The eggshell cuticle is the proteinaceous outermost layer of the eggshell which regulates water exchange and protects against entry of micro-organisms. Outer eggshell and cuticle protein was extracted from domestic chicken. The aim of the research is to find out the effect of the treated and untreated nano particles of egg shells with micro wave cold plasma on the effectiveness of E. coli (negative bacteria) that infect the skin and measure the diameter of bacterial inhibition zone, the eggshell has been prepared by a chemical method (sol gel) and measure the level of acidity and the PH is neutral. The result of Atomic Force Microscope (AFM) shows that the particles diameters become smaller with nano-particles solution than for egg shell tests (powder). The nano-particles solution for egg shell exposed to cold plasma. The microwave plasma used in this search has voltage "175v" and the gas flow at " 5liter /min " and frequency (2.45 GHz) at the room temperature for five minutes’ time. The E. coli bacteria were treated with nano-particles solution egg shells. The results indicated that after 24 hours of placing the bacteria in the incubator, the inhibition zone for the nano-particles eggshell that for the is greater than that for nano-particles eggshell exposed to the plasma, and in both cases there were a good response.