Corneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications

Marisa Meloni,1 Giampiero Cattaneo,2 Barbara De Servi11VitroScreen In Vitro Research Laboratories, 2Thea Farma, Milan, ItalyBackground: By using a biologically relevant and sensitive three-dimensional model of human corneal epithelium and multiple endpoint analysis, assessment of the potential for e...

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Autores principales:	Meloni M, Cattaneo G, De Servi B
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2012
Materias:	Ophthalmology RE1-994
Acceso en línea:	https://doaj.org/article/506c7bc83cdf4da1873d3c58641197dc
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spelling	oai:doaj.org-article:506c7bc83cdf4da1873d3c58641197dc2021-12-02T07:48:55ZCorneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications1177-54671177-5483https://doaj.org/article/506c7bc83cdf4da1873d3c58641197dc2012-09-01T00:00:00Zhttp://www.dovepress.com/corneal-epithelial-toxicity-of-antiglaucoma-formulations-in-vitro-stud-a10908https://doaj.org/toc/1177-5467https://doaj.org/toc/1177-5483Marisa Meloni,1 Giampiero Cattaneo,2 Barbara De Servi11VitroScreen In Vitro Research Laboratories, 2Thea Farma, Milan, ItalyBackground: By using a biologically relevant and sensitive three-dimensional model of human corneal epithelium and multiple endpoint analysis, assessment of the potential for eye irritation and long-term compatibility of four registered ophthalmological preparations, ie, Timolabak&reg;, Timoptol&reg;, Nyogel&reg;, and Timogel&reg;, was performed. This approach enables classification of the potential for irritation, discriminating between mildly irritant and non-irritant ocular substances.Methods: The exposure protocol included two time periods, ie, 24 hours (acute application) and 72 hours (repeated applications twice daily). This approach allows assessment of not only the acute reaction but also possible recovery, as well as mimicking the potential cumulative effects associated with long-term application. Using benzalkonium chloride (BAK) 0.01% as a positive control, the following parameters were quantified: cellular viability by MTT test, histological analysis by hematoxylin and eosin staining, passive release of interleukin-1a by enzyme-linked immunosorbent assay, and OCLN gene expression by quantitative real-time polymerase chain reaction.Results: Cell viability was reduced to under the 50% cutoff value after acute exposure (24 hours) to BAK 0.01%, and after repeated application (72 hours) of Timoptol and Nyogel. Histological analysis after acute exposure showed signs of superficial damage with all formulations, and severe changes after repeated applications of Timoptol, BAK 0.01%, and Nyogel. Timolabak and Timogel did not significantly alter the morphology of the human corneal epithelial cells after the different exposure times. Interleukin-1&alpha; release was greater than that for the negative control (&gt;20 pg/mL) and the positive control (BAK 0.01%), Nyogel, and Timoptol treatments and not different after treatment with Timolabak and Timogel. Expression of OCLN, a sign of epithelial barrier impairment, was only significantly upregulated at 24 hours by BAK 0.01%, suggesting a toxic reaction at the ocular surface. OCLN was also overexpressed after repeated application of Nyogel and Timogel.Conclusion: Overall, the multiple endpoint analysis approach allows classification of these products according to decreasing order of irritation potential as follows: BAK 0.01%, Timoptol, Nyogel, Timogel, and Timolabak.Keywords: occludin, gene expression, interleukin-1&alpha;, eye compatibility, human corneal epitheliumMeloni MCattaneo GDe Servi BDove Medical PressarticleOphthalmologyRE1-994ENClinical Ophthalmology, Vol 2012, Iss default, Pp 1433-1440 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Ophthalmology RE1-994
spellingShingle	Ophthalmology RE1-994 Meloni M Cattaneo G De Servi B Corneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications
description	Marisa Meloni,1 Giampiero Cattaneo,2 Barbara De Servi11VitroScreen In Vitro Research Laboratories, 2Thea Farma, Milan, ItalyBackground: By using a biologically relevant and sensitive three-dimensional model of human corneal epithelium and multiple endpoint analysis, assessment of the potential for eye irritation and long-term compatibility of four registered ophthalmological preparations, ie, Timolabak&reg;, Timoptol&reg;, Nyogel&reg;, and Timogel&reg;, was performed. This approach enables classification of the potential for irritation, discriminating between mildly irritant and non-irritant ocular substances.Methods: The exposure protocol included two time periods, ie, 24 hours (acute application) and 72 hours (repeated applications twice daily). This approach allows assessment of not only the acute reaction but also possible recovery, as well as mimicking the potential cumulative effects associated with long-term application. Using benzalkonium chloride (BAK) 0.01% as a positive control, the following parameters were quantified: cellular viability by MTT test, histological analysis by hematoxylin and eosin staining, passive release of interleukin-1a by enzyme-linked immunosorbent assay, and OCLN gene expression by quantitative real-time polymerase chain reaction.Results: Cell viability was reduced to under the 50% cutoff value after acute exposure (24 hours) to BAK 0.01%, and after repeated application (72 hours) of Timoptol and Nyogel. Histological analysis after acute exposure showed signs of superficial damage with all formulations, and severe changes after repeated applications of Timoptol, BAK 0.01%, and Nyogel. Timolabak and Timogel did not significantly alter the morphology of the human corneal epithelial cells after the different exposure times. Interleukin-1&alpha; release was greater than that for the negative control (&gt;20 pg/mL) and the positive control (BAK 0.01%), Nyogel, and Timoptol treatments and not different after treatment with Timolabak and Timogel. Expression of OCLN, a sign of epithelial barrier impairment, was only significantly upregulated at 24 hours by BAK 0.01%, suggesting a toxic reaction at the ocular surface. OCLN was also overexpressed after repeated application of Nyogel and Timogel.Conclusion: Overall, the multiple endpoint analysis approach allows classification of these products according to decreasing order of irritation potential as follows: BAK 0.01%, Timoptol, Nyogel, Timogel, and Timolabak.Keywords: occludin, gene expression, interleukin-1&alpha;, eye compatibility, human corneal epithelium
format	article
author	Meloni M Cattaneo G De Servi B
author_facet	Meloni M Cattaneo G De Servi B
author_sort	Meloni M
title	Corneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications
title_short	Corneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications
title_full	Corneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications
title_fullStr	Corneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications
title_full_unstemmed	Corneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications
title_sort	corneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications
publisher	Dove Medical Press
publishDate	2012
url	https://doaj.org/article/506c7bc83cdf4da1873d3c58641197dc
work_keys_str_mv	AT melonim cornealepithelialtoxicityofantiglaucomaformulationsinvitrostudyofrepeatedapplications AT cattaneog cornealepithelialtoxicityofantiglaucomaformulationsinvitrostudyofrepeatedapplications AT deservib cornealepithelialtoxicityofantiglaucomaformulationsinvitrostudyofrepeatedapplications
_version_	1718399153613045760

Corneal epithelial toxicity of antiglaucoma formulations: in vitro study of repeated applications

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