The purpose of this study were to prepare and characterize nonionic nano-vesicles prepared from various types (span20, span 60, span 80, brij 72, glyceryl monostearate and polyethylene glycol-6 stearate). The vesicles were prepared by film hydration followed by high pressure homogenization techniques. Physicochemical properties were investigated by varying molar ratios, total lipids, negative charge inducers, membrane modifiers, electrolytes, and pH of aqueous phases. Results showed that nonionic vesicle approximately 100 to 200 nm depending on surfactant types could be prepared with narrow size distribution (PDV<0.3). The vesicle sizes were incresed with increasing cholesterol molar ratios and total lipid concentrations. Mixing nonionic nano-vesicles containing 0, 5, 10 mol% oleic acid as negative charge inducer with osmotic agent, KCl solution at 60, 40 and 80 mM caused significantly increase in vesicle size and aggreation. Rigid nano-vesicles are observed when using precirol ATO5 concentration < 25 mol%. Vesicles were fromed independent of pH of aqueous solutions but agrregations were observed at pH 3 and 5. Differential scanning calorimetry thermograms revealed that excipients used in production of nano-vesicles were presented in amorphous form. These results suggested that a proper selection of excipient can lead to successful encapsulation of active ingredients for drug and cosmetic application.