<p>In present work, silver nanoparticles were generated in situ within a cotton matrix by bio-reduction method using infused lemon leaves through aqueous AgNO₃ solution at ambient conditions. The prepared Nanocomposite Cotton Fabrics (NCCFs) with “in situ” generated AgNPs had been characterized by studies of SEM, FTIR spectroscopy, X-ray diffraction, Thermal and Antibacterial tests. The FTIR spectra of pristine Cotton fabric, Matrix and NCCFs were pronounced similar peak positions except variation in intensity of the peaks. The SEM analysis revealed the presence of in situ generated AgNPs in the NCCFs. The generated silver nanoparticles exhibit the crystallinity within NCCFs. The prepared NCCFs exhibited good thermal stability and antibacterial activity against Gram negative as well as Gram positive bacteria. Hence, prepared NCCFs can be utilized as medicated dressing materials and preparation method is proven as cost effective.</p> Graphical abstract <p></p>