This study offers a new wound dressing by immobilization of amino acid residues on partially carboxymethylated cotton nonwoven fabrics (CM-CN). To improve the absence of capacity to protect the open wound from infection, firstly, a usual cotton nonwoven fabric was chemically modified by utilizing a pad-dry-cure technique including a carboxymethylation procedure. Subsequently, the chicken feather wastes as a natural source were used for the cationization of CM-CN by the same method. Keratin successfully was extracted from chicken feathers using both chemical and enzymatic processes. Physicochemical and biological properties of the prepared samples were evaluated by FTIR, SEM, mechanical properties such as tensile, bending test and wrinkle recovery, biodegradation, permeability (air and water vapor), and MTT assay for cell viability and proliferation. The tensile strength increased to 200%, the air permeability almost doubled, and the greatest cell growth was observed for modified samples. According to the obtained results, enzymatic extraction was influential and the samples coated with enzyme-extracted keratin showed enhanced properties suitable for wound dressing applications.