Coffea bengalensis for Antiwrinkle and Skin Toning Applications

Sep 1, 2011 | Contact Author | By: Marida Bimonte, Antonietta Carola, Annalisa Tito, Ani Barbulova and Francesca Carucci, Arterra Bioscience; Fabio Apone and Gabriella Colucci, Arterra Bioscience and Biolab; Irene Monoli and Mirna Cucchiara, INTERCOS SpA; and Jacqueline Hill, CRB SA
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Title: Coffea bengalensis for Antiwrinkle and Skin Toning Applications
Coffea bengalensis stem cell extractx antiwrinkle activityx hydrationx collagen synthesisx adipocyte differentiationx
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Keywords: Coffea bengalensis stem cell extract | antiwrinkle activity | hydration | collagen synthesis | adipocyte differentiation

Abstract: Coffea species have been widely evaluated for effects including oxidative stress protection and immunomodulation. The present study therefore evaluates the cosmetic activity of an extract obtained from cultured Coffea bengalensis stem cells. Results showed the extract imparted hydration activity in the keratinocytes, stimulated collagen synthesis in the fibroblasts and enhanced adipocyte differentiation starting from mesenchymal precursor cells.

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Coffea bengalensis for Antiwrinkle and Skin Toning Applications

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The aging process leads to a loss of firmness and elasticity in the skin, causing tissues to collapse and the subsequent production of wrinkles. This wrinkle formation involves dramatic structural alterations in the epidermis, dermis and hypodermis. For example, alterations of the hydration state and water-holding capacity of the epidermis have been found to contribute significantly to the formation of wrinkles1 and can easily be associated with a deficiency of water mostly due to the drying activity of environmental factors such as UV, light, heat and pollution.2

The epidermis is mainly composed of keratinocytes, which proliferate in the stratum basale (SB), differentiate and migrate toward the skin surface. The outermost layer of the epidermis, the stratum corneum (SC), is mostly composed of flattened and cornified keratinocytes, which produce high amounts of keratin during the differentiation process. Dead keratinocytes are held together by junctions and microfibrils composed of different insoluble proteins that are essential for the skin’s mechanical integrity and waterproof properties.

Involucrin (INV) and filaggrin (FLG) are two important proteins in the epidermis, and both contribute to the structural integrity of the SC.3 Moreover, FLG is a source of natural moisturizing factors (NMF), as it generates precursors of the NMF upon hydrolysis.4 Another important factor that plays a key role in skin hydration and integrity of the epidermis is aquaporin (AQP); specifically, aquaporin 3 (AQP3), located at the plasma membrane in the SB and stratum spinosum, where it plays an essential role in water transport.5

In the dermis, the fibroblasts are mainly responsible for the production of all the components of the extra-cellular matrix (ECM), such as collagen, elastin, hyaluronan and proteoglycans. The formation of wrinkles is associated with changes in the elastic and supporting proprieties of the ECM, mainly due to the denaturation of the collagen fibers, and/or to an unbalanced ratio of collagen synthesis and degradation.6 Visible signs of aging also appear as the adipocytes present in the hypodermis cannot fill in the skin and compensate for damages in the outer layers. Adipocytes provide volume and help add thickness and suppleness to the skin.7 In time, there is a decline in the capacity of pre-adipocytes to differentiate and of fat cells to create new lipids. This contributes to a decrease in fat mass and to alterations in fat tissue function.8

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Table 1. Organoleptic characteristic of Coffea stem cell extract, in aqueous solution at 5%, to pH variations

Table 1. Organoleptic characteristic of Coffea stem cell extract, in aqueous solution at 5%, to pH variations

The extract also has been shown to remain stable at pH ranges from 4.0 to 8.0, as illustrated in Table 1.

Figure 1. Gene expression analysis of filaggrin (FLG), involucrin (INV) and aquaporin-3 (AQP3) in keratinocytes treated with C. bengalensis cell extract

Figure 1. Gene expression analysis of filaggrin (FLG), involucrin (INV) and aquaporin-3 (AQP3) in keratinocytes treated with <em>C. bengalensis</em> cell extract

As shown in Figure 1, at a concentration of 0.05%, the extract increased the expression of INV and FLG by 40% and 42%, respectively.

Figure 2. Production of new collagen (type I and III) in fibroblasts after treatment with C. bengalensis cell extract and ascorbate

Figure 2. Production of new collagen (type I and III) in fibroblasts after treatment with <em>C. bengalensis</em> cell extract and ascorbate

After treatment with the extract at a concentration of 0.03%, the production of new collagen (pro-collagen) of type I and III increased by 80% and 45%, respectively, suggesting it could also have a firming activity in the dermis (see Figure 2).

Figure 3. Differentiation of hMSC into adipocytes after treatment with C. bengalensis cell extract, and with nicotinamide (positive control)

Figure 3. Differentiation of hMSC into adipocytes after treatment with <em>C. bengalensis</em> cell extract, and with nicotinamide (positive control)

The values obtained illustrated that the cell extract had a strong stimulatory effect on adipocyte differentiation, indicating its potential capacity to counteract the collapse of tissues caused by aging (see Figure 3).

Figure 4. The Coffea bengalensis cell extract activates mechanisms associated with hydration, firming and replumping in the skin.

Figure 4. The <em>Coffea bengalensis</em> cell extract activates mechanisms associated with hydration, firming and replumping in the skin.

The in vitro results thus far suggest that C. bengalensis cell extract acts independently on each of the three main layers forming the skin, activating mechanisms of hydration, firming and replumping, which are all important to reducing the visible signs of aging (see Figure 4).

Figure 5. Production of: a) ROS in fibroblasts treated with Coffea bengalensis cell extract and with ascorbate, and b) ATP in fibroblasts treated with Coffea bengalensis cell extract

Figure 5. Production of: a) ROS in fibroblasts treated with <em>Coffea bengalensis</em> cell extract and with ascorbate, and b) ATP in fibroblasts treated with <em>Coffea bengalensis</em> cell extract

Moreover, it produced a 35% increase of ATP in the cells, as shown in Figure 5.

Footnotes (CT1109 Apone)

a The GenElute Mammalian Total RNA Purification Kit is a product of Sigma-Aldrich Inc., St. Louis, USA.
b QuantumRNATM 18S internal standard kit is manufactured by Ambion, Austin, Texas, USA.
c The Geliance 200 Imaging system is manufactured by PerkinElmer Inc., Waltham, Ma., USA.
d The primary goat monoclonal antibodies raised against pro-collagen type I and type III used for this study are manufactured by Santa Cruz Biotechnology Inc., Santa Cruz, Calif., USA.
e The AdipoRed reagent used for this study is a product of Lonza, Basel, Switzerland.
f The CellTiter-Glo Reagent used for this study is a product of Promega.
g The Victor 3 plate reading instrument is manufactured by PerkinElmer Inc., Waltham, Ma., USA.
h CMDCFDA is a product of Invitrogen, Carlsbad, Calif., USA.
j The EnVision device is manufactured by PerkinElmer Inc., Waltham, Ma., USA.

Formula 1. Restoring cream

Formula 1. Restoring cream

Two examples of formulations that include the C. bengalensis stem cell ex- tract at 0.5% are shown in Formulas 1 and 2.

Formula 2. Lip contour

Formula 2. Lip contour

Two examples of formulations that include the C. bengalensis stem cell ex- tract at 0.5% are shown in Formulas 1 and 2.

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