Recent Developments in Nano-Formulations Against Diabetes.

Diabetes mellitus (DM) is a life-threatening metabolic syndrome, but patient compliance is poor due to the pain and inconvenience caused by the subcutaneous injection of insulin and other macromolecular diabetic therapies. Current challenges in DM management are to optimize the use of available therapies and reduce complications. For clinical improvements, future therapies need to be easier to use, achieving tighter glycemic control, better safety profiles, and reduced manufacturing costs. The medical applications of nanotechnology are enormous and have been proven to be the best approach to improve compliance and clinical efficacy by overturning biopharmaceutical obstacles. Nanoformulations enhance the properties of conventional drugs and are specific to the targeted delivery site. The aim of the present review is to provide an overview of the application of nano-formulations in diabetes management. We analyze the current state of most of the available approaches which are in various stages of research and development. Herein, we review the developing role of nanotechnology in diabetes management and focus on the technologies that we feel are most likely to have an impact.

Nanomicelles: Types, properties and applications in drug delivery.

Nanomicelles are self-assembling nanosized (usually with particle size within a range of 10 to 100 nm) colloidal dispersions with a hydrophobic core and hydrophilic shell. Owing to its size, solubility, customised surface or its exposure to the environment, nanomicelles show some unique or novel characteristics, which makes it multifunctional and thus makes its use indispensable in biomedical application and various other fields. This review presents the unique properties of nanomicelles that makes it different from other particles and paves its way to be used as drug delivery agent and many other biological uses or applications. It also emphasises on the drug encapsulation ability of the nanomicelles and different technique of drug loading and delivery along with its advantages and disadvantages.

Relative larvicidal property of common oxide nanostructures against Culex quinquefasciatus.

Common oxide nanostructures such as silicon-di-oxide, magnesium-oxide, zinc-oxide, and copper-oxide (CuO) having useful functional and bioactive properties have been synthesised and characterised. All these nanostructures have been found to be larvicidal towards Culex quinquefasciatus mosquito especially against lower instars in comparison with higher instars in 48 h. Only, CuO is larvicidal against late instar stages after 48 h. Moreover, CuO is larvicidal against first instar stages after 24 h (LC50 157 mg/l). However, none of these nanostructures are pupicidal. Post mortality larval morphology was found to be distorted under bright field microscopy and scanning electron microscopy images of affected larval surface appeared to be rough and uneven. Fluorescent images showed that nanostructures infiltrated inside visceral organs of larvae. Nanostructures also caused tissue oxidative stress in larvae. These results indicate that above stated oxide nanostructures are effective larvicidal agents against early instar stages of Culex larvae.

Silymarin-Loaded, Lactobionic Acid-Conjugated Porous PLGA Nanoparticles Induce Apoptosis in Liver Cancer Cells.

HepG2 cells (HCC), characterized by epithelial-like morphology, high proliferation rates, and nontumorigenicity, require cost-effective and efficient treatment. Silymarin, a flavonoid extract of Silybum marianum, is effective in the treatment of HCC. Here, we have reported a comparative anticancer study of the well-characterized nanoformulations of lactobionic acid-adorned porous PLGA-encapsulated silymarin (LA-PLGA-Sil) with only porous PLGA-encapsulated silymarin (PLGA-Sil) against HepG2 cells. Treatment of HepG2 cells with LA-PLGA-Sil produced a significant deterioration in cell viability at an essentially low dose as compared with PLGA-Sil, due to the adorned lactobionic acid moiety, which results in better targeting. p53, a tumor suppressor gene, essentially initiates apoptosis in cells procuring wild-type p53 (p53 +/+). In our report, treatment of HepG2 cells (p53 +/+) with LA-PLGA-Sil activated p53, which in turn inhibited the proliferation of cells by instigating cell-cycle arrest and apoptosis in a concentration-dependent manner and simultaneously stabilized the nuclear translocation of NFκB-p65. To explore the effect of LA-PLGA-Sil on the expression of microRNA, we observed that LA-PLGA-Sil markedly upregulated the miR-29b in human HCC cells. Reactivation of the p53 gene by miR-29b targeted Bcl-2 and triggered the sequential activation of mediators such as proapoptotic Bax protein, release of cytochrome c, and the activation of caspase proteins (caspase-3 and caspase-9). Furthermore, the overexpression of NFκB-p65 in HepG2 cells reversed the repression, and this stabilization effect of LA-PLGA-Sil on the nuclear translocation of p65 led to the significant downregulation of miR-29b and successively decreased the p53 expression in LA-PLGA-Sil-treated cells, thereby providing a survival mechanism to HepG2. In entirety, our study demonstrated the extensive potential of LA-PLGA-Sil to instigate the cell death of HepG2 cells via apoptosis by targeting the miR-29b/p53 axis through the stabilization of NFκB. It also impaired the migratory activity of HepG2 cells and thereby furnished a comprehensive way to HCC therapeutic treatment.

