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The preservative polyquaternium-1 increases cytoxicity and NF-kappaB linked inflammation in human corneal epithelial cells.

Paimela T, Ryhänen T, Kauppinen A, Marttila L, Salminen A, Kaarniranta K - Mol. Vis. (2012)

Bottom Line: Interestingly, Travatan and Systane Ultra activated NF-κB and elevated the secretion of inflammation markers IL-6 by 3 to eightfold and IL-8 by 1.5 to 3.5 fold, respectively, as analyzed with ELISA.Eye drops containing PQ-1 evoke cytotoxicity and enhance the NF-κB driven inflammation reaction in cultured HCE-2 cells.Our results indicate that these harmful effects of ocular solutions preserved with PQ-1 should be further evaluated in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Institute of Clinical Medicine, University of Eastern Finland, Kuopio, Finland.

ABSTRACT

Purpose: In numerous clinical and experimental studies, preservatives present in eye drops have had detrimental effects on ocular epithelial cells. The aim of this study was to compare the cytotoxic and inflammatory effects of the preservative polyquaternium-1 (PQ-1) containing Travatan (travoprost 0.004%) and Systane Ultra eye drops with benzalkonium chloride (BAK) alone or BAK-preserved Xalatan (0.005% latanoprost) eye drops in HCE-2 human corneal epithelial cell culture.

Methods: HCE-2 cells were exposed to the commercial eye drops Travatan, Systane Ultra, Xalatan, and the preservative BAK. Cell viability was determined using colorimetric MTT (3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and by release of lactate dehydrogenase (LDH). Induction of apoptosis was measured with a using a colorimetric caspase-3 assay kit. DNA binding of the nuclear factor kappa B (NF-κB) transcription factor, and productions of the proinflammatory cytokines, interleukins IL-6 and IL-8, were determined using an enzyme-linked immunosorbent assay (ELISA) method.

Results: Cell viability, as measured by the MTT assay, declined by up to 50% after exposure to Travatan or Systane Ultra solutions which contain 0.001% PQ-1. BAK at 0.02% rather than at 0.001% concentration evoked total cell death signs on HCE-2 cells. In addition, cell membrane permeability, as measured by LDH release, was elevated by sixfold with Travatan and by a maximum threefold with Systane Ultra. Interestingly, Travatan and Systane Ultra activated NF-κB and elevated the secretion of inflammation markers IL-6 by 3 to eightfold and IL-8 by 1.5 to 3.5 fold, respectively, as analyzed with ELISA.

Conclusions: Eye drops containing PQ-1 evoke cytotoxicity and enhance the NF-κB driven inflammation reaction in cultured HCE-2 cells. Our results indicate that these harmful effects of ocular solutions preserved with PQ-1 should be further evaluated in vitro and in vivo.

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Related in: MedlinePlus

Interleukin-8 secretion from HCE-2 cells analyzed by ELISA. Columns represent the amount of IL-8 pg/ml (mean±SD).. One-Way ANOVA, followed by Dunnett’s post hoc test, evaluated the statistical differences (n=6, *0.01<p≤0.05, **0.001<p≤0.01, ***p≤0.001, ns=not significant). Experiments were repeated three times.
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f5: Interleukin-8 secretion from HCE-2 cells analyzed by ELISA. Columns represent the amount of IL-8 pg/ml (mean±SD).. One-Way ANOVA, followed by Dunnett’s post hoc test, evaluated the statistical differences (n=6, *0.01<p≤0.05, **0.001<p≤0.01, ***p≤0.001, ns=not significant). Experiments were repeated three times.

Mentions: Travatan, Systane Ultra and BAK 0.001% increased IL-6 levels already after a 5 min exposure when compared to the controls (Figure 4). There was approximately three times more IL-6 released into the culture medium of the Travatan and Systane Ultra-treated cells. After 15 min of exposure to Travatan and Systane Ultra, the levels were 8 times higher than the controls. For comparison, BAK 0.001% increased the amount of IL-6 by only about twofold in response to 15 min´ treatment. The 30 min exposure amplified the levels even more in Systane Ultra (~10×) and in BAK 0.001%-treated (~4×) samples, whereas in Travatan exposed samples the IL-6 levels were the same as in the 15 min samples. The response of interleukin-8 was similar to that seen with IL-6 (Figure 5), although the elevations were not as dramatic (with Travatan ~50%–300%, Systane Ultra ~0%–350% and no elevation with BAK 0.001%). NF-κB levels were increased statistically significantly in the Systane Ultra (15 and 30 min exposures) and Travatan (30 min exposure) treated cells (Figure 6). BAK 0.001% did not have any effect on the NF-κB expression.


