primary human corneal epithelial cells hcecs  (ATCC)

Journal: EMBO Molecular Medicine

Article Title: Corneal biomechanical cues mediated by PAI-2: the origin of PM2.5-induced corneal disease

doi: 10.1038/s44321-025-00341-0

Figure Lengend Snippet: ( A ) Immunofluorescence staining for phosphorylated non-muscle myosin IIa of HCECs exposed for different times to PM2.5 (scale bar, 75 μm) ( n = 3 biological replicates). ( B ) Cellular tensile stress of HCECs exposed to PM2.5 (with or without blebb) for 3 h was measured by traction force microscopy ( n = 40 for control, n = 32 for PM, n = 25 for PM+blebb). ( C ) Cellular stiffness of HCECs exposed to PM2.5 for 3 h measured by atomic force microscopy ( n = 8). ( D ) Cellular viability of HECEs in the presence of RI or blebb ( n = 3 biological replicates). ( E ) Impression cytology was used to evaluate rat corneal epithelial cell circularity in two groups (scale bar, 25 μm). n = 5 (PBS), 9 (PM). ( F ) PM2.5 exposure leads to the activated biomechanical response at the subclinical stage in humans, rats, and HCECs. This curve provides a conceptual illustration of the correlation between biomechanical response and PM2.5-induced corneal disease progression; it does not reflect precise numerical changes. Data in ( A – E ) are graphed as mean ± standard deviation with individual values shown as dots or circles. Statistical analysis was conducted using the unpaired t test in ( A – E ). The P values are labeled in the figure. .

Article Snippet: Human corneal epithelial cell lines , ATCC , PCS-700-010.

Techniques: Immunofluorescence, Staining, Microscopy, Control, Biomarker Discovery, Standard Deviation, Labeling

Journal: EMBO Molecular Medicine

Article Title: Corneal biomechanical cues mediated by PAI-2: the origin of PM2.5-induced corneal disease

doi: 10.1038/s44321-025-00341-0

Figure Lengend Snippet: ( A ) PAI-2 staining in rat cornea after short-term (2-day) PM2.5 exposure (scale bar, 25 μm). The while arrow refers to the rat corneal superficial epithelial cells. n = 4. ( B ) The mRNA level of PAI-2 in rat corneas after short-term (2-day) PM2.5 exposure. n = 4. ( C ) The protein level of PAI-2 in HCECs after short-term (3-h) PM2.5 exposure ( n = 3 biological replicates). ( D ) The viability of NC and KO after 24-h PM2.5 treatment ( n = 3 biological replicates). ( E ) The heatmap showed the different expression profiles in three groups. ( F ) The Venn diagram demonstrated the common DEGs between the two sets of comparisons. ( G ) GO analysis revealed biological process, cellular component, and molecular function enriched in DEGs. ( H ) KEGG pathway analysis of DEGs. ( I ) PAI-2 is associated with several cellular mechanics-related processes, which may regulate PM2.5-induced cellular mechanical response. Data in ( A – D ) are graphed as mean ± standard deviation with individual values shown as circles. Statistical analysis was conducted using the unpaired t test in ( A – D ). The P values are labeled in the figure. .

Article Snippet: Human corneal epithelial cell lines , ATCC , PCS-700-010.

Techniques: Staining, Expressing, Standard Deviation, Labeling

Journal: EMBO Molecular Medicine

Article Title: Corneal biomechanical cues mediated by PAI-2: the origin of PM2.5-induced corneal disease

doi: 10.1038/s44321-025-00341-0

Figure Lengend Snippet: ( A ) The workflow of extracellular PAI-2 detection in rat tears, HCEC conditioned medium, and human tears. For the details of the panel study in humans, participants completed 24 h of continuous personal air monitoring followed by a questionnaire survey, tear sample collection, and ophthalmic examination. Tear samples were then sent to a laboratory to detect PAI-2 content. n = 28. ( B ) The PAI-2 concentration in rat tears upon PM2.5 exposure for different times was detected via ELISA. n = 5 (PBS group), 6 (PM2.5 group). ( C ) The PAI-2 level in HCEC conditioned medium upon 24-h PM2.5 exposure was detected via ELISA ( n = 7 biological replicates). ( D – I ) Spearman analysis was performed to separately define the correlation between PM2.5 concentration and ocular surface disease index/eyesight/tear film break-up time/tear secretion/corneal epithelial defect/PAI-2 content in tears. n = 28. Individual values are shown as scattered dots. ( J ) PIPs of PM2.5 constituents on the PAI-2 level calculated by the BKMR model. ( K ) Univariate exposure−response curves for PM2.5 constituents with PIPs above 0.9, while all other PM2.5 elemental constituents were at their median concentrations (shown with credible interval). Data in ( B , C ) are graphed as mean ± standard deviation with individual values shown as circles. Statistical analysis was conducted using the unpaired t test in ( B , C ). The P values are labeled in the figure. .

