In this work, we developed novel nanocomposite three-dimensional (3D) scaffolds composed of chitosan (CTS), halloysite nanotubes (HNTs) and silver nanoparticles (AgNPs) with enhanced antimicrobial activity and fibroblast cell compatibility for their potential use in wound dressing applications. A stock CTS-HNT solution was obtained by mixing water-dispersed HNTs with CTS aqueous-acid solution, and then, AgNPs, in different concentrations, were synthesized in the CTS-HNT solution a CTS-mediated reduction method. Finally, freeze-gelation was used to obtain CTS-HNT-AgNP 3D porous scaffolds (sponges). Morphology analysis showed that synthesized AgNPs were spherical with an average diameter of 11 nm. HNTs' presence did not affect the AgNPs morphology or size but improved the mechanical properties of the scaffolds, where CTS-HNT sponges exhibited a 5 times larger compression stress than bare-CTS sponges. AgNPs in the scaffolds further increased their mechanical strength in correlation to the AgNP concentration, and conferred them improved antibacterial activity against Gram-negative and Gram-positive bacteria, inhibiting the planktonic proliferation and adhesion of bacteria in a AgNP concentration depending on manner. cell viability and immunofluorescence assays exhibited that human fibroblast (HF) culture was supported by the sponges, where HF retained their phenotype upon culture on the sponges. Present CTS-HNT-AgNP sponges showed promising mechanical, antibacterial and cytocompatibility properties to be used as potential scaffolds for wound dressing applications.