title: In vitro and in vivo biocompatibility studies of a recombinant analogue of spidroin 1 scaffolds.
abstract: The goal of this study was to generate porous scaffolds from the genetically engineered protein, an analogue of Nephila clavipes spidroin 1 (rS1/9) and to assess the properties of new rS1/9 scaffolds essential for bioengineering. The salt leaching technique was used to make the rS1/9 scaffolds of interconnected macroporous structure with spontaneously formed micropores. The tensile strength of scaffolds was 18 ± 5 N/cm(2). Scaffolds were relatively stable in a phosphate buffer but degraded in oxidizing environment after 11 weeks of incubation. Applicability of the recombinant spidroin 1 as a substrate for cell culture was demonstrated by successful 3T3 cells growth on the surface of rS1/9 films (270 ± 20 cells/mm(2) vs. 97 ± 8 cells/mm(2) on the glass surface, p < 0.01). The 3T3 fibroblasts readily proliferated within the rS1/9 scaffold (from initially plated 19 ± 2 cells/mm(3) to 3800 ± 304 cells/mm(3) after 2 weeks). By this time, cells were uniformly distributed between the surface and deeper layers (27% ± 8% and 33% ± 4%, respectively; p > 0.05), whereas the initial distribution was 58% ± 7% and 11% ± 8%, respectively; p < 0.05). The rS1/9 scaffolds implanted subcutaneously into Balb/c mice were well tolerated. Over a 2-month period, the scaffolds promoted an ingrowth of de novo formed vascularized connective tissue elements and nerve fibers. Thus, scaffolds made of the novel recombinant spidroin 1 analogue are potentially applicable in tissue engineering.
authors.affiliation: Faculty of Biology, Lomonosov Moscow State University, Moscow, Russian Federation. mmoisenovich@mail.ru
authors.forename: M M
authors.initials: MM
authors.lastname: Moisenovich
authors.forename: O L
authors.initials: OL
authors.lastname: Pustovalova
authors.forename: A Yu
authors.initials: AY
authors.lastname: Arhipova
authors.forename: T V
authors.initials: TV
authors.lastname: Vasiljeva
authors.forename: O S
authors.initials: OS
authors.lastname: Sokolova
authors.forename: V G
authors.initials: VG
authors.lastname: Bogush
authors.forename: V G
authors.initials: VG
authors.lastname: Debabov
authors.forename: V I
authors.initials: VI
authors.lastname: Sevastianov
authors.forename: M P
authors.initials: MP
authors.lastname: Kirpichnikov
authors.forename: I I
authors.initials: II
authors.lastname: Agapov
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D005349:Fibroins / Q000737:chemistry* / Q000235:genetics; D008422:Materials Testing; D051379:Mice; D008807:Mice, Inbred BALB C; D016062:Porosity; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D001672:Biocompatible Materials; D011994:Recombinant Proteins; C086087:spidroin 1; D005349:Fibroins