Neural differentiation directed by self-assembling peptide scaffolds presenting laminin-derived epitopes

We prepared biofunctionalized matrices for cell growth using (RADA)(3)IKVAV(RADA)(3) ((Arg-Ala-Asp-Ala)(3)-Ile-Lys-Val-Ala-Val-(Arg-Ala-Asp-Ala)(3)) and (RADA)(4)-IKVAV ((Arg-Ala-Asp-Ala)(4)-Ile-Lys-Val-Ala-Val), self-assembling peptides with a laminin-derived sequence inserted between and attached terminally to the repeats of RADA, respectively. The material cell interactions were investigated with PC12, a cell line commonly used as a model for studying neural differentiation. The behavior of PC12 and especially the neural differentiation was guided by the presence of IKVAV. Furthermore, the cell material interactions were dependent on the culture dimensionality and the position of IKVAV in the self-assembling peptide template. In the two-dimensional (2-D) culture, matrices containing IKVAV stimulated significantly longer neurite outgrowths from PC12 cells than did (RADA)(4). More pronounced effect was observed in (RADA)(3)IKVAV(RADA)(3) than in (RADA)(4)IKVAV. In the three-dimensional (3-D) culture, neurite outgrowth was not observed in the biofunctionalized matrices. Instead, cells displayed higher proliferation rate and survived longer culture time than in the 2-D culture, with such enhancement being most significant in (RADA)(3)IKVAV(RADA)(3). Despite the lack of differentiation phenotype, the cells grown in 3-D biofunctionalized matrices were primed for differentiation, as evident by enhanced neurite outgrowth, increased neurite networking, and up-regulated expression of differentiation markers upon their reintroduction to the 2-D culture condition on petri dish. With the ease of incorporating biofunctional epitopes, and the flexibility to support either 2-D or 3-D culture, self-assembling peptides provide versatile scaffolds to study the multiple facets of biomaterial cell interactions. (C) 2010 Wiley Periodicals, Inc. J Biomed Mater Res 94A: 688499, 201