Nowadays, modern functional wound dressings have become prominence due to their biocompatible, biodegradable, and non-toxic nature and similarity with ECM of skin. Environmentally friendly halloysite nanotubes (HNTs) are gained attention due to their tubular structure that allow encapsulation of various agents into these tubes to enhance antimicrobial activities in a sustained manner as well as with enhanced mechanical properties for the development of antimicrobial nanocomposites. In this thesis, bilayer wound dressing was fabricated by using ZeinVancomycin loaded HNTs nanofibers as upper layer to mimic ECM of skin and provide antibacterial protection, and chitosan sponge as bottom layer to absorb the excess exudates of wound, provide gas transmission and facilitate the migration of inflammatory and fibroblast cells into the healing wounds. The morphology of nanofibers and encapsulation efficiency of Vancomycin into HNTs are optimized to achieve similar fiber structure with skin tissue, and to obtain sustained release of the drug. Chemical interaction between Vancomycin-HNT and Zein-HNT were characterized by FT-IR. The surface charge of drug loaded and unloaded HNTs was determined by zeta potential analysis. Bilayer sponges were characterized by SEM, FT-IR, porosity, mechanical properties, contact angle, water vapor transmission rate, swelling, degradation, in vitro drug release, and antimicrobial activity analyzes. The diameter of drug loaded HNT-Zein nanofibers were found in the range of native skin collagen fibril (203± 0,05- 225±0,06 nm). The water vapor permeability of the wound dressings is found in the appropriate range for wound healing (2833- 2490 g/m2 day). Bilayer dressings containing two different drug:HNTs ratios reached 78% cumulative release at the end of 14 days, however rand the release medium showed antimicrobial activity. In conclusion, the developed drug-loaded bilayer mat has been found as a potential candidate for wound dressings applications to treat the chronic infections.