Bailiang Wang

Surface modification of intraocular lenses with hyaluronic acid and lysozyme for the prevention of endophthalmitis and posterior capsule opacification

Posterior capsule opacification is one of the complications of cataract surgery caused by the adhesion and reproduction of residual human lens epithelial cells (HLECs) on the posterior capsule. Infectious endophthalmitis is another common complication due to bacterial colonization and biofilm formation. Hyaluronic acid–lysozyme (HA–lysozyme) composite coating was covalently grafted on the surface of PMMA intraocular lenses by reaction with N-hydroxysuccinimide (NHS) and 1-etil-3-(3-dimetilaminopropil) carbodiimida (EDC). Measurements of water contact angle and AFM revealed that the coating created a highly hydrophilic surface with low roughness. Adherences of S. aureus and HLECs on PMMA with HA or HA–lysozyme coating were significantly reduced due to the hydrophilic property of HA. Bactericidal activity of HA–lysozyme coatings, measured by SEM and LIVE/DEAD bacterial viability kit, was remarkably effective against S. aureus owing to the component of lysozyme. The achievement of the HA–lysozyme composite coating with anti-adhesive and antibacterial properties can potentially be widely used on surface modification of PMMA intraocular lenses and other biomedical implants.

A self-defensive antibacterial coating acting through the bacteria-triggered release of a hydrophobic antibiotic from layer-by-layer films

Drug delivery systems play important roles in the construction of antibacterial coatings on the surfaces of biomaterials. However, excessive release of antibiotics in the environment can lead to the generation of resistant bacteria. A methoxy poly(ethylene glycol)-poly(ε-caprolactone)-chitosan (MPEG-PCL-CS) block polymer was prepared through covalent grafting of CS onto MPEG-PCL. MPEG-PCL-CS micelles were prepared and showed a high load capacity for the hydrophobic antibiotic triclosan (TCA) (∼5 wt%). Multilayer films were constructed through self-assembling TCA/MPEG-PCL-CS cationic micelles with poly(acrylic acid) (PAA). Transmission and scanning electron microscopy analyses confirmed the presence of micelles on the surface (20-40 nm). As barriers for the antibiotic, the (TCA/MPEG-PCL-CS)/PAA multilayer films contained a high load of TCA (255 μg cm-2). Importantly, the multilayer films showed both bacteria-triggered and pH-responsive release properties and can be used as self-defensive antibacterial coatings. Bacterial adhesion caused a local acidic environment and altered the permeability of the multilayer films, promoting drug release. Both in vitro and in vivo antibacterial tests indicated a high bactericidal activity of drug-loaded multilayer films against both E. coli and S. aureus.

Direct Loading and Tunable Release of Antibiotics from Polyelectrolyte Multilayers To Reduce Bacterial Adhesion and Biofilm Formation.

Bacteria adhesion on the surface of biomaterials and following biofilm formation are important problems in biomedical applications. The charged antibiotics with small molar mass can hardly deposit alternately with polymers into multilayered films to load the drug. Herein, the (poly(acrylic acid)-gentamicin/poly(ethylenimine))n ((PAA-GS/PEI)n) multilayer film was designed and constructed via a layer-by-layer self-assembly method. Low molar mass GS cations were first combined with polyanion PAA and self-assembled with PEI to form multilayer films showing exponential growth behavior. The GS dosage could be adjusted by changing the layer number of films. Furthermore, the thermal cross-linking method was used to control the release rate of GS in PBS buffer. Owing to the diffusion of GS, a zone of inhibition of about 7.0 mm showed the efficient disinfection activity of the multilayer film. It could also be seen from the biofilm inhibition assay that the multilayer film effectively inhibited bacterial adhesion and biofilm formation. As the drug loading dosage was 160 μg/cm(2), the multilayer films showed very low cytotoxicity against human lens epithelial cells. The present work provides an easy way to load GS into multilayer films which can be applied to surface modification of implants and biomedical devices.

Surface-initiated RAFT polymerization of p (MA POSS-co-DMAEMA+) brushes on PDMS for improving antiadhesive and antibacterial properties

ABSTRACT Poly (dimethylsiloxane) (PDMS) silicones, as a common intraocular lenses material, often lead to the happen of posterior capsule opacification due to the adhesion and reproduction of residual human lens epithelial cells (HLECs) on the posterior capsule after surgery. Bacterial infection is a potentially catastrophic complication of cataract surgery, which occurs within several days after implantation of the lens. A novel poly (methacrylisobutyl polyhedral oligomeric silsesquioxane-co-2-(dimethylamino)-ethyl methacrylate) (p (MA POSS-co-DMAEMA)) brush was synthesized by reversible addition–fragmentation chain-transfer (RAFT) polymerization. After being quaternized treatment by 1-bromo-heptane, p (MA POSS-co-DMAEMA+) brushes with bactericidal function were obtained. The optical transmission of the brushes functionalized PDMS was better than p (DMAEMA+) coating modified PDMS, which may be due to the similar chemical composition of MA POSS and PDMS. Measurements of water contact angle, spectroscopic ellipsometry, and atomic force microscope were used to characterize hydrophilicity, thickness, and morphology of the brushes. Results revealed that brushes rendered PDMS surface more hydrophilicity and higher roughness. Adherences of bovine serum albumin and HLECs on the p (MA POSS-co-DMAEMA+) brushes were significantly reduced due to the hydrophilic property and cytotoxicity of the brushes. Bactericidal activity of the brushes, measured by shake-flask culture and LIVE/DEAD bacterial viability kit staining methods, was remarkably effective against S. aureus owing to the component of p (DMAEMA+). GRAPHICAL ABSTRACT

