Echogenic Gold Nanorod Incorporated Hybrid Poly(2-oxazoline) Nanocapsules for Real-Time Ultrasound/Fluorescent Imaging and Targeted Cancer Theranostics.

In recent years, various nanocarrier systems have been explored to enhance the targeting of cancer cells by improving the ligand-receptor interactions between the nanocarrier and cancer cells for selective cancer cell imaging and targeted delivery of anticancer drugs. Herein, we report multifunctional hydrogen-bonded multilayer nanocapsules functionalized with both folic acid-derived quantum dots (FAQDs) and gold nanorods (AuNRs) for targeted cancer therapy and cancer cell imaging using fluorescence microscopy and medical-range ultrasound imaging systems. The encapsulation efficiency of nanocapsules was found to be 49% for 5-fluorouracil (5-FU). The release percentage reached a plateau at 37% after 1 h at pH 7.4 and increased to 57% after 3 h when the release pH was decreased to pH 5.5 (i.e., the pH of the tumor environment). Under ultrasound irradiation, the release was significantly accelerated, with a total release of 52% and 68% after only 6 min at pH 7.4 and pH 5.5, respectively. While the sonoporation process plays an important role in anticancer activity experiments under ultrasound exposure by generating temporary pores, the targeting ability of FAQDs brings the capsules closer to the cell membrane and improves the cellular uptake of the released drug, thereby increasing local drug concentration. In vitro cytotoxicity experiments with HCT-116 and HEp-2 cells demonstrated anticancer activities of 96% and 98%, respectively. The nanocapsules showed enhanced ultrasound scattering signal intensity and bright spots under ultrasound exposure, most likely caused by high scattering ability and internal reflections of preloaded AuNRs in the interior structure of the nanocapsules. Hence, the demonstrated nanocapsule system not only has the potential to be used as an integrated system for early- stage detection and treatment of cancer cells but also has the ability for live tracking and imaging of cancer cells while undergoing treatment with chemotherapy and radiation therapy.

Co-encapsulation and release of apigenin and ascorbic acid in polyelectrolyte multilayer capsules for targeted polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS), a prevalent endocrine disorder in women of reproductive age, is linked to hormonal imbalances and oxidative stress. Our study investigates the regenerative potential of apigenin (AP, hydrophobic) and ascorbic acid (AC, hydrophilic) encapsulated within poly (allylamine hydrochloride) and dextran sulfate (PAH/DS) hollow microcapsules for PCOS. These microcapsules, constructed using a layer-by-layer (LbL) assembly, are found to be 4 ± 0.5 µm in size. Our research successfully demonstrates the co-encapsulation of AP and AC in a single PAH/DS system with high encapsulation efficiency followed by successful release at physiological conditions by CLSM investigations. In vitro tests with testosterone-treated CHO cells reveal that the dual-drug-loaded PAH/DS capsules effectively reduce intracellular ROS levels and apoptosis and offering protection. In an in-vivo zebrafish model, these capsules demonstrate active biodistribution to targeted ovaries and reduce testosterone levels through radical scavenging. Histopathological examinations show that the injected dual-drug-loaded PAH/DS microcapsules assist in the development of ovarian follicles in testosterone-treated zebrafish. Hence, this dual-drug-loaded system, capable of co-encapsulating two natural compounds, effectively interacts with ovarian cells, reducing cellular damage and normalizing PCOS conditions.

Rapid and sensitive electrochemical detection of oxidized form of glutathione in whole blood samples using Bi-metallic nanocomposites.

