To use nano-structured materials as a novel cell therapeutic agent, we have devised a novel method for the fabrication of nano-scaled 3D scaffolds consisting of heparin/poly(ethylenimine) (PEI) nanoparticles in a thermo-reversible hydrogel, attached via a layer-by-layer system. Bioassay results showed significant difference in DNA amount between groups. Specifically, groups with heparin/PEI nanoparticles had almost twice the glycosaminoglycan content per construct starting at day 7 as compared to controls. Gene expression of total collagen was evident in groups containing heparin/PEI nanoparticles.