Bionanocomposite membranes based on polyurethane (PU), extracellular matrix (EM), and silver nanoparticles (AgNPs) were prepared by applying both solvent casting method and electrospinning/electrospraying method. PU-EM-Ag compositions were electrospun/electrosprayed onto PU membrane to realize improved biocompatible biomaterials. Surface morphological characteristics and wettability properties were investigated by SEM and AFM techniques and water contact angle measurements. Water contact angle depends on surface chemistry and the two methods employed for preparation of biomembranes as well as roughness of the membrane surfaces. Rheological study brings information on electrospinability of the polymer solutions/dispersions. Silver nanoparticles greatly influence the electrospinability of the polymer dispersions because of the increase in dynamic viscosity with the increasing silver content. Native PU and PU incorporated with low contents of AgNPs less than 0.3 % show high cell proliferation and good biocompatibility. The electrospun PU-EM-Ag nanobiocomposite membranes bring the advantage of using of low amounts of bioactive and biocidal components. The obtained silver nanobiocomposite membranes possess good bioactivity and non-cytotoxicity necessary for biomedical device applications. The obtained nanobiocomposite membranes are expected to find application for medical devices such as urinary catheters, wound dressings, etc.