Photo-Controlled Gating of Selective Bacterial Membrane Interaction and Enhanced Antibacterial Activity for Wound Healing.

Reversible biointerfaces are essential for on-demand molecular recognition to regulate stimuli-responsive bioactivity such as specific interactions with cell membranes. The reversibility on a single platform allows the smart material to kill pathogens or attach/detach cells. Herein, we introduce a 2D-MoS2 functionalized with cationic azobenzene that interacts selectively with either Gram-positive or Gram-negative bacteria in a light-gated fashion. The trans conformation (trans-Azo-MoS2 ) selectively kills Gram-negative bacteria, whereas the cis form (cis-Azo-MoS2 ), under UV light, exhibits antibacterial activity against Gram-positive strains. The mechanistic investigation indicates that the cis-Azo-MoS2 exhibits higher affinity towards the membrane of Gram-positive bacteria compared to trans-Azo-MoS2 . In case of Gram-negative bacteria, trans-Azo-MoS2 internalizes more efficiently than cis-Azo-MoS2 and generates intracellular ROS to kill the bacteria. While the trans-Azo-MoS2 exhibits strong electrostatic interactions and internalizes faster into Gram-negative bacterial cells, cis-Azo-MoS2 primarily interacts with Gram-positive bacteria through hydrophobic and H-bonding interactions. The difference in molecular mechanism leads to photo-controlled Gram-selectivity and enhanced antibacterial activity. We found strain-specific and high bactericidal activity (minimal bactericidal concentration, 0.65 µg/ml) with low cytotoxicity, which we extended to wound healing applications. This methodology provides a single platform for efficiently switching between conformers to reversibly control the strain-selective bactericidal activity regulated by light.

2D-MoS2-supported copper peroxide nanodots with enhanced nanozyme activity: application in antibacterial activity.

Peroxidase (POD)-like nanozymes are an upcoming class of new-generation antibiotics that are efficient for broad-spectrum antibacterial action. The POD-like activity employs the generation of reactive oxygen species (ROS), which have been utilized for bactericidal action. However, their intrinsic low catalytic activity and stability limit their bactericidal properties. In this study, we prepared a MoS2-based nanocomposite with copper peroxide nanodots (MoS2@CP) to achieve pH-dependent light-induced nanozyme-based antibacterial action. It has shown superior peroxidase and antibacterial activity at low pH. The mechanism behind the enhanced POD-like activity and high antibacterial activity was established. The mechanistic pathway involves estimating ROS generation, membrane depolarization, inner membrane permeabilization, metal ion release, and the effect of NIR on photothermal and photodynamic activities. Overall, our work highlighted the combinatorial approach for eradicating bacterial infections using enzyme-based antibacterial agents.

Post-functionalization of sulfur quantum dots and their aggregation-dependent antibacterial activity.

Sulfur quantum dots (SQDs) have emerged as an intriguing class of luminescent nanomaterial due to their exceptional physiochemical and optoelectronic properties. However, their biomedical application is still in its infancy due to the limited scope of their surface functionalization. Herein, we explored the surface functionalization of SQDs through different thiol ligands with tuneable functionality and tested their antibacterial efficacy. Notably, very high antibacterial activity of functionalized SQDs (10-25 ng ml-1) was noted, which is 105 times higher compared to that of nonfunctionalized SQDs. Moreover, a rare phenomenon of the reverse trend of antibacterial activity through surface modification was observed, with increasing surface hydrophobicity of various nanomaterials as the antibacterial activity increased. However, we also noted that as the surface hydrophobicity increased, the SQDs tended to exhibit a propensity for aggregation, which consequently decreased their antibacterial efficacy. This identical pattern was also evident in in vivo assessments. Overall, this study illuminates the importance of surface modifications of SQDs and the role of surface hydrophobicity in the development of antibacterial agents.

Solvent Induced Conversion of a Self-Assembled Gyrobifastigium to a Barrel and Encapsulation of Zinc-Phthalocyanine within the Barrel for Enhanced Photodynamic Therapy.

A rare gyrobifastigium architecture (GB) was constructed by self-assembly of a tetradentate donor (L) with PdII acceptor in DMSO. The GB was converted to its isomeric tetragonal barrel (MB) upon treatment with water. The hydrophobic cavity of MB has been explored for the encapsulation of zinc-phthalocyanine (ZnPc), which is an excellent photosensitizer for photodynamic therapy (PDT). However, the poor water-solubility and aggregation tendency are the main reasons for the suboptimal PDT performance of free ZnPc in the aqueous medium. Effective solubilization of ZnPc in an aqueous medium was achieved by encapsulating it in the cavity of MB. The inclusion complex (ZnPc⊂MB) showed enhanced singlet oxygen generation in water. Higher cellular uptake and anticancer activity of the ZnPc⊂MB compared to free ZnPc on HeLa cells indicate that encapsulation of ZnPc in an aqueous host is a potential strategy for enhancement of its PDT activity in water.