Development of antibacterial and high light transmittance bulk materials: Incorporation and sustained release of hydrophobic or hydrophilic antibiotics

Infection associated with medical devices is one of the most frequent complications of modern medical biomaterials. Bacteria have a strong ability to attach on solid surfaces, forming colonies and subsequently biofilms. In this work, a novel antibacterial bulk material was prepared through combining poly(dimethyl siloxane) (PDMS) with either hydrophobic or hydrophilic antibiotics (0.1-0.2 wt%). Scanning electron microscopy, water contact angle and UV-vis spectrophotometer were used to measure the changes of surface topography, wettability and optical transmission. For both gentamicin sulfate (GS) and triclosan (TCA), the optical transmission of the PDMS-GS and PDMS-TCA blend films was higher than 90%. Drug release studies showed initial rapid release and later sustained release of GS or TCA under aqueous physiological conditions. The blend films demonstrated excellent bactericidal and sufficient biofilm inhibition functions against Gram-positive bacteria (Staphylococcus aureus, S. aureus) measured by LIVE/DEAD bacterial viability kit staining method. Kirby-Bauer method showed that there was obvious zone of inhibition (7.5-12.5mm). Cytocompatibility assessment against human lens epithelial cells (HLECs) revealed that the PDMS-GS blend films had good cytocompatibility. However, the PDMS-TCA blend films showed certain cytotoxicity against HLECs. The PDMS-0.2 wt% GS blend films were compared to native PDMS in the rabbit subcutaneous S. aureus infection model. The blend films yielded a significantly lower degree of infection than native PDMS at day 7. The achievement of the PDMS-drug bulk materials with high light transmittance, excellent bactericidal function and good cytocompatibility can potentially be widely used as bio-optical materials.

Construction of a temperature-responsive terpolymer coating with recyclable bactericidal and self-cleaning antimicrobial properties

Once a biomedical implant is implanted into a human body, proteins and bacteria can easily colonize the implant, and subsequently, a biofilm can grow on the surface. A biofilm can protect the inhabiting bacteria against macrophages and neutrophil cell attack from the host immune system. The most important issue for artificial antibacterial surfaces is the accumulation of the bacteria corpse after they are killed by contact, which promotes further adhesion of bacteria and biofilm formation. Therefore, we constructed a novel multifunctional bactericidal and fouling release antibacterial surface through the combination of temperature-responsive N-vinylcaprolactam (VCL), hydrophilic 2-methacryloyloxyethyl phosphorylcholine (MPC) and a bactericidal quaternary ammonium salt (2-(dimethylamino)-ethyl methacrylate (DMAEMA+)). The terpolymer coating was prepared through surface-initiated reversible addition-fragmentation chain-transfer (RAFT) polymerization and characterized using water contact angle measurements, atomic force microscopy and spectroscopic ellipsometry. At a temperature above the lower critical solution temperature (LCST), the P(VCL-co-DMAEMA+-co-MPC) terpolymer coating was in a compressed and hydrophobic state with low moisture content, which displayed bactericidal efficiency against Gram-positive Staphylococcus aureus. The coating could be switched into a relatively hydrophilic surface at a temperature below the LCST, which showed self-cleaning properties against both bacteria and bovine serum albumin. The functionalized surface showed good biocompatibility against human lens epithelial cells as evaluated by morphology studies and activity measurements.

Fabrication of nonfouling, bactericidal, and bacteria corpse release multifunctional surface through surface-initiated RAFT polymerization

