DELIVERING EFFECTIVENESS ENHANCING THE GROWTH OF NATURAL EYELASHES IN COLOR COSMETICS FORMULATION: THE MECHANISM OF PLANT-BASED ACTIVE INGREDIENTS WITH IN VITRO AND IN VIVO CORRELATION
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Presented by: Milene Haraguchi Padilha
INTRODUCTION Few research has been done about the human eyelash, however, recent discoveries on the role of eyelashes and its distinctive characteristics have led to an increased scientific interest. Prominent eyelashes are often considered a sign of beauty and can be associated with increased levels of attractiveness, confidence, and well-being. Mascara aims to improve the esthetic appearance temporarily, providing darken, lengthen, and thicken eyelashes by using a combination of waxes, pigments, and resins. Little is known about the mechanism of action of natural plant-based active ingredients and its effectiveness enhancing the growth and volume increase of natural eyelashes in color cosmetics formulation. To delivery these innovative performances while still maintaining the mascara must-haves was a challenge accepted.
OBJECTIVE This work aims to study the effectiveness and to evaluate the correlation between in vitro and in vivo methods in the ability to provide eyelash growth of an oil/wax-in water emulsion formulation of mascara. The proposed mechanisms by which the topical application of natural-based ingredients enhances eyelash growth and volume is also approached.
MATERIALS AND METHODS The following natural plant-based active ingredients were evaluated in this study: a composition of Arginine/Glycine Soybean Germ Extract/Triticum Vulgare Germ Extract/Scutellaria Baicalensis Root Extract (active A) and a composition of Hydrolyzed Ceratonia Siliqua Seed Extract/Hydrolyzed Soy Protein/Zea Mays Starch/Amaranthus Hypochondriacus Leaf Extract (active B). Baicalin was elected as a model for in vitro validation and a water-based formulation, composed by texture modifiers, emollients, emulsifier, film formers, pigments with the active ingredients, were used for the in vivo study.
In vitro tests were performed to evaluate the effect of baicalin on primary human keratinocytes and dermal papilla cell spheroids. Cells in monolayer or three-dimensional model were treated with 0.032% active in culture medium and gene expression for B-catenin, BCL-2, VEGF and CK15 was quantified by qPCR. In addition, complementary efficacy studies were carried out with the objective of confirming the capacity for growth and volume increase of natural eyelashes through an color cosmetic formulation, considering the use of active ingredient A and B with established concentrations.
The in vivo study were conducted with 30 female participants and the color cosmetic formulation was applied by each of the participants for 45 days. Facial region images were captured using the Reveal Imagner® equipment (Canfield), in a standardized way, before applications (D0) and after 45 days of product application (D45), to determine the size and volume of eyelash. The product results were calculated through the treatment of the measurement area in pixels, in relation to time.
RESULTS AND DISCUSSION With the in vitro tests performed, there was a significant increase in the expression of the B-catenin and BCL-2 genes, related to the maintenance and prolongation of the anagen phase, and an increase in VEGF, a growth factor that induces vascularization follicular tissue helping to prevent apoptosis. In addition, treatment with the active induced an increase in CK15, a biomarker of stem cell potential, in neopapiles, demonstrating the potential of the active in aging prevention and hair growth.
Finally, in the in vivo test, the product promoted a statistically significant increase of eyelashes length and volume after 45 days of use, providing an increase of 8% eyelash growth and 9% increase of volume, supporting the benefits promised by the product, expected by the consumers and supporting its value proposition. The product perception evaluation is also a strong indication of the fulfillment of consumer acceptability, presenting highly favorable purchase intent results in 87% and proof of the eyelash growth action in 80%.
The innovative mechanism through the combination of biofunctional actives evaluated, prolongs the anagen phase inducing growth, in addition to strengthening its anchorage in the follicle, preventing hair loss by reducing the telogen phase and protecting its integrity. These are known mechanisms for hair, however, have been now innovatively proven for eyelashes as well.
CONCLUSION Eyelashes have an important role in determing beauty, and as such, prominent eyelashes are a highly sought-after attribute.
The combination of in vitro and in vivo tests demonstrates the action of the natural plant-based actives present in the color cosmetic formulation, to promote growth and improve the volume of eyelashes through the prolongation of anagen phase and maintenance of the stem cell potential of follicular cells.
OBJECTIVE This work aims to study the effectiveness and to evaluate the correlation between in vitro and in vivo methods in the ability to provide eyelash growth of an oil/wax-in water emulsion formulation of mascara. The proposed mechanisms by which the topical application of natural-based ingredients enhances eyelash growth and volume is also approached.
MATERIALS AND METHODS The following natural plant-based active ingredients were evaluated in this study: a composition of Arginine/Glycine Soybean Germ Extract/Triticum Vulgare Germ Extract/Scutellaria Baicalensis Root Extract (active A) and a composition of Hydrolyzed Ceratonia Siliqua Seed Extract/Hydrolyzed Soy Protein/Zea Mays Starch/Amaranthus Hypochondriacus Leaf Extract (active B). Baicalin was elected as a model for in vitro validation and a water-based formulation, composed by texture modifiers, emollients, emulsifier, film formers, pigments with the active ingredients, were used for the in vivo study.
In vitro tests were performed to evaluate the effect of baicalin on primary human keratinocytes and dermal papilla cell spheroids. Cells in monolayer or three-dimensional model were treated with 0.032% active in culture medium and gene expression for B-catenin, BCL-2, VEGF and CK15 was quantified by qPCR. In addition, complementary efficacy studies were carried out with the objective of confirming the capacity for growth and volume increase of natural eyelashes through an color cosmetic formulation, considering the use of active ingredient A and B with established concentrations.
The in vivo study were conducted with 30 female participants and the color cosmetic formulation was applied by each of the participants for 45 days. Facial region images were captured using the Reveal Imagner® equipment (Canfield), in a standardized way, before applications (D0) and after 45 days of product application (D45), to determine the size and volume of eyelash. The product results were calculated through the treatment of the measurement area in pixels, in relation to time.
RESULTS AND DISCUSSION With the in vitro tests performed, there was a significant increase in the expression of the B-catenin and BCL-2 genes, related to the maintenance and prolongation of the anagen phase, and an increase in VEGF, a growth factor that induces vascularization follicular tissue helping to prevent apoptosis. In addition, treatment with the active induced an increase in CK15, a biomarker of stem cell potential, in neopapiles, demonstrating the potential of the active in aging prevention and hair growth.
Finally, in the in vivo test, the product promoted a statistically significant increase of eyelashes length and volume after 45 days of use, providing an increase of 8% eyelash growth and 9% increase of volume, supporting the benefits promised by the product, expected by the consumers and supporting its value proposition. The product perception evaluation is also a strong indication of the fulfillment of consumer acceptability, presenting highly favorable purchase intent results in 87% and proof of the eyelash growth action in 80%.
The innovative mechanism through the combination of biofunctional actives evaluated, prolongs the anagen phase inducing growth, in addition to strengthening its anchorage in the follicle, preventing hair loss by reducing the telogen phase and protecting its integrity. These are known mechanisms for hair, however, have been now innovatively proven for eyelashes as well.
CONCLUSION Eyelashes have an important role in determing beauty, and as such, prominent eyelashes are a highly sought-after attribute.
The combination of in vitro and in vivo tests demonstrates the action of the natural plant-based actives present in the color cosmetic formulation, to promote growth and improve the volume of eyelashes through the prolongation of anagen phase and maintenance of the stem cell potential of follicular cells.