Article Snippet: Human corneal epithelial cell lines , ATCC , PCS-700-010.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Labeling

Journal: EMBO Molecular Medicine

Article Title: Corneal biomechanical cues mediated by PAI-2: the origin of PM2.5-induced corneal disease

doi: 10.1038/s44321-025-00341-0

Figure Lengend Snippet: ( A ) The corneal epithelium defect was examined by slit lamp examination using fluorescein sodium staining under cobalt-blue light. The red arrows pointed to the corneal epithelium defect. The fluorescein sodium staining grades were evaluated. n = 6. ( B ) H&E staining showed the morphological changes in the rat cornea, lens, and retina with different treatments (scale bar, 50 μm). The corneal epithelial thickness was measured based on H&E staining. n = 4 (PBS), 4 (PM), 4 (PM + LNP), 3 (PM + LNP-siNC), 4 (PM + LNP-siPAI-2). ( C ) Immunofluorescence staining of rat cornea for PAI-2 and LC3B (scale bar, 50 μm). n = 3. The fluorescence intensities of PAI-2 and LC3B were calculated based on immunofluorescence staining. ( D ) Immunohistochemistry showed the expression level of IL-1a of rat corneas in different groups (scale bar, 50 μm). n = 4 (PBS), 3 (PM), 3 (PM + LNP), 3 (PM + LNP-siNC), 4 (PM + LNP-siPAI-2). The average optical density of IL-1a in rat corneal epithelium was measured based on immunohistochemistry. ( E ) The rat model workflow and therapeutic mechanism diagram of the LNP-siPAI-2 system. LNP-siPAI-2 delivers siPAI-2 to rat corneal epithelial cells, thereby targeting and reducing PAI-2 expression levels and inhibiting autophagy and IL-1a to attenuate corneal epithelial cell damage. At the same time, the reduction in corneal damage reduces the inflammatory response of the intraocular lens and retina, thus realizing the protection of the whole eye by the LNP-siPAI-2 system. Data in ( A – D ) are graphed as mean ± standard deviation with individual values shown as circles, squares, or triangles. Statistical analysis was conducted using the unpaired t test in ( A – D ). The P values are labeled in the figure. .

Article Snippet: Human corneal epithelial cell lines , ATCC , PCS-700-010.

Techniques: Staining, Immunofluorescence, Fluorescence, Immunohistochemistry, Expressing, Standard Deviation, Labeling

Journal: EMBO Molecular Medicine

Article Title: Corneal biomechanical cues mediated by PAI-2: the origin of PM2.5-induced corneal disease

doi: 10.1038/s44321-025-00341-0

Figure Lengend Snippet: ( A ) DLin-MC3-DMA LNPs can deliver the load (luciferase mRNA) into the corneal epithelium of rats (scale bar, 75 μm). n = 4. ( B ) Detection of PAI-2 knockdown efficiency by LNP (siPAI-2) ( n = 3 biological replicates). ( C ) Immunohistochemistry showed the expression level of IL-1a in rat lens and retinas of different groups (scale bar, 50 μm). n = 4 (PBS), 3 (PM), 3 (PM + LNP), 3 (PM + LNP-siNC), 4 (PM + LNP-siPAI-2). The red boxed refer to the rat lens epithelial cells. The average optical density of IL-1a in rat lens and retinas of different groups. ( D ) The retinal thickness based on H&E staining in Fig. . n = 4 (PBS), 3 (PM), 4 (PM + LNP), 3 (PM + LNP-siNC), 4 (PM + LNP-siPAI-2). Data in ( A – D ) are graphed as mean ± standard deviation with individual values shown as circles, squares, or triangles. Statistical analysis was conducted using the unpaired t test in ( A – D ). The P values are labeled in the figure.

Article Snippet: Human corneal epithelial cell lines , ATCC , PCS-700-010.

Techniques: Luciferase, Knockdown, Immunohistochemistry, Expressing, Staining, Standard Deviation, Labeling

Journal: Journal of Pharmaceutical Analysis

Article Title: Ursolic acid ameliorates ocular surface dysfunction in dry eye via targeting EGFR/RAS/RAF/MAP2K1/MAPK1 pathway

doi: 10.1016/j.jpha.2025.101294

Figure Lengend Snippet: Ursolic acid (UA) effectively restores human corneal epithelial cells (HCEs) homeostasis under hypertonic stress (HS). (A) Two-dimensional (2D) and 3D chemical structures of UA. (B) Schematic representation of the in vitro study design. (C) Relative viability of HCEs treated with UA for 24 or 48 h. (D) Half maximal inhibitory concentration (IC 50 ) value of UA on HCEs after 48 h. (E) Relative cell viability of HCEs treated with varying UA concentrations under hyperosmotic conditions (500 mOsM) for 24 h. (F) Expression levels of interleukin-6 ( IL-6 ), IL-17 , and IL-13 in HCEs under hyperosmotic conditions after treatment with different UA concentrations for 24 h. (G) Representative images (left) and fluorescence intensity quantification (right) of TdT-mediated dUTP nick-end labeling (TUNEL) staining in HCEs treated with UA under hyperosmotic conditions for 24 h ( n = 4 per group). (H) Representative images (left) and fluorescence intensity quantification (right) of Ki67 staining in HCEs treated with UA under hyperosmotic conditions for 24 h ( n = 3 per group). Data are presented as mean ± standard error of the mean (SEM). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 for comparisons with the model group; # P < 0.05, ## P < 0.01, and ### P < 0.001 for comparisons of relative cell viability between 24 and 48 h treatments. CCK-8: Cell Counting Kit-8; qRT-PCR: quantitative reverse transcription polymerase chain reaction; HS: hypertonic stress; LC-UA: low concentration of UA; MC-UA: medium concentration of UA; HC-UA: high concentration of UA; DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: Human corneal epithelial cells (HCEs) were obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA) and cultured in a 5% CO 2 incubator at 37 °C.

Techniques: In Vitro, Concentration Assay, Expressing, Fluorescence, End Labeling, TUNEL Assay, Staining, CCK-8 Assay, Cell Counting, Quantitative RT-PCR, Reverse Transcription, Polymerase Chain Reaction