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Since the number of antibiotic-resistant bacterial infections is growing and cases are getting worse every year, the search for new alternative bactericidal wound dressing treatments is becoming crucial. Within this context, the use of polysaccharides from plants and seeds in innovative biopolymer technologies is of key importance. In this work, bio-nano-composite guar gum/polyvinyl alcohol (PVOH) membranes loaded with aluminum-doped zinc oxide nanoparticles were produced via electrospinning. Citric acid was added to the mixture to increase spinnability. However, depending on the pH, zinc oxide nanoparticles are partially dissociated, decreasing their bactericidal efficiency. Thus, a second successful alkaline thermo-chemical regrowth step was added to the process to treat the obtained fibers. This alkaline thermo-chemical treatment reconstituted both the nanoparticles and their bactericidal properties. The Staphylococcus aureus antibacterial assay results show that the membranes obtained after the alkaline thermo-chemical treatment presented a 57% increase in growth inhibition.