We report a facile one-pot synthesis of bimetallic nickel-gold (Ni-Au) nanocomposite for ultra-sensitive and selective electrochemical detection of oxidized glutathione (GSSG) by electrochemical deposition on fluorine doped tin oxide (FTO) substrate. The electrodeposition of Ni-Au nanocomposite on FTO was confirmed by various characterization techniques such as field emission scanning electron microscopy (FE-SEM), X-ray diffractometer (XRD) and Fourier transform infra-red (FTIR) spectroscopy. The cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) was utilized for the electrochemical characterization of glutathione reductase (GR)/Ni-Au/FTO working electrode at each stage of modification. The GR enzyme immobilized on the Ni-Au/FTO working electrode via glutaraldehyde cross-linking exhibited excellent selectivity against GSSG in the presence of nicotinamide adenine dinucleotide phosphate (NADPH). The immobilized GR enzyme breaks down the GSSG to reduced glutathione (GSH) and converting NADPH to NADP+ whereby generating an electron for the electrochemical sensing of GSSG. The synergistic behavior of bimetals and good electro-catalytic property of the fabricated sensor provided a broad linear detection range from 1 fM to 1 µM with a limit of detection (LOD) of 6.8 fM, limit of quantification (LOQ) of 20.41 fM and sensitivity of 0.024 mA/µM/cm2. The interference with other molecules such as dopamine, glycine, ascorbic acid, uric acid and glucose was found to be negligible due to the better selectivity of GR enzyme towards GSSG. The shelf-life and response time of the fabricated electrode was found to be 30 days and 32 s, respectively. The real sample analysis of GSSG in whole blood samples showed average recovery percentage from 95 to 101% which matched well with the standard calibration plot of the fabricated sensor with relative standard deviation (RSD) below 10%.

Synthesis of Ag nanoparticles for selective dual detection of glutathione and dopamine using N, N-dimethyl-p-phenylenediamine mediated colorimetric probe.

We report a simple and easy method to synthesize Ag nanoparticles (Ag NPs) and demonstrate its potential for the detection of glutathione (GSH) and dopamine (DA) via colorimetric assay. The Ag NPs were found to be monodispersed and spherical with a size of 5 ± 2 nm. The X-ray diffraction (XRD) and high resolution transmission electron microscopy (HR-TEM) investigations revealed the formation of crystalline Ag NPs. The colour of N, N-dimethyl-p-phenylenediamine assay changed from dark pink to colourless when the concentration of GSH was increased from 1 to 40 µM. Notably, the suspension colour changed from dark pink to blue when a similar set of experiments were performed with DA. The UV/Visible and interference experiments of Ag NPs exhibited excellent sensitivity and selectivity against both GSH and DA even after the addition of 40 µM of different interference biomolecules. The calculated limit of detection (LOD) was 141 and 245 nM for GSH and DA, respectively. The real-time analysis with serum samples showed satisfactory recovery percentages of >95 and 80-90% for GSH and DA, respectively. Hence, the Ag NPs reported here have huge potential to serve as a sensitive and selective colorimetric sensor for the detection of GSH and DA for diverse applications ranging from catalysis to cancer therapy and theranostics.

Lysozyme microspheres incorporated with anisotropic gold nanorods for ultrasound activated drug delivery.

We report on the fabrication of lysozyme microspheres (LyMs) incorporated with gold nanorods (NRs) as a distinctive approach for the encapsulation and release of an anticancer drug, 5-Fluorouracil (5-FU). LyMs with an average size of 4.0 ± 1.0 µm were prepared by a sonochemical method and characterized using scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Fourier-transform infrared spectroscopy (FTIR). The LyMs were examined using hydrophobic (nile red) as well as hydrophilic (trypan blue) dyes under confocal laser scanning microscopy (CLSM) to obtain information about the preferential distribution of fluorescent molecules. Notably, the fluorescent molecules were accumulated in the inner lining of LyMs as the core was occupied with air. The encapsulation efficiency of 5-FU for LyMs-NR was found to be ∼64%. The drug release from control LyMs as well as LyMs incorporated with NRs was investigated under the influence of ultrasound (US) at 200 kHz. The total release for control LyMs and LyMs incorporated with gold NRs was found to be ∼70 and 95% after 1 h, respectively. The density difference caused by NR incorporation on the shell played a key role in rupturing the LyMs-NR under US irradiation. Furthermore, 5-FU loaded LyMs-NR exhibited excellent anti-cancer activity against the THP-1 cell line (∼90% cell death) when irradiated with US of 200 kHz. The enhanced anti-cancer activity of LyMs-NR was caused by the transfer of released 5-FU molecules from bulk to the interior of the cell via temporary pores formed on the surface of cancer cells, i.e., sonoporation. Thus, LyMs-NR demonstrated here has a high potential for use as carriers in the field of drug delivery, bio-imaging and therapy.