Vitamin C/Stearic Acid Hybrid Monolayer Adsorption at Air-Water and Air-Solid Interfaces.

Because of the antioxidant activity of vitamin C (Vit C) polar heads, they can be used as a protective agent for fatty acids. Hence, the study on the growth of Vit C/stearic acid (SA) mixed binary films at air-water interface (known as Langmuir monolayer) and air-solid interface (known as Langmuir-Blodgett films) is of paramount interest. Although Vit C is situated at subsurface beneath SA molecules and interacts via hydrogen bonding between the hydroxyl groups of Vit C and SA, several Vit C molecules may infiltrate within SA two-dimensional matrix at the air-water interface. The increased mole fraction of Vit C (0.125-0.5) and the reduction of temperature (from 22 to 10 °C) of the subphase water result in an increase in the amount of adsorbed Vit C at the air-water interface. The surface pressure (π)-area (A) isotherms illustrate that such inclusion of Vit C provokes a spreading out of Vit C/SA binary monolayers, which leads to an alteration of different physicochemical parameters such as elasticity, Gibbs free energy of mixing, enthalpy, entropy, interaction energy parameter, and activity coefficient. However, being polar in nature, the transfer of pure Vit C on substrates gets affected. It can be transferred onto substrate by mixing suitably with SA as confirmed by infrared spectra. Their structures, extracted X-ray reflectivity, and atomic force microscopy (topography and phase imaging) are found to be strongly dependent on the nature of the substrate (hydrophilic and hydrophobic).

Chitosan-fatty acid interaction mediated growth of Langmuir monolayer and Langmuir-Blodgett films.

The interaction of chitosan with bio-membranes, which plays important role in deciding its use in biological applications, is realized by investigating the interaction of chitosan with stearic acid (fatty acid) in Langmuir monolayers (at air-water interface) and Langmuir-Blodgett (LB) films (after transferring it onto solid substrate). It is found from the pressure-area isotherms that the chitosan insertion causes an expansion of chitosan-fatty acid hybrid monolayers, which reduces the elasticity and make the film heterogeneous. It is likely that at low surface pressure chitosan is situated at the interface, interacting with stearic acid molecules via electrostatic and hydrophobic interactions whereas at high pressure chitosan mainly located at subsurface beneath stearic acid molecules. In the latter case the interaction is predominantly electrostatic yielding very small contribution to the surface pressure. The reduction of temperature of the subphase water allows more number of chitosan molecules to reach surface to increase the pressure/interaction. On the other hand, although pure chitosan is found difficult to relocate on the substrate from air-water interface due to its hydrophilic-like nature, it alongside stearic acid (amphiphilic molecules) can be transferred onto substrate using LB technique as evident from infrared spectra. Their out-of-plane and in-plane structures, as extracted from two complementary surface sensitive techniques- X-ray reflectivity and atomic force microscopy, are found strongly dependent on the chitosan mole fraction and the deposition pressure. These analysis of the film-structure will essentially allow one to model the system better and provide better insight into the interaction.

