Tissue regeneration supported by biocompatible scaffolds faces delay in regeneration due to Staphylococcal attack. This study was conducted to develop a chitosan based tissue regeneration scaffold with antistaphylococcal property. Chitosan membrane loaded with different concentrations of green tea aqueous extracts {0.016mg (GTM 1), 0.02mg (GTM 2), 0.1mg (GTM 3), 0.5mg (GTM 4), 1.0mg (GTM 5), 1.5mg (GTM 6) & 2.0mg (GTM 7) extract/0.86mg membrane} were prepared and morphological properties, degradation and antistaphylococcal activity were examined. Microbial growth inhibition zone diameter in solid agar medium was significantly higher (P<0.05) for GTM-7, when compared to other concentrations. The inhibition of Staphylococcus aureus in the presence of GTM 7 was analysed in liquid media and the antibacterial property was evaluated by spread plate and turbidity measurement. This study revealed that chitosan membrane loaded with green tea aqueous extract showed a better activity against Staphylococcus aureus and therefore, it can be used as a scaffold for tissue regeneration. Film solubility test performed in PBS, pH 7.4 showed that the solubility of GTM 7 was significantly increased when compared to unloaded chitosan membrane. The scanning electron microscopic image of GTM 7 showed a rough and uneven surface morphology due to the incorporation of green tea components. This study showed a remarkable lethal action against the nosocomial infecting agent during wound healing with the help of green tea aqueous extract loaded chitosan membrane. This will have a good therapeutic significance in fast tissue regeneration.