Nanofibrous biocomposite scaffolds of chitosan (CS), PVA, and hydroxyapatite (HA) were prepared by electrospinning. The scaffolds were characterized by FTIR, SEM, TEM, and XRD techniques. Tensile testing was used for the characterization of mechanical properties. Mouse fibroblasts (L929) attachment and proliferation on the nano-fibrous scaffold were investigated by MTT assay and SEM observation. FTIR, TEM, and XRD results showed the presence of nanoHA in the scaffolds. The scaffolds have porous nanofibrous morphology with random fibers in the range of 100-700 nm diameters. The CS/PVA (90/10) fibrous matrix (Without HA) showed a tensile strength of 3.1 +/- 0.2 MPa and a tensile modulus 10 I MPa with a strain at failure of 21.1 +/-0.6%. Increase the content of HA up to 2% increased the ultimate tensile strength and tensile modulus, but further increase HA LIP to 5-10% caused the decrease of tensile strength and tensile modulus. The attachment and growth of mouse fibroblast was on the Surface of nanofibrous structure, and cells' morphology characteristics and viability were unaffected. A combination of nanofibrous CS/PVA and HA that mimics the nanoscale features of the extra cellular matrix Could be promising for application as scaffolds for tissue regeneration, especially in low or nonload bearing areas. (C) 2008 Wiley Periodicals, Inc. J Appl Polym Sci 110: 3328-3335, 2008