RESULTS
The effect of the antioxidant matrix on viability of human dermal fibroblasts after 48 hours is depicted in Figure 1. Viability of untreated cells has been set equal to 100% and the corresponding effect of treatment with the antioxidant matrix is expressed in relation to that.As the lowest concentrations assessed eg, 0.001%, 0.005%, and 0.01%, did not affect viability of human dermal fibroblasts after 48 hours below a range of 80%, the named concentrations have been used for testing the ability of Berrimatrix to protect from IRA-induced up-regulation of MMP1.Next, the ability of the antioxidant matrix to protect from infrared A induced up-regulation of MMP1 was studied (Figure 2). Infrared A irradiation in a dose of 360J/cm2 significantly induced MMP1 up-regulation by 6.1±0.31 (mean±SE). The application of the antioxidant matrix alone did not significantly affect basal MMP1 expression (ANOVA on ranks SNK, P less than 0.05). The fact that the basal MMP1 expression of Berrimatrix-treated fibroblast followed an inverse dose-dependency might be explained by the antioxidants included in the blueberry extract itself or the additional compounds included in this antioxidant matrix. Accordingly, antioxidants such as ascorbate, but also flavonoids and many polyphenols, can undergo rapid oxidation in cell culture media resulting in formation of H2O2 and other reactive oxygen species.7 The latter may lead to up-regulation of genes also modulated by reactive oxygen species. The cell culture medium itself does not promote further oxidation because no iron is included.Pre-incubation with the antioxidant matrix prior to infrared A irradiation (IRA) resulted in a significant decrease of MMP1 expression by all three doses selected. The highest dose of the antioxidant matrix employed (0.01%) inhibited IRA-induced MMP1 up-regulation by 68%. Lower doses such as 0.001% decreased MMP1 up-regulation by 55%. And 0.005% Berrimatrix resulted in an inhibition of 48%.
CONCLUSION
A dose of 0.01% Berrimatrix was able to inhibit IRA-induced MMP-1 expression by 68%, while lower concentrations resulted in inhibition of 48% for 0.005% and 55% for 0.001%.
DISCLOSURES
This study was funded by Airelle Skincare, LLC. Katherine Wilkens and Kasey D'Amato are employed by Airelle Skincare, LLC. Dr Krutmann has served as a consultant for Airelle Skincare, LLC and Dr Krutmann received honoraria for work on this supplement.
REFERENCES
- Schroeder P, Lademann J. Darvin ME, Stege H, Marks C, Bruhnke S, Krutmann J. Infrared radiation-induced matrix metalloproteinase in human skin. Implications for protection. J Invest Dermatol 2008;128:2491-2497
- Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63
- Grether-Beck S. Felsner I. Brenden H. Krutmann J. Mitochondrial cytochrome C release mediates ceramide-induced AP-2 activation and gene expression in human cells. J Biol Chem 2003;278:47498-47507
- Knaapen AM, Albrecht C, Becker CA, Hohr D, Winzer A, Haenen G, Borm PJA, Schins RFP. DNA damage in lung epithelial cells isolated from rats exposed to quartz: role of surface reactivity and neutrophilic inflammation. Carcinogenesis 2002;23:1111-1120
- Grether-Beck S, Mu?hlberg K, Brenden H, Felsner I, Brynjólfsdóttir A, Einarsson S, Krutmann J. In-vitro and in-vivo assessment of silica mud and microalgae extracts for their effects on skin barrier function and prevention of skin aging. Exp Dermatol 2008;17:771-779
- Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2006; 25:402-408
- Halliwell B. Oxidative stress in cell culture: an under-appreciated problem? FEBS Letters 2003;540:3-6
AUTHOR CORRESPONDENCE
Katherine Wilkens MPAP katherine.wilkens@airelleskin.com