Ligand-Mediated Exfoliation and Antibacterial Activity of 2H Transition-Metal Dichalcogenides.

Transition-metal dichalcogenides (TMDs) exists mainly in two polymorphs, namely, 1T (metallic) and 2H (semiconducting). To tailor the characteristics and practical utility of TMDs for different applications, functionalization is essential. In our earlier studies, we have shown that functionalized 1T and 2H MoS2 exhibit exceptionally high antibacterial activity. The functionalization and related biological applications of other 1T (chemically exfoliated) TMDs were reported, but regarding other 2H TMDs, the functionalization and antibacterial activity are not explored yet. Hence, here we prepared functionalized 2H TMDs such as WS2, WSe2, and MoSe2 other than MoS2 by using a positively charged thiolate surfactant ligand. Further, functionalized 2H TMDs were utilized for antibacterial activity against Gram-positive and Gram-negative bacteria for a comparative antibacterial analysis. Interestingly, we found disparity in activity among the functionalized 2H TMDs, that is, MoS2 shows higher activity than WS2 followed by MoSe2 and WSe2. The intracellular reactive oxygen species measurement was found to be in the order MoS2 > WS2 > MoSe2 > WSe2, which is solely responsible for variation in the activity of functionalized TMDs. These results indicate that the easy functionalization of all types TMDs by using thiol ligand and importance of core material should be considered while designing functionalized material for specific applications.

Ligand Exchange on MoS2 Nanosheets: Applications in Array-Based Sensing and Drug Delivery.

Two-dimensional MoS2 nanosheets (2D-MoS2) have been widely used in many biological applications due to their distinctive physicochemical properties. Further, the development of surface modification using thiolated ligands allows us to use them for many specific applications. But the effect of possible ligand exchange on 2D-MoS2 has never been explored, which can play an important role in diverse biological applications. In this study, we have observed the ligand-exchange phenomenon on 2D-MoS2 in the presence of different thiolated ligands. The initial study proceeded with boron-dipyrromethene (BODIPY) functionalized MoS2 with different concentrations of glutathione (GSH), which is the most abundant thiol species in the cytoplasm of various cancer cells. It was found that in the presence of GSH the fluorescence of BODIPY can be regenerated, which is time and concentration dependent. We have also examined this phenomenon with different thiol ligands and transition-metal dichalcogenides (TMDs). We observed a variable rate of ligand exchange in different solvents, surface functionality, and receptor environments that helped us to construct sensor arrays. Interestingly, a ligand-exchange process was not observed in the presence of dithiols. Further, this concept was applied to a cancerous cell line for in vitro delivery. We found that BODIPY-functionalized 2D-MoS2 undergoes thiol exchange by intracellular GSH and subsequently enhanced the fluorescence in the cytoplasm of cancer cells. This strategy can be applied to the development of 2D-TMD-based materials for various biological applications related to ligand exchange.

Shifting the Triangle-Square Equilibrium of Self-Assembled Metallocycles by Guest Binding with Enhanced Photosensitization.

Shifting a triangle-square equilibrium in one direction is an important problem in supramolecular self-assembly. Reaction of a benzothiadiazole-based diimidazole donor with a cis-Pt(II) acceptor yielded an equilibrium mixture of a triangle ([C18H24N10O6S1Pt1]3≡ PtMCT) and a square ([C18H24N10O6S1Pt1]4≡ PtMCS). We report here the shifting of such equilibrium toward a triangle using a guest (pyrene aldehyde, G1). While both benzothiadiazole and pyrene aldehyde can form reactive oxygen species (ROS) in organic solvents, their therapeutic use in water is restricted due to aqueous insolubility. The enhanced water solubility of the benzothiadiazole unit and G1 by macrocycle formation and host-guest complexation, respectively, enabled enhanced ROS generation by the host-guest complex (G1' ⊂ PtMCT) in water (G1' = hydrated form of G1). The guest-encapsulated metallacycle (G1' ⊂ PtMCT) has shown synergistic antibacterial activity compared to the mixture of macrocycles upon white-light irradiation due to enhanced ROS generation. The mechanism for such enhanced activity was established by measuring the oxidative stress and relative internalization of PtMCs and G1' ⊂ PtMCT.