Tailored-CuO-nanowire decorated with folic acid mediated coupling of the mitochondrial-ROS generation and miR425-PTEN axis in furnishing potent anti-cancer activity in human triple negative breast carcinoma cells.

Metal oxide nanoparticles are the forthcoming anti-tumor therapeutics and provide a versatile platform in the development of therapeutic approaches for drug-resistant cancers such as triple negative breast cancer (TNBC). Copper oxide nanoparticles have been characterized as anti-cancer agents but its toxicity has been a matter of concern. Herein, we have developed a targeted CuO Nanowire fabricated with Folic acid (CuO-Nw-FA) that enables enhanced cellular uptake in TNBC cells without imparting significant toxicity in normal cellular system. In the present study, we enumerated that CuO-Nw-FA caused mitochondrial-dependent apoptosis in MDAMB-231 cells. Furthermore, CuO-Nw-FA mediated cytosolic retardation of NF-κB favoured inactivation of miR-425 and henceforth activated PTEN to induce apoptosis in TNBC cells. Simultaneously, CuO-Nw-FA also restricted the in-vitro cell migration through the miR-425/PTEN axis via pFAK. Studies extended to ex-ovo and in-vivo mice models further validated the efficacy of CuO-Nw-FA. Additionally, the accumulations of nanoparticles in tumor as well as different organs in mice were examined by in-vivo biodistribution and ex-vivo optical imaging studies. Thus our results cumulatively propose that CuO-Nw-FA cross-talks two distinct signalling pathways to induce apoptosis and retard migration in TNBC cells and raises the possibility for the use of CuO-Nw-FA as a potent anti-tumor agent.

Simple synthesis of biocompatible biotinylated porous hexagonal ZnO nanodisc for targeted doxorubicin delivery against breast cancer cell: In vitro and in vivo cytotoxic potential.

Targeted drug delivery with porous materials features great promise as improved therapeutic potential for treatment of various diseases. In the present study we have attempted a microwave synthesis of porous hexagonal nanodisc of zinc oxide (PZHD) for the first time and its subsequent targeted delivery to breast cancer cells, MCF7. PZHD has been fabricated suitably with 3-aminopropyltriethoxysilane to impart additional stability and surface amines to anchor site directing ligand NHS-biotin. Biotinylated scaffold showed targeted delivery of anticancer drug doxorubicin and pH triggered release to MCF 7 cells with preferential distribution on specified domain. A detailed in vitro cytotoxicity study was associated with it to evaluate the mode of action of Dox loaded PZHD on MCF-7 cells by means of cell cycle analysis, apoptosis assays, Western blot and immuno-fluorescence image analysis. The efficacy of the Dox loaded PZHD was further validated from our in vivo tumor regression studies. Finally, the whole study has been supported by in vitro and in vivo bio-safety studies which also signified its biocompatibility with real time applications. To the best of our knowledge this is the first effort to use biotinylated PZHD for targeted delivery of doxorubicin within MCF 7 cells with a detailed study of its mechanistic application. This study might thus hold future prospects for therapeutic intervention for treatment of cancer.

PEGylated-thymoquinone-nanoparticle mediated retardation of breast cancer cell migration by deregulation of cytoskeletal actin polymerization through miR-34a.

