Supramolecular self-assemblies of engineered polyethylenimines as multifunctional nanostructures for DNA transportation with excellent antimicrobial activity.

In this study, indole-3-butanoic acid (IBA), a biologically and environmentally safe entity, has been grafted onto low and high molecular weight (1.8 and 25 kDa) polyethylenimines (PEI) mainly through primary amines to obtain amphiphilic indole-3-butanoyl-polyethylenimines (IBPs). Two series of IBPs (IBP1.8 and IBP25) were prepared which, on self-assembly in aqueous medium, yielded multifunctional nanomicellar structures (IBP1.8 and IBP25) capable of transporting genetic material in vitro and exhibiting other biological activities. Physicochemical characterization showed the size of IBP1.8 and IBP25 nanostructures in the range of ~332-234 nm and ~283-166 nm, respectively, with zeta potential varying from ~+29-17 mV and ~+37-25 mV. DNA release assay demonstrated higher release of plasmid DNA from IBP nanostructures as compared to native PEIs. Cytotoxicity showed a decreasing pattern with increasing degree of grafting of IBA onto PEIs making these nanostructures non-toxic. pDNA complexes of these nanostructures (both IBPs1.8 and IBPs25) displayed considerably higher transfection efficiency, however, IBP1.8/pDNA complexes performed much better (~7-9 folds) as compared to native PEI/pDNA and Lipofectamine/pDNA complexes on mammalian cells. CLSM analysis revealed that these complexes entered nucleus in sufficient amounts suggesting higher uptake and efficient internalization of the complexes. Besides, these supramolecular nanostructures not only exhibited excellent antimicrobial potential (MIC ~49-100 µg/ml) against clinical as well as resistant pathogenic strains but also found to possess antioxidant property. Overall, the projected low molecular weight PEI-based vectors could serve as more effective multifunctional nanomaterials having promising potential for future gene therapy applications with capability to provide protection against other bacterial infections.

Unveiling the cytotoxic and anti-proliferative potential of green-synthesized silver nanoparticles mediated by Colletotrichum gloeosporioides.

Fungal endophytes are a putative source of bioactive metabolites that have found significant applications in nanomedicine due to their metabolic versatility. In the present study, an aqueous extract of the fungal endophyte, Colletotrichum gloeosporioides associated with a medicinal plant Oroxylum indicum, has been used for the fabrication of green silver nanoparticles (CgAgNPs) and further evaluated their cytotoxic and anti-proliferative activity. Bioanalytical techniques including UV-Vis spectral analysis revealed a sharp band at 435 nm and functional molecules from the aqueous extract involved in the synthesis of CgAgNPs were evidenced through FTIR. Further, the crystalline nature of CgAgNPs was determined through XRD analysis and microscopy techniques including AFM, TEM and FESEM demonstrated the spherical shape of CgAgNPs exhibiting a crystalline hexagonal lattice and the size was found to be in the range of 9-29 nm. The significant cytotoxic potential of CgAgNPs was observed against breast cancer cells, MDA-MB-231 and MCF-7 with IC50 values of 18.398 ± 0.376 and 38.587 ± 1.828 µg mL-1, respectively. The biochemical study revealed that the treatment of MDA-MB-231 and MCF-7 cells with CgAgNPs reduces glucose uptake, suppresses cell proliferation, and enhances LDH release, indicating reduced cell viability and progression. Moreover, our research revealed differential expression of genes associated with apoptosis, cell cycle inhibition and metastasis suppression, evidencing anti-proliferative activity of CgAgNPs. The main objective of the present study is to harness anti-breast cancer activity of novel biogenic nanoparticles synthesized using the aqueous extract of O. indicum associated C. gloeosporioides and study the underlying mechanistic pathway exerted by these mycogenic nanoparticles.

Sustainable Synthesis of Novel Green-Based Nanoparticles for Therapeutic Interventions and Environmental Remediation.

