Treating infectious diseases with current available antimicrobial drugs is extremely difficult due to biofilms that act as

barriers and reduce the concentration of antimicrobial agents that reach the bacteria embedded in the biofilms. In this

study, we hypothesized that extracellular polymeric substances (EPS)-binding liposomes anchor to biofilm matrices and

sterically block the communication between bacteria, leading to biofilm inhibition. A 16-mer peptide, which binds to hyaluronic

acid as one of the EPS, was covalently conjugated to PEG (polyethylene glycol)-lipid for producing EPS-binding

liposomes. The effect of the liposomes on inhibiting or eradicating biofilm formation was investigated, compared to the

bare liposomes. Dynamic light scattering (DLS) measurement results showed that the EPS-binding liposomes and bare

liposomes have a particle size of < 200 nm and nearly neutral zeta potential. The molecular interaction of EPS extracted

from S. aureus biofilm with EPS-binding liposomes and free EPS-binding peptides was determined using isothermal titration

calorimetry (ITC) and the result revealed that EPS-binding liposome (Ka ~ 4.82 × 105) has better affinity than the free

EPS-binding peptides (Ka ~ 1.79 × 103). The minimal biofilm inhibitory concentration (MBIC) assay showed EPS-binding

liposomes have a better biofilm inhibition effect, in a dose-dependent manner, compared to the bare liposomes and free

EPS-binding peptides. Physical disruption and blocking chemical communication via biofilm binding are likely a key

mechanism behind the effectiveness of EPS-binding liposomes in biofilm inhibition although further study is needed.