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[Antioxidant effects of mast cell inhibitors in a human conjunctival cell line]
C Debbasch1, P J Pisella, P Rat
1Unité de Pharmaco-Toxicologie Cellulaire, CHNO des Quinze-Vingts, 28, rue de Charenton, 75012 Paris.
Purpose:
The aim of this study was to evaluate in vitro antioxidant effects of two mast cells inhibitors.
Methods:
Cytotoxicity tests were done on a continuous human conjunctival cell line using microplate cold light cytofluorimetry. Membrane integrity (neutral red test), DNA condensation (Hoechst 33342 test), and reactive oxygen species (ROS) production (dichlorofluoresceine diacetate and hydroethidine tests) were evaluated on living cells treated with sodium cromoglycate and N-acetyl-aspartyl glutamic acid (NAAGA) preserved (benzalkonium chloride: BAC at 0.01%) and unpreserved after 60 minutes of treatment or 60 minutes and 24 hours of cell recovery. They were tested pure and at a 1/10 dilution. ROS production was also evaluated after a 60 minute pretreatment with antiallergic drugs and a 15-minute treatment with BAC, according to previous experiments performed on BAC showing its ROS production properties.
Results:
No cytotoxicity was observed with the unpreserved formulations of antiallergic drugs. An apoptotic phenomenom was suggested with preserved drugs after a 1-hour treatment, whereas a necrotic mechanism appeared after a 24-hour cell recovery period. A ROS production decrease was observed with the two preserved and unpreserved drugs tested (p<0.001 compared to BAC) even if it was significantly higher with cromoglycate formulations. A ROS production decrease also was detected after a pretreatment with antiallergic drugs and treatment with BAC (p<0.001 compared to BAC alone).
Conclusion:
In vitro, no cytotoxicity was found with the two unpreserved mast cell inhibitors tested. An antioxidant effect also was observed with these two molecules; sodium cromoglycate appeared to be the best free radical scavenger.
Insights
Two mast cell inhibitors, sodium cromoglycate and N-acetyl-aspartyl glutamic acid (NAAGA), show antioxidant effects in vitro. Unpreserved formulations were non-cytotoxic, with sodium cromoglycate demonstrating superior free radical scavenging.
Area of Science:
- Ophthalmology
- Cell Biology
- Pharmacology
Background:
- Mast cells play a key role in allergic responses.
- Mast cell inhibitors are used to treat ocular allergies.
- Preservatives like benzalkonium chloride (BAC) can induce oxidative stress.
Purpose of the Study:
- To evaluate the in vitro antioxidant effects of two mast cell inhibitors: sodium cromoglycate and N-acetyl-aspartyl glutamic acid (NAAGA).
- To assess the cytotoxicity and reactive oxygen species (ROS) production of preserved and unpreserved formulations.
Main Methods:
- Cytotoxicity was assessed using neutral red and Hoechst 33342 tests on a human conjunctival cell line.
- ROS production was measured using dichlorofluoresceine diacetate and hydroethidine assays.
- Cells were treated with pure or diluted drugs, with or without BAC, and evaluated at different time points.
Main Results:
- Unpreserved antiallergic drug formulations showed no cytotoxicity.
- Preserved drugs induced apoptosis after 1 hour and necrosis after 24 hours.
- Both preserved and unpreserved drugs significantly reduced ROS production compared to BAC alone, with cromoglycate showing a greater effect.
Conclusions:
- Unpreserved mast cell inhibitors (sodium cromoglycate, NAAGA) are non-cytotoxic in vitro.
- These inhibitors exhibit antioxidant properties, reducing ROS production.
- Sodium cromoglycate is identified as a potent free radical scavenger.