Raspberry Stem Cell Extract to Protect Skin from Inflammation and Oxidative Stress

Jul 1, 2010 | Contact Author | By: Ani Barbulova, Annalisa Tito, Antonietta Carola, Marida Bimonte, Francesco De Laurentis, Pasquale D’Ambrosio, Fabio Apone and Gabriella Colucci, Arterra Bioscience; Irene Monoli and Mirna Cucchiara, INTERCOS SpA; Jacqueline Hill, CRB SA
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Title: Raspberry Stem Cell Extract to Protect Skin from Inflammation and Oxidative Stress
Rubus idaeus stem cellsx anti-inflammationx oxidative stress protectionx
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Keywords: Rubus idaeus stem cells | anti-inflammation | oxidative stress protection

Abstract: In the present article, the authors evaluate a wild red raspberry (Rubus idaeus) extract for its beneficial compounds and potential applications in skin care. From leaf explants, raspberry stem cells were obtained to develop an extract rich in antioxidants, anthocyanins and phenolics. This extract exhibited anti-inflammatory and antioxidant activities.

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A Barbulova et al, Raspberry stem cell extract to protect skin from inflammation and oxidative stress, Cosm & Toil 125(7) 38-47 (Jul 2010)

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Skin cells, in particular keratinocytes and fibroblasts, are the cells most exposed to environmental stressors such as UV light, smog, toxins, cigarette smoke and pollution. Once these cells are damaged, skin loses firmness and vigor, becoming weaker and more vulnerable to diseases. Skin inflammation is indeed one of the most serious consequences of environmental stress. Although inflammation is the first line of defense against microorganisms and external assault, it can also lead to visible signs of aging. In fact, during the inflammatory process, free radicals are generated, blood flow increases and immune cells are attracted by chemical signals to the site of injury.

In typical cases, when the inflammatory response is no longer required, the physiological events are rapidly terminated. However, in the case of prolonged and persistent environmental stress, this termination process fails, leading to chronic inflammation characterized by the continuous production of free radicals and further causing DNA damage, wrinkles and skin aging.

DNA integrity is the most important prerequisite for the fulfillment of vital cellular functions: The loss of functionally important genes compromises the health of cells and accelerates the skin aging process. Recent studies have demonstrated that in the complex scenario of signal transduction mechanisms activated to protect cells, there are elements that play pivotal roles to guarantee DNA integrity. Among these elements is the family of GADD45 proteins, which assists cells by preventing accumulated mutations in the nuclear DNA. These proteins act as stress sensors and are able to mediate a variety of interactions among different cellular components, triggering processes of DNA repair and protection.

Other elements that maintain DNA integrity and stability are sirtuins—proteins that regulate cell longevity. Sirtuins are present in all multicellular organisms and regulate the activity of transcription factors and DNA-binding proteins, acting as nucleus stabilizing factors.


Lab Practical: Using Rubus idaeus Extract

  • The obtained Rubus idaeus extract is a hydrosoluble, odorless, pale yellow powder. 
  • The extract is soluble in aqueous medium, glycerin and ethanol and is stable at pH 4.0-8.0. 
  • Once dissolved, the extract can be incorporated during cooling or at the end since it can withstand temperatures of up to 70°C.
  • Due to its hydrophilic nature, the extract is not recommended for anhydrous systems. 
  • Dilutions of 1.0% and 0.5% are recommended.

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Table 1. Compatibility of Rubus idaeus extract

Table 1. Compatibility of <em>Rubus idaeus</em> extract

The described ingredient can be easily incorporated in polar or weakly polar solvents, which represent the majority of ingredients used in cosmetic formulations.

Figure 1. Antioxidant profile of the Rubus stem cell extract

Figure 1. Antioxidant profile of the <em>Rubus</em> stem cell extract as determined by MS/LC

The extract obtained was analyzed by mass spectrometry liquid chromatography (MS-LC) as described, which showed the extract to be rich in phenolic acids and flavonoids, the most abundant being ferulic acid and quercitin ramnoside.

