3.8.1. Antimicrobial Activities

The disc diffusion method has been used to evaluate the antibacterial activities of the composite hydrogels, which have been assessed against Gram-positive (*Staphylococcus aureus* (*S. aureus*)) and Gram-negative (*Escherichia coli* (*E. coli*) and *Pseudomonas aeruginosa* (*P. aeruginosa*)) bacterial strains. The bacterial strains were provided by ATCC, USA. The molten agar was poured on the Petri dishes and allowed to cool at ambient temperature and bacterial media was applied via cotton swab. Then, the hydrogels (70 μL) were placed on bacterial cultured Petri dishes using a micropipette. The Petri dishes were incubated at 37 ◦C for 24 h, and the diameter of zone inhibition was measured in millimeters (mm).

#### 3.8.2. Anticancer Analysis

The cytotoxicity of the materials was investigated against the U87 cell line by MTT assay to determine cell viability and cell morphology under in vitro conditions. The purpose of this study is to determine the anticancer behavior of composite hydrogels and drug-loaded composite hydrogels [20].