Thymoquinone (TQ), a major active constituent of black seeds of Nigella sativa, has potential medical applications including spectrum of therapeutic properties against different cancers. However, little is known about their effect on breast cancer cell migration, which is the cause of over 90% of deaths worldwide. Herein, we have synthesized TQ-encapsulated nanoparticles using biodegradable, hydrophilic polymers like polyvinylpyrrolidone (PVP) and polyethyleneglycol (PEG) to overcome TQ's poor aqueous solubility, thermal and light sensitivity as well as consequently, minimal systemic bioavailability which can greatly improve the cancer treatment efficiency. Sizes of synthesized TQ-Nps were found to be below 50 nm and they were mostly spherical in shape with smooth surface texture. Estimation of the zeta potential also revealed that all the three TQ-Nps were negatively charged which also facilitated their cellular uptake. In the present investigation, we provide direct evidence that TQ-Nps showed more efficiency in killing cancer cells as well as proved to be less toxic to normal cells at a significantly lower dose than TQ. Interestingly, evaluation of the anti-migratory effect of the TQ-Nps, revealed that PEG4000-TQ-Nps showed much potent anti-migratory properties than the other types. Further studies indicated that PEG4000-TQ-Nps could significantly increase the expression of miR-34a through p53. Moreover, NPs mediated miR-34a up-regulation directly down-regulated Rac1 expression followed by actin depolymerisation thereby disrupting the actin cytoskeleton which leads to significant reduction in the lamellipodia and filopodia formation on cell surfaces thus retarding cell migration. Considering the biodegradability, non-toxicity and effectivity of PEG4000-TQ-Nps against cancer cell migration, TQ-Nps may provide new insights into specific therapeutic approach for cancer treatment.

Copper nanoparticle (CuNP) nanochain arrays with a reduced toxicity response: a biophysical and biochemical outlook on Vigna radiata.

Copper deficiency or toxicity in agricultural soil circumscribes a plant's growth and physiology, hampering photochemical and biochemical networks within the system. So far, copper sulfate (CS) has been used widely despite its toxic effect. To get around this long-standing problem, copper nanoparticles (CuNPs) have been synthesized, characterized, and tested on mung bean plants along with commercially available salt CS, to observe morphological abnormalities enforced if any. CuNPs enhanced photosynthetic activity by modulating fluorescence emission, photophosphorylation, electron transport chain (ETC), and carbon assimilatory pathway under controlled laboratory conditions, as revealed from biochemical and biophysical studies on treated isolated mung bean chloroplast. CuNPs at the recommended dose worked better than CS in plants in terms of basic morphology, pigment contents, and antioxidative activities. CuNPs showed elevated nitrogen assimilation compared to CS. At higher doses CS was found to be toxic to the plant system, whereas CuNP did not impart any toxicity to the system including morphological and/or physiological alterations. This newly synthesized polymer-encapsulated CuNPs can be utilized as nutritional amendment to balance the nutritional disparity enforced by copper imbalance.

Microwave synthesis of ZnO@mSiO2 for detailed antifungal mode of action study: understanding the insights into oxidative stress.

A simple chemical method has been devised for deliberate incorporation of zinc oxide nanoparticles (ZNPs) within mesoporous nanosilica (mSiO2) matrix to yield zinc oxide nanoparticles embedded in mesoporous nanosilica (ZnO@mSiO2). ZnO@mSiO2 inhibited the growth of four strains of fungi in a dose dependant manner. A series of biochemical assays revealed generation of oxidative stress from ZnO@mSiO2 for such biocidal response. We proposed transient superoxide and its subsequent conversion to H2O2 played a pivotal role behind such biocidal response as revealed from our systematic evaluation. This resulted morphological alteration of fungi through increase in number of facets, in correlation we found up-regulation in oxidative stress related genes. Bioavailability within the fungal sample was confirmed from microscopic, spectroscopic, biophysical techniques. Protein carbonylation of fungal species was the chemical outcome of such above mentioned stress and quantified by high performance liquid chromatography (HPLC) via subsequent hydrazone derivatization. Several in vitro and in vivo evaluations revealed the biocompatibility of ZnO@mSiO2. Altogether this report claims a new biocidal agent with a detailed mode of action focusing on the origin and quantification of oxidative stress through biophysical and biochemical techniques for the first time for real time applications.

Manganese nanoparticles: impact on non-nodulated plant as a potent enhancer in nitrogen metabolism and toxicity study both in vivo and in vitro.

