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Witch hazel ( Hammamelis virginiana) bark is a rich source of both condensed and hydrolizable oligomeric tannins. From a polyphenolic extract soluble in both ethyl acetate and water, we have generated fractions rich in pyrogallol-containing polyphenols (proanthocyanidins, gallotannins, and gallates). The mixtures were highly active as free radical scavengers against ABTS, DPPH (hydrogen donation and electron transfer), and HNTTM (electron transfer). They were also able to reduce the newly introduced TNPTM radical, meaning that they included some highly reactive components. Witch hazel phenolics protected red blood cells from free radical-induced hemolysis and were mildly cytotoxic to 3T3 fibroblasts and HaCat keratinocytes. They also inhibited the proliferation of tumoral SK-Mel 28 melanoma cells at lower concentrations than grape and pine procyanidins. The high content in pyrogallol moieties may be behind the effect of witch hazel phenolics on skin cells. Because the most cytotoxic and antiproliferative mixtures were also the most efficient as electron transfer agents, we hypothesize that the final putative antioxidant effect of polyphenols may be in part attributed to the stimulation of defense systems by mild prooxidant challenges provided by reactive oxygen species generated through redox cycling.

There is a great interest in characterizing the biological properties of natural compounds obtained from plants, especially polyphenols. We studied the structure-activity-cytotoxicity relationships of polyphenolic fractions obtained from grape pomace and pine bark. These fractions contained similar polymerised flavonoids but different percentages of pyrogallol groups that confer on them different biological properties. The human keratinocyte cell line HaCaT and the mouse fibroblast cell line 3T3 were used to study the cytotoxicity of the different fractions after 24, 48, and 72 h of exposure. Antioxidant activity of the fractions was evaluated by measuring the inhibition of hemolysis mediated by AAPH. Our results demonstrate that the polyphenolic fractions studied show high antioxidant capacity in a concentration range that is not harmful to normal human cells. Pine fractions presented slightly lower antioxidant activity than grape fractions but are less cytotoxic. This data provides useful information to help design safe antioxidant products that act without altering critical cell functions.

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