Dolichos Cell Culture Extract for Protection Against UV Damage

Nov 15, 2013 | Contact Author | By: Marida Bimonte, Annalisa Tito, Antonietta Carola and Ani Barbulova, Arterra Bioscience, Naples, Italy; Fabio Apone and Gabriella Colucci, Vitalab srl and Arterra Bioscience srl, Naples, Italy; Mirna Cucchiara, Intercos SpA, Milan, Italy; and Jacqueline Hill, CRB SA, Puidoux, Switzerland
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Title: Dolichos Cell Culture Extract for Protection Against UV Damage
Dolchios cell culture extractx UV protectionx anti-inflammatoryx antioxidantx
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Keywords: Dolchios cell culture extract | UV protection | anti-inflammatory | antioxidant

Abstract: Dolichos biflorus cell culture extract is evaluated here for its effects on skin cells. Results indicate the extract provides strong anti-inflammatory, antioxidant and protective properties for skin, and the ability to repair UV-induced damage. Further, this work reinforces that cell culture systems are efficient biofactories to produce natural and effective compounds.

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Marida Bimonte et al., e-pub ahead of print, available at: www.cosmeticsandtoiletries.com/formulating/category/suncare/Testing-emDolichosem-Cell-Culture-Extract-for-Protection-Against-UV-Damage-231978261.html; in print, Cosm & Toil 129(3) 46 (April 2014)

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The Dolichos species have been reported to benefit human health with regard to the protection of blood cell progenitors and anti-hepatotoxic, hypo-lipidemic, anti-nephrotoxic and free radical-scavenging activities. Dolichos biflorus (D. biflorus), in particular, is a legume crop with high nutritional value in terms of proteins, vitamins and minerals. For this reason, its beans have been widely used as a dietary source worldwide. In relation, recent studies have shown that plant cell culture extracts in general are rich sources for compounds with a wide range of efficacy and benefits for cosmetic formulations, including phenols, flavonoids and their derivatives capable of neutralizing free radical damage. Compared with plants grown in fields, plant cell cultures prove useful in developing extracts with multiple specific activities on skin cells and desired characteristics, thanks to their totipotency, safety and biosustainability. Here, the authors evaluate a D. biflorus cell culture extract for its effects on skin cells. Results indicate the extract provides strong anti-inflammatory, antioxidant and protective properties for skin, and the ability to repair UV-induced damage. Further, the work presented reinforces cell culture systems as efficient biofactories to produce natural and effective compounds. Skin Aging To investigate the effects of the extract on skin, it is important to first consider mechanisms of the skin aging process. This complex progression is characterized by dramatic morphological changes of the cells and results in wrinkle formation, loss of elasticity and dryness. These effects are mainly determined by chronological aging and photoaging, and amplied by other environmental assaults, such as free radicals, extreme temperatures and pollutants. Prolonged and chronic UV exposure can cause alterations and damage to the various skin cell types—i.e., epidermal keratinocytes, melanocytes, dermal fibroblasts and even cells of the immune system, such as macrophages and neutrophils, which can transiently reside in the skin. These alterations lead to an increase of inflammation, connective tissue degradation and oxidative stress, all accompanied by a decrease in cellular metabolism and functionality. Keratinocytes, which constitute the outermost skin layer, are the most exposed to UV light and thus the most affected by this type of stress. UV light, particularly UVB, induces keratinocytes to express cytokines, which are small proteins that act as inter-cellular messengers and trigger the inflammatory response. During the inflammatory process, blood flow increases and immune cells are attracted by chemical signals to the site of injury, causing skin redness and erythema. Moreover, cytokines induce the dermal fibroblasts to secrete matrix metalloproteases (MMPs) and elastase, which digest collagen and elastin fibers, respectively. As a consequence, the skin loses fitness and vigor, leading to wrinkle formation and sagging, the most visible signs of skin aging.

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Figure 1. Antioxidant profile

Figure 1. Antioxidant profile

Antioxidant profile of the hydrosoluble extract obtained from Dolichos cell cultures as determined by MS/LC analysis

Figure 2. Gene expression analysis of inflammatory cytokines

Figure 2. Gene expression analysis of inflammatory cytokines

Gene expression analysis of inflammatory cytokines (IL-1β, IL-6 and IL-8) in epidermal keratinocytes under UVB stress, treated with Dolichos cell extract and the drug TO9001317 (positive control)

Figure 3. Gene expression analysis of the MMPs

Figure 3. Gene expression analysis of the MMPs

Gene expression analysis of MMP-1 and MMP-3 in dermal fibroblasts under UVA stress, treated with Dolichos cell extract and retinoic acid (positive control)

Figure 4. Comet assays in epidermal keratinocytes

Figure 4. Comet assays in epidermal keratinocytes.

Comet assays in epidermal keratinocytes; after treatment with the reagents indicated, the cells were a) either UVB-irradiated or b) stressed with H2O2.

Figure 5. Common deletion assay

Figure 5. Common deletion assay

Common deletion assay in UV-irradiated epidermal keratinocytes treated with Dolichos cell extract, vitamin E and Coenzyme Q (both as positive controls)

Figure 6. ATP synthesis assay

Figure 6. ATP synthesis assay

ATP synthesis assay in UV-irradiated epidermal keratinocytes treated with Dolichos cell extract and a water-soluble analog of vitamin E (positive control)

Figure 7. Proteasome activity in UV-irradiated epidermal keratinocytes treated with Dolichos cell extract

Figure 7. Proteasome activity in UV-irradiated epidermal keratinocytes treated with <em>Dolichos</em> cell extract

At the highest concentration of 0.01%, the extract completely eliminated the reduction in activity caused by both UVA and UVB, suggesting a strong effect on preserving proteasome functionality.

Figure 8. In vivo results on UV-induced erythema

Figure 8. In vivo results on UV-induced erythema

The extract inhibited the formation of the UV-induced erythema by 23.94% compared with untreated areas, and by 12.79% compared with placebo-treated area.

Figure 9. In vivo results on erythema redness caused by UV radiation

Figure 9. In vivo results on erythema redness caused by UV radiation

Results indicated that D. biflorus cell culture extract reduced the erythema redness caused by UV radiation by 7.56% after 2 hr and 12.98% after 4 hr.

Footnotes [Apone 128(11)]

a The Gamborg B5 salts used to prepare the medium were obtained from Duchefa Biochemie BV, www.duchefa-biochemie.nl.

b Miracloth is a product of EMD Millipore, www.emdmillapore.com.

c Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) is a registered trademark of F. Hoffmann-La Roche Ltd., www.roche.com.

d Ambion QuantumRNA 18S is a product of Life Technologies Corp., www.lifetechnologies.com.

e The Geliance 200 is a device from PerkinElmer, www.perkinelmer.com.

f The QIAamp DNA Mini Kit is a product of Qiagen, www.quiagen.com.

g The CellTiter-Glo Reagent is a product of Promega Corp., www.promega.com.

h The Victor 3, and
j EnVision instruments are devices from PerkinElmer, www.perkinelmer.com.

k The T0901317 agonist is a product of Sigma-Aldrich, www.sigmaaldrich.com.

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