DOI 10.1515/hsz-2012-0202 Biol. Chem. 2012; 394(1): 113–123 Radoslaw Szmyd a , Anna Grazyna Goralczyk a , Lukasz Skalniak, Agnieszka Cierniak, Barbara Lipert, Francesca Larese Filon, Matteo Crosera, Julia Borowczyk, Eliza Laczna, Justyna Drukala, Andrzej Klein and Jolanta Jura* Effect of silver nanoparticles on human primary keratinocytes Abstract: Silver nanoparticles (AgNPs) have many biologi- cal applications in biomedicine, biotechnology and other life sciences. Depending on the size, shape and the type of carrier, AgNPs demonstrate different physical and chemical properties. AgNPs have strong antimicrobial, antiviral and antifungal activity, thus they are used extensively in a range of medical settings, particularly in wound dressings but also in cosmetics. This study was undertaken to examine the potential toxic effects of 15 nm polyvinylpyrrolidone- coated AgNPs on primary normal human epidermal keratinocytes (NHEK). Cells were treated with different concentrations of AgNPs and then cell viability, metabolic activity and other biological and biochemical aspects of keratinocytes functioning were studied. We observed that AgNPs decrease keratinocyte viability, metabolism and also proliferatory and migratory potential of these cells. Moreover, longer exposure resulted in activation of caspase 3/7 and DNA damage. Our studies show for the first time, that AgNPs may present possible danger for primary keratinocytes, concerning activation of genotoxic and cytotoxic processes depending on the concentration. Keywords: caspase activation; cell viability; MAP kinase activation; migration; primary keratinocytes; silver nanoparticles. a These authors contributed equally to this work. *Corresponding author: Jolanta Jura, Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland, e-mail: jolanta.jura@uj.edu.pl Radoslaw Szmyd : Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland Anna Grazyna Goralczyk: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland Lukasz Skalniak: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland Agnieszka Cierniak: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland Barbara Lipert: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland Francesca Larese Filon: Unit of Occupational Medicine, University of Trieste, 34129 Trieste, Italy Matteo Crosera: Department of Chemical and Pharmaceutical Sciences, University of Trieste, 34127 Trieste, Italy Julia Borowczyk: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland Eliza Laczna: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland Justyna Drukala: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland Andrzej Klein: Department of General Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland Introduction Nanoparticles are of great scientific interest because of the wide variety of potential applications. There is a growing list of reports concerning applications of nanoparticles in biomedicine, biotechnology and cosmetology. Because of strong antimicrobial, antiviral and antifungal activity (Edwards-Jones, 2009), silver nanoparticles (AgNPs) have been used extensively in medical and healthcare sectors (Benn and Westerhoff, 2008; Li et al., 2011; Teow et al., 2011). They are used as anti-pathogenic additives in prod- ucts such as wound dressings, surgical instruments, clean- ing products, cosmetics and cloths. Nanoparticles incorpo- rated to these products come in a direct contact with skin and may affect the biology of epidermal cells (Chen and Schluesener, 2008; Ahamed et al., 2010; Teow et al., 2011). Although the data on the effect of AgNPs on various cell types are often not consistent, it is now clear that dif- ferent surface chemistry and size of nanoparticles are some of the most important parameters, which must be included when considering their unique properties. For example, Liu and co-workers (2010) tested the impact of 5, 20 and 50 nm AgNPs on cell lines derived from different human organs Authenticated | jolanta.jura@uj.edu.pl author's copy Download Date | 12/6/12 10:10 AM