An electrospun hydrophilic non-water-soluble biocompatible polylactic acid (PLA) nonwoven material was used as a delivery system for propolis ethanolic extract (PEE) and silver nanoparticles (AgNPs) that are known for their established antiseptic and antimicrobial activity. Combination of PEE and AgNPs in a single product should provide efficient antimicrobial protection and improved wound healing. Evaluations of PEE and AgNPs on morphology of electrospun materials, release kinetics of AgNPs and phenolic compounds, antibacterial properties, and cytotoxicity of electrospun PLA materials were performed. The presence of PEE or/and AgNPs resulted in denser mats formed by thicker PLA fibers. The average diameter of PLA microfibers was 168±29 nm. The average diameter of microfibers increased to 318±40 and 370±30 nm when 10 wt% and 20 wt% ethanol were added, respectively. Addition of 10 wt% or 20 wt% PEE increased the diameter to 282±25 and 371±25 nm, respectively. Suspension of AgNPs also caused the formation of thicker microfibers with 254±25 nm diameter. Electrospun PLA microfibers with PEE maintained viability of HaCaT cells. Testing of antimicrobial activity confirmed the ability of AgNPs containing PLA electrospun materials to inhibit the growth of microorganisms.