Biocompatible crosslinked nanofiber mats of chitosan (CS)/poly(vinyl alcohol) (PVA) were fabricated using electrospinning technique. CS and PVA blends (7, 9 and 11 weight %) of prepared keeping ratios of CS: PVA to 1:4 and was crosslinked with tetraethoxysilane (TEOS). Fourier transform infrared (FUR) analysis confirmed the existence of inter- and intramolecular hydrogen bonding between polymer chains and the development of siloxane linkage within the nanofibers. X-ray diffractometry (XRD) analysis showed an increase in the crystallinity of electrospun nanofibers after crosslinking as compared to the uncrosslinked nanofibers and with an increase in CS/PVA content. Scanning electron microscopy (SEM) micrographs exhibited the formation bead-free fibers at higher polymer concentration. The average size of the nanofibers was found in the range of 40 to 100 nm. The concentration and crosslinker content affected the mechanical and thermal properties of the nanofibers. The crosslinker has increased the tensile strength (TS) values upto 120 % and Young's modulus by 71 % as compared to the uncrosslinked nanofibers while elongation at break was decreased in all nanofibers. The cell viability of the nanofibers was investigated by employing human cancerous bone cells (MG63). The obtained results showed that cancerous bone cells were not proliferated in the presence of nanofibers and the growth of the cells was inhibited confirming the worth of CS/PVA nanofibers against cancerous bone cells. This inherent behaviour can be exploited as base material for anticancer biomedical applications.