Comparative Toxicity of Tetracaine, Proparacaine and Cocaine Evaluated with Primary Cultures of Rabbit Corneal Epithelial Cells

1994 ◽  
Vol 58 (4) ◽  
pp. 469-478 ◽  
Author(s):  
Roberta L. Grant ◽  
Daniel Acosta
Keyword(s):  
Epithelial Cells ◽  
Primary Cultures ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial ◽  
Comparative Toxicity

scholarly journals Characterisation of the prereceptor regulation of glucocorticoids in the anterior segment of the rabbit eye

2006 ◽  
Vol 190 (2) ◽  
pp. 483-493 ◽  
Author(s):  
Claire U Onyimba ◽  
Neelima Vijapurapu ◽  
S John Curnow ◽  
Pamela Khosla ◽  
Paul M Stewart ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Corneal Epithelium ◽  
Ciliary Body ◽  
Primary Cultures ◽  
Anterior Segment ◽  
Body Tissue ◽  
Corneal Epithelial Cell ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial ◽  
Rabbit Eye

The prereceptor regulation of glucocorticoids (GCs) by 11β-hydroxysteroid dehydrogenase type-1 (11β-HSD1), a bidirectional isozyme that interconverts active (cortisol) and inactive (cortisone) GCs, is an established determinant of GC function in tissues such as liver, adipose and bone. Although the therapeutic use of GCs is abundant in ophthalmic practice, where GC interactions with nuclear receptors modulate gene transcription, the prereceptor regulation of endogenous cortisol is not well described in ocular tissues. Recent descriptive studies have localised 11β-HSD1 to the human corneal epithelium and non-pigmented epithelium (NPE) of the ciliary body, indicating a link to corneal epithelial physiology and aqueous humour production. In this study, we characterise the functional aspects of the autocrine regulation of GCs in the anterior segment of the rabbit eye. Using our in-house generated primary antibody to human 11β-HSD1, immunohistochemical analyses were performed on paraffin-embedded sections of whole New Zealand white albino rabbits, (NZWAR) eyes. As in human studies, 11β-HSD1 was localised to the corneal epithelium and the NPE. No staining was seen in the albino ‘pigmented’ ciliary epithelium. Specific enzyme assays for oxo-reductase (cortisone→cortisol) and dehydrogenase (cortisol→cortisone) activity indicated predominant 11β-HSD1 oxo-reductase activity from both the intact ciliary body tissue (n=12, median 2.1 pmol/mg per h and range 1.25–2.8 pmol/mg per h; P=0.006) and primary cultures of corneal epithelial cells (n=12, median 3.0 pmol/mg per h and range 1.0–7.4 pmol/mg per h, P=0.008) compared with dehydrogenase activity (median 1.0 pmol/mg per h and range 0.5–2.0 pmol/mg per h; median 0.5 pmol/mg per h and range 0.25–1.9 pmol/mg per h respectively). These findings were supported by expression of 11β-HSD1 protein as visualised by Western blotting of ciliary body tissue and immunocytochemistry of corneal epithelial cells. Reduction of corneal epithelial cell proliferation was seen after primary cultures were co-incubated with cortisol and cortisone. 11β-HSD1 activity was not demonstrated in naïve conjunctival fibroblasts or corneal stromal keratocytes. Our results indicate that the distribution of 11β-HSD1 in the rabbit resembles that of the human eye and activates cortisone to cortisol in both corneal and uveal tissues. The NZWAR provides a suitable in vivo model for the further evaluation of 11β-HSD1 activity in the eye, especially its role in corneal epithelial and ciliary body physiology.

scholarly journals Expression of immune response genes in human corneal epithelial cells interacting with Aspergillus flavus conidia

BMC Genomics ◽  
2022 ◽  
Vol 23 (1) ◽  
Author(s):  
Divya Arunachalam ◽  
Shruthi Mahalakshmi Ramanathan ◽  
Athul Menon ◽  
Lekshmi Madhav ◽  
Gopalakrishna Ramaswamy ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cell Line ◽  
Differentially Expressed Genes ◽  
Primary Cultures ◽  
Differentially Expressed ◽  
Corneal Epithelial Cell ◽  
Epithelial Cell Line ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial ◽  
Human Corneal Epithelial Cells

