Three dimensional (3D) scaffolds provide the necessary support for cells to attach, proliferate and differentiate, and define the overall shape of the tissue engineered transplant. In this study, 3D honeycomb patterned chitosan/poly (L-lactic acid) composite scaffolds fabricated by an easy manipulated technique with good mechanical property and cytocompatability, as demonstrated by a previous study. Here we investigated further the in vitro cytocompatibility and spine regeneration in vivo by implanting the construct into male white rabbits for 4 and 8weeks. Results showed that such a honeycomb patterned scaffolds have a good cytocompatibilty. Also, the rabbit spinal defect was perfectly restored. These findings supported that such a 3D honeycomb patterned scaffold is an ideal candidate for the tissue engineering scaffold.