The development and application of antibacterial film were highly anticipated to prevent food spoilage caused by bacteria. In this investigation, antibacterial and antioxidant functionalized gelatin-based film was formed with the incorporation of oregano essential emulsion Pickering emulsion (OPE). ε-Polylysine-Carboxymethyl Chitosan nanoparticles (CMCS-ε-PL) composed of different mass ratios of CMCS and ε-PL were orchestrated by electrostatic forces and hydrogen bonding, which effectively acted as a stabilizer for OPE. The design of different mass ratios of CMCS and ε-PL in CMCS-ε-PL has a deep effect on the structure and functional properties of OPE and film. It successfully improved the encapsulation efficiency of OPE from 49.52 % to 79.83 %. With the observation of AFM images, the augmentation of surface roughness consequent to OPE incorporation can be relieved by the increased contention of ε-PL in CMCS-ε-PL. Meanwhile, the mechanical properties, barrier properties, anti-oxidation, and antibacterial properties of the films were improved with the incorporation of the above OPE. In particular, a synergistic antibacterial activity between ε-PL and OEO in the film was demonstrated in this study and the mechanism of enhanced antibacterial activity was elucidated by examining the integrity of bacteria cell membrane. The film unequivocally demonstrated its ability to appreciably prolong the shelf life of both beef and strawberries with excellent antioxidant and antibacterial properties.

In this study, an efficient synthetic strategy and potential route to obtain a photo-reactive silver-containing cytosine-functionalized polypropylene glycol polymer (Ag-Cy-PPG) was developed by combining a hydrophilic oligomeric polypropylene glycol (PPG) backbone with dual pH-sensitive/photo-reactive cytosine-silver-cytosine (Cy-Ag-Cy) linkages. The resulting photo-responsive Ag-Cy-PPG holds great promise as a multifunctional biomedical material that generates spherical-like nanogels in water; the nanogels exhibit high antibacterial activity and thus may significantly enhance the efficacy of antibacterial treatment. Due to the formation of photo-dimerized Cy-Ag-Cy cross-linkages after UV irradiation, Ag-Cy-PPG converts into water-soluble cross-linked nanogels that possess a series of interesting chemical and physical properties, such as intense and stable fluorescence behavior, highly sensitive pH-responsive characteristics, on/off switchable phase transition behavior, and well-controlled release of silver ions (Ag+) in mildly acidic aqueous solution. Importantly, antibacterial tests clearly demonstrated that irradiated Ag-Cy-PPG nanogels exhibited strong antibacterial activity at low doses (MIC values of < 50 μg/mL) against gram-positive and gram-negative bacterial pathogens, whereas non-irradiated Ag-Cy-PPG nanogels did not inhibit the viability of bacterial pathogens. These results indicate that irradiated Ag-Cy-PPG nanogels undergo a highly sensitive structural change in the bacterial microenvironment due to their relatively unstable π-conjugated structures (compared to non-irradiated nanogels); this change results in a rapid structural response that promotes intracellular release of Ag+ and induces potent antibacterial ability. Overall, this newly created metallo-supramolecular system may potentially provide an efficient route to dramatically enhance the therapeutic effectiveness of antibacterial treatments.

To improve the action of already in use antibiotics or new antimicrobial agents against different bacteria, the development of effective combinations of antimicrobial peptides (AMPs) with enzymes that can quench the quorum (QQ) sensing of bacterial cells was undertaken. Enzymes hydrolyzing N-acyl homoserine lactones (AHLs) and peptides that are signal molecules of Gram-negative and Gram-positive bacterial cells, respectively, were estimated as \"partners\" for antibiotics and antimicrobial peptides in newly designed antimicrobial-enzymatic combinations. The molecular docking of six antimicrobial agents to the surface of 10 different QQ enzyme molecules was simulated in silico. This made it possible to choose the best variants among the target combinations. Further, bacterial cellulose (BC) was applied as a carrier for uploading such combinations to generally compose prototypes of effective dressing materials with morphology, providing good absorbance. The in vitro analysis of antibacterial activity of prepared BC samples confirmed the significantly enhanced efficiency of the action of AMPs (including polymyxin B and colistin, which are antibiotics of last resort) in combination with AHL-hydrolyzing enzymes (penicillin acylase and His6-tagged organophosphorus hydrolase) against both Gram-negative and Gram-positive cells.