The advancement in nanotechnology has completely revolutionized various fields, including pharmaceutical sciences, and streamlined the potential therapeutic of many diseases that endanger human life. The synthesis of green nanoparticles by biological processes is an aspect of the newly emerging scientific field known as "green nanotechnology". Due to their safe, eco-friendly, nontoxic nature, green synthesis tools are better suited to produce nanoparticles between 1 and 100 nm. Nanoformulation of different types of nanoparticles has been made possible by using green production techniques and commercially feasible novel precursors, such as seed extracts, algae, and fungi, that act as potent reducing, capping, and stabilizing agents. In addition to this, the biofunctionalization of nanoparticles has also broadened its horizon in the field of environmental remediation and various novel therapeutic innovations including wound healing, antimicrobial, anticancer, and nano biosensing. However, the major challenge pertaining to green nanotechnology is the agglomeration of nanoparticles that may alter the surface topology, which can affect biological physiology, thereby contributing to system toxicity. Therefore, a thorough grasp of nanoparticle toxicity and biocompatibility is required to harness the applications of nanotechnology in therapeutics.

White-Light-Responsive Prussian Blue Nanophotonic Particles for Effective Eradication of Bacteria and Improved Healing of Infected Cutaneous Wounds.

The increasing burden of cutaneous wound infections with drug-resistant bacteria underlines the dire need for novel treatment approaches. Here, we report the preparation steps, characterization, and antibacterial efficacy of novel chitosan-coated Prussian blue nanoparticles loaded with the photosensitizer fluorescein isothiocyanate-dextran (CHPB-FD). With excellent photothermal and photodynamic properties, CHPB-FD nanoparticles can effectively eradicate both Gram-positive methicillin-resistant Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa in vitro and in vivo. The antibacterial efficacy of CHPB-FD nanophotonic particles further increases in the presence of white light. Using a bacteria-infected cutaneous wound rat model, we demonstrate that CHPB-FD particles upregulate genes involved in tissue remodeling, promote collagen deposition, reduce unwanted inflammation, and enhance healing. The light-responsive CHPB-FD nanophotonic particles can, therefore, be potentially used as an economical and safe alternative to antibiotics for effectively decontaminating skin wounds and for disinfecting biomedical equipment and surfaces in hospitals and other places.

An injectable self-assembling hydrogel based on RGD peptidomimetic ß-sheets as multifunctional biomaterials.

Ability of the cells to adhere to an extracellular material is central to successful tissue genesis. Arg-Gly-Asp (RGD) sequences found in extracellular matrix proteins are well known for cell adhesion, however, enzymatic degradation and lack of specificity have limited their widespread use. Besides, a multifunctional material with inherent antimicrobial ability would help in invigorating the practical tissue engineering applications. Here, we report novel modified RGD (MR) and RGD mimic [R(K)] peptides (MOH and MNH2) which were synthesized post-in-silico screening, based on their interactions with integrin protein αVß3 using HEX 8.0 docking server. These mimics, containing hydrophobic Phe-Phe (FF) moiety which has been specifically introduced to initiate the self-assembling process of ß-sheet structures, were characterized thoroughly using various physicochemical and spectroscopic techniques. Under physiological conditions, these mimetics displayed thixotropic behavior rendering them highly suitable as injectable hydrogels having an added advantage of site-specific targeting abilities. Electron microscopy further revealed the formation of nanofibers upon self-assembly of these peptides. Besides, enhanced cell adhesiveness by these peptides compared to the commercial Poly l-lysine coated surfaces as well as the inherent antimicrobial potential against both sensitive and antibiotic-resistant pathogens (Methicillin-resistant Staphylococcus aureus and multi-drug resistant Salmonella enteritidis) substantiated the applicability of these unique injectable hydrogels wherein the porous fibrous framework offered a favorable environment for drug entrapment and 3D cell culture. Altogether, these properties render these novel RGD mimic peptides as promising multifunctional candidates for various tissue regenerative applications.

In vitro evaluation of antioxidant and radioprotective properties of a novel extremophile from mud volcano: implications for management of radiation emergencies.