Figure 2. Gene expression analysis of iNOS2 and COX2 in macrophages

Figure 2. Gene expression analysis of iNOS2 and COX2 in macrophages

The induction of these genes caused by the bacterial inflammatory agent lipopolysaccharide (LPS) was significantly attenuated by the presence of the raspberry stem cell extract, suggesting a potential anti-inflammatory effect on skin.

Figure 3. Total antioxidant power of Rubus idaeus stem cell extract

Figure 3. Total antioxidant power of <em>Rubus idaeus</em> stem cell extract as measured by a) TAC and b) ORAC assays

The results of the TAC and ORAC assays as measured by a) TAC and b) ORAC assays; the extract shows excellent antioxidant power—higher than that calculated for resveratrol, which also is known for good antioxidant and cell protection properties.

Figure 4. Production of ROS in NIH-3T3 cells

Figure 4. Production of ROS in NIH-3T3 cells

Production of ROS in NIH-3T3 cells treated with varying amounts of Rubus idaeus stem cell extract and in comparison with ascorbate

Figure 5. Gene expression analysis of a) GADD45α or b) SIRT1 genes

Figure 5. Gene expression analysis of a) GADD45α or b) SIRT1 genes

Gene expression analysis of a) GADD45α or b) SIRT1 genes in NIH-3T3 cells treated with Rubus idaeus stem cell extract, compared with resveratrol and ascorbate

Figure 6. Comet assay

Figure 6. Comet assay

Treatment with the extract clearly produced a protective effect on DNA from H2O2-induced damage.

Footnotes [Barbulova 125(7)]

a Gamborg B5 medium, a product of Duchefa Biochemie, was used for this study.

b Miracloth fabric is product of Calbiochem.

c The HPLC apparatus equipped with two Micropumps Series 200 used for this study is manufactured by Perkin Elmer, Shellton, CT, USA.

d UV/VIS series detector 200 used for this study is manufactured by Perkin Elmer, Shellton, CT, USA.

e The Prodigy ODS3 100 Å column used for this study is manufactured by Phenomenex, CA, USA.

f The API 3000 triple quadrupole mass spectrometer used for this study is product of Applied Biosystems, Canada.

g The Supelcosil LC-18A HPLC Column used for this study is manufactured by Supelco, Bellefonte, PA, USA.

h Trolox is a product of Hoffman-LaRoche.

j The resveratrol used for this study is a product of Sigma-Aldrich.

k The non-enzymatic detaching solution used for this study is a product of Sigma-Aldrich.

Formula 1. Hydrating emulsion

Cetearyl Alcohol, 2.500% w/w
Hydrogenated Olive Oil (and) Olea Europaea Fruit Oil (and) Olea Europaea Oil Unsaponifiables (and) Shea Butter, 4.500
Vitamin E Acetate, 0.200
Water (Aqua), qs to 100.00
Butylene Glycol (and) Glycerin, 6.500
Xanthan Gum, 0.250
Glyceryl Stearate (and) Cetearyl Alcohol (and) Potassium Behenoyl Hydroxyproline (and) Water (Aqua) (and) Behenic Acid (and) Phenoxyethanol (and) Rosa Centifolia Flower Extract (Roseflor 22 HYP; CRB), 6.500
Acrylates/C10-30 Alkyl Acrylate Crosspolymer, 0.200
Sodium Hydroxide, qs to pH 6.0
Preservative, qs
Rubus Idaeus Stem Cell Extract (Cellintegrity, Arterra Bioscience/CRB), 1.000

Formula 2. Skin care serum

Water (Aqua), qs to 100.00%
Disodium EDTA, 0.100
Glycerin (and) Butylene Glycol, 6.500
Xanthan Gum, 0.200
Carbomer, 0.200
Potassium Cetyl Phosphate, 0.250
Glyceryl Stearate (and) PEG-100 Stearate, 1.500
Cetyl Alcohol, 1.000
Vegetable Oil, 1.000
Coco-caprylate (and) Caprate (and) Ethylhexylpalmitate (and) Octyldodecanol, 10.000
Antioxidant, 0.020
Dimethicone, 2.000
Sodium Hydroxide, qs to pH 6.5
Preservatives, qs
Rubus Idaeus Stem Cell Extract (Cellintegrity, Arterra Bioscience/CRB), 0.500

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