Infections after surgery or endophthalmitis are potentially blinding complications caused by bacterial adhesion and subsequent biofilm formation on the intraocular lens. Neither single-function anti-adhesion surface nor contacting killing surface can exhibit ideal antibacterial function. In this work, a novel (2-(dimethylamino)-ethyl methacrylate-co-2-methacryloyloxyethyl phosphorylcholine) (p (DMAEMA-co-MPC)) brush was synthesized by “grafting from” method through reversible–addition fragmentation chain transfer polymerization. 1-Bromoheptane was used to quaternize the p (DMAEMA-co-MPC) brush coating and to endow the surface with bactericidal function. The success of the surface functionalization was confirmed by atomic force microscopy, water contact angle, and spectroscopic ellipsometry. The quaternary ammonium salt units were employed as efficient disinfection that can eliminate bacteria through contact killing, whereas the 2-methacryloyloxyethyl phosphorylcholine units were introduced to suppress unwanted nonspecific adsorption. The functionalized poly(dimethyl siloxane) surfaces showed efficiency in reducing bovine serum albumin adsorption and in inhibiting bacteria adhesion and biofilm formation. The copolymer brushes also demonstrated excellent bactericidal function against gram-positive (Staphylococcus aureus) bacteria measured by bacteria live/dead staining and shake-flask culture methods. The surface biocompatibility was evaluated by morphology and activity measurement with human lens epithelial cells in vitro. The achievement of the p (DMAEMA+-co-MPC) copolymer brush coating with nonfouling, bactericidal, and bacteria corpse release properties can be used to modify intraocular lenses.

Construction of High Drug Loading and Enzymatic Degradable Multilayer Films for Self-Defense Drug Release and Long-Term Biofilm Inhibition

Bacterial infections and biofilm formation on the surface of implants are important issues that greatly affect biomedical applications and even cause device failure. Construction of high drug loading systems on the surface and control of drug release on-demand is an efficient way to lower the development of resistant bacteria and biofilm formation. In the present study, (montmorillonite/hyaluronic acid-gentamicin)10 ((MMT/HA-GS)10) organic/inorganic hybrid multilayer films were alternately self-assembled on substrates. The loading dosage of GS was as high as 0.85 mg/cm2, which could be due the high specific surface area of MMT. The obtained multilayer film with high roughness gradually degraded in hyaluronidase (HAS) solutions or a bacterial infection microenvironment, which caused the responsive release of GS. The release of GS showed dual enzyme and bacterial infection responsiveness, which also indicated good drug retention and on-demand self-defense release properties of the multilayer films. Moreover, the GS release responsiveness to E. coli showed higher sensitivity than that to S. aureus. There was only ∼5 wt % GS release from the film in PBS after 48 h of immersion, and the amount quickly increased to 30 wt % in 105 CFU/mL of E. coli. Importantly, the high drug dosage, smart drug release, and film peeling from the surface contributed to the efficient antibacterial properties and long-term biofilm inhibition functions. Both in vitro and in vivo antibacterial tests indicated efficient sterilization function and good mammalian cell and tissue compatibility.

Bacterial infection microenvironment-responsive enzymatically degradable multilayer films for multifunctional antibacterial properties

The adhesion of bacteria is one of the most important stages in biofilm formation and bacterial infection. Once bacteria have adhered to a biomaterial surface, it is hard to eliminate them, and bacterial growth and infection are inevitable. In the current study, we have designed and constructed enzymatically degradable (hyaluronic acid/chitosan)n-(hyaluronic acid/polylysine)n ((HA/CHI)n-(HA/PLL)n) composite multilayer films via a layer-by-layer self-assembly method. Spectroscopic ellipsometry and scanning electron microscopy cross section measurements showed the exponential growth behavior of (HA/CHI)10-(HA/PLL)10 multilayer films (∼2 μm). The increased secretion of hyaluronidase and chymotrypsin in the bacterial infection microenvironment led to the fast degradation of the outer (PLL/HA)n multilayer films in the first 24 h. Enzymatic degradation of the multilayer films efficiently reduced the adhesion of both Staphylococcus aureus and Escherichia coli (>99%). Bacterial live/dead staining demonstrated the bactericidal action of the remaining bottom (CHI/HA)n multilayer films against the two kinds of bacteria. In vivo subcutaneous tests on New Zealand white rabbits, wound appearance and histopathology analysis showed that the implantation of composite multilayer film-modified PDMS promoted wound healing and the materials demonstrated a self-defense antibacterial effect. The material demonstrated both anti-adhesive and bactericidal properties and could be used to modify biomedical implants.

Recent developments in smart antibacterial surfaces to inhibit biofilm formation and bacterial infections

Since their development over 70 years, antibiotics are still the most effective strategy to treat bacterial biofilms and infections. However, the overuse of antibiotics in human healthcare and industrial applications has resulted in the development of serious antibiotic-resistant bacteria. Therefore, alternative ways to prevent bacteria attachment and biofilm formation are urgently needed. Recently, mediated biofilm formation processes and smart antibacterial surfaces have emerged as promising strategies to prevent and treat bacterial infections. This review discusses the recent progress in biofilm interference and smart antibacterial surfaces. Smart antibacterial and anti-biofilm surfaces should be responsive to the bacterial infection environment, switchable between various antibacterial functions and have a special bio-inspired structure and function. The major topics discussed are: (i) smart anti-biofilm surfaces via the prevention of biofilm formation or promoting mature biofilm dissolution, (ii) smart materials for reversible killing and/or release of bacteria, (iii) smart surfaces responsive to bacterial infection microenvironments or external stimuli and (iv) bio-inspired surfaces with antifouling and bactericidal properties.