Comparison of preservative-induced toxicity on monolayer and stratified Chang conjunctival cells

Yanochko, Gina M.; Khoh‐Reiter, Su; Evans, Mark; Jessen, Bart

doi:10.1016/j.tiv.2010.02.001

Cited by 16 publications

(10 citation statements)

References 29 publications

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“…25 Similar results were found in a study evaluating the preservative-induced toxicity on monolayer and stratified air-lifted cultures, Chang conjunctival cells. 15 These findings suggest that multilayer cultures are more realistic models that should be preferentially chosen for in vitro ocular surface toxicology.…”

Section: Discussionmentioning

confidence: 97%

“…In an attempt to resemble ocular surface epithelium, air-lifting 3-dimensional (3D) cultures of corneal and conjunctival epithelial cells have been established in the last years. [13][14][15] In this study, we investigated the effect on cell viability of several common anti-allergic drugs, some of them containing BAC as preservative, as well as the effect of the preservative alone, analyzing differences in response of monolayer and stratified cell cultures. For these experiments, a human corneal-limbal epithelial cell line that can stratify in culture medium rather than at an air interface 16 was used.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

In VitroEffects of Preserved and Unpreserved Anti-Allergic Drugs on Human Corneal Epithelial Cells

Guzmán-Aránguez

Calvo

Ropero

et al. 2014

Journal of Ocular Pharmacology and Therapeutics

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Section: Discussionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

In VitroEffects of Preserved and Unpreserved Anti-Allergic Drugs on Human Corneal Epithelial Cells

Guzmán-Aránguez

Calvo

Ropero

et al. 2014

Journal of Ocular Pharmacology and Therapeutics

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“…This study compared the in vitro toxic effects of five preservatives belonging to different chemical classes, by flow cytometry, a standard assay. The existing information about the comparative effects of these preservatives on cytotoxicity and induction of apoptosis, necrosis and genotoxicity is very scarce [25, 26] and most of the available data related to preservatives used in ocular medications [27, 28]. This investigation was intended to reduce this gap, allowing a better knowledge on cosmetic preservatives cytotoxicity and genotoxicity potentials.…”

Section: Discussionmentioning

confidence: 99%

In vitro induction of apoptosis, necrosis and genotoxicity by cosmetic preservatives: application of flow cytometry as a complementary analysis by NRU

Carvalho¹,

Menezes²,

Letenski³

et al. 2011

Intern J of Cosmetic Sci

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Preservatives are used in cosmetics to prevent microbial contamination; however, some preservatives are not free of allergenic and cytotoxic potential. Allergenicity and cytotoxicity potential values are major aspects of preservative safety, which determine limitations and maximum concentration dose in a cosmetic product. The purpose of this study was to investigate and compare the in vitro apoptosis, necrosis and genotoxicity-inducing potential of five different types of preservatives: Phenoxyethanol (PE), Propylparaben (PP), Methylparaben (MP), Benzyl Alcohol (BA) and Ethylhexyl Glycerine (EG). In vitro experiments were carried out on human dermal fibroblasts by a quantitative flow cytometry method, using specific cell markers (Annexin V, Propidium Iodide and H2AX). We compared the resulting cell viability by means of neutral red uptake (NRU) and established the IC(50) . Our results showed that PE, PP, MP and BA have similar cytotoxic mechanisms (high apoptosis and necrosis levels only at the test concentration of 1%), whereas EG showed only an apoptosis pathway. For genotoxicity, both parabens yielded the highest values. Results obtained by flow cytometry for necrosis were comparable to those produced by NRU; however, NRU does not distinguish apoptosis from necrosis. We propose that flow cytometry is a more sophisticated methodology for understanding the cytotoxic mechanisms of cosmetic preservatives and can be used to complement the NRU.

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“…This is likely due to the very high cleansing action of SLS and its protein solubility. Yanochko et al (2010) used monolayer culture and a three-dimensional model (stratified culture model) to examine the IC 50 (applied concentration-cytotoxicity curve) following application of BC for the same exposure time, and reported that with monolayer culture, cells were impacted at a concentration 1/100 that of the three-dimensional model. The IC 50 of SLS was reported to be 44.67 μg/mL by cytotoxicity evaluation in a 24 hr exposed monolayer culture that used the MTT incorporation method with keratinocyte (Sanchez et al, 2004); this was 1/156-1/248 that observed in the present examination.…”

Section: Comparison Of Cmc and Ic 50mentioning

confidence: 99%

Predicting the results of a 24-hr human patch test for surfactants: utility of margin-setting in a reconstructed human epidermis model

et al. 2019

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To predict the results of a 24-hr closed human patch test, we previously recommended the use of in vitro test with a reconstructed human epidermis (RhE) model adopted in OECD TG 439, and proposed the margin method, which includes evaluation of twice the concentration to avoid a false positive for surfactants. Therefore, in this study, we used LabCyte EPI-MODEL as a RhE model, and confirmed the reproducibility of this method using five surfactants, including benzalkonium chloride (BC), sodium lauryl sulfate (SLS), and lauryl betaine (LB), for which false negative results have previously been reported, and three different surfactants. For all surfactants, prediction of patch test results using a margin of two revealed that human tests could be performed safely, confirming the utility of the margin method. In addition, we examined the relationship with critical micellar concentration (CMC). The IC 50 for cell viability in the RhE model for three types of surfactants (BC, SLS, and LB) was 2.7-to 49.7-times the CMC. Therefore, the range of concentrations in which tests were performed with the present method was within the range of concentrations with high cleansing. Furthermore, we examined the relationship between cell viability and release of the inflammatory mediator interleukin-1α (IL-1α). IL-1α release was associated with cell viability, supporting the results of the human patch test. that the result of a patch test is "non-irritant" if an in vitro test performed with a concentration twice as high as the test concentration has a survival rate exceeding 50%

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Comparison of preservative-induced toxicity on monolayer and stratified Chang conjunctival cells

Cited by 16 publications

References 29 publications

In VitroEffects of Preserved and Unpreserved Anti-Allergic Drugs on Human Corneal Epithelial Cells

In VitroEffects of Preserved and Unpreserved Anti-Allergic Drugs on Human Corneal Epithelial Cells

In vitro induction of apoptosis, necrosis and genotoxicity by cosmetic preservatives: application of flow cytometry as a complementary analysis by NRU

Predicting the results of a 24-hr human patch test for surfactants: utility of margin-setting in a reconstructed human epidermis model

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