Objective: To examine how three types of AOs affected fibroblasts and whether AOs could induce the synthesis of Col-I and Col-Ill, components of human dermis.
Methods: Three types of alginate oligosaccharides (AOs) were prepared from sodium alginate after treatment with alginate lyase with various time periods (6, 12, and 24 hours, called "Type 1, Type 2, Type 3" respectively) and their effects on collagen expression in HS27 human dermal fibroblasts were examined. Cytotoxicity of alginate oligosaccharides was confirmed by MTT assay. Also expression levels at mRNA and protein levels were determined by reverse tran-scription-PCR (RT-PCR) and western blot, respectively. Results: AOs do not show any cellular toxicity, so they were applied directly to the cells. Treatment with Type 3 AOs resulted in the highlighted expression level of collagen type I (Col-I) mRNA while exposure with Type 1 AOs lead to the highest expression of Collagen type III (Col-Ill) mRNA in HS27 fibroblasts. In addition, the mRNA expression level of matrix metalloprotein-ase-1 (MMP-1), an enzyme involved in collagen degradation, was found to increase in cells treated with AOs at low concentrations but decrease in cells treated with AOs at higher concentrations. Moreover, mRNA expression level of tissue inhibitor metalloproteinase-1 (TIMP-1), an antagonist of MMP-1, was found to be increased by AOs in a concentration-dependent manner.
Conclusion: These results suggest that this AOs might have potential to prevent skin aging by promoting collagen synthesis through the inhibition of collagen degrading enzyme.
Alginate oligosaccharide Collagen type I Collagen type III Matrix metalloproteinase-1 Tissue inhibitor metalloproteinase-1
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This work was carried out with the support of “Cooperative Research Program for agriculture Science & Technology Development (Project No: PJ01267701)” funded by Rural Development Administration, Republic of Korea. The authors are grateful for their support.
This article does not contain any studies with human participants or animals performed by any of the authors.
Riita, R., Mataleena, P. & Arja, J. Increased expression of collagen type and in human skin as a consequence of radiotherapy. Arch. Dermatol. Res.294, 178–184 (2002).CrossRefGoogle Scholar
Talwar, H., Griffiths, C, Fisher, G., Hamilton, T. & Voorhees, J. Reduced type I and type III procollagens in photodamaged adult human skin. J. Invest. Dermatol.105, 285–290 (1995).CrossRefGoogle Scholar
Wlaschek, M. et al. Solar UV irradiation and dermal photoaging. J. Photochem. Photobiol. B63, 41–51 (2001).CrossRefGoogle Scholar
Enjoji, M., Kotoh, K., Iwamoto, H., Nakamuta, M. & Nawata, H. Self-regulation of type I collagen degradation by collagen-induced production of matrix metallo-proteinase-1 on cholangiocarcinoma and hepatocellular carcinoma cells. In Vitro Cell. & Dev. Biol. Anim.36, 71–73 (2000).CrossRefGoogle Scholar
Fisher, G., Talwar, H., Lin, J. & Voorhees, J. Molecular Mechanisms of Photoaging in Human Skin In Vivo and Their Prevention by All - Trans Retinoic Acid. J. Photochem. Photobiol.A Chem.69, 154–157 (1999).CrossRefGoogle Scholar
Chernoff, R. Micronutrient requirements in older women-. Am. J. Clin. Nutr.81, 1240S–1245S (2005).CrossRefGoogle Scholar
Wendt, M., Soparkar, C, Louie, K, Basinger, S. & Gross, R. Ascorbate stimulates type I and type III collagen in human Tenon’s fibroblasts. J. Glaucoma6, 402–407 (1997).CrossRefGoogle Scholar
Lee, K. & Mooney, D. Alginate: properties and biomedical applications. Progr. Polym. Sci.37, 106–126 (2012).CrossRefGoogle Scholar
Otterlei, M. et al. Induction of cytokine production from human monocytes stimulated with alginate. J. Immunother.10, 286–291 (1991).CrossRefGoogle Scholar
Zimmermann, U. et al. Production of mitogen - contamination free alginates with variable ratios of man-nuronic acid to guluronic acid by free flow electrophoresis. Electrophoresis13, 269–274 (1992).CrossRefGoogle Scholar
De La Motte, C. et al. Platelet-derived hyaluronidase 2 cleaves hyaluronan into fragments that trigger mono-cyte-mediated production of proinflammatory cytokines. Am. J. Pathol.174, 2254–2264 (2009).CrossRefGoogle Scholar
De la Motte, C. Hyaluronan in intestinal homeostasis and inflammation: implications for fibrosis. American journal of physiology. Am. J. Physiol. Gastrointest. Liver Physiol.301, G945–G949 (2011).CrossRefGoogle Scholar
Papakonstantinou, E., Roth, M. & Karakiulakis, G. Hyaluronic acid: A key molecule in skin aging. Dermatoendocrinol. 4, 253–258 (2012).CrossRefGoogle Scholar
Tolg, C. et al. A RHAMM mimetic peptide blocks hyaluronan signaling and reduces inflammation and fibrogenesis in excisional skin wounds. Am. J. Pathol.181, 1250–1270 (2012).CrossRefGoogle Scholar
Taylor, K. et al. Recognition of hyaluronan released in sterile injury involves a unique receptor complex dependent on Toll-like receptor 4, CD44, and MD-2. J. Biol. Chem.282, 18265–18275 (2007).CrossRefGoogle Scholar
Leung, A., Crombleholme, T. & Keswani, S. Fetal wound healing: implications for minimal scar formation. Curr. Opin. Pediatr.24, 371 (2012).CrossRefGoogle Scholar
Buchanan, E., Longaker, M. & Lorenz, H. Fetal skin wound healing. Adv. Clin. Chem.48, 137–161 (2009).CrossRefGoogle Scholar
West, D., Shaw, D., Lorenz, P., Adzick, N. & Longaker, M. Fibrotic healing of adult and late gestation fetal wounds correlates with increased hyaluronidase activity and removal of hyaluronan. Int. J. Biochem. Cell Biol.29, 201–210 (1997).CrossRefGoogle Scholar
Toole B. Hyaluronan: from extracellular glue to pericellular cue. Nat. Rev. Cancer4, 528 (2004).CrossRefGoogle Scholar
Dubois, M., Gilles, K., Hamilton, J., Rebers, Pt. & Smith, F Colorimetric method for determination of sugars and related substances. Anal. Chem.28, 350–356 (1956).CrossRefGoogle Scholar
Mosmann, T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods65, 55–63 (1983).CrossRefGoogle Scholar
Tsoureli-Nikita, E, Watson, R. & Griffiths, C. Photoageing: the darker side of the sun. Photochem. Photobiol. Sci.5, 160–164 (2006).CrossRefGoogle Scholar
Ferrari, M., Fornasiero, M. & Isetta, A. MTT colorimetric assay for testing macrophage cytotoxic activity in vitro. J. Immunol. Methods131, 165–172 (1990).CrossRefGoogle Scholar