Sounak Dutta

Dipeptide-based low-molecular-weight efficient organogelators and their application in water purification.

The development of new low-molecular-weight gelators for organic solvents is motivated by several potential applications of gels as advanced functional materials. In the present study, we developed simple dipeptide-based organogelators with a minimum gelation concentration (MGC) of 6-0.15 %, w/v in aromatic solvents. The organogelators were synthesized using different L-amino acids with nonpolar aliphatic/aromatic residues and by varying alkyl-chain length (C-12 to C-16). The self-aggregation behavior of these thermoreversible organogels was investigated through several spectroscopic and microscopic techniques. A balanced participation of the hydrogen bonding and van der Waals interactions is crucial for efficient organogelation, which can be largely modulated by the structural modification at the hydrogen-bonding unit as well as by varying the alkyl-chain length in both sides of the hydrophilic residue. Interestingly, these organogelators could selectively gelate aromatic solvents from their mixtures with water. Furthermore, the xerogels prepared from the organogels showed a striking property of adsorbing dyes such as crystal violet, rhodamine 6G from water. This dye-adsorption ability of gelators can be utilized in water purification by removing toxic dyes from wastewater.

Counterion dependent hydrogelation of amino acid based amphiphiles: switching from non-gelators to gelators and facile synthesis of silver nanoparticles

With the growing importance of hydrogels in scientific applications, the search for low molecular weight hydrogelators (LMWH), with simultaneous logical structural correlation is continuously increasing. In the present work, counterion variation of amino acid based amphiphiles was done to qualitatively evaluate its contribution towards hydrogelation. Further changes in the molecular skeleton of the amino acid amphiphile were done along with counterion variation to establish the importance of π–π interactions of aromatic planar ring in hydrogelation. An efficient conversion of a non-gelator to gelator molecule was achieved simply by changing the counterions to aromatic carboxylates. Role of the counterion in the mechanism of gelation process through the self-assembly of amino acid based amphiphiles has been discussed. The formation of supramolecular structures during hydrogelation was investigated by FESEM, CD, FT-IR, luminescence, 2D-NOESY and rheological studies. Interestingly, the L-tryptophan containing amphiphile hydrogelators were further utilized for synthesis of Ag nanoparticles under mild conditions without any need for high temperature, alkaline medium and external reducing agent. The nanoparticles obtained were characterized by UV-Vis, TEM, AFM and XRD experiments.

Counterion-induced modulation in the antimicrobial activity and biocompatibility of amphiphilic hydrogelators: influence of in-situ-synthesized Ag-nanoparticle on the bactericidal property.

The necessity for the development of new antimicrobial agents due to the ever increasing threat from microbes is causing a rapid surge in research. In the present work, we have shown the efficient antimicrobial activity of a series of amino acid-based hydrogelating amphiphiles through alteration in their counterion. The subtle variation in the counterion from chloride to various organic carboxylates had a significant impact on the antimicrobial properties with notable improvement in biocompatibility toward mammalian cells. Incorporation of a hydrophobic moiety in the counterion augmented the antibacterial property of the amphiphilic hydrogelator as minimum inhibitory concentration (MIC) against the Gram-positive bacterial strain, Bacillus subtilis decreased up to 5-fold (with respect to the chloride) in the case of n-hexanoate. These counterion-varied amphiphilic hydrogelators were also found to be effective against fungal strains (Candida albicans and Saccharomyces cerevisiae) where they exhibited MICs in the range of 1.0-12.5 μg/mL. To widen the spectrum of antibacterial activity, particularly against Gram-negative bacteria, silver nanoparticles (AgNPs) were synthesized in situ within the supramolecular assemblies of the carboxylate hydrogelators. These AgNP-amphiphile soft-nanocomposites showed bactericidal property against both Gram-positive and Gram-negative bacteria. Encouragingly, these carboxylate hydrogelators showed superior biocompatibility toward mammalian cells, HepG2 and NIH3T3, as compared to the chloride analogue at a concentration range of 10-200 μg/mL. Importantly, the AgNP composites also showed sufficient viability to mammalian cells. Because of the intrinsic hydrogelation ability of these counterion-varied amphiphiles, the resulting soft materials and the nanocomposites could find applications in biomedicine and tissue engineering.

