Artificial Skin – Culturing of Different Skin Cell Lines for Generating an Artificial Skin Substitute on Cross-Weaved Spider Silk Fibres

Background: In the field of Plastic Reconstructive Surgery the development of new innovative matrices for skin repair is in urgent need. The ideal biomaterial should promote attachment, proliferation and growth of cells. Additionally, it should degrade in an appropriate time period without releasing harmful substances, but not exert a pathological immune response. Spider dragline silk from Nephila spp meets these demands to a large extent. Methodology/Principal Findings: Native spider dragline silk, harvested directly out of Nephila spp spiders, was woven on steel frames. Constructs were sterilized and seeded with fibroblasts. After two weeks of cultivating single fibroblasts, keratinocytes were added to generate a bilayered skin model, consisting of dermis and epidermis equivalents. For the next three weeks, constructs in co-culture were lifted on an originally designed setup for air/liquid interface cultivation. After the culturing period, constructs were embedded in paraffin with an especially developed program for spidersilk to avoid supercontraction. Paraffin cross-sections were stained in Haematoxylin & Eosin (H&E) for microscopic analyses. Conclusion/Significance: Native spider dragline silk woven on steel frames provides a suitable matrix for 3 dimensional skin cell culturing. Both fibroblasts and keratinocytes cell lines adhere to the spider silk fibres and proliferate. Guided by the spider silk fibres, they sprout into the meshes and reach confluence in at most one week. A well-balanced, bilayered cocultivation in two continuously separated strata can be achieved by serum reduction, changing the medium conditions and the cultivation period at the air/liquid interphase. Therefore spider silk appears to be a promising biomaterial for the enhancement of skin regeneration

Antibacterial Activity of Positive and Negative Polarity Low-voltage Pulsed Current (LVPC) on Six Typical Gram-positive and Gram-negative Bacterial Pathogens of Chronic Wounds

The positive effect of electrical stimulation (ES) on wound healing has been shown in vitro and in vivo. On the basis of increased blood flow, protein denaturation, and stimulation of cellular defense, an antibacterial effect of ES is to be expected. Although the antibacterial effect of ES already has been demonstrated in vitro, little attention has been paid to the direct antibacterial effect of changing polarity of the applied current. The aim of this study was to investigate the antibacterial effect of positive and negative monophasic low-voltage pulsed current on typical Gram-positive and Gram-negative pathogens of chronic wounds. Using the Dermapulse®-System, three Gram-negative (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae) and three Gram-positive (Staphylococcus aureus, Staphylococcus epidermidis, Escherichia faecium) organisms were tested against positive and negative polarity low voltage pulsed current. All tested organisms were significantly reduced by ES. The reduction differed significantly between positive polarity and control and negative polarity and control, with the highest log10 reduction factor (RF) achieved with positive polarity. Using positive polarity, the maximum RF was measured for E. coli (median log10 RF 0.83; 25th percentile 0.59, 75th percentile 0.98) and the lowest for S. epidermidis (median log10 RF 0.20; 25th percentile 0.17, 75th percentile 0.24). Yet, there was no significant difference with positive ES against Gram-positive or Gram-negative organisms