MRSA infections are a major global healthcare problem associated with high morbidity and mortality. The application of novel materials in antibiotic delivery has efficiently contributed to the treatment of MRSA infections. The aim of the study was to develop novel hyaluronic acid-oleylamine (HA-OLA) conjugates with 25-50% degrees of conjugation, for application as a nano-drug carrier with inherent antibacterial activity. The biosafety of synthesized novel HA-OLA conjugates was confirmed by in vitro cytotoxicity assay. Drug carrying ability of HA-OLA conjugates was confirmed by 26.1-43.12% of vancomycin (VCM) encapsulation in self-assembled polymersomes. These polymersomes were dispersed in nano-sized range (196.1-360.9 nm) with a negative zeta potential. Vancomycin loaded polymersomes were found to have spherical and bilayered morphology. The VCM loaded polymersomes displayed sustained drug release for 72 h. In vitro studies showed moderate antibacterial activity for HA-OLA conjugates against both S. aureus and MRSA with minimum inhibitory concentration (MIC) of 500 μg/mL. The VCM loaded HA-OLA polymersomes displayed four-fold lower MIC (1.9 μg/mL) than free VCM (7.8 μg/mL) against MRSA. Furthermore, synergism was observed for VCM and HA-OLA against MRSA. Flow cytometry showed 1.8-fold higher MRSA cell death in the population for VCM loaded polymersomes relative to free drug, at concentration of 1.95 μg/mL. Bacterial cell morphology showed that the drug loaded polymersomes had stronger impact on MRSA membrane, compared to free VCM. These findings suggest that, HA-OLA conjugates are promising nano-carriers to function as antibiotic delivery vehicles for the treatment of bacterial/MRSA infections.

Novel antimicrobials with well biocompatibility are urgently needed for human public health protecting. Here, the silk fibroin (SF) nanofibrous mats coated with carboxymethyl chitosan (CMC), both extracted from natural polymers, were fabricated by combining electrospinning and electrostatic layer-by-layer (LBL) self-assembly techniques. The amphoteric CMC could be easily adsorbed on the surface of SF nanofibers due to the electrostatic interaction, which were a highly efficient and \"green\" route for the surface modification of SF mats. The mats after LBL procedure exhibited better hydrophilicity and stronger mechanical properties. The experimental results demonstrated that the LBL modified mats not only kept well biocompatibility but also obtained extremely enhanced antibacterial activity. More importantly, the mats displayed better bacterial inhibition with the increased CMC layers. LBL10 and LBL10.5 owned the antibacterial activity of more than 67% at the bacterial concentrations of 106 cfu ml-1 after 24 h cultivation, which implied that these novel natural polymer-based materials could be utilized as wound dressings for clinical skin and tissue regeneration, especially for infected wounds.

The aim of the present study was to formulate a nanosuspension (FA-NS) of fusidic acid (FA) to enhance its aqueous solubility and antibacterial activity. The nanosuspension was characterized using various in vitro, in silico, and in vivo techniques. The size, polydispersity index, and zeta potential of the optimized FA-NS were 265 ± 2.25 nm, 0.158 ± 0.026, and -16.9 ± 0.794 mV, respectively. The molecular dynamics simulation of FA and Poloxamer-188 showed an interaction and binding energy of -74.42 kJ/mol and -49.764 ± 1.298 kJ/mol, respectively, with van der Waals interactions playing a major role in the spontaneous binding. There was an 8-fold increase in the solubility of FA in a nanosuspension compared to the bare drug. The MTT assays showed a cell viability of 75-100% confirming the nontoxic nature of FA-NS. In vitro antibacterial activity revealed a 16- and 18-fold enhanced activity against Staphylococcus aureus (SA) and methicillin-resistant SA (MRSA), respectively, when compared to bare FA. Flowcytometry showed that MRSA cells treated with FA-NS had almost twice the percentage of dead bacteria in the population, despite having an 8-fold lower MIC in comparison to the bare drug. The in vivo skin-infected mice showed a 76-fold reduction in the MRSA load for the FA-NS treated group compared to that of the bare FA. These results show that the nanosuspension of antibiotics can enhance their solubility and antibacterial activity simultaneously.