pH-Triggered conversion of soft nanocomposites: in situ synthesized AuNP-hydrogel to AuNP-organogel

Amino acid based amphiphilic gelators (carboxylate salts) were employed for the in situ synthesis of gold nanoparticles (GNPs) in hydrogel networks at room temperature without using any external reducing or capping agents for the development of AuNP-hydrogel soft composite. Synthesized AuNP-hydrogel composites were then successfully converted to AuNP-organogel composites simply by lowering the pH of the aqueous medium, as the hydrogelating amphiphilic carboxylates were transformed to corresponding carboxylic acids that are efficient organogelators. These water insoluble carboxylic acids spontaneously moved from the aqueous phase to the nonpolar organic media (toluene) along with the synthesized GNPs to form the AuNP-organogel composite. The phase transfer of the GNPs from a hydrogel network to an organogel network was investigated by UV-Vis spectroscopy, field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM) and X-ray diffraction (XRD) studies. Supramolecular networks of both the gels played a crucial role in stabilization of the GNPs. Fluorescence spectroscopy was used to investigate the mechanistic detail of the in situ GNP synthesis. The characterizations indicated the formation of spherical and uniform sized GNPs and even phase transfer of the nanoparticles did not result in change of shape or size of the nanoparticles. Rational designing of the gelator/nongelator molecules helped us to recognize the key structural components required for the efficient synthesis and stabilization of the GNPs in both the phases. Rheological study suggested that the hydrogel-GNP composites possess improved viscoelastic property than the native hydrogel.

Imidazolium Bromide-Based Ionic Liquid Assisted Improved Activity of Trypsin in Cationic Reverse Micelles

The present work reports the imidazolium-based ionic liquids (ILs) assisted enhancement in activity of water-pool solubilized enzyme trypsin in cationic reverse micelles of CTAB. A set of imidazolium ILs (1-alkyl-3-methyl imidazolium bromides) were prepared with varying lengths of their side arm which results in the differential location of these organic salts in the reverse micelles. The different ILs offered varied activating effects on the biocatalyst. The activity of trypsin improved approximately 30-300% in the presence of 0.1-10 mM of different ILs in reverse micelles of CTAB. Trypsin showed approximately 300% (4-fold) increment in its activity in the presence of IL 2 (1-ethyl-3-methyl imidazolium bromide, EMIMBr) compared to that observed in the absence of IL in CTAB reverse micelles. The imidazolium moiety of the IL, resembling the histidine amino acid component of the catalytic triad of hydrolases and its Br(-) counterion, presumably increases the nucleophilicity of water in the vicinity of the enzyme by forming a hydrogen bond that facilitates the enzyme-catalyzed hydrolysis of the ester. However, the ILs with increasing amphiphilic character had little to no effect on the activity of trypsin due to their increased distance from the biocatalyst, as they tend to get localized toward the interfacial region of the aggregates. Dynamic light scattering experimentation was carried out in the presence of ILs to find a possible correlation between the trypsin activity and the size of the aggregates. In concurrence with the observed highest activity in the presence of IL 2, the circular dichroism (CD) spectrum of trypsin in CTAB reverse micelles doped with IL 2 exhibited the lowest mean residue ellipticity (MRE), which is closest to that of the native protein in aqueous buffer.