The preservative polyquaternium-1 increases cytoxicity and NF-kappaB linked inflammation in human corneal epithelial cells.

Paimela T, Ryhänen T, Kauppinen A, Marttila L, Salminen A, Kaarniranta K - Mol. Vis. (2012)

Interleukin-8 secretion from HCE-2 cells analyzed by ELISA. Columns represent the amount of IL-8 pg/ml (mean±SD).. One-Way ANOVA, followed by Dunnett’s post hoc test, evaluated the statistical differences (n=6, *0.01<p≤0.05, **0.001<p≤0.01, ***p≤0.001, ns=not significant). Experiments were repeated three times.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3351434&req=5

f5: Interleukin-8 secretion from HCE-2 cells analyzed by ELISA. Columns represent the amount of IL-8 pg/ml (mean±SD).. One-Way ANOVA, followed by Dunnett’s post hoc test, evaluated the statistical differences (n=6, *0.01<p≤0.05, **0.001<p≤0.01, ***p≤0.001, ns=not significant). Experiments were repeated three times.
Mentions: Travatan, Systane Ultra and BAK 0.001% increased IL-6 levels already after a 5 min exposure when compared to the controls (Figure 4). There was approximately three times more IL-6 released into the culture medium of the Travatan and Systane Ultra-treated cells. After 15 min of exposure to Travatan and Systane Ultra, the levels were 8 times higher than the controls. For comparison, BAK 0.001% increased the amount of IL-6 by only about twofold in response to 15 min´ treatment. The 30 min exposure amplified the levels even more in Systane Ultra (~10×) and in BAK 0.001%-treated (~4×) samples, whereas in Travatan exposed samples the IL-6 levels were the same as in the 15 min samples. The response of interleukin-8 was similar to that seen with IL-6 (Figure 5), although the elevations were not as dramatic (with Travatan ~50%–300%, Systane Ultra ~0%–350% and no elevation with BAK 0.001%). NF-κB levels were increased statistically significantly in the Systane Ultra (15 and 30 min exposures) and Travatan (30 min exposure) treated cells (Figure 6). BAK 0.001% did not have any effect on the NF-κB expression.

Bottom Line: Interestingly, Travatan and Systane Ultra activated NF-κB and elevated the secretion of inflammation markers IL-6 by 3 to eightfold and IL-8 by 1.5 to 3.5 fold, respectively, as analyzed with ELISA.Eye drops containing PQ-1 evoke cytotoxicity and enhance the NF-κB driven inflammation reaction in cultured HCE-2 cells.Our results indicate that these harmful effects of ocular solutions preserved with PQ-1 should be further evaluated in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Institute of Clinical Medicine, University of Eastern Finland, Kuopio, Finland.

ABSTRACT

Purpose: In numerous clinical and experimental studies, preservatives present in eye drops have had detrimental effects on ocular epithelial cells. The aim of this study was to compare the cytotoxic and inflammatory effects of the preservative polyquaternium-1 (PQ-1) containing Travatan (travoprost 0.004%) and Systane Ultra eye drops with benzalkonium chloride (BAK) alone or BAK-preserved Xalatan (0.005% latanoprost) eye drops in HCE-2 human corneal epithelial cell culture.

Methods: HCE-2 cells were exposed to the commercial eye drops Travatan, Systane Ultra, Xalatan, and the preservative BAK. Cell viability was determined using colorimetric MTT (3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and by release of lactate dehydrogenase (LDH). Induction of apoptosis was measured with a using a colorimetric caspase-3 assay kit. DNA binding of the nuclear factor kappa B (NF-κB) transcription factor, and productions of the proinflammatory cytokines, interleukins IL-6 and IL-8, were determined using an enzyme-linked immunosorbent assay (ELISA) method.

Results: Cell viability, as measured by the MTT assay, declined by up to 50% after exposure to Travatan or Systane Ultra solutions which contain 0.001% PQ-1. BAK at 0.02% rather than at 0.001% concentration evoked total cell death signs on HCE-2 cells. In addition, cell membrane permeability, as measured by LDH release, was elevated by sixfold with Travatan and by a maximum threefold with Systane Ultra. Interestingly, Travatan and Systane Ultra activated NF-κB and elevated the secretion of inflammation markers IL-6 by 3 to eightfold and IL-8 by 1.5 to 3.5 fold, respectively, as analyzed with ELISA.

Conclusions: Eye drops containing PQ-1 evoke cytotoxicity and enhance the NF-κB driven inflammation reaction in cultured HCE-2 cells. Our results indicate that these harmful effects of ocular solutions preserved with PQ-1 should be further evaluated in vitro and in vivo.

Show MeSH
Related in: MedlinePlus