JOURNAL OF MEDICINAL FOOD J Med Food 12 (2) 2009, 429–434 © Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition DOI: 10.1089/jmf.2008.0123 Cell Viability of Normal Human Skin Fibroblast and Fibroblasts Derived from Granulation Tissue: Effects of Nutraceuticals M.H. Borawska, 1 S.K. Czechowska, 1 R. Markiewicz, 1 A. Hayirli, 2 E. Olszewska, 3 and K. Sahin 4 Departments of 1 Bromatology and 3 Otolaryngology, Medical University of Bialystok, Bialystok, Poland; 2 Department of Animal Nutrition and Nutritional Disorders, Faculty of Veterinary Medicine, Atatürk University, Erzurum; and 4 Department of Animal Nutrition, Faculty of Veterinary Medicine, Firat University, Elazig, Turkey ABSTRACT The effects of lycopene, genistein, and epigallocatechin-3-gallate (EGCG) on cell viability were tested in vitro using a normal human skin fibroblast (NHSF) cell line (CRL-1474) and granulation tissue fibroblasts (GTFs) obtained from a patient with middle ear cholesteatoma. Cell cultures were added with lycopene (1, 5, and 10 M), genistein (1, 5, 10, 25, and 50 M), and EGCG (1, 5, 10, 25, and 50 M) and their respective control cultures were established by adding 5 mL/L tetrahydrofuran (THF), 5 mL/L dimethyl sulfoxide (DMSO), and 5 mL/L DMSO. A colorimetric assay was employed for de- termining cell viability using thiazolyl blue tetrazolium bromide. Cell viability was expressed as a percentage of the control. Data were analyzed using two-way analysis of variance separately for each compound. Lycopene addition decreased viabil- ity of NHSFs and GTFs compared with THF addition (64.1%, 60.5%, and 100%, respectively, P  .0001). Genistein addi- tion also increased viability of both NHSFs and GTFs compared with DMSO addition (P  .02). Increasing EGCG concen- tration tended to cause a linear increase in viability of NHSFs but did not alter viability of GTFs (P  .10). Our data suggest that genistein and EGCG but not lycopene could help maintaining or improving skin health through enhancing viability of skin fibroblasts. KEY WORDS: • epigallocatechin-3-gallate • fibroblast viability • functional food • genistein • lycopene • skin health 429 INTRODUCTION P HYTOCHEMICALS ARE NON-NUTRITIVE plant chemicals that play an important role in protecting tissues from haz- ardous compounds and preventing diseases due to possess- ing antioxidant, anti-inflammatory, and antineoplastic prop- erties. 1 Lycopene in tomatoes, 2–4 genistein in soya, 5–8 and epigallocatechin-3-gallate (EGCG) in green tea 9–12 are some well-known phytochemicals. There is a growing concern that global warming is also adversely affecting skin health. 7 Numerous studies have demonstrated tissue damage in skin resulting from exposures to environmental pollution 13 and ultraviolet radiation 14 that induce skin carcinoma, aging (e.g., wrinkling, roughness), and other skin lesions such as sunburn, erythema, edema, hyperpigmentation, and cutaneous immunosuppression. Photooxidation causes connective tissue damage via various cellular events that involve activation of angiogenic factors, growth factors, and cytokine receptors and degradation of collagen in keratinocytes and dermal cells, which in turn lead to downstream signal transduction through activation of mitogen-activated protein kinase pathways and induce ex- pression of matrix metalloproteinase-1, collagenase, and heme oxygenase-1 mRNAs. 15–17 Moreover, elevation in re- active oxygen species initiates lipid peroxidation that is linked directly or indirectly to aging, allergies, and carcino- genesis. 18,19 The protective and antioxidant effects of lycopene, genis- tein, and EGCG have been extensively studied in cells sub- jected to damage and cancerous cell lines. Lycopene, 3,20 genistein, 21,22 and EGCG 23,24 have photoprotective effects on skin cells through various paths, which are associated with their high reductive powers to quench singlet molecu- lar oxygen and peroxyl radicals, to induce immunoregula- tory agents (e.g., interleukins, T cells, angiogenic factors) and phosphorylation of mitogen-activated protein kinase cell signaling pathways, and to inhibit protein tyrosine kinase and up-regulation of matrix metalloproteinase-1. This in vitro experiment was conducted to determine the effects of lycopene, genistein, and EGCG on viability of normal hu- man skin fibroblasts (NHSFs) (CRL-1474) and granulation tissue fibroblasts (GTFs) obtained from a patient with cholesteatoma. Manuscript received 8 May 2008. Revision accepted 9 September 2008. Address correspondence to: Kazim Sahin, D.V.M., Ph.D., Professor of Nutrition, Faculty of Veterinary Medicine, Firat University, Elazig 23119, Turkey, E-mail: nsahinkm@ya- hoo.com