Gram-selective antibacterial activity of mixed-charge 2D-MoS2.

The development of nanomaterial-based antibiotics can be the most potent alternative due to the increasing resistance against conventional antibiotics. However, one of the important parameters in the development of antibacterial agents is their Gram selectivity, which has been seldomly explored in the case of nano-antibiotics. The multimodal action of surface-functionalized nanomaterials can exhibit strain selectivity and enhanced antibacterial activity. Herein, we designed a Gram-selective antibacterial system based on two-dimensional molybdenum disulphide (2D-MoS2) functionalized with different proportions of positively and negatively charged ligands. Two representative ESKAPE pathogenic strains, i.e., Gram-positive methicillin-resistant Staphylococcus aureus (MRSA) and Gram-negative Pseudomonas aeruginosa (P. aeruginosa) were considered to evaluate the selective antibacterial activity. The mechanistic insight behind selectivity was established by evaluating the degree of membrane depolarization together with oxidative stress. The selective generation of intracellular reactive oxygen species (ROS) together with membrane depolarization contributed to the selective killing of the pathogenic bacteria. Gram selectivity was achieved by simply controlling the surface functionality based on the different cell wall compositions and structures of bacterial strains. The interplay between polyvalent electrostatic and non-covalent interactions was mainly responsible for damaging the cell membrane. Furthermore, to establish the antibacterial mechanism, we performed extracellular and intracellular reactive oxidative stress, membrane depolarization and permeabilization assays. In summary, we prepared simple and efficient Gram-selective 2D-MoS2-based antibacterial agents, which can be extended to other nano-antibiotic systems.

Self-Assembled PtII8 Metallosupramolecular Tubular Cage as Dual Warhead Antibacterial Agent in Water.

Development of photosensitizer-based self-assembled metallosupramolecular architectures with important applications is an emerging trend in supramolecular chemistry. In this study, we report a new benzothiadiazole-based tetra-pyridyl ligand (L), which upon self-assembly with a cis-block 90° Pt(II) acceptor generated an unprecedented tetrafacial Pt(II)8 photoactive tubular molecular cage (PMB1). This cage could bring an extraordinary photosensitizer, benzothiadiazole, into water which is otherwise insoluble. PMB1 is fluorescent and shows photogeneration of singlet oxygen in an aqueous medium. These features make PMB1 a potent antimicrobial agent in water in both the presence and absence of light. In comparison to its building blocks and water-soluble alkylated charged ligand ([LMe4][4NO3]), the cage shows much enhanced photoinduced antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) as a representative of Gram-positive bacteria and Pseudomonas aeruginosa (PA) as a representative of Gram-negative bacteria. PMB1 is successful at inactivating the bacterial growth via both photoactivation of molecular oxygen and membrane depolarization mechanisms, thus proving to be a dual warhead. Inactivation of bacteria in water using such a supramolecular architecture is noteworthy and can shed light on the generation of new antimicrobial supramolecular systems.

In Situ Synthesis of Amino Acid Functionalized Carbon Dots with Tunable Properties and Their Biological Applications.

Emissive carbon dots (C-Dots) are known for displaying versatile properties, which have been widely utilized in many applications such as bioimaging, light-emitting devices, and photocatalysis. Further functionalization can tune their physical and chemical properties, which play the key role in many biologically and chemically relevant applications. Amino acids provide an attractive means for introducing functionality with a structural diversity. In this work, water-soluble emissive C-Dots were synthesized from pyrolysis of citric acid in the presence of various amino acids under hydrothermal conditions. We established that the diverse functionality-bearing polyamides and the polyester backbone of C-Dots are the origin of optical properties with a high quantum yield. The amino acid based C-Dots were systematically characterized using various analytical methods to confirm the core structure as well as the functionality. The results show that, depending on functionalization, the quantum yield can be varied and enhanced up to 62% and surface charge and hydrophobicity can be tuned. Most importantly, we observed a correlation between the quantum yield and properties of the side chain residue of amino acids such as hydrophobicity index and volume. Furthermore, it was found that the synthesized, diversely functionalized C-Dots exhibit no cellular toxicity and can be used for cell imaging. Due to variable surface functionality, we have also applied these C-Dots for array-based protein sensing. Overall, based on this reported method, we can easily tune the optical as well as surface properties of C-Dots, which will be suitable for many future applications.