Development of nanostructured lipid carriers as a promising tool for methotrexate delivery: physicochemical and in vitro evaluation.

The aim of the present study is to fabricate the stable nanostructured lipid carriers (NLCs) using biocompatible excipients for the encapsulation of Methotrexate (MTX), a chemotherapeutic agent for breast cancer treatment. MTX has restricted clinical applications owing to its low solubility, non-specific targeting and adverse side effects. Glyceryl Monostearate (GMS) and Miglyol 812 (MI1) were chosen as solid and liquid lipids, respectively, for the fabrication of NLCs, and the influence of variation of solid and liquid composition was investigated. The prepared NLCs exhibited long-term stability and spherical shape morphology as characterized by electron microscopy. The internal structure of fabricated NLCs was arranged into cubic crystalline as confirmed by small-angle X-ray scattering (SAXS) analysis. MTX's encapsulation efficiency of ∼85 ± 0.9%. and sustained in vitro release of MTX ∼ 52% ± 3.0 in 24 h was achieved. Classical molecular dynamics (MD) simulations were performed to study the structural stability of the MTX encapsulated NLCs. Hemolysis carried out on the NLCs showcased the biosafety of the formulation under the tolerance limit (<10%). Further, the MTT assay demonstrates that MTX-loaded NLCs exhibited toxicity against HeLa and MCF-7 cell lines as compared to blank NLCs. The finding demonstrates NLCs as promising vehicles for MTX delivery to address cancer.Communicated by Ramaswamy H. Sarma.

Development of Phosphatidylcholine/Tween 80 based biocompatible clove oil-in-water nanoemulsion as a green nanocarrier for controlled herbicide delivery.

Recently, the development of ecofriendly and biocompatible agrochemical delivery systems has garnered widespread attention because of their great potential in sustainable agri-food applications. Atrazine (ATZ) is a globally used herbicide used to control weeds, but it suffers from poor aqueous solubility, poor efficacy, and environmental loss. Herein, we report a novel, eco-friendly and biocompatible clove oil-based nanoemulsion as a green nanocarrier to enhance the solubility, bioavailability, and control release of ATZ. Food grade surfactants, Tween 80 and Phosphatidylcholine (PC) were used to formulate clove oil nanoemulsion with size <200 nm using ultrasonic emulsification technique, without any use of organic solvent. The ATZ encapsulation efficiency in NEm was greater than 95%. DLS confirms the nanosize (106 nm) and monodispersity of NEm. HRTEM reveals the spherical morphology of the nanodroplets. FTIR and DSC confirm the successful incorporation of ATZ inside the NEm oil droplet core. ATZ loaded NEm showed excellent thermal and storage stability, low Ostwald ripening rate, slow and sustained herbicide release behavior, which is of vital importance for an herbicide formulation. The release rate was better than commercial ATZ and free ATZ formulations. Results from herbicidal activity assays demonstrate that ATZ NEm exhibited excellent herbicidal activity even at low concentrations as compared to commercial ATZ analogs. In consideration of biocompatible excipients, free of organic solvent, and a simple fabrication process, ATZ loaded clove oil NEm can hold great potential in weed control and sustainable agri-food applications.

Combined delivery of TLR2 and TLR7 agonists by Nanostructured lipid carriers induces potent vaccine adjuvant activity in mice.

Toll-like receptor (TLR) agonists are promising adjuvants and the combination of TLR agonists enhance immune responses by providing synergistic immune activity via triggering different signalling pathways. However, systematic cytotoxicity due to the immediate release of such immune potentiators from the site of injection hampers its clinical performance. Nanostructured lipid carriers (NLCs) offer a possibility to incorporate multiple TLR agonists with high encapsulation efficiency and slow drug release. Herein, we synthesized NLCs from didodecyldimethylammonium bromide (D12DAB) and oleic acid and used these to co-encapsulate a Pam2CS derivative (T-2, TLR2 agonist) with an imidazoquinoline derivative (T-7, TLR7 agonist) as a combination vaccine adjuvant. Hydrodynamic diameter and zeta potential of the prepared NLCs were found to be in the range of 200-500 nm and 23-27 mV, respectively. Spherical shape and size of prepared NLCs were also assessed through Field Emission Scanning Electron Microscopy (FE-SEM) and Transmission Electron Microscopy (TEM) analysis. In-vitro release studies of T-7 demonstrated sustained release and the addition of lipopeptide T-2 augmented encapsulation efficiency (from 84 to 92.9%) with a slight trigger in the release percentage. All NLC formulations were screened in TLR2/1, TLR2/6, TLR7 and TLR8 reporter cell lines and loaded NLC formulation showed high TLR2 and TLR7 agonistic activity. Adjuvant potency was evaluated through intramuscular immunization of female C57BL/6 mice with recombinant hepatitis B surface antigen and influenza hemagglutinin protein. T-2 and T-7 loaded NLCs induced good protective efficacy in mice challenged with a lethal dose of influenza virus.

