Aim: The study aims to determine the optimal concentration of Aloe vera that demonstrates the highest antibacterial efficacy against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Methods: Aloe vera gel at concentrations of 20, 40, and 80% were tested for antibacterial activity against Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli using the agar diffusion method, with Ciprofloxacin as a control. Mueller Hinton Agar (MHA) was prepared, sterilized, poured into Petri plates, and swabbed with the bacteria. Wells were created in the agar, and zones of inhibition were measured. Data was analyzed using Statistical Package for the Social Sciences (SPSS) Software version 23.0 with a post hoc test. Results: Aloe vera gel at 20% inhibited only Escherichia coli, while 40 and 80% concentrations showed broad antibacterial activity against Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli. Ciprofloxacin was effective against all tested bacteria. Around 40 and 80% of Aloe vera gel were significantly more effective against Pseudomonas aeruginosa and Staphylococcus aureus compared to 20% (p < 0.05). Conclusion: Aloe vera gel exhibited antibacterial properties against Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli, at higher concentrations of 40 and 80%. Aloe vera exhibited distinct antibacterial properties when compared to Ciprofloxacin, indicating its potential as an alternative therapy and in facilitating wound healing. Clinical significance: Aloe vera gel, rich in vitamins, minerals, enzymes, and amino acids, exhibits antibacterial, anti-inflammatory, and wound-healing properties. Clinically, this study suggests Aloe vera gel could serve as a natural alternative or adjunct to antibiotics, potentially improving patient outcomes by reducing reliance on traditional antibiotics and lowering the risk of adverse effects and antibiotic resistance.