In situ synthesized Ag nanoparticle in self-assemblies of amino acid based amphiphilic hydrogelators: development of antibacterial soft nanocomposites

The present work reports the development of a new class of antibacterial soft-nanocomposites by in situ synthesis of silver nanoparticle (AgNP) within the supramolecular self-assemblies of amino acid (tryptophan/tyrosine) based amphiphilic hydrogelators. Interestingly, the nanoparticle synthesis does not require the use of any external reducing/stabilizing agents. The nanocomposites were characterized by UV-vis spectra, transmission electron microscopy (TEM) images, X-ray diffraction spectroscopy (XRD) and thermo gravimetric analysis (TGA). Encouragingly, these soft nanocomposites showed excellent antibacterial activity against both Gram-positive and Gram-negative bacteria whereas the amphiphiles alone were lethal only toward Gram-positive bacteria. Judicious combination of bactericidal AgNP within the self-assemblies of inherently antibacterial amphiphilic gelators led to the development of soft nanocomposites effective against both type of bacteria. The head group charge and structure of the amphiphiles were altered to investigate their important role on the synthesis and stabilization of AgNP and also in modulating the antibacterial activity of the nanocomposites. The antibacterial activities of soft nanocomposites comprising amphiphiles with cationic head group were found to be more efficient than the anionic soft nanocomposites. Interestingly, these nanocomposites have shown considerable biocompatibility to mammalian cell, NIH3T3. Furthermore, the well-known tissue engineering scaffold, agar-gelatin film infused with these soft nanocomposites allowed normal growth of mammalian cells on its surface while being lethal toward both Gram-positive and Gram-negative bacteria.

Structure and properties of cholesterol-based hydrogelators with varying hydrophilic terminals: biocompatibility and development of antibacterial soft nanocomposites.

The present work demonstrates a rational designing and synthesis of cholesterol-based amino acid containing hydrogelators with the aim to improve the biocompatibility of these amphiphilic molecules. A thorough structure-property correlation of these hydrogelators was carried out by varying the hydrophilic terminal from a neutral amine to a quaternized ammonium chloride. The amphiphiles having a cationic polar head as the hydrophilic domain and cholesterol as the hydrophobic unit showed better water gelation efficiency (minimum gelation concentration (MGC) ∼0.9-3.1%, w/v) than the analogous free amines. Presumably, the additional ionic interactions for the quaternized amphiphiles might have played the crucial role in gelation as counterions also got involved in hydrogen bonding with solvent molecules. Hence, the attainment of desired hydrophilic-lipophilic balance (HLB) of hydrophobic cholesterol in combination with the appropriate hydrophilic terminal led to the development of efficient hydrogels. Microscopic investigations revealed the formation of various supramolecular morphologies of hydrogels due to the variation in the molecular structure of the amphiphile. Spectroscopic investigations showed the involvement of hydrogen-bonding, hydrophobic, and π-π interactions in the self-assembled gelation. Importantly, biocompatibility of all the cholesterol-based hydrogelators tested against human hepatic cancer derived HepG2 cells showed increased cell viability than the previously reported alkyl-chain-based amphiphilic hydrogelators. In order to incorporate broad spectrum antibacterial properties, silver nanoparticles (AgNPs) were synthesized in situ within the hydrogels using sunlight. The amphiphile-AgNP soft nanocomposite exhibited notable bactericidal property against both gram-positive and gram-negative bacteria.

Covalently functionalized single-walled carbon nanotubes at reverse micellar interface: a strategy to improve lipase activity.

The present work reports covalent functionalization of single-walled carbon nanotubes (f-SWNTs) to introduce hydrophilicity to the otherwise amphiphobic nanotubes. The charge and spacer length of the functional moiety were varied by using quaternized ethylene diamine, 6-aminocaproate, quaternized (ethylenedioxy)bis(ethylamine), and a poly(ethylene glycol) (PEG) unit (f-SWNT-1 to f-SWNT-4, respectively). These f-SWNTs with varying degrees of hydrophilicity were incorporated within cetyltrimethyl ammonium bromide (CTAB) reverse micelles to develop stable self-assembled nanohybrids. An optimum hydrophilicity on the SWNT surface led to interfacial localization of f-SWNTs resulting in the augmentation of space at the interface. A surface-active enzyme, lipase, localized at this enhanced interface of f-SWNT-containing CTAB reverse micelles exhibited significant activation (2.5-fold) compared to that in the absence of the nanoconstructs. This improvement in lipase activity was mainly due to the smooth occupancy of lipase and also presumably because of the increase in the concentrations of both substrate and the enzyme at the augmented interface. Interestingly, the f-SWNTs that activate lipase in reverse micelles deactivate the same enzyme in water. The dispersion of f-SWNTs in water and its matching integration at the interface of reverse micelles were confirmed through transmission electron microscopic (TEM) investigations. The interfacial localization of these nanoconstructs was also established from the distinct fluorescence behavior of a hydrophobic fluorescent probe, fluorescein isothiocyanate (FITC), adsorbed onto the f-SWNT surface. In concurrence with the observed lipase activity, the corresponding changes in the enzyme conformation within f-SWNTs integrated reverse micelle as well as in aqueous medium were studied by circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy.

