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Metal/metal oxide nanoparticles have long been used as an antibacterial substitute, but fabrication of an effective carrier or delivery matrix for achieving a sustain release profile with high bactericidal efficacy alongwith good cytocompatibility is still an unresolved challenge. Herein, the study demonstrates a facile and unique route to fabricate a hierarchical nanobiocomposite with effective loading of ZnO/silver nanoparticles (Ag–NPs) in order to attain excellent bactericidal efficacy with good and sustainable release profile. Surface functionalized eggshell membranes (ESM) were deployed as three-dimensional loading matrices for efficient loading of ZnO/Ag–NPs. A simple sonochemical guided approach was adopted to synthesize ZnO nanoflakes in situ onto the microfibrous ESM and decorate it with Ag–NPs to fabricate a nanobiocomposite. Microstructural analysis confirms successful anchorage of ZnO nanoflakes and Ag–NPs on microfibrous eggshell membrane thus reinstating hierarchical morphology of the nanobiocomposites. FT-IR spectra confirms the biochemical composition whereas XPS analysis ratifies the interaction between ZnO and Ag–NPs further substantiating metallic state of Ag. ICP-MS studies affirms excellent and sustainable release profile of nanoparticles from the nanobiocomposites. Owing to the synergistic activity of ZnO/Ag–NPs, the nanobiocomposites demonstrated exceptional bactericidal activity against Gram-negative, E. coli or P. aeruginosa, and Gram-positive, S. aureus or B. subtilis, bacterial cells. Moreover, inherent antibacterial property of microfibrous natural ESM contributes positively toward the overall bactericidal activity. Further, a direct exposure of nanobiocomposites with NIH 3T3 cells revealed the biocompatible nature of developed matrices. Prolonged exposure also indicated that the 3T3 cells tend to adhere onto the microfibrous nanobiocomposite without any observable deformation in cellular morphology. The architectural tribology and excellent bactericidal performance of the nanobiocomposites along with its cytocompatible nature manifests its application as an alternate platform for varying biomedical applications.

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