Most of the bacteria are on the verge of becoming resistant to available potential antibiotics. Novel approaches to combat these drug resistant bacteria are turning out to be crucial. This study aimed to synthesize novel fatty acid based cationic amphiphiles (FCA) that would serve as nano-drug carrier having intrinsic antibacterial activity. Three fatty acids oleic acid, linoleic acid and linolenic acid based cationic amphiphiles were synthesized and evaluated for antibacterial activity and cytotoxicity. The application in the delivery of vancomycin (VCM) was demonstrated using oleic based cationic amphiphilic (OCA). OCA was self-assembled in aqueous media to prepare VCM loaded OCA vesicles. The particle size, polydispersity index, zeta potential and entrapment efficiency were found to be 132.9 ± 2.5 nm, 0.167 ± 0.02, 18.9 ± 1.2 mV and 61.24 ± 1.8% respectively. The images from transmission electron microscopy (TEM) revealed that the vesicles were spherical and bilayered. The release of VCM from OCA vesicles was sustained throughout the studied period of 72 h. From in vitro studies, a significant antibacterial activity was observed for all three FCAs and it was found that, VCM loaded OCA vesicles displayed indifference and synergism against Gram positive methicillin susceptible and resistant staphylococcus aureus respectively (MRSA). In contrast to minimum inhibitory concentration (MIC) of VCM against Gram negative Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aeruginosa), the synthesized FCAs were more potent against both the strains, further there was no synergism observed against either of the strains when VCM was encapsulated in OCA vesicles. The synergism against MRSA was further confirmed in in vivo studies using mouse infection model. These findings therefore suggest that, FCAs can make promising nano-carrier systems for the delivery of antibiotics to treat infections caused by multi drug resistant bacteria.

The cytotoxicity of 2D graphene-based nanomaterials (GBNs) is highly important for engineered applications and environmental health. However, the isotropic orientation of GBNs, most notably graphene oxide (GO), in previous experimental studies obscured the interpretation of cytotoxic contributions of nanosheet edges. Here, we investigate the orientation-dependent interaction of GBNs with bacteria using GO composite films. To produce the films, GO nanosheets are aligned in a magnetic field, immobilized by cross-linking of the surrounding matrix, and exposed on the surface through oxidative etching. Characterization by small-angle X-ray scattering and atomic force microscopy confirms that GO nanosheets align progressively well with increasing magnetic field strength and that the alignment is effectively preserved by cross-linking. When contacted with the model bacterium Escherichia coli, GO nanosheets with vertical orientation exhibit enhanced antibacterial activity compared with random and horizontal orientations. Further characterization is performed to explain the enhanced antibacterial activity of the film with vertically aligned GO. Using phospholipid vesicles as a model system, we observe that GO nanosheets induce physical disruption of the lipid bilayer. Additionally, we find substantial GO-induced oxidation of glutathione, a model intracellular antioxidant, paired with limited generation of reactive oxygen species, suggesting that oxidation occurs through a direct electron-transfer mechanism. These physical and chemical mechanisms both require nanosheet penetration of the cell membrane, suggesting that the enhanced antibacterial activity of the film with vertically aligned GO stems from an increased density of edges with a preferential orientation for membrane disruption. The importance of nanosheet penetration for cytotoxicity has direct implications for the design of engineering surfaces using GBNs.