A laminin and nerve growth factor-laden three-dimensional scaffold for enhanced neurite extension.

Published

Journal Article

Agarose hydrogel scaffolds were engineered to stimulate and guide neuronal process extension in three dimensions in vitro. The extracellular matrix (ECM) protein laminin (LN) was covalently coupled to agarose hydrogel using the bifunctional cross-linking reagent 1,19- carbonyldiimidazole (CDI). Compared to unmodified agarose gels, LN-modified agarose gels significantly enhanced neurite extension from three-dimensionally (3D) cultured embryonic day 9 (E9) chick dorsal root ganglia (DRGs), and PC 12 cells. After incubation of DRGs or PC 12 cells with YIGSR peptide or integrin beta1 antibody respectively, the neurite outgrowth promoting effects in LN-modified agarose gels were significantly decreased or abolished. These results indicate that DRG/PC 12 cell neurite outgrowth promoting effect of LN-modified agarose gels involves receptors for YIGSR/integrin beta1 subunits respectively. 1,2-bis(10, 12-tricosadiynoyl)-sn-glycero-3-phosphocholine (DC(8,9)PC)-based lipid microcylinders were loaded with nerve growth factor (NGF), and embedded into agarose hydrogels. The resulting trophic factor gradients stimulated directional neurite extension from DRGs in agarose hydrogels. A PC 12 cell-based bioassay demonstrated that NGF-loaded lipid microcylinders can release physiologically relevant amounts of NGF for at least 7 days in vitro. Agarose hydrogel scaffolds may find application as biosynthetic 3D bridges that promote regeneration across severed nerve gaps.

Full Text

Duke Authors

Cited Authors

  • Yu, X; Dillon, GP; Bellamkonda, RB

Published Date

  • August 1999

Published In

Volume / Issue

  • 5 / 4

Start / End Page

  • 291 - 304

PubMed ID

  • 10477852

Pubmed Central ID

  • 10477852

Electronic International Standard Serial Number (EISSN)

  • 1557-8690

International Standard Serial Number (ISSN)

  • 1076-3279

Digital Object Identifier (DOI)

  • 10.1089/ten.1999.5.291

Language

  • eng