Mung bean plants were grown under controlled conditions and supplemented with macro- and micronutrients. The objective of this study was to determine the response of manganese nanoparticles (MnNP) in nitrate uptake, assimilation, and metabolism compared with the commercially used manganese salt, manganese sulfate (MS). MnNP was modulated to affect the assimilatory process by enhancing the net flux of nitrogen assimilation through NR-NiR and GS-GOGAT pathways. This study was associated with toxicological investigation on in vitro and in vivo systems to promote MnNP as nanofertilizer and can be used as an alternative to MS. MnNP did not impart any toxicity to the mice brain mitochondria except in the partial inhibition of complex II-III activity in ETC. Therefore, mitochondrial dysfunction and neurotoxicity, which were noted by excess usage of elemental manganese, were prevented. This is the first attempt to highlight the nitrogen uptake, assimilation, and metabolism in a plant system using a nanoparticle to promote a biosafe nanomicronutrient-based crop management.

Mechanical downsizing of a gadolinium(III)-based metal-organic framework for anticancer drug delivery.

A Gd(III) -based porous metal-organic framework (MOF), Gd-pDBI, has been synthesized using fluorescent linker pDBI (pDBI=(1,4-bis(5-carboxy-1H-benzimidazole-2-yl)benzene)), resulting in a three-dimensional interpenetrated structure with a one-dimensional open channel (1.9×1.2 nm) filled with hydrogen-bonded water assemblies. Gd-pDBI exhibits high thermal stability, porosity, excellent water stability, along with organic-solvent and mild acid and base stability with retention of crystallinity. Gd-pDBI was transformed to the nanoscale regime (ca. 140 nm) by mechanical grinding to yield MG-Gd-pDBI with excellent water dispersibility (>90 min), maintaining its porosity and crystallinity. In vitro and in vivo studies on MG-Gd-pDBI revealed its low blood toxicity and highest drug loading (12 wt %) of anticancer drug doxorubicin in MOFs reported to date with pH-responsive cancer-cell-specific drug release.

High throughput electron transfer from carbon dots to chloroplast: a rationale of enhanced photosynthesis.

A biocompatible amine functionalized fluorescent carbon dots were developed and isolated for gram scale applications. Such carbogenic quantum dots can strongly conjugate over the surface of the chloroplast and due to that strong interaction the former can easily transfer electrons towards the latter by assistance of absorbed light or photons. An exceptionally high electron transfer from carbon dots to the chloroplast can directly effect the whole chain electron transfer pathway in a light reaction of photosynthesis, where electron carriers play an important role in modulating the system. As a result, carbon dots can promote photosynthesis by modulating the electron transfer process as they are capable of fastening the conversion of light energy to the electrical energy and finally to the chemical energy as assimilatory power (ATP and NADPH).

Photochemical modulation of biosafe manganese nanoparticles on Vigna radiata: a detailed molecular, biochemical, and biophysical study.

Manganese (Mn) is an essential element for plants which intervenes mainly in photosynthesis. In this study we establish that manganese nanoparticles (MnNP) work as a better micronutrient than commercially available manganese salt, MnSO4 (MS) at recommended doses on leguminous plant mung bean (Vigna radiata) under laboratory condition. At higher doses it does not impart toxicity to the plant unlike MS. MnNP-treated chloroplasts show greater photophosphorylation, oxygen evolution with respect to control and MS-treated chloroplasts as determined by biophysical and biochemical techniques. Water splitting by an oxygen evolving complex is enhanced by MnNP in isolated chloroplast as confirmed by polarographic and spectroscopic techniques. Enhanced activity of the CP43 protein of a photosystem II (PS II) Mn4Ca complex influenced better phosphorylation in the electron transport chain in the case of MnNP-treated chloroplast, which is evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and corresponding Western blot analysis. To the best of our knowledge this is the first report to augment photosynthesis using MnNP and its detailed correlation with different molecular, biochemical and biophysical parameters of photosynthetic pathways. At effective dosage, MnNP is found to be biosafe both in plant and animal model systems. Therefore MnNP would be a novel potential nanomodulator of photochemistry in the agricultural sector.

Entomotoxicity and biosafety assessment of PEGylated acephate nanoparticles: a biologically safe alternative to neurotoxic pesticides.