A thermophilic bacterium, designated as RH 127, was isolated from mud volcano (Baratang Islands) of Andaman region, India (12°07'N 92°47'E/12.117°N 92.783°E) for the first time. Biochemical tests and 16S rRNA gene sequencing indicate that it belongs to the genus Geobacillus. The strain showed 98% confirmed 16S rRNA gene sequence homology with Geobacillus toebii. The bacteria was extracted in various solvent systems and three different fractions prepared. In the present study, antioxidant and radioprotective activity of extracts (INM-7860, INM-7861, and INM-7862) of bacterium G. toebii (strain RH 127) were evaluated. The fractions were evaluated for their introspective comparison of the relative antioxidant efficiency. The antioxidative activities, DPPH radical scavenging effects, hydroxyl radical scavenging effects, membrane protection, antihemolytic activity, and linoleic acid degradation efficacies were assayed. INM-7861 and INM-7862 activated NF-κB expression, as evidenced by reporter assay studies, and thereby contributed to overall radioprotective effect. INM-7862 exhibited best results. This study explicitly shows that the extracts of G. toebii have immense potential as a radiation countermeasure agent.

Robust dual modality antibacterial action using silver-Prussian blue nanoscale coordination polymer.

We report the synthesis of novel silver-doped Prussian blue nanoscale coordination polymers (SPB NCPs), for dual modality photothermal ablation and oxidative toxicity in bacterial cells. The comparison of SPB NCPs (having Fe-CN-Ag bonds) with the conventionally used Prussian blue nanoscale coordination polymers (PB NCPs, having Fe-CN-Fe bonds) was investigated in terms of their physical and therapeutic properties. It was observed that both PB and SPB NCPs have similar physical dimensions, crystalline phase and optical properties. Both these NCPs showed robust photothermal effect by heat generation (hyperthermia) upon exposure to red laser light. However, among the two, only SPB NCP showed oxidase-like activity by generating H2O2 in aqueous medium, presumably due to its silver content. In vitro antibacterial studies revealed that the SPB NCPs, but not PB NCPs, show inherent toxicity towards bacteria with an IC50 value close to 2.5 µg/ml. It can be inferred that this toxicity is oxidative in nature, as a result of the oxidase-like behaviour shown by SPB NCPs. Furthermore, light activation resulted in substantial additional antibacterial effect (photothermal toxicity) in bacterial cells treated with SPB NCPs. In comparison, marginal additional photothermal toxicity was observed in PB NCP-treated bacteria. Thus, we conclude that the combination of dual modality oxidative and photothermal toxicities demonstrated by SPB NCPs, but not by control PB NCPs, makes the former promising antibacterial agents at low dosages.

Dual Modality FeS Nanoparticles with Reactive Oxygen Species-Induced and Photothermal Toxicity toward Pathogenic Bacteria.

Bacterial infections pose a major threat to human health, primarily because of the evolution of mutated strains that are resistant to antibiotic treatment. As a viable alternative, several nanoparticles have emerged as attractive antibacterial agents. Herein, we report the development of iron sulfide (FeS) nanoparticles that show dual-modality therapy: namely reactive oxygen species (ROS)-induced toxicity and red-laser induced photothermal therapy. The aqueous synthesized nanoparticles have been characterized based on their size, shape, crystallinity, and magnetic and optical properties. These nanoparticles showed sustained release of Fe2+ ions in an aqueous dispersion. They also have a high absorption cross-section in the visible and near infra-red regions and could be excited by a continuous wave diode laser of wavelength 635 nm leading to significant hyperthermia. Nanoparticle treatment, followed by light irradiation, led to significant cell death in two ghastly pathogenic bacterial strains. Stepwise enhancement of intrabacterial ROS levels, as a result of nanoparticle treatment followed by light activation, has been identified as the primary antibacterial mechanism.

Fabrication of cationic nanostructures from short self-assembling amphiphilic mixed α/ß-pentapeptide: Potential candidates for drug delivery, gene delivery, and antimicrobial applications.