Reply to the 'Comment on "Physicochemical stimuli as tuning parameters to modulate the structure and stability of nanostructured lipid carriers and release kinetics of encapsulated antileprosy drugs"' by J. Kang and A. M. Kang, J. Mater. Chem. B, 2020, 8, DOI.

In a recent comment [J. Kang and A. M. Kang, J. Mater. Chem. B, 2020, 8, DOI: 10.1039/D0TB01160F] on our article [R. Kanwar, M. Gradzielski, S. Prevost, G. Kaur, M. S. Appavou and S. K. Mehta, J. Mater. Chem. B., 2019, 7, 6539], the authors tried to demonstrate that the binding phenomenon between drug-loaded nanostructured lipid carriers and the protein BSA was subjected to enthalpy-entropy compensation, rather than being entropy-driven. The outcome, however, remains the same in spite of the complicated analysis attempted. While we appreciate their effort to extract more information from our data, we have a number of remarks on their process.

Physicochemical stimuli as tuning parameters to modulate the structure and stability of nanostructured lipid carriers and release kinetics of encapsulated antileprosy drugs.

To unveil the effect of electrolyte concentration, pH and polymer addition on Tween 80 stabilized nanostructured lipid carriers (NLCs, based on dialkyldimethylammonium bromides DxDAB and Na oleate), an in-depth scattering analysis was performed. Dynamic and static light scattering (DLS/SLS) and small-angle neutron scattering (SANS) techniques along with zeta potential studies were exploited to understand the structural evolution and physical stability of NLCs. In these experiments, we varied the salt concentration, pH, and the admixture of Pluronic F127 in order to elucidate their effect on NLC morphologies. In most cases, two populations of different sizes are present which differ by one order of magnitude. The antileprosy drugs (ALD) Rifampicin and Dapsone were encapsulated in NLCs and the vector properties were assessed for a series of DxDAB (where x = 12, 14, 16 and 18) NLCs. The influence of composition on the entrapment and release behavior of NLCs was investigated: The size of NLCs correlates with the release rate of the incorporated drug. The interaction of drug-loaded NLCs with bovine serum albumin was studied to understand the release of ALD in the plasma.

Green Nanotechnology-Driven Drug Delivery Assemblies.

Green nanotechnology incorporates the principles of green chemistry and green engineering to fabricate innocuous and eco-friendly nanoassemblies to combat the problems affecting the human health or environment. Subsequently, amalgamation of green nanotechnology with drug delivery area has actually commenced a new realm of "green nanomedicine". The burgeoning demand for green nanotechnology-driven drug delivery systems has led to the development of different types of delivery devices, like inorganic (metallic) nanoparticles, quantum dots, organic polymeric nanoparticles, mesoporous silica nanoparticles, dendrimers, nanostructured lipid carriers, solid lipid nanoparticles, etc. The present article deals with a brief account of delivery devices produced from green methods and describes site-specific drug delivery systems (including their pros and cons) and their relevance in the field of green nanomedicine.

Experimental validation of biocompatible nanostructured lipid carriers of sophorolipid: Optimization, characterization and in-vitro evaluation.

