Many bacteria live as biofilms to cope with unfavourable surroundings. Biofilms start from (i) a planktonic stage, (ii) initial adhesion to surfaces and (iii) formation of sessile micro-colonies that secrete extracellular polymeric substance (EPS), leading to bacterial resistance to antibiotics. Antimicrobial peptides (AMPs) are extensively studied with regard to planktonic bacteria but much less so with regard to biofilm formation. In the present study, we investigated how the above three steps are affected by the properties of the AMPs using a series of peptides composed of six lysines and nine leucines, which differ in their sequences and hence their biophysical properties. Treatment with bactericidal peptides at non-inhibitory concentrations resulted in reduced biofilm growth, for some starting from 25 nM which is 0.2 and 0.4% of their minimum inhibitory concentration (MIC 6.3 and 12.5 μM, respectively), continuing in a dose-dependent manner. We suggest that reduced bacterial adhesion to surfaces and decreased biofilm growth are due to the peptide's ability to coat either the biomaterial surface or the bacterium itself. Degradation of established biofilms by bactericidal and non-bactericidal peptides, within 1 h of incubation, occurs by either killing of embedded bacteria or detachment of live ones. In addition to shedding light on the mechanism of biofilm inhibition and degradation, these data may assist in the design of anti-biofilm AMPs.
- antibiotic resistance
- anti-biofilm activity
- antimicrobial peptides
- Pseudomonas aeruginosa
Abbreviations: AMP, antimicrobial peptide; CD, circular dichroism; CFU, colony-forming units; CV, Crystal Violet; DMF, dimethyl formamide; EPS, extracellular polymeric substance; FTIR, Fourier transform infrared spectroscopy; GdnHCl, guanidine hydrochloride; LB, Luria–Bertani; LPS, lipopolysaccharide; MBHA, methylbenzhydrylamine; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; RP-HPLC, reverse-phase HLPC; TFA, trifluoroacetic acid
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