Abstract
BackgroundIn 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 FindingsNative 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/SignificanceNative 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.
Highlights
The regeneration of skin poses an important challenge for plastic surgery
Additional adhesion-points were given to Mouse embryonic fibroblasts (MEF) cells, which were able to colonise the interspace along the spider silk fibres
Single culturing of fibroblasts or keratinocytes on spider silk frames
Summary
The regeneration of skin poses an important challenge for plastic surgery. Due to its glandular- and barrier- functions, skin protects from chemical and physical environmental effects and fulfils important tasks in the regulation of homeostasis [1]. Damages to the integrity or the loss of large portions of the skin, e.g. caused by burns, may result in a significant disability or even signify a lifethreatening situation. The ‘‘gold standard’’ for burn wound closure is the application of split-thickness autografts, harvested from uninjured areas. In severely burned patients with extensively injured areas, not enough donor sites are available to provide sufficient autografts. The ideal biomaterial should promote attachment, proliferation and growth of cells. 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
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