Controlled decoration of nanoceria on the surface of MoS2nanoflowers to improve the biodegradability and biocompatibility inDrosophila melanogastermodel.

In recent years, nanozymes based on two-dimensional (2D) nanomaterials have been receiving great interest for cancer photothermal therapy. 2D materials decorated with nanoparticles (NPs) on their surface are advantageous over conventional NPs and 2D material based systems because of their ability to synergistically improve the unique properties of both NPs and 2D materials. In this work, we report a nanozyme based on flower-like MoS2nanoflakes (NFs) by decorating their flower petals with NCeO2using polyethylenimine (PEI) as a linker molecule. A detailed investigation on toxicity, biocompatibility and degradation behavior of fabricated nanozymes in wild-typeDrosophila melanogastermodel revealed that there were no significant effects on the larval size, morphology, larval length, breadth and no time delay in changing larvae to the third instar stage at 7-10 d for MoS2NFs before and after NCeO2decoration. The muscle contraction and locomotion behavior of third instar larvae exhibited high distance coverage for NCeO2decorated MoS2NFs when compared to bare MoS2NFs and control groups. Notably, the MoS2and NCeO2-PEI-MoS2NFs treated groups at 100µg ml-1covered a distance of 38.2 mm (19.4% increase when compared with control) and 49.88 mm (no change when compared with control), respectively. High-resolution transmission electron microscopy investigations on the new born fly gut showed that the NCeO2decoration improved the degradation rate of MoS2NFs. Hence, nanozymes reported here have huge potential in various fields ranging from biosensing, cancer therapy and theranostics to tissue engineering and the treatment of Alzheimer's disease and retinal therapy.

Synthesis and characterization of MXene (Ti3C2Tx)/Iron oxide composite for ultrasensitive electrochemical detection of hydrogen peroxide.

Due to the widespread usage of hydrogen peroxide (H2O2) in various consumer and industrial products (Examples: fuel cells and antibacterial agents), it became important to accurately detect H2O2 concentration in environmental, medical and food samples. Herein, titanium carbide Ti3C2Tx (MXene) was synthesized by using Ti, Al and C powders at high-temperature. Then, nanocrystalline iron oxide (α-Fe2O3) was obtained from a single solid-phase method. Using Ti3C2Tx and Fe2O3 powders, Ti3C2Tx-Fe2O3 nanocomposite was prepared by ultrasonication. As-synthesized, Ti3C2Tx-Fe2O3 composite had been characterized by UV-Visible (UV-Vis), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and Raman spectroscopy. The Fe2O3 nanoparticles (NPs) were decorated on the surface of Ti3C2Tx as observed by high resolution scanning electron microscopy (HR-SEM) and high resolution transmission electron microscopy (HR-TEM). The Ti3C2Tx nanosheets were formed with the average size of 400-500 nm. HR-SEM images of α-Fe2O3 showed that the coral-like particles with the average length ~5 µm were obtained. The electrochemical properties of the individual (Ti3C2Tx and α-Fe2O3) and composite materials (Ti3C2Tx-Fe2O3) were investigated by cyclic voltammetry (CV). Ti3C2Tx-Fe2O3 nanocomposite modified electrode had exhibited potent electro-catalytic activity for H2O2 reduction by reducing the overpotential about 320 mV and a linear response was recorded from 10 nM to 1 µM H2O2. The optimization of various parameters such as material composition ratio, amount of catalyst, effects of pH, scan rate and interference effects with other biomolecules were carried out. In addition, the kinetic parameters such as rate constant, diffusion coefficient and the active surface area of the electrodes were calculated. Moreover, the Ti3C2Tx-Fe2O3 composite modified electrode was used successfully to detect H2O2 in food and urine samples. We believe that Ti3C2Tx-Fe2O3 composite based materials could be used for the fabrication of non-enzymatic H2O2 sensors for medical diagnosis, food safety and environmental monitoring applications.

Multilayer capsules encapsulating nimbin and doxorubicin for cancer chemo-photothermal therapy.

