613 CENTELLA ASIATICA EXTRACTS IN SKIN REPAIR PRODUCTS article describes the results of the clinical and laboratory tests made with Blastoactiva™, formulated with two main active constituents from C asiatica: asiaticoside and asiatic acid, which contribute to enhance skin repair and regeneration. Thus, based on the properties of asiaticoside and asiatic acid, the cosmetic indications of Blastoactiva™ include the repair of skin damaged by dryness (xerosis), superficial acne scarring, and superficial aggression produced by physical agents (e.g., cold, wind, pollution, solar exposure), and cosmetic treatments (e.g., laser, peeling, depilation and epilation, tattoos, and phototherapy sessions). METHODS AND RESULTS IN VITRO AND IN VIVO EVALUATION OF TOLERABILITY In vitro evaluation of eye irritant potential. The in vitro evaluation of the eye irritant potential of Blastoactiva™ was carried out based on a test involving application of the product in the chorioallantoic membrane (CAM) of 10-day incubated chick embryos. This technique constitutes an alternative to animal testing and is similar to that described by Luepke and Kemper (28). Use was made of four white Leghorn chicken eggs (40–70 g) incubated during 10 d under controlled temperature (37.8 ± 1°C) and relative humidity conditions (50–60%) on oscillating plates in the vertical position with the egg air chamber facing upwards. After breaking and sectioning the shell, 300 µL of the product was deposited onto the CAM with a micropipette. After 20 s of contact, the CAM was cleansed with 5 mL of isotonic saline solution. The following characteristics were analyzed over the subsequent 5 min: appearance of hyperemia (appearance of visible capillaries or dilatation and reddening of existing capillaries), hemorrhage (diffuse, in layers or punctate), and coagulation (opacity and/or thrombosis) after ≤30 s, between 30 s and ≤2 min, and 2 min and ≤5 min after contact with the CAM. The cases of hyperemia, hemorrhage, and coagulation were summed for each egg with a total score of 0 to 21 and classified as: 1 is practically nonirritating ≥1 to 5 is slightly irritating ≥5 and 9 is moderately irritating and ≥9 is irritating. Negative and positive controls were included in the form of a 0.8% NaCl solution and different concentrations of sodium lauryl sulfate, respectively. The results obtained in vitro, with a mean (± standard deviation) of 0 ± 0 for hyperemia, hemorrhage, and coagulation phenomena, showed the product to cause practically no eye irritation. Allergenic potential. The allergenic potential of Blastoactiva™ was studied in 50 volunteers of either gender and between 18 and 70 yr of age with a Fitzpatrick I–V skin phototype and sensitive skin. For this purpose, use was made of the Human Repeat Insult Patch Test (HRIPT) described by Marzulli in 1973 (29). The study was approved by the evaluation committee of EVIC Romania (Bucharest, Romania) (registry CI-313/08). In accordance with standard practice of the HRIPT, the volunteers were exposed to the study product using adhesive patches (Finn Chamber standard®, Merck spol s.r.o., Bratislava, The Slovak Republic) with an aluminum cap containing 20 μL (∼20 mg) of Blastoactiva™. The patches were affixed to a zone of the skin with hypoallergenic adhesive tape (Scanpor® [Norgesplaster, Granlivegen 21, NO-4707 Vennesla Norway]) with an internal diameter of 8 mm and surface area of 50 mm2). Another 20 additional products were tested in parallel in the same way.

614 JOURNAL OF COSMETIC SCIENCE During the first 3 weeks (induction or sensitization phase), the patches were applied intermittently on days D1, D3, D5, D8, D10, D12, D15, D17, and D19, with removal after 48 h of contact of those patches that had been placed on days D3, D5, D10, D12, D17, and D19, and after 72 h of those placed on days D8, D15, and D22. This period was followed by a 15-day resting phase. Finally, the patch with Blastoactiva™ was again placed on day D37 (challenge phase), and was removed 48, 72, and 96 h after application (days D39, D40, and D41). The product was applied onto the skin of the dorsal region. Test reading was performed by the dermatologist based on inspection of the study zone for 15 min (or longer if alterations were observed). The appearance of a positive test includes erythema, edema, vesicles, blisters, papules, dryness, skin coloring, and a soapy effect (reddened, glass-like appearance). The scale of the International Contact Dermatitis Research Group was used to evaluate the capacity of the product to cause allergic contact dermatitis, based on the following grades: ?+ (doubtful reaction), + (erythema, infiltration, minimum papule), ++ (erythema, papules, vesicles), and +++ (blisters). Likewise, the participants were questioned about possible reactions. No skin reaction of any kind was observed in either the induction phase or in the challenge phase. Blastoactiva™, therefore, can be regarded as a product for topical use with low clinical potential to cause allergic contact dermatitis. Phototoxic potential. The phototoxic potential of Blastoactiva™ was studied in 12 healthy volunteers (nine women and three men) between 18 and 65 yr of age (with a mean age of 48.3) with a Fitzpatrick I–III skin phototype and no skin alterations capable of influencing the test results. The participants gave written informed consent to participation, and the study protocol was approved by the evaluation committee of EVIC Hispania, Centro Experimental de Evaluación Cutánea (Barcelona, Spain) (registry CEN-41-09). The test involved the application with occlusive patches of 0.2 mL of the product onto two 4 cm2 zones on the dorsal region between the waist and scapula, while a third zone without product was used as control. The patches were removed after 24 h. Following visual inspection of the three zones and having discarded the presence of allergic contact dermatitis according to the International Contact Dermatitis Research Group scale, one of the treated areas and the control zone were irradiated with filtered light (with a wavelength of 320–400 nm) equivalent to 10.0 J/cm2 of ultraviolet irradiation. Skin reaction in the irradiated zones was assessed after 24, 48, and 72 h. No phototoxic reactions were observed in the area treated with Blastoactiva™ and irradiated, in the treated and nonirradiated zone, or in the irradiated control zone. The tested product therefore can be considered to lack phototoxic capacity. Blastoactiva™ is therefore a cosmetic product that has shown negligible risk of eye irritation in vitro, no allergic contact dermatitis, and no phototoxic effects, and can be regarded as safe. CLINICAL EFFICACY AND ACCEPTABILITY Study in healthy adult volunteers. Three clinical efficacy variables were evaluated in 36 healthy volunteers exposed to Blastoactiva™: anti-irritant action, skin barrier function repair, and epidermal cell renewal capacity. The volunteers served as their own control. The participants gave written informed consent to participation, and the study protocol (N°: 08-0319/1) was approved by the evaluation committee of EVIC Hispania, Centro