Acidic sophorolipid and antimicrobial peptide based formulation as antimicrobial and antibiofilm agents

Antimicrobial peptides (AMPs) are considered promising candidates to treat various infections in soft tissues and skin. However, no effective treatment based on AMPs has been reached to clinics due to their instability in serum and wounds. Biosurfactants such as acidic sophorolipids (ASLs) of very high concentrations (equal or above 5 mg/mL) have been demonstrated to be antimicrobial agents, however these concentrations might induce cytotoxic effects to human cells. Here, we have demonstrated the synergistic antimicrobial effect of ASL nanoparticles (NPs) and LL37 peptides (below their minimum inhibitory concentrations; MICs) to eradicate Gram-positive and Gram-negative bacteria in human serum (HS) and in the presence of trypsin. The formulations containing ASL NPs (500 mu g/mL) and LL37 peptides (15-25 mu g/mL) effectively kill wide strains of bacteria in 5 % HS and the presence of trypsin. Moreover, the combination of ASL NPs (500 mu g/mL) and LL37 peptides (15 mu g/mL) prevents the formation of S. aureus biofilm and eradicates the one-day old biofilm. Importantly, the combination of ASL NPs and LL37 peptides severely damages the cell membrane of Escherichia coli (E. coli) as shown by atomic force microscopy (AFM). The combination of ASL NPs and LL37 peptides rapidly damages the outer (OM) and inner membrane (IM) of E. coli, while ASL NPs (1000 mu g/mL) alone slowly compromise the integrity of the bacterial membrane. Importantly, the combination of ASL NPs and LL37 peptides is biocompatible to human keratinocyte cells (HaCaTs) and human umbilical vein endothelial cells (HUVECs), and induces the expression of antiinflammatory cytokine in macrophages. Overall, ASL NPs in combination with LL37 peptides might be developed as an effective topical formulation to prevent bacterial infections in the skin.