Layer-by-layer (LbL) assembled poly(allylamine hydrochloride) (PAH) and poly(methacrylic acid) (PMA) microcapsules were designed to incorporate gold nanorods (NRs) and co-encapsulate and release two drugs for cancer therapy. Calcium carbonate (CaCO3) microparticles modified with preformed NRs were used as sacrificial templates for the fabrication of hollow PAH/PMA/NR capsules incorporated with NRs. The hollow capsules were found to be 4.5 ± 0.5 µm in size and appeared with uniformly distributed NRs in the interior of the capsules. The morphology of the capsules transformed from pore free continuous structure to porous structure under laser light irradiation at 808 nm and 0.5 W cm-2. The encapsulation experiments showed that the hydrophilic drug (doxorubicin hydrochloride, Dox) was encapsulated in the interior of the capsules while the hydrophobic drug (nimbin, NB) was entrapped in the porous polymeric network of the layer components. The encapsulation efficiency was found to be 30% for both Dox and NB. The release experiments showed an initial burst release followed by sustained release up to 3 h. Notably, the release was completed within 30 min under NIR irradiation at 808 nm. The estimated IC50 values against THP-1 cells were 75 and 1.8 µM for NB and Dox, respectively. The dual drug loaded capsules showed excellent anticancer activity against THP-1 cells under NIR light exposure in in-vitro experiments. Thus, such remotely addressable dual-drug loaded capsules with the provision for encapsulation of natural drugs demonstrate high potential for use as theranostics in cancer therapy.

Hydrogen-Bonded Multilayer Thin Films and Capsules Based on Poly(2-n-propyl-2-oxazoline) and Tannic Acid: Investigation on Intermolecular Forces, Stability, and Permeability.

In recent years, hydrogen-bonded multilayer thin films and capsules based on neutral and nontoxic building blocks have been receiving interest for the design of stimuli-responsive drug delivery systems and for the preparation of thin-film coatings. Capsule systems made of tannic acid (TA), a natural polyphenol, as a hydrogen bonding donor and poly(2-n-propyl-2-oxazoline) (PnPropOx), a polymer with lower critical solution temperature around 25 °C, as a hydrogen bonding acceptor are advantageous over other conventional hydrogen-bonded systems because of their high stability in physiological pH range, biocompatibility, good renal clearance, stealth behavior, and stimuli responsiveness for temperature and pH. In this work, investigations on the interactive forces in TA/PnPropOx capsule formation, film thickness, stability, and permeability are reported. The multilayer thin films were assembled on quartz substrates, and the layer-by-layer film growth was investigated by UV-vis spectroscopy, atomic force microscopy, and profilometry. Hollow capsules were fabricated by sequential coating of TA and PnPropOx onto CaCO3 colloidal particles, followed by template dissolution with a 0.2 M ethylenediaminetetraacetic acid solution. The obtained capsules and multilayer thin films were found to be stable over a wide pH range of 2-9. It is found that both hydrogen bonding and hydrophobic interactions are responsible for the enhanced stability of the capsules at higher pH range. Swelling followed by dissolution of the capsules was observed at a pH value lower than 2, while the capsules undergo shrinking at a pH value higher than 8 and finally transform into a particle-like morphology before dissolution. The TA/PnPropOx capsules reported here could be used as a temperature-responsive drug delivery system in controlled drug delivery applications.

Reusable Hollow Polymer Microreactors Incorporated with Anisotropic Nanoparticles for Catalysis Application.

We report a methodology to encapsulate gold nanorods (AuNRs) and gold bipyramids (AuBPs) into polyelectrolyte capsules for catalytic application. Microreactors (capsules with encapsulated NRs or BPs) were fabricated by sequential deposition of poly(allylamine hydrochloride) and dextran sulfate on modified sacrificial template, followed by core dissolution. AuNRs and AuBPs of size 25-30 nm were successfully encapsulated in the fabricated polyelectrolyte capsules and were stable and distributed uniformly in the interior. Fabricated microreactors were investigated as catalysts for the reduction of p-nitrophenol to p-aminophenol in the presence of sodium borohydride in aqueous phase. Reaction parameters such as order, conversion, and rate constants were estimated for microreactors and compared to free anisotropic nanoparticles in suspension. The reaction rate was higher for NRs in both free and capsule forms compared to BPs. Microreactors demonstrated excellent catalytic activity even after three times of use. Such capsules have high potential for use as microreactors in applications such as catalysis, drug delivery, imaging, and cancer chemo-photothermal therapy.

