Antibacterial activity and cell viability of hyaluronan fiber with silver nanoparticles.

Silver has been used since time immemorial in different chemical form to treat burns, wounds and several different infections caused by pathogenic bacteria, advancement of biological process of nanoparticles synthesis is evolving into a key area of nanotechnology. The current study deals with the green synthesis, characterization, and evaluation of the biological activity and cell viability of hyaluronan fibers with incorporated silver nanoparticles (HA-Ag NPs). Hyaluronan fiber was prepared by the dissolving of sodium hyaluronate (HA) in aqueous alkaline solution to prepare a transparent solution, which was used for the preparation of fibers by a wet-spinning technique. Consequently, hyaluronan fiber was used as capping and stabilizing agent for the preparation of fibers with silver nanoparticles. HA-Ag NPs were confirmed by transmission electron microscopy, dynamic light scattering, UV/VIS spectroscopy, scanning electron microscopy, energy-dispersive X-ray spectroscopy, thermal analysis, nuclear magnetic resonance, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. HA-Ag NPs showed high antibacterial activity of against Staphylococcus aureus and Escherichia coli. Cell viability tests indicated that hyaluronan, hyaluronan fibers and hyaluronan fibers with silver nanoparticles were non-toxic on the cell growth. Two different particles size of Ag NPs (10, 40 nm) had not any toxicity till the concentration limit. These tests were performed using mouse fibroblast cell line 3T3.

Hyaluronan minimizes effects of UV irradiation on human keratinocytes.

Exposure to ultraviolet (UV) irradiation has detrimental effects on skin accompanied by the increased metabolism of hyaluronan (HA), a linear polysaccharide important for the normal physiological functions of skin. In this study, the modulation of human keratinocyte response to UVB irradiation by HA (970 kDa) was investigated. Immortalized human keratinocytes (HaCaT) were irradiated by a single dose of UVB and immediately treated with HA for 6 and 24 h. The irradiation induced a significant decrease in the gene expression of CD44 and toll-like receptor 2 6 h after irradiation. The expressions of other HA receptors, including toll-like receptor 4 and the receptor for HA-mediated motility, were not detected in either the control or UVB-irradiated or HA-treated HaCaT cells. UVB irradiation induced a significant decrease in the gene expression of HA synthase-2 and hyaluronidase-2 6 h after irradiation. The expressions of HA synthase-3 and hyaluronidase-3 were not significantly modulated by UV irradiation. Interestingly, HA treatment did not significantly modulate any of these effects. In contrast, HA significantly suppressed UVB-induced pro-inflammatory cytokine release including interleukin-6 and interleukin-8. Similarly, HA treatment reduced the UVB-mediated production of transforming growth factor ß1. HA treatment also significantly reduced the UV irradiation-mediated release of soluble CD44 into the media. Finally, HA partially, but significantly, suppressed the UVB-induced decrease in cell viability. Data indicate that HA had significant protective effects for HaCaT cells against UVB irradiation.

The comparison of impedance-based method of cell proliferation monitoring with commonly used metabolic-based techniques.

OBJECTIVES: Determination of cell numbers is a crucial step in studies focused on cytokinetics and cell toxicity. The impedance-based analysis employing electronic sensor array system xCELLigence System allowing label-free dynamic monitoring of relative viable adherent cell amounts was compared with the most utilized methods for relative quantification of viable cell numbers based on a determination of cellular metabolism. DESIGN: Colorimetric assay based on reduction of tetrazolium salt (MTT) by mitochondrial enzymes and chemiluminiscent assay based on intracellular adenosine triphosphate (ATP) determination were compared with the impedance-based system. Cell morphology was compared by microscopic evaluation. Normal human epidermal keratinocytes (NHEK) and normal human dermal fibroblasts (NHDF), together with 3T3 mouse fibroblast and HaCaT keratinocyte cell lines were employed. RESULTS: The progress of cell growth curves obtained by different methods during 72 hours reflected cell type and cell seeding densities. The impedance-based method was found to be applicable for the determination of the cell proliferation of 3T3 fibroblasts, HaCaT and NHDF, since the comparison of this method with ATP and MTT determinations showed a comparable results. In contrast, the proliferation of NHEK measured by the impedance-based method did not correlate with other methodological approaches. This could be accounted to the specific morphological appearance of these cells. CONCLUSION: The study shows the impedance-based detection of viable adherent cells is a valuable approach for cytokinetics and pharmacological studies. However, the specific morphological characteristics of cell lines have to be considered employing this method for determination of cell proliferation without using other reference methods.