This is a report of an experimental study on a nanoencapsulation of the organophosphate acephate. Acephate was encapsulated in polyethylene glycol, using a simple, easy-to-replicate method that required no special equipment or conditions. The nanoencapsulation (nanoacephate) was characterized and its bioefficacy as compared to the regular commercial acephate was tested. The biosafety of the new compound was also tested on a murine model. Our new nanoencapsulation scored over the regular variety on all counts. It was found to successfully incorporate the active pesticidal component, acephate and this compound retained greater functional integrity over time as a nanoencapsulation. It was significantly more efficacious than the regular variety. It was biosafe when tested on murine model. We have reason to believe that this nanoencapsulation would allow the use of an organophosphate in a more targeted manner, thereby making it a cost-effective and eco-friendly alternative to the regular variety in use now.

Luminescent S-doped carbon dots: an emergent architecture for multimodal applications.

A facile route has been developed to synthesise and isolate sulphur doped fluorescent carbon dots for the first time. Such carbogenic quantum dots exhibit a wide band gap of 4.43 eV with a high open circuit voltage (VOC) of 617 mV along with a fill factor (FF) as high as 37%, using phenyl-C60-butyric acid methyl ester (PCBM) as the electron transporting layer. Besides the wide band gap, which is useful in the fabrication of solar cells, sulphur modified carbon dots also exhibit a high fluorescence quantum yield of 11.8% without any additional surface passivation, producing a unique fluorescent probe for further applications. In addition, the particles have a strong tendency to interact with the surface of gold nanoparticles and produce a thin fluorescent layer over their surfaces. Moreover, as they are completely biocompatible in nature, the highly fluorescent S-doped carbon dots have a strong potential for use in bioimaging applications. Interestingly, owing to the presence of oxygen and sulphur functionality, the highly negatively charged particles can easily bind with positively charged DNA-PEI complexes, simply by mixing them, and after interaction with DNA, bright blue fluorescence has been observed under an excitation wavelength of 405 nm .

Biochemical-, biophysical-, and microarray-based antifungal evaluation of the buffer-mediated synthesized nano zinc oxide: an in vivo and in vitro toxicity study.

Here we describe a simple, novel method of zinc oxide nanoparticle (ZNP) synthesis and physicochemical characterization. The dose-dependent antifungal effect of ZNPs, compared to that of micronized zinc oxide (MZnO), was studied on two pathogenic fungi: Aspergillus niger and Fusarium oxysporum. Superoxide dismutase (SOD) activity, ascorbate peroxidase activity, catalase activity, glutathione reductase (GR) activity, thiol content, lipid peroxidation, and proline content in ZNP-treated fungal samples were found to be elevated in comparison to the control, which strongly suggested that the antifungal effect of ZNPs was due to the generation of reactive oxygen species (ROS). Protein carbonylation, another marker of oxidative stress, was also evaluated by the dinitrophenyl hydrazine (DNPH) binding assay and Fourier transform infrared (FTIR) spectral analysis followed by Western blot and microarray analysis of fungal samples to confirm ROS generation by ZNPs. Micrographic studies for the morphological analysis of fungal samples (ZNP-treated and a control) exhibited an alteration in fungal morphology. The bioavailability of ZNPs on fungal cell was confirmed by energy-dispersive X-ray (EDX) analysis followed by high-resolution transmission electron microscopy (HR-TEM) and confocal microscopic analysis of the fungal samples. In vivo acute oral toxicity, acetylcholine esterase activity, and a fertility study using a mice model were also investigated for ZNPs. The long-term toxicity of ZNPs through intravenous injection was evaluated and compared to that of MZnO. The in vitro comparative toxicity of ZNPs and MZnO was evaluated on MRC-5 cells with the help of water-soluble tetrazolium (WST-1) and lactate dehydrogenase (LDH) assays. These results suggested that ZNPs could be used as an effective fungicide in modern medical and agricultural sciences.