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Effects of Cigarette Smoke (CS) From Control And Antioxidant Modified Cigarette Filters on Cytotoxicity of Fetal Fibroblasts and Human Lymphocytes

Theodore Hersh, MD, MACG, Abraham Z. Reznick, PhD & Rafael Nagler, DMD, PhD
Thione International, Inc., Atlanta, GA & Technion Institute of Technology, Haifa, Israel
Presented at the American Academy for Advancement of Science Annual Meeting, February 2002

Reactive free radicals derived from the burning of cigarettes contribute to tobacco related diseases, including heart and lung disease and cancer. Antioxidants suppress oxidative stress in smokers and help protect cells. Previous studies have shown that thiol antioxidants neutralize oxidants and aldehydes in CS. This study compares cytotoxicity in fetal fibroblasts and in lymphocytes from exposure to CS of control cigarettes and from same brand with an antioxidant complex incorporated in the filter.

Methods: A smoking device was used to allow CS to be collected and placed on a confluent cell line of WI-38 fetal fibroblasts and on human lymphocytes. Fibroblasts were monitored via Alamar Blue determining cell viability as ability of functioning mitochondria to reduce the dye. Lymphocyte survival was assessed by the Trypan Blue exclusion test using haemocytometer counting. The test cigarette was prepared with an antioxidant complex in a liposome composed of L-glutathione, N-acetyl-L-cysteine and L-selenomethionine incorporated in the plasticizer application during the manufacture of the filter. Control and test cigarettes had the same tobacco rod.

Results: Fibroblast viability from CS exposure averaged 60% and 77% at 24 and 48 hours compared to 95% and 100%, respectively, for the antioxidant treated CS. Lymphocyte survival averaged 65%, 40%, 20% and 8% at 20, 40, 60 and 80 minutes of exposure to the smoke compared to 100%, 82%, 40% and 15% survival of lymphocytes, respectively, for the antioxidant treated CS. Conclusions: An antioxidant complex incorporated in the filter of a cigarette decreases acute cell mortality of fetal fibroblasts and of human lymphocytes after exposure to CS. This antioxidant application in a cigarette filter represents a method to reduce toxicity to cells of inhaled oxidants and aldehydes in cigarette smoke.

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