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Antioxidants Protect Fibroblasts From Cytotoxicity Of Cigarette Smoke

Theodore Hersh, MD; Wendy Barkin
Thione International, Inc.
Presented by title; American Academy Of Dermatology, July 2001

The clinical entity "Smoker's Face" results from tissue damage by free radicals (FR) generated by cigarette smoke. FR derived from burning of tobacco contribute to tobacco related diseases both to primary and secondary smokers emanating from these FR and other toxins in mainstream and sidestream smoke, respectively. This study was designed to assess viability of cells in culture exposed to untreated and antioxidant treated cigarette smoke to evaluate the protective effect of antioxidants for "Smoker's Face."

Method: A smoking device was used to allow smoke from a single cigarette to be bubbled through cell culture media, which is then placed on confluent cell lines. The survival of the fibroblasts was monitored for up to 48 hours via Alamar Blue, determining cell viability as ability of "living" mitochondria to oxidize the dye. The tests were repeated placing various single or combined synergistic antioxidants in the cell culture media and coated within a filter prior to introduction of the smoke. Antioxidants tested included glutathione/selenium complex, antioxidant enzymes, grape seed and green tea extracts.

Results: With control smoke, cell viability averaged 60% in fetal fibroblasts, 52% in 26 yr. old and 10% in 83yr. old dermal fibroblasts compared to 84%, 80% and 78%, respectively, with GSH/Se antioxidant treated smoke. Addition of selective antioxidants and enzymes increased cell viability to approximately 100%.

Tobacco gas phase smoke results in significant cell death, greatly accentuated in older cell lines. Synergistic antioxidants yielded reduced cytotoxicity compared to a single antioxidant. Antioxidants reduce cigarette smoke FR and significantly reduce acute mortality in both young and senescent cells. Topical antioxidants may help protect smokers from developing the cutaneous changes which result in "Smoker's Face" from tobacco FR damage.

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