For bone regeneration from KUSA-A1 oesteoblast cells (KUSA), chitosan (CS) scaffolds possessing different surface properties, sponge-type (CSS) and nonwoven-type (CSNW), were manufactured. Surface area and pore size of CSNW were larger than those of CSS. On the other hand, the pore volume of CSNW was smaller than that of CSS. Cell attachment evaluation showed CSNW was more adequate then CSS, and this was attributed to the large surface area. For in vivo investigation, KUSA were seeded into CS scaffolds in wells followed by a week of cell culture. Obtained CS scaffolds with KUSA were implanted on the subcutaneous tissue of BALB/C nude mice. After surgery, implanted scaffolds were harvested and assayed by immunological staining. Network stability of CSS was better than that of CSNW, even if CSS scaffolds were destroyed between 4 and 6 weeks. Calcification was observed after 4 and 8 weeks for CSNW and CSS, respectively.