Fibroblast Medium, FM, Catalog #2301) were purchased from sciencell (sciencell, Carlsbad, CA, United States).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Comparison