The antioxidant activity and cellular effects of the bioactive components of Flavonix® Cytoflamm Gel used in wound treatment

Abstract

Wound healing is one of the most complex processes in the human body consisting of several different phases namely inflammatory phase, proliferative phase, remodeling phase and contraction phase (Jurjus et al., 2007). Bioactive ingredients of Cytoflamm Gel (FCG), a complementary wound healing product are honey (H), oleuropein (OL), witch hazel (WH), xylitol (X), vitamin E (VE), aloe vera (AV) and farnesol (Fa). The aim of this study was to evaluate the antioxidant properties and cellular protective effects of FCG. This required the testing of each bioactive ingredient as well as related to antioxidant effects the interaction that occurs between ingredients. This was achieved by measuring the antioxidant content and activity (chemical and cellular) of FCG and each ingredient at the concentrations found in FCG. Two groups of ingredients related to concentration were identified, major (H, OL and WH) and minor (X, VE, AV, Fa). A strategy was developed to determine the type of interaction; synergistic, additive or antagonistic that occurs between ingredients. For all samples, the total polyphenolic and flavonoid content (TPC and TFC) was determined. Antioxidant activity was measured using the 2,2'-diphenyl-2-picrylhydrazyl (DPPH), trolox equivalent antioxidant capacity (TEAC) both single electron transfer assays and the hydrogen atom transfer assay, as well as the oxygen radical antioxidant capacity (ORAC) assay. Cellular antioxidant protective effect/s was determined in the SC-1 cell line with the dichlorofluorescein diacetate assay with 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) as source of oxidative damage. The high antioxidant content of FCG was due to major components OL and H contributing 84.08% and 81.70% respectively to TPC and TFC. VE, OL and H, contributed 44.56%, 32.06% and 11.56% (DPPH assay) and 56.44%, 15.12% and 13.80% (TEAC assay) respectively to antioxidant activity. With the ORAC assay, H, OL and WH were found to contribute equally 29.97%, 29.91% and 29.67% with a contribution of only 7.59% by VE. Strong synergism was found between H+OL, H+WH and OL+WH. In the in vitro SC-1 cell model, FCG, the antioxidant ingredients and mixtures showed significant cellular protective effects especially in combination with VE. Strong synergism was found between VE+OL, VE+OL+H and VE+OL+H+WH, indicating that effects may be related to antioxidant type and concentration although in some instances the effect of WH was antagonistic. In conclusion, FCG and bioactive ingredients have significant levels of antioxidant activity and cellular protection against oxidative damage and this is due to synergism between antioxidant ingredients especially VE, OL and H.