Various methods to combine hyaluronic acid and antimicrobial peptides coatings and evaluation of their antibacterial behaviour.

To prevent bacterial adhesion and contamination, biomaterials exhibiting both antiadhesive and biocidal properties are the most promising way. However, control of the properties combination is not so easy due, in particular, to antagonist mechanisms. Antibacterial surfaces against Staphylococcus epidermidis adhesion were here elaborated by using both nisin grafting and repelling polysaccharide coating. We evaluated two strategies aiming to improve the antimicrobial peptide (AMP) immobilization parameters (i.e., the accessibility and/or local density) in order to obtain the best antimicrobial activity on surfaces. We thus (i) grafted the nisin on a surface previously coated with hydrolyzed hyaluronic acid (HA) (to decrease the length of the polysaccharide chains) or (ii) coupled nisin and HA in solution before grafting this complex on surfaces. XPS analysis pointed out a lower amount of nisin on the surface for both approaches compared to the immobilization of nisin on native HA. However, an antibacterial activity was maintained, probably due to a higher local density of the AMP when surfaces were modified with hydrolyzed hyaluronic acid, leading to a better combination of antiadhesive-biocidal properties. Microscopy fluorescent observations demonstrated that accumulation of dead cells was also avoided by some coatings architecture.

Elaboration of antibacterial plastic surfaces by a combination of antiadhesive and biocidal coatings of natural products.

Antibacterial polyolefins surfaces, combining biocidal and antiadhesive properties, were elaborated by a covalent grafting of antimicrobial peptides (AMPs), able to kill adherent bacteria, on a pre-immobilized hyaluronic acid (HA) layer, able to repel the micro-organisms. Different HA activation rate for its immobilization, were used to change HA layer morphology and number of residual free carboxylic acid functions for AMPs grafting. Based on adhesion tests on Staphylococcus epidermidis and microscopy fluorescent observations, the presence of the two combined properties was shown to be depended on the HA activation rate. Thus, the best addition effect was observed for an AMP grafting on a surface based on a high HA activation, data pointing out a decrease of the bacterial adhesion up to 99.8% and a perturbation of the bacterial membrane integrity of adhered bacteria. On the contrary, a decrease of the antibacterial activity was observed for an AMP grafting on a surface based on a low HA activation.