Modification of fluorous substrates with oligo(ethylene glycol) via "click" chemistry for long-term resistance of cell adhesion.

In this work perfluorinated substrates fabricated from SiO2 glass slides are modified with oligo(ethylene glycol) (OEG) units for long-term resistance of cell adhesion purposes, based on fluorous interactions and click chemistry. Specifically, fluorous substrates, prepared by treatment of glass slides with 1H, 1H, 2H, 2H-perfluorodecyltrimethoxysilane (FAS17), were coated with ethynyl-OEG-C8F17, followed by covalent attachment of an azido-OEG via copper-catalyzed azide-alkyne cycloaddition (CuAAC) "click" reaction. We demonstrate that the resultant surface avoid fibrinogen adsorption and resisted cell adhesion for over 14days. X-ray photoemission spectroscopy (XPS) analysis and contact angle goniometry measurements confirm the presence of the OEG molecules on the fluorous substrates. Bright field optical images show total absence of 3T3 fibroblast cells on the OEG modified fluorinated substrate for 1 and 5days, and a remarkably decrease of cell adhesion at 14days.

Immobilization of antimicrobial peptide IG-25 onto fluoropolymers via fluorous interactions and click chemistry.

We report a practical method for biofunctionalization of fluoropolymers based on noncovalent, fluorous interactions and click chemistry that allows incorporation of biomolecules under physiological solutions. We demonstrate the method by immobilization of an antimicrobial peptide (AMP) on fluorous thin films and fluorosilicone contact lens. The fluorous surfaces were dip-coated with fluorous-tagged oligo(ethylene) chain terminated with a reactive group, such as an alkynyl group. This simple step generates a "clickable" surface. The noncovalent fluorous interaction was strong enough to allow subsequent covalent attachment of IG-25, a truncated version of the most extensively studied human AMP LL-37. The attachment was through copper-catalyzed click reaction between the alkynyl group on the surface and the azido-OEG tag at the N-terminus of IG-25. In comparison to surfaces presenting IG-25 randomly bound via carbodiimide chemistry, the surfaces presenting IG-25 tethering to the surface at the N-terminus via click chemistry displayed higher antibacterial activities against an ocular pathogen Pseudomonas aeruginosa (strain PA-O1).

Antimicrobial PVK:SWNT nanocomposite coated membrane for water purification: performance and toxicity testing.

This study demonstrated that coated nitrocellulose membranes with a nanocomposite containing 97% (wt%) of polyvinyl-N-carbazole (PVK) and 3% (wt%) of single-walled carbon nanotubes (SWNTs) (97:3 wt% ratio PVK:SWNT) achieve similar or improved removal of bacteria when compared with 100% SWNTs coated membranes. Membranes coated with the nanocomposite exhibited significant antimicrobial activity toward Gram-positive and Gram-negative bacteria (≈ 80-90%); and presented a virus removal efficiency of ≈ 2.5 logs. Bacterial cell membrane damage was considered a possible mechanism of cellular inactivation since higher efflux of intracellular material (Deoxyribonucleic acid, DNA) was quantified in the filtrate of PVK-SWNT and SWNT membranes than in the filtrate of control membranes. To evaluate possible application of these membrane filters for drinking water treatment, toxicity of PVK-SWNT was tested against fibroblast cells. The results demonstrated that PVK-SWNT was non toxic to fibroblast cells as opposed to pure SWNT (100%). These results suggest that it is possible to synthesize antimicrobial nitrocellulose membranes coated with SWNT based nanocomposites for drinking water treatment. Furthermore, membrane filters coated with the nanocomposite PVK-SWNT (97:3 wt% ratio PVK:SWNT) will produce more suitable coated membranes for drinking water than pure SWNTs coated membranes (100%), since the reduced load of SWNT in the nanocomposite will reduce the use of costly and toxic SWNT nanomaterial on the membranes.

Graphene nanocomposite for biomedical applications: fabrication, antimicrobial and cytotoxic investigations.

Materials possessing excellent bacterial toxicity, while presenting low cytotoxicity to human cells, are strong candidates for biomaterials applications. In this study, we present the fabrication of a nanocomposite containing poly(N-vinylcarbazole) (PVK) and graphene (G) in solutions and thin films. Highly dispersed PVK-G (97-3 w/w%) solutions in various organic and aqueous solvents were prepared by solution mixing and sonication methods. The thermal properties and morphology of the new composite were analyzed using thermal gravimetry analysis (TGA) and atomic force microscopy (AFM), respectively. PVK-G films were immobilized onto indium tin oxide (ITO) substrates via electrodeposition. AFM was used to characterize the resulting topography of the nanocomposite thin films, while cyclic voltammetry and UV-vis were used to monitor their successful electrodeposition. The antimicrobial properties of the electrodeposited PVK-G films and solution-based PVK-G were investigated against Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis). Microbial growth after exposure to the nanocomposite, metabolic assay and live-dead assay of the bacterial solutions exposed to PVK-G presented fewer viable and active bacteria than those exposed to pure PVK or pure graphene solutions. The PVK-G film inhibited about 80% of biofilm surface coverage whereas the PVK- and G-modified surfaces allowed biofilm formation over almost the whole coated surface (i.e. > 80%). The biocompatibility of the prepared PVK-G solutions on NIH 3T3 cells was evaluated using the MTS cell proliferation assay. A 24 h exposure of the PVK-G nanocomposite to the NIH 3T3 cells presented ~80% cell survival.