Nanoceria decorated flower-like molybdenum sulphide nanoflakes: an efficient nanozyme for tumour selective ROS generation and photo thermal therapy.

We report nanoceria (NCeO2) decorated flower-like MoS2 nanoflakes as a nanozyme for cancer photothermal therapy (PTT). They exhibited enzyme-like activity for selectively killing tumor cells by ROS induction. The NCeO2 decoration significantly improved the photoconversion efficiencies (PCEs) of MoS2 nanoflakes when a NCeO2 concentration of ≤0.5 mg mL-1 was used for synthesis. The novel material demonstrated here showed high photostability and PCE, without any systemic toxicity for cancer PTT.

Two-dimensional cancer theranostic nanomaterials: Synthesis, surface functionalization and applications in photothermal therapy.

In recent years, novel two-dimensional (2D) nanomaterials are of great interest for diverse potential applications such as device fabrication, energy storage, sensing and theranostics because of their superlative physical features namely, large surface area, minimal thickness, tunable composition and easier surface modification methods. Rapid exploration in design and fabrication of 2D nano-structures have opened new avenue for cancer theranostics as it can encapsulate group of cancer cells and inflict major damage with great specificity in a non-invasive manner. Among the reported 2D materials such as graphene and its derivatives, metallic compounds, transition metal dichalcogenides (TMDC), black phosphorous and MXenes (e.g., carbides, nitrides, or carbonitrides), 2D nanomaterials based on graphene and TMDCs have gathered most of the limelight in this field due to their easily tunable properties. In this review, we summarize recent progress in the design of 2D theranostic nanomaterials, functionalization methods and their applications in photothermal therapy (PTT) as well as synergistic cancer therapy. We have also addressed the different modes of cellular entry of 2D nanomaterials into tumor cells based on their unique structural properties and investigated different methodologies to enhance PTT effect by optimizing the physico-chemical properties of the 2D sheets. Recent progress on in vitro and in vivo short and long term biocompatibility, immunotoxicity and excretion of the decorated structure is also highlighted. Investigation of the interaction of 2D nanomaterial with hematological factors such as RBC and WBC is of paramount importance as they are key indicators in in vivo responses, and this investigation will give a better solution for overcoming direct inflammation and infection related issues of the animal system. Besides, investigations on addressing the ways to incorporate polymer linkers and drug conjugates on to the surface of 2D materials, multiplexing capability, and the influence of surface functionalization on PTT effect is vital for future developments in clinical level diagnosis and cancer therapy. Finally, we conclude our opinion on current challenges and future prospective on meeting the various demands of advanced cancer imaging and therapies.

Polyelectrolyte capsules preloaded with interconnected alginate matrix: An effective capsule system for encapsulation and release of macromolecules.

In recent years, the design of stimuli-responsive hollow polymeric capsules is of tremendous interest for the scientific community because of the broad application of these capsules in the biomedical field. The use of weak polyelectrolytes as layer components for capsule fabrication is especially interesting as it results in hollow capsules that show unique release characteristics under physiological conditions. In this work, a methodology to prepare sub-micron sized alginate doped calcium carbonate (CaCO3) particles through controlled precipitation in the presence of alginate is reported. Hollow capsules obtained by Layer-by-Layer (LbL) assembly of poly(allylamine hydrochloride) (PAH) and poly(methacrylic acid) (PMA) are showing an interconnected alginate matrix in the interior of the capsules. Investigations showed that the presence of alginate matrix enhances the encapsulation of cationic molecules (e.g. doxorubicin hydrochloride) manifold by charge controlled attraction mechanism. Capsule permeability investigated by confocal laser scanning microscopy revealed that the transformation from an open state to closed state is accompanied by an intermediate state where capsules are neither open nor closed. Furthermore, time dependent study indicated that the encapsulation process is linear as a function of time. The cell viability experiments demonstrated excellent biocompatibility of hollow capsules with mouse embryonic fibroblast cells. Anticancer investigations showed that DOX loaded capsules have significant anti-proliferative characteristics against HeLa cells. Such capsules have high potential for use as drug carrier for cationic drugs in cancer therapy.