title: Artificial skin--culturing of different skin cell lines for generating an artificial skin substitute on cross-weaved spider silk fibres.
abstract: 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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Plastic, Hand, and Reconstructive Surgery, Medical School Hannover, Hannover, Germany. HannaFW@web.de
authors.forename: Hanna
authors.initials: H
authors.lastname: Wendt
authors.forename: Anja
authors.initials: A
authors.lastname: Hillmer
authors.forename: Kerstin
authors.initials: K
authors.lastname: Reimers
authors.forename: Joern W
authors.initials: JW
authors.lastname: Kuhbier
authors.forename: Franziska
authors.initials: F
authors.lastname: Schäfer-Nolte
authors.forename: Christina
authors.initials: C
authors.lastname: Allmeling
authors.forename: Cornelia
authors.initials: C
authors.lastname: Kasper
authors.forename: Peter M
authors.initials: PM
authors.lastname: Vogt
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002478:Cells, Cultured; D018920:Coculture Techniques; D005347:Fibroblasts / Q000166:cytology; D015603:Keratinocytes / Q000166:cytology; D047011:Silk / Q000737:chemistry*; D012867:Skin / Q000737:chemistry*; D017768:Skin, Artificial; D013112:Spiders; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D001672:Biocompatible Materials; D047011:Silk