Abstract : Nowadays, synthesis of plant based nanoparticles were gaining importance in the field of medicines for efficient drug delivery to cure many diseases. In the present study iron nanoparticles were synthesized with waste pista shell extracts and were subjected to various characterization studies such as UV-Vis spectrophotometer and FTIR. Antibacterial activity of Fe3O4-NP doped pista extract was anlysed by agar well diffusion method against E.coli, Pseudomonas aeruginosa, Bacillus subtlis and Staphylococcus aureus with gentamicin (20µl) as reference drug. Maximum zone of inhibition was found to be in Staphylococcus aureus with a diameter of 19mm and minimum zone of inhibition was found in Pseudomonas aeruginosa of 16mm. Escherichia coli was found to have 18mm and Bacillus subtilis to be 17mm during the period of study. Later Fe3O4-NP was doped onto cotton fabric of size 7 mm x 7 mm as per standard methods. Fe3O4-NP doped pista extract cotton fabric was characterized using SEM and EDAX for confirmation of size and chemical compounds. Swabbed petri plates with following bacterial cultures such as E.coli, Pseudomonas aeruginosa, Bacillus subtlis and Staphylococcus aureus were prepared as per standard procedures for antibacterial analysis by placing Fe3O4-NP doped pista extract cotton fabric onto the developed cultures, respectively and incubated for 24 hrs at 370C. E.coli registered the maximum zone of inhibition of 22mm followed by S. aureus with 20mm, P. aeruginosa with 19mm, 17mm by B. subtilis, respectively during the period of study. Cytotoxicity studies with vero cells lines revealed that 10µg/mL, 20µg/mL and 30µg/mL approximately 90% of cells remained viable with 97.78%, 95.21% and 93.29%, respectively. In vitro scratch wound healing assay was carried out as per standard procedure and calculated the scratch closure rate (SCR) which was found to be 20.52% at 24 hrs and 77.36% at 48 hrs of incubation.