title: Electrospun, biofunctionalized fibers as tailored in vitro substrates for keratinocyte cell culture.
abstract: Cell adhesion preventing fiber surfaces were tailored differently with bioactive peptides (a fibronectin fragment (GRGDS), a collagen IV fragment (GEFYFDLRLKGDK) and a combination of both) to provide an artificial extracellular matrix as a substrate for HaCaT keratinocyte cell culture. Therefore, a polymer blend containing a six-arm star-shaped statistical copolymer of ethylene oxide and propylene oxide in the ratio 80:20 (NCO-sP[EO-co-PO]) and poly-[D,L-(lactide-co-glycolide)] (PLGA) was electrospun. The resulting fibers were biofunctionalized and investigated as in vitro substrates using the HaCaT kerationcyte cell line. Appropriate surface chemistry on these electrospun fibers proved to prevent adhesion of keratinocytes, while additional immobilization of certain peptide sequences induced cell adhesion. These specific fibers enable investigation of immobilized active molecules and the subsequent cellular response to the scaffold. HaCaT keratinocytes were found to selectively adhere to those fibers modified with either collagen IV segment GEFYFDLRLKGDK or a mixture of the two peptide sequences GEFYFDLRLKGDK and GRGDS (1:1). However, the synergistic effects of both (the fibronectin fragment and the collagen IV fragment) seem to significantly increase the numbers of adherent keratinocytes.
authors.affiliation: DWI e.V. and Institute of Technical and Macromolecular Chemistry, RWTH Aachen Pauwelsstr. 8, D-52056 Aachen, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Grafahrend
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Heffels
authors.forename: Martin
authors.initials: M
authors.lastname: Möller
authors.forename: Doris
authors.initials: D
authors.lastname: Klee
authors.forename: Jürgen
authors.initials: J
authors.lastname: Groll
mesh_terms: D000595:Amino Acid Sequence; D001672:Biocompatible Materials / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques; D002460:Cell Line; D048430:Cell Shape / Q000187:drug effects; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D013379:Substrate Specificity
chemical_list: D001672:Biocompatible Materials; D009842:Oligopeptides; D010455:Peptides; C047981:arginyl-glycyl-aspartic acid