Abstract Background Aspergillus flavus, one of the causative agents of human fungal keratitis, can be phagocytosed by human corneal epithelial (HCE) cells and the conidia containing phagosomes mature into phagolysosomes. But the immunological responses of human corneal epithelial cells interacting with A. flavus are not clear. In this study, we report the expression of immune response related genes of HCE cells exposed to A. flavus spores using targeted transcriptomics. Methods Human corneal epithelial cell line and primary cultures were grown in a six-well plate and used for coculture experiments. Internalization of the conidia was confirmed by immunofluorescence microscopy of the colocalized endosomal markers CD71 and LAMP1. Total RNA was isolated, and the quantity and quality of the isolated RNA were assessed using Qubit and Bioanalyzer. NanoString nCounter platform was used for the analysis of mRNA abundance using the Human Immunology panel. R-package and nSolver software were used for data analysis. KEGG and FunRich 3.1.3 tools were used to analyze the differentially expressed genes. Results Different morphotypes of conidia were observed after 6 h of coculture with human corneal epithelial cells and found to be internalized by epithelial cells. NanoString profiling showed more than 20 differentially expressed genes in immortalized human corneal epithelial cell line and more than ten differentially expressed genes in primary corneal epithelial cells. Distinct set of genes were altered in their expression in cell line and primary corneal epithelial cells. KEGG pathway analysis revealed that genes associated with TNF signaling, NF-KB signaling, and Th17 signaling were up-regulated, and genes associated with chemokine signaling and B cell receptor signaling were down regulated. FunRich pathway analysis showed that pathways such as CDC42 signaling, PI3K signaling, and Arf6 trafficking events were activated by the clinical isolates CI1123 and CI1698 in both type of cells. Conclusions Combining the transcript analysis data from cell lines and primary cultures, we showed the up regulation of immune defense genes in A. flavus infected cells. At the same time, chemokine signaling and B cell signaling pathways are downregulated. The variability in the expression levels in the immortalized cell line and the primary cultures is likely due to the variable epigenetic reprogramming in the immortalized cells and primary cultures in the absence of any changes in the genome. It highlights the importance of using both cell types in host-pathogen interaction studies.

Corneal epithelial-specific cell surface antigen recognized by a monoclonal antibody

Development ◽  
1986 ◽  
Vol 94 (1) ◽  
pp. 163-172
Author(s):  
Marian G. Langer ◽  
C. V. Sundarraj ◽  
Nirmala Sundarraj
Keyword(s):  
Epithelial Cells ◽  
Cell Surface ◽  
Primary Cultures ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Cell ◽  
Immunoperoxidase Technique ◽  
Igg Antibody ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial ◽  
Human Corneal Epithelial Cells

Monoclonal antibodies, specific against cell surface differentiation antigens of human corneal epithelial cells, were developed using epithelial cells resected from human corneas as the immunogens. One of these antibodies reacted specifically with corneal epithelial cells and not with epithelial cells of other tissues when tested by an indirect immunoperoxidase technique. Nonidet P-40 extracts of different subcellular fractions of human corneal epithelial cells were tested for their reactivity against this antibody using an enzyme-linked immunosorbent assay. The results indicated that the antigen recognized by this antibody is associated with the plasma membrane. This was further verified by immuno-electron-microscopic analysis using ferritin-conjugated anti-mouse IgG antibody. This antigen was not detectable in the corneal epithelial cells in primary cultures nor in the epithelial cells from early stages of developing cornea (12 to 18 weeks in utero) but was present in the epithelial cells in the corneas of an 8-month-old infant. Therefore, this surface-associated antigen identified in the present study is a developmentally regulated marker of human corneal epithelium.

scholarly journals Nuclear Ferritin Protects DNA From UV Damage in Corneal Epithelial Cells

1998 ◽  
Vol 9 (5) ◽  
pp. 1037-1051 ◽  
Author(s):  
Cindy X. Cai ◽  
David E. Birk ◽  
Thomas F. Linsenmayer
Keyword(s):  
Epithelial Cells ◽  
Primary Cultures ◽  
Cell Types ◽  
Dna Strand Breaks ◽  
Uv Damage ◽  
Corneal Epithelial Cells ◽  
Free Radical Damage ◽  
Corneal Epithelial ◽  
Strand Breaks ◽  
Dna Strand

Previously, we identified the heavy chain of ferritin as a developmentally regulated nuclear protein of embryonic chicken corneal epithelial cells. The nuclear ferritin is assembled into a supramolecular form indistinguishable from the cytoplasmic form of ferritin found in other cell types and thus most likely has iron-sequestering capabilities. Free iron, via the Fenton reaction, is known to exacerbate UV-induced and other oxidative damage to cellular components, including DNA. Since corneal epithelial cells are constantly exposed to UV light, we hypothesized that the nuclear ferritin might protect the DNA of these cells from free radical damage. To test this possibility, primary cultures of cells from corneal epithelium and stroma, and from skin epithelium and stroma, were UV irradiated, and DNA strand breaks were detected by an in situ 3′-end labeling method. Corneal epithelial cells without nuclear ferritin were also examined. We observed that the corneal epithelial cells with nuclear ferritin had significantly less DNA breakage than other cell types examined. Furthermore, increasing the iron concentration of the culture medium exacerbated the generation of UV-induced DNA strand breaks in corneal and skin fibroblasts, but not in the corneal epithelial cells. Most convincingly, corneal epithelial cells in which the expression of nuclear ferritin was inhibited became much more susceptible to UV-induced DNA damage. Therefore, it seems that corneal epithelial cells have evolved a novel, nuclear ferritin-based mechanism for protecting their DNA against UV damage.

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