This study focuses on fabricating sodium nitroprusside-releasing chitosan-based (CS/SNP) nanofibers through electrospinning. Based on SEM images and FTIR spectroscopy, the application of one-step photocrosslinking significantly improved the stability of the nanofibers in aqueous environments. SEM images showed that the porous nanofibrous structure was maintained for up to 24 h. Incorporating SNP into the nanofibrous scaffolds demonstrated an enhanced biocompatibility with osteogenic cells. The cell viability increased with rising SNP content. Enhanced cell attachment, spreading, and proliferation were also observed through fluorescence microscope images of CS/SNP nanofibers, followed by the positive regulation of Smad and Runx2 pathway. The crosstalk between CS and NO reduced the possible toxicity and enhanced the osteoinductivity of NO. Furthermore, the nanofibers contributed to escalated osteogenic differentiation and mineralization, as evidenced by heightened expression of osteogenic markers such as alkaline phosphatase (ALP) expression and calcium deposition. Overall, the photo-crosslinked electrospun CS/SNP nanofibers demonstrate substantial potential as scaffolds in the field of bone tissue engineering