Several phenolic compounds were identified in C. sativa leaf extract obtained under optimized conditions, namely, phenolic acids (chlorogenic acid and ellagic acid) and flavonoids (rutin, isoquercitrin and hyperoside) (fig. 3). Their amounts are expressed in table 2. The major component was found to be rutin. DPPH scavenging activity has been found for these phenolic compounds [13,35,36], and thus their putative contribution to the free radical scavenging activity of the whole extract might be inferred. No skin adverse effects were previously reported for these phenolic com- pounds. In fact, a preventive effect against photo-oxidative stress induced by UVA radiation has been described for rutin, the major phenolic compound found in this extract [37]. Cytotoxicity studies performed with McCoy mouse fibroblast cells also showed that at a concentration of 500 μg/ml, rutin failed to produce any overt signs of toxicity [38]. In accordance, a comparative study between the cyto- tocixity of nine flavonoids towards cultured human normal cells showed that rutin was among the least toxic compounds [39]. Despite the absence of reports of adverse effects of C. sativa leaves or of the polyphenolic compounds found in its composition, safety can not be assumed, and suitable tolerance tests should be carried out. Patch testing after a single application is a widely used procedure to evaluate acute irritant reactions. Initially, evaluation of irritancy testing was based on visual scoring only. This type of evaluation, although subjective, can be a sensitive, reliable and repro- ducible method [40]. Several bioengineering techniques have been developed to provide objective and quantitative data. Although they present significant advantages over visual assessment, non-invasive evaluation methods of skin irrita- tion measure only one of its particular aspects. TEWL is considered to be the first choice to evaluate slight skin reactions, as it detects disrupted epidermal barrier that results in a higher loss of transepidermal water [41]. However, water barrier function and water evaporation from the skin is neither felt nor seen and, therefore, findings based on TEWL measurements are indirect and not automatically clinically relevant [15]. Erythema, sometimes associated with infiltration and sometimes with superficial erosion of the epithelium, is the main feature of acute inflammatory reaction. The evaluation of irritant reactions was carried out with a combined approach in order to cover the different aspects of skin irritation.
Several phenolic compounds were identified in C. sativa leaf extract obtained under optimized conditions, namely, phenolic acids (chlorogenic acid and ellagic acid) and flavonoids (rutin, isoquercitrin and hyperoside) (fig. 3). Their amounts are expressed in table 2. The major component was found to be rutin. DPPH scavenging activity has been found for these phenolic compounds [13,35,36], and thus their putative contribution to the free radical scavenging activity of the whole extract might be inferred. No skin adverse effects were previously reported for these phenolic com- pounds. In fact, a preventive effect against photo-oxidative stress induced by UVA radiation has been described for rutin, the major phenolic compound found in this extract [37]. Cytotoxicity studies performed with McCoy mouse fibroblast cells also showed that at a concentration of 500 μg/ml, rutin failed to produce any overt signs of toxicity [38]. In accordance, a comparative study between the cyto- tocixity of nine flavonoids towards cultured human normal cells showed that rutin was among the least toxic compounds [39]. Despite the absence of reports of adverse effects of C. sativa leaves or of the polyphenolic compounds found in its composition, safety can not be assumed, and suitable tolerance tests should be carried out. Patch testing after a single application is a widely used procedure to evaluate acute irritant reactions. Initially, evaluation of irritancy testing was based on visual scoring only. This type of evaluation, although subjective, can be a sensitive, reliable and repro- ducible method [40]. Several bioengineering techniques have been developed to provide objective and quantitative data. Although they present significant advantages over visual assessment, non-invasive evaluation methods of skin irrita- tion measure only one of its particular aspects. TEWL is considered to be the first choice to evaluate slight skin reactions, as it detects disrupted epidermal barrier that results in a higher loss of transepidermal water [41]. However, water barrier function and water evaporation from the skin is neither felt nor seen and, therefore, findings based on TEWL measurements are indirect and not automatically clinically relevant [15]. Erythema, sometimes associated with infiltration and sometimes with superficial erosion of the epithelium, is the main feature of acute inflammatory reaction. The evaluation of irritant reactions was carried out with a combined approach in order to cover the different aspects of skin irritation.
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