Synthesis of modified guanidine-based polymers and their antimicrobial activities revealed by AFM and CLSM

Modified guanidine-based polymers with chain extension were synthesized by condensation and cross-linking polymerizations in an attempt to increase molecular weight and charge density of the antimicrobial polymers. The antimicrobial activity and the corresponding mechanisms were investigated by several approaches. The results indicated that the antimicrobial activities of the modified guanidine-based polymer, based on the minimum inhibition concentration (MIC) against E.coli, varied with alkyl monomer ratios. UV absorption at 260 nm further quantified the amount of intracellular components leaked into bacteria suspension. The UV absorption measurements were also used to monitor inhibition processes dynamically. It was found that the modified guanidine-based polymer inhibited the growth of bacteria by causing membrane compromised and intracellular leaked. Dual fluorescent dyes were used to stain all bacteria including the dead ones, which enabled us to utilize CLSM to visualize the viability of bacteria in the presence of various modified guanidine-based polymers without causing any damage. The morphologies of bacteria untreated and treated with modified guanidine-based polymer were observed using an atomic force microscope (AFM), which further demonstrated the damage of E.coli membrane and the leakage of intracellular component induced by the modified guanidine-based polymers.