Toxicity of a polymer-graphene oxide composite against bacterial planktonic cells, biofilms, and mammalian cells.

It is critical to develop highly effective antimicrobial agents that are not harmful to humans and do not present adverse effects on the environment. Although antimicrobial studies of graphene-based nanomaterials are still quite limited, some researchers have paid particular attention to such nanocomposites as promising candidates for the next generation of antimicrobial agents. The polyvinyl-N-carbazole (PVK)-graphene oxide (GO) nanocomposite (PVK-GO), which contains only 3 wt% of GO well-dispersed in a 97 wt% PVK matrix, presents excellent antibacterial properties without significant cytotoxicity to mammalian cells. The high polymer content in this nanocomposite makes future large-scale material manufacturing possible in a high-yield process of adiabatic bulk polymerization. In this study, the toxicity of PVK-GO was assessed with planktonic microbial cells, biofilms, and NIH 3T3 fibroblast cells. The antibacterial effects were evaluated against two Gram-negative bacteria: Escherichia coli and Cupriavidus metallidurans; and two Gram-positive bacteria: Bacillus subtilis and Rhodococcus opacus. The results show that the PVK-GO nanocomposite presents higher antimicrobial effects than the pristine GO. The effectiveness of the PVK-GO in solution was demonstrated as the nanocomposite "encapsulated" the bacterial cells, which led to reduced microbial metabolic activity and cell death. The fact that the PVK-GO did not present significant cytotoxicity to fibroblast cells offers a great opportunity for potential applications in important biomedical and industrial fields.

Antimicrobial applications of electroactive PVK-SWNT nanocomposites.

The antibacterial properties of a nanocomposite containing an electroactive polymer, polyvinyl-N-carbazole (PVK) (97 wt %), and single-walled carbon nanotubes (SWNT) (3 wt %) was investigated as suspensions in water and as thin film coatings. The toxic effects of four different PVK-SWNT (97:3 wt %) nanocomposite concentrations (1, 0.5, 0.05, and 0.01 mg/mL) containing 0.03, 0.015, 0.0015, and 0.0003 mg/mL of SWNT, respectively, were determined for planktonic cells and biofilms of Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis). The results showed that the nanocomposite PVK-SWNT had antibacterial activity on planktonic cells and biofilms at all concentration levels. Higher bacterial inactivation (94% for E. coli and 90% for B. subtilis) were achieved in planktonic cells at a PVK-SWNT concentration of 1 mg/mL. Atomic force microscopy (AFM) imaging showed significant reduction of biofilm growth on PVK-SWNT coated surfaces. This study established for the first time that the improved dispersion of SWNTs in aqueous solutions in the presence of PVK enhances the antimicrobial effects of SWNTs at very low concentrations. Furthermore, PVK-SWNT can be used as an effective thin film coating material to resist biofilm formation.

Antimicrobial graphene polymer (PVK-GO) nanocomposite films.

The first report on the fabrication and application of a nanocomposite containing poly-N-vinyl carbazole (PVK) polymer and graphene oxide (GO) as an antimicrobial film was demonstrated. The antimicrobial film was 90% more effective in preventing bacterial colonization relative to the unmodified surface. More importantly, the nanocomposite thin film showed higher bacterial toxicity than pure GO-modified surface.

"Click" immobilization on alkylated silicon substrates: model for the study of surface bound antimicrobial peptides.

We describe an effective approach for the covalent immobilization of antimicrobial peptides (AMPs) to bioinert substrates via Cu(I) -catalyzed azide-alkyne cycloaddition (CuAAC). The bioinert substrates were prepared by surface hydrosilylation of oligo(ethylene glycol) (OEG) terminated alkenes on hydrogen-terminated silicon surfaces. To render the OEG monolayers "clickable", mixed monolayers were prepared using OEG-alkenes with and without a terminal alkyne protected by a trimethylgermanyl (TMG) group. The mixed monolayers were characterized by X-ray photoelectron spectroscopy (XPS), elliposometry and contact angle measurement. The TMG protecting group can be readily removed to yield a free terminal alkyne by catalytic amounts of Cu(I) in an aqueous media. This step can then be combined with the subsequent CuAAC reaction. Thus, the immobilization of an azide modified AMP (N3-IG-25) was achieved in a one-pot deprotection/coupling reaction. Varying the ratio of the two alkenes in the deposition mixture allowed for control over the density of the alkynyl groups in the mixed monolayer, and subsequently the coverage of the AMPs on the monolayer. These samples allowed for study of the dependence of antimicrobial activities on the AMP density. The results show that a relative low coverage of AMPs (∼1.6×10(13) molecule per cm(2)) is sufficient to significantly suppress the viability of Pseudomonas aeruginosa, while the surface presenting the highest density of AMPs (∼2.8×10(13) molecule per cm(2)) is still cyto-compatible. The remarkable antibacterial activity is attributed to the long and flexible linker and the site-specific "click" immobilization, which may facilitate the covalently attached peptides to interact with and disrupt the bacterial membranes.

Functionalization of fluorous thin films via "click" chemistry.

The first covalent modification of thin films non-covalently immobilized via fluorous interactions was demonstrated with "click" reactions in 70-80% yields.