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We report on the layer-by-layer design principles of poly(methacrylic acid) (PMAA) ultrathin hydrogel coatings that release antimicrobial agents (AmAs) in response to pH variations. The studied AmAs include gentamicin and an antibacterial cationic peptide L5. Adipic acid dihydrazide (AADH) is a cross-linker which, relative to ethylenediamine (EDA), increases the hydrogel hydrophobicity and introduces centers for hydrogen bonding to AmAs. AmA retention in AADH-cross-linked hydrogels in high-salt solutions was enhanced while AmA release at low pH was suppressed. L5 retains its antibacterial activity toward planktonic Staphylococcus epidermidis after release from PMAA hydrogels in response to pH decreases in the surrounding medium due to bacterial growth. Staphylococcus epidermidis adhesion and colonization was almost completely inhibited by L5 loading of hydrogels. The AmA-releasing and AmA-retaining properties of these hydrogel coatings provide new opportunities to study the fundamental mechanisms of AmA-coating-bacteria interactions and develop a new class of clinically relevant antibacterial coatings for medical devices.

A hydrogen-bonded layer-by-layer (LbL) technique was used to build multilayers of neutral, temperature-responsive polymers such as poly(N-isopropylacrylamide) (PNIPAM), poly(N-vinylcaprolactam) (PVCL), poly(vinyl methyl ether) (PVME), or poly(acrylamide) (PAAm) with a polycarboxylic acid such as poly(acrylic acid) (PAA), poly(methacrylic acid) (PMAA), or poly(ethacrylic acid) (PEAA). For all multilayers involving temperature-responsive polymers, the temperature used during or after self-assembly had a significant effect on film stability with pH changes. The proximity of the self-assembly or post-self-assembly temperature to the critical temperature of phase separation of a neutral polymer from solution resulted in a higher pH stability of multilayers. However, for polymers with a lower critical solution temperature (LCST) such as PNIPAM, PVCL, or PVME within PNIPAM/PMAA, PVCL/PMAA, or PVME/PMAA multilayers, the critical pH of film disintegration (pH(crit)) increased in the temperature range from 10 to 37 degrees C, whereas for polymer films with an upper critical solution temperature (UCST), such as PAAm within PAAm/PMAA, the film showed the opposite trend. Using a hydrogen-bonded polyvinylpyrrolidone (PVPON)/PMAA system, which is not responsive to temperature changes, we constructed hybrid films with lower [PNIPAM/PMAA](n) and higher [PVPON/PMAA](m) strata and obtained free-floating [PVPON/PMAA](m) films by temperature-triggered dissolution of the PNIPAM/PMAA layers at a constant pH value. The kinetics of [PVPON/PMAA](m) film release was strongly dependent on the number of bilayers within the PNIPAM/PMAA stratum, indicating significant interpenetration between PNIPAM/PMAA and PVPON/PMAA bilayers. Importantly, the use of PEAA instead of PAA or PMAA in film assembly enabled the construction of hydrogen-bonded LbL films that can be released by applying temperature as a trigger at near-physiological pH values. This feature makes such release layers attractive candidates for future tissue engineering applications.

We report pH/bacteria-responsive nanocomposite coatings with multiple mechanisms of antibacterial protection that include the permanent retention of antimicrobials, bacteria-triggered release of antibiotics and bacteria-induced film swelling. A novel small-molecule-hosting film was constructed using layer-by-layer deposition of montmorillonite (MMT) clay nanoplatelets and polyacrylic acid (PAA) components, both of which carry a negative charge at neutral pH. The films were highly swollen in water, and they exhibited major changes in swelling as a function of pH. Under physiologic conditions (pH 7.5, 0.2 M NaCl), hydrogel-like MMT/PAA films took up and sequestered B45% of the dry film matrix mass of the antibiotic gentamicin, causing dramatic film deswelling. Gentamicin remained sequestrated within the films for months under physiologic conditions and therefore did not contribute to the development of antibiotic resistance. When challenged with bacteria (Staphylococcus aureus, Staphylococcus epidermidis or Escherichia coli), the coatings released PAA-bound gentamicin because of bacteria-induced acidification of the immediate environment, whereas gentamicin adsorbed to MMT nanoplatelets remained bound within the coating, affording sustained antibacterial protection. Moreover, an increase in film swelling after gentamicin release further hindered bacterial adhesion. These multiple bacteria-triggered responses, together with nontoxicity to tissue cells, make these coatings promising candidates for protecting biomaterial implants and devices against bacterial colonization. INTRODUCTIONPolymer nanocomposites uniquely combine the properties of inorganic and organic components that is central to the development of novel advanced materials. The naturally occurring smectite clays, which are chemically and thermally stable and biocompatible, are attractive nanocomposites. Exfoliated clay nanoplatelets strongly impact the mechanical, transport, optical, fire retardant and gas barrier properties of nanocomposite materials. 1,2 For many biomedical applications, however, hydrophilic rather than hydrophobic nanocomposites are of great interest, and responsive properties are essential. 3 Reports on responsive hydrophilic nanocomposites are rare, and those few known to us advantageously use the complementary properties of temperature/pH-responsive polymers and clay nanosheets to yield environmentally controlled free-standing materials. 4 A promising way to leverage the favorable properties of hydrophilic-responsive nanocomposites on surfaces is to use the layer-by-layer (LbL) technique to construct 'smart' surface coatings. 5 To construct clay-containing LbL films, electrostatic interactions of positively charged polymers with negatively charged basal planes of silicate nanosheets, 6,7 neutral polymer/nanoplatelet hydrogen bonding 8 or a combination of the two have been explored. 9

We developed a highly efficient, biocompatible surface coating that disperses bacterial biofilms through enzymatic cleavage of the extracellular biofilm matrix. The coating was fabricated by binding the naturally existing enzyme dispersin B (DspB) to surface-attached polymer matrices constructed via a layer-by-layer (LbL) deposition technique. LbL matrices were assembled through electrostatic interactions of poly(allylamine hydrochloride) (PAH) and poly(methacrylic acid) (PMAA), followed by chemical crosslinking with glutaraldehyde and pH triggered removal of PMAA, producing a stable PAH hydrogel matrix used for DspB loading. The amount of DspB loaded increased linearly with the number of PAH layers in surface hydrogels. DspB was retained within these coatings in the pH range from 4 to 7.5. DspB-loaded coatings inhibited biofilm formation by two clinical strains of Staphylococcus epidermidis. Biofilm inhibition was ≥ 98% compared to mock-loaded coatings as determined by CFU enumeration. In addition, DspB-loaded coatings did not inhibit attachment or growth of cultured human osteoblast cells. We suggest that the use of DspB-loaded multilayer coatings presents a promising method for creating biocompatible surfaces with high antibiofilm efficiency, especially when combined with conventional antimicrobial treatment of dispersed bacteria.