Receptor mediated targeting of EGF-conjugated alginate-PAMAM nanoparticles to lung adenocarcinoma: 2D/3D in vitro and in vivo evaluation.

Carboplatin (cis-diamine (1,1-cyclobutandicarboxylaso)­platinum (II)) is a second-generation antineoplastic drug, which is widely used for chemotherapy of lung, colon, breast, cervix, testicular and digestive system cancers. Although preferred over cisplatin due to the lower incidence of nephrotoxicity and ototoxicity, efficient carboplatin delivery remains as a major challenge. In this study, carboplatin loaded alginate- poly(amidoamine) (PAMAM) hybrid nanoparticles (CAPs) with mean sizes of 192.13 ± 4.15 nm were synthesized using a microfluidic platform, then EGF was conjugated to the surface of CAPs (EGF-CAPs) for the receptor-targeted delivery. Hence, increased FITC+ cell counts were observed in A549 spheroids after EGF-CAP treatment compared to CAP in the 3D cellular uptake study. As such, the cytotoxicity of EGF-CAP was approximately 2-fold higher with an IC50 value of 35.89 ± 10.37 µg/mL compared to the CAPs in A549 spheroids. Based on in vivo experimental animal model, anti-tumor activities of the group treated with CAP decreased by 61 %, whereas the group treated with EGF-CAP completely recovered. Additionally, EGF-CAP application was shown to induce apoptotic cell death. Our study provided a new strategy for designing a hybrid nanoparticle for EGFR targeted carboplatin delivery with improved efficacy both in vitro and in vivo applications.

Continuous Microfluidic Production of Citrem-Phosphatidylcholine Nano-Self-Assemblies for Thymoquinone Delivery.

Lamellar and non-lamellar liquid crystalline nanodispersions, including liposomes, cubosomes, and hexosomes are attractive platforms for drug delivery, bio-imaging, and related pharmaceutical applications. As compared to liposomes, there is a modest number of reports on the continuous production of cubosomes and hexosomes. Using a binary lipid mixture of citrem and soy phosphatidylcholine (SPC), we describe the continuous production of nanocarriers for delivering thymoquinone (TQ, a substance with various therapeutic potentials) by employing a commercial microfluidic hydrodynamic flow-focusing chip. In this study, nanoparticle tracking analysis (NTA) and synchrotron small-angle X-ray scattering (SAXS) were employed to characterize TQ-free and TQ-loaded citrem/SPC nanodispersions. Microfluidic synthesis led to formation of TQ-free and TQ-loaded nanoparticles with mean sizes around 115 and 124 nm, and NTA findings indicated comparable nanoparticle size distributions in these nanodispersions. Despite the attractiveness of the microfluidic chip for continuous production of citrem/SPC nano-self-assemblies, it was not efficient as comparable mean nanoparticle sizes were obtained on employing a batch (discontinuous) method based on low-energy emulsification method. SAXS results indicated the formation of a biphasic feature of swollen lamellar (Lα) phase in coexistence with an inverse bicontinuous cubic Pn3m phase in all continuously produced TQ-free and TQ-loaded nanodispersions. Further, a set of SAXS experiments were conducted on samples prepared using the batch method for gaining further insight into the effects of ethanol and TQ concentration on the structural features of citrem/SPC nano-self-assemblies. We discuss these effects and comment on the need to introduce efficient microfluidic platforms for producing nanocarriers for delivering TQ and other therapeutic agents.

Synthesis of Resveratrol Loaded Hybrid Silica-PAMAM Dendrimer Nanoparticles With Emphases on Inducible Nitric Oxide Synthase and Cytotoxicity.

Resveratrol is a naturally occurring polyphenolic compound exhibiting therapeutic activities. However, the stability can be altered by UV light, pH and changes in temperature. Encapsulation would be an ideal strategy to improve the stability and bioavailability. Thus, trans-resveratrol (Res) was encapsulated within hybrid nanoparticles consisted with silica and G4 polyamidoamine dendrimer (PAMAM) by sol-gel method. The diameters of synthesized nanoparticles (NPs) were at a range of 212-574 nm and the encapsulation efficiency was 86 %. RAW 264.7 murine macrophage cell line induced with endotoxin/lipopolysaccharide was treated with free resveratrol and Res-loaded NPs for assessing inhibition of inducible nitric oxide synthase (iNOS), where IC50 values of free resveratrol and Res-loaded NPs were 122.68 µM and 249.74 µM. As for cytotoxicity, IC50 values of free resveratrol were found as 176.57 µM and 201.54 µM for MCF-7 and MDA-MB-231 cells, whereas 197.16 µM and 219.07 µM for Res-loaded NPs for the respective cell lines. Overall, sol-gel technique proved to be an ideal technology as can be carried out under mild conditions and Res-loaded NPs have potential to be utilized in the industry.

Advances in microfluidic synthesis and coupling with synchrotron SAXS for continuous production and real-time structural characterization of nano-self-assemblies.

Microfluidic platforms have become highly attractive tools for synthesis of nanoparticles, including lipid nano-self-assemblies, owing to unique features and at least three important aspects inherent to miniaturized micro-devices. Firstly, the fluids flow under controlled conditions in the microchannels, providing well-defined flow profiles and shorter diffusion lengths that play important roles in enhancing the continuous production of lipid and polymer nanoparticles with relatively narrow size distributions. Secondly, various geometries adapted to microfluidic device designs can be utilized for enhancing the colloidal stability of nanoparticles and improving their drug loading. Thirdly, microfluidic devices are usually compatible with in situ characterization methods for real-time monitoring of processes occurring inside the microchannels. This is unlike conventional nanoparticle synthesis methods, where a final solution or withdrawn aliquots are separately analysed. These features inherent to microfluidic devices provide a tool-set allowing not only precise nanoparticle size control, but