Pyridinium based amphiphilic hydrogelators as potential antibacterial agents

Summary The numerous applications of hydrogelators have led to rapid expansion of this field. In the present work we report the facile synthesis of amphiphilic hydrogelators having a quaternary pyridinium unit coupled to a hydrophobic long alkyl chain through an amide bond. Different amphiphiles with various hydrophobic chain length and polar head groups were rationally designed and synthesized to develop a structure-property relation. A judicious combination of hydrophilic and hydrophobic segments led to the development of pyridinium based amphiphilic hydrogelators having a minimum gelation concentration of 1.7%, w/v. Field emission scanning electronic microscopy (FESEM), atomic force microscopy (AFM), photoluminescence, FTIR studies, X-ray diffraction (XRD) and 2D NOESY experiments were carried out to elucidate the different non-covalent interactions responsible for the self-assembled gelation. The formation of three-dimensional supramolecular aggregates originates from the interdigitated bilayer packing of the amphiphile leading to the development of an efficient hydrogel. Interestingly, the presence of the pyridinium scaffold along with the long alkyl chain render these amphiphiles inherently antibacterial. The amphiphilic hydrogelators exhibited high antibacterial activity against both Gram-positive and Gram-negative bacteria with minimum inhibitory concentration (MIC) values as low as 0.4 μg/mL. Cytotoxicity tests using MTT assay showed 50% NIH3T3 cell viability with hydrogelating amphiphile 2 up to 100 μg/mL.

Biotinylated amphiphile-single walled carbon nanotube conjugate for target-specific delivery to cancer cells

The present work reports the specific targeting of cancerous cells using a non-covalently water dispersed nanoconjugate of biotinylated amphiphile-single walled carbon nanotube (SWNT). The fundamental approach involves incorporation of the biotin into the architecture of the carbon nanotube (CNT) dispersing agent to develop a multifaceted delivery vehicle having a high colloidal stability, substantial cell viability and targeted specificity towards cancer cells. A three way functionalization strategy was employed to introduce a C-16 hydrophobic segment, polyethylene glycol hydrophilic fragment and biotin as the target-specific unit at the -OH, -COOH and -NH2 terminals of l-tyrosine, respectively. The newly developed neutral amphiphile exhibited an efficient SWNT dispersion (72%) in water, significant viability of different mammalian cells (Hela, HepG2, CHO and HEK-293) up to 48 h and also media stability. Most importantly, the biotinylated amphiphile-SWNT dispersion successfully transported the fluorescently labelled Cy3-oligoneucleotide (loaded on the surface of CNT) inside the cancerous Hela, HepG2 cells after 3 h of incubation, in contrast to CHO and HEK-293 cells (devoid of overexpressed biotin receptors). The presence of the biotin moiety in the cellular transporters facilitated the internalization of cargo due to the overexpressed biotin receptors in the cancer cells. Importantly, this nanohybrid was also capable of specifically transporting the anticancer drug doxorubicin to cancer cells, which led to the significant killing of Hela cells compared to the normal CHO cells. Thus, the receptor-mediated specific transportation of cargo into cancer cells was possible only due to the biotinylated CNT dispersing agent. To the best of our knowledge this is the first reported amino acid based biotinylated small amphiphilic molecule that non-covalently dispersed SWNTs and the corresponding nanoconjugate showed excellent cell viability, antibiofouling properties and the desired target-specific drug delivery.