To date, the potential of sophorolipids (an important class of glycolipids) has been exploited solely as amphipathic molecules but their ability to formulate lipid nanoparticles has never been explored. In this report, for the first time, lipid nanostructures coated with polysorbates (Tweens) were formulated by a hot dispersion method. By varying the amount of lipid, type of surfactant, and alcohol, dilution ratio etc., the formulation was optimized with respect to its stability, which is a central aspect of their potential applications. Their comprehensive physicochemical characterization was done using static and dynamic light scattering (SLS, DLS), small angle neutron scattering (SANS), zeta-potential, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and atomic force microscopy (AFM) techniques. Further hemolysis study was conducted to understand the in-vitro cytotoxicity levels of the lipidic nanoparticles prior to its application as a potent drug delivery device for countermanding the problems associated with challenging tuberculosis and leprosy drug-Rifampicin. Attaining high entrapment efficiency and sustained release from the developed carrier, further interaction with bovine serum albumin was investigated, to understand the in-vivo behavior of the nanostructured lipid carriers (NLCs).

Effect of lipid chain length on nanostructured lipid carriers: Comprehensive structural evaluation by scattering techniques.

The objective of the present investigation was to evaluate nano-dispersed systems of differently chained lipids (solid phases) using various scattering techniques. Nanostructured lipid carriers (NLCs) were fabricated by employing the microemulsification methodology in which dialkyldimethyl ammonium bromide (DxDAB) of different alkyl chain length (x = 12, 14, 16, 18) and oleic acid were used as the solid lipid and liquid lipid, respectively. For the first time, the effect of DxDAB as a function of the chain length of the double alkyl chain on the structural characteristics of NLCs was investigated. Dynamic light scattering (DLS) and static light scattering (SLS) results showed that a small but systematic size increase occurs with increase in the chain length 'x' of the DxDAB from 12 to 16, yet D18DAB based NLCs exhibited the smallest size irrespective of its higher hydrophobicity. Small angle neutron scattering (SANS) analysis revealed the structural make up of NLCs having spherical nanoparticles and triaxial ellipsoidal core-shell micelles in the system. In-vitro cytotoxicity evaluation indicated that toxicity is simply concentration-dependent phenomena and NLCs with less than 5 mg/mL are preferred for better in-vivo tolerance.

Biomimetic Solid Lipid Nanoparticles of Sophorolipids Designed for Antileprosy Drugs.

The objective of the present work was to develop solid lipid nanoparticles (SLNs) as drug-encapsulating structures by the solvent injection method. In this report, for the first time the inherent potential of lactonic sophorolipid (glycolipid) was exploited to formulate SLNs. A range of different Pluronic copolymers were screened by dynamic and static light scattering with the aim of obtaining most stable SLNs. To comprehend the structure of the SLNs, techniques such as transmission electron microscopy, differential scanning calorimetry, Fourier transform infrared spectroscopy, and X-ray diffraction were employed. A clear correlation between the type of Pluronic and size and stability of the SLNs could be drawn. The vector properties of the formed SLNs were assessed for both the encapsulated hydrophobic drugs-rifampicin and dapsone. To elucidate the transport mechanism of drug release, kinetic modeling was carried out on the drug release profiles. The promising results of sophorolipid-based SLNs have actually established a new arena beneath the significantly developed field of SLNs.

Revealing the potential of Didodecyldimethylammonium bromide as efficient scaffold for fabrication of nano liquid crystalline structures.

To exploit the potential of Didodecyldimethylammonium bromide (D12DAB) as a core lipidic constituent, an attempt was made to fabricate and optimize cationic nanostructured lipid carriers (cNLCs) using a cost-effective microemulsification methodology. Designed composition was optimized by studying the effect of different microemulsion components on D12DAB cNLCs characteristics. ​Spherical shaped D12DAB cNLCs were obtained with an average size of ∼160 nm and zeta potential of +30.2 mV. Differential Scanning Calorimetry (DSC) depicted the presence of thermotropic character, whereas polarized optical microscopy confirmed the mesophase like behavior of D12DAB based cNLCs. In addition, hemolysis analysis revealed that the toxicity was concentration dependent as LC50 was reached at a concentration of 50 µg/mL of cNLCs. This class of cNLCs is expected to become a potent candidate for a broad spectrum of medicaments as carriers, targeting for pharmaceutical and medicinal purposes, due to the combination of a hard lipid with a soft lipid, where the liquid crystalline structure of the lipid co-exists.