Lepidium perfoliatum seed gum (LPSG) biopolymer was co-electrospun in a hybrid nanofiber (HNF) structure using polyvinyl alcohol and polycaprolactone polymers. The obtained results showed the improvement in the hydrophilicity of HNFs with an increase in the LPSG content. In-vitro assay showed two stages of degradation of HNFs (34% and 4% weight loss within 24 h and 14 days, respectively). The antibacterial assay also showed the antibacterial activity of HNFs against Grampositive and Gram-negative bacteria species. The biocompatibility of the LPSG-based nanofibers was also justified by the MTT assay against L929 fibroblast cells (97.4% cell viability).