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The antibacterial sponges with high macroporosity, high interconnectivity and high biocompatibility is a significant concern for wound healing applications. In this work, novel Aloe vera (AV) based sponges were developed via subsequent lyophilization with further chemical crosslinking throughout the double network sponges. Single network was composed of gelatin-sodium alginate (G-SA) while the double network using gelatin-sodium alginate- sodium hyaluronate (G-SA-HA) was produced with the addition of hyaluronic acid solution into the gelatin-sodium alginate matrix. Lastly, Aloe vera as the bioactive agent was fabricated throughout the gelatin-sodium alginate-sodium hyaluronate matrix. The AV-based sponges demonstrated large pores with high interconnectivity. The swelling degree of the AV-based sponges were higher than that of G-SA and G-SA-HA sponges. The release of AV from the sponges reached an equilibrium value after 24 h showing a more controlled release at pH 5.5 than at pH 7.4. AV-based sponges showed antibacterial activity against Staphylococcus aureus, Enterococcus faecalis, Streptococcus pneumoniae and Bacillus cereus and displayed any cytotoxicity against Mesenchymal stem cells. Scientific Projects Coordination Unit of Hitit UniversityHitit University [MUH19001.18.001]; Hitit University Scientific and Tecnical Application and Research Center The financial support from the Scientific Projects Coordination Unit of Hitit University under the project number MUH19001.18.001 is gratefully acknowledged. The authors are also greateful to the support of Hitit University Scientific and Tecnical Application and Research Center. WOS:000608662700001 2-s2.0-85100181974