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Category: Cytotoxicity

Human corneal epithelial cell viability is important in evaluating of toxic effects of drugs, new chemical entities (NCEs) and excipients in ocular drops.

SUMMARY

Cells

Human corneal epithelial cells1 (HCE-T, RIKEN, Japan)

Exposure

Cells are exposed to study compounds at different concentrations for selected times (e.g. 5, 15, 30 or 60 min)

Positive control

Benzalkonium chloride (BAC) at concentrations 0 – 0.02%2

Indicator and detection

Indicator: resazurin (7-Hydroxy-3H-phenoxazin-3-one-10-oxide) dye is irreversibly reduced to highly fluorescent metabolite resorufin (7-Hydroxy-3H-phenoxazin-3-one);2

Detection: after 2 h exposure to resazurin, the amount of resorufin is measured at 560 nm (ex), 590 nm (em) by using Cytation 3 multi-mode reader (BioTek Instruments, Winooski, VT, USA).2

Please contact us, if you would like to receive more information: info@experimentica.com

IMAGES

HCE-T cell viability after exposure to clinically approved drug formulations

Expression of tight junction proteins occludin (left) and ZO-1 (right) in human corneal epithelial cells (HCE-T) cultured on chamber slide (Lab-Tek™ Chamber Slide System, Nunc™, Thermo Fisher Scientific, Waltham, MA, USA) detected by immunofluorescence staining (green). Nuclei were stained with DAPI (blue). Scale bar 50 µm.

Cell viability of approved drugs (bimatoprost, latanoprost). HCE-T cell viability after 10 min exposure to ophthalmic drugs and excipients in ocular drops. Data are presented as mean±SEM. BAC, benzalkonium chloride was used as a positive control. Statistical significance of differences was tested by one-way ANOVA followed by Dunnett’s multiple comparisons test by using GraphPad Prism 6.07 software (San Diego, CA, USA). Statistical significance was set at P<0.05. Studies were designed and funded by Experimentica Ltd.

Cell viability of approved drugs (tafluprost). HCE-T cell viability after 10 min exposure to ophthalmic drugs and excipients in ocular drops. Data are presented as mean±SEM. PS80, polysorbate 80. No statistically significant cytotoxicity of tafluprost/PS80 and PS80 were found. Studies were designed and funded by Experimentica Ltd.

Cell viability of approved drugs (latanoprost, travoprost). HCE-T cell viability after 10 min exposure to ophthalmic drugs and excipients in ocular drops. Data are presented as mean±SEM. MGHS40, Macrogolglycerol hydroxystearate 40. No statistically significant cytotoxicity of latanoprost/MGHS40, travoprost/MGHS40 and MGSH40 were found. Studies were designed and funded by Experimentica Ltd.

SELECTED PUBLICATIONS

  • Hakkarainen JJ, Reinisalo M, Ragauskas S, Seppänen A, Kaja S, Kalesnykas G. Acute cytotoxic effects of marketed ophthalmic formulations on human corneal epithelial cells. Int J Pharm. 2016;511(1):73-78. doi: 10.1016/j.ijpharm.2016.06.135.

REFERENCES

  1. Araki-Sasaki K, Ohashi Y, Sasabe T, Hayashi K, Watanabe H, Tano Y, Handa H. An SV40-immortalized human corneal epithelial cell line and its characterization. Invest Ophthalmol Vis Sci 1995,36:614–621.
  2. Hakkarainen JJ, Reinisalo M, Ragauskas S, Seppänen A, Kaja S, Kalesnykas G. Acute cytotoxic effects of marketed ophthalmic formulations on human corneal epithelial cells. Int J Pharm. 2016;511(1):73-78. doi: 10.1016/j.ijpharm.2016.06.135.

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