The present article describes designing and fabrication of nanostructures from a mixed α/ß-pentapeptide, Lys-ßAla-ßAla-Lys-ßAla, which majorly contains non-natural ß-alanine residues in the backbone with two α-lysine residues at 1- and 4-positions. The amphiphilic pentapeptide showed the ability to self-assemble into cationic nanovesicles in an aqueous solution. The average size of peptide nanostructures was found to be ~270 nm with a very high cationic charge of ~+40 mV. TEM micrographs revealed the average size of the same nanostructures ~80 nm bearing vesicular morphology. CD and FTIR spectroscopic studies on self-assembled pentapeptide hinted at random coil conformation which was also correlated with conformational search program using Hyper Chem 8.0. The pentapeptide nanostructures were then tested for encapsulation of hydrophobic model drug moieties, L-Dopa, and curcumin. Transfection efficiency of the generated cationic nanostructures was evaluated on HEK293 cells and compared the results with those obtained in the presence of chloroquine. The cytotoxicity assay performed using MTT depicted ~75-80% cell viability. The obtained nanostructures also gave positive results against both Gram-negative and Gram-positive bacterial strains. Altogether the results advocate the promising potential of the pentapeptide foldamer, H-Lys-ßAla-ßAla-Lys-ßAla-OEt, for drug and gene delivery applications along with the antimicrobial activity.

Amphiphilic azobenzene-neomycin conjugate self-assembles into nanostructures and transports plasmid DNA efficiently into the mammalian cells.

The present study demonstrates the use of self-assembled nanostructures of cationic amphiphilic azobenzene-neomycin (a small molecule) conjugate, Azo-Neo, as delivery vector for plasmid DNA. These nanostructures efficiently condensed nucleic acid and formed more compact nanoassemblies. DLS analysis showed size and zeta potential of the resulting Azo-Neo/pDNA nanoassemblies ∼153.7nm and +7.26mV, respectively. The nanoassemblies were characterized by physicochemical techniques and evaluated for its toxicity and ability to deliver nucleic acid therapeutics. The flow cytometry results on MCF-7 and HEK293T cells revealed that Azo-Neo/pDNA nanoassemblies transfected ∼31% and 23% cells, respectively, at a w/w ratio of 250, while the standard transfection reagent, bPEI/pDNA complex, could transfect only ∼21% and 29% cells, respectively, at its best w:w ratio of 2.3. MTT and hemolysis assays showed the non-toxic nature of the projected nanoassemblies and nanostructures, respectively, at various concentrations. Further, Azo-Neo nanostructures showed efficient antibacterial activity against different strains, laboratory strain of Staphylococcus aureus (MTCC 740) as well as MRSA strains (Staphylococcus aureus ATCC 33591, ATCC 43300 and ATCC 700699). These results ensure the great potential of these nanostructures in gene delivery and antimicrobial applications.

Multifunctional self-assembled cationic peptide nanostructures efficiently carry plasmid DNA in vitro and exhibit antimicrobial activity with minimal toxicity.

In this study, a modified dehydropeptide, Boc-FΔF-εAhx-OH, was conjugated with an aminoglycoside antibiotic, neomycin, to construct a multifunctional conjugate, Pep-Neo. The amphiphilic conjugate (Pep-Neo) was able to self-assemble into cationic nanostructures in an aqueous solution at low concentrations. Nanostructure formation was evidenced by TEM and dynamic light scattering analyses. The average hydrodynamic diameter of the self-assembled Pep-Neo nanostructures was found to be ∼279 nm with a zeta potential of +28 mV. The formation of nanostructures with a hydrophobic core and cationic hydrophilic shell resulted in an increased local concentration of cationic charge (ca. in 50% aqueous methanol, i.e. disassembled structure, zeta potential decreased to +17.6 mV), leading to efficient interactions with negatively charged plasmid DNA (pDNA). The size and zeta potential of the resulting Pep-Neo/pDNA complex were found to be ∼154 nm and +19.4 mV, respectively. Having been characterized by physicochemical techniques, the complex was evaluated for its toxicity and ability to deliver nucleic acid therapeutics. The flow cytometry results on MCF-7 cells revealed that Pep-Neo/pDNA complex transfected ∼27% cells at a w/w ratio of 66.6 while the standard transfection reagent, Lipofectamine, could transfect only ∼15% cells. MTT and hemolysis assays showed the non-toxic nature of the projected conjugate at various concentrations. Further, these nanostructures were shown to encapsulate hydrophobic drugs in the core. Finally, Pep-Neo nanostructures showed efficient antibacterial activity against different strains of Gram-positive and -negative bacteria. Interestingly, unlike neomycin, which is highly effective against Gram-negative bacteria, these nanostructures showed considerably high efficiency against Gram-positive strains, highlighting the promising potential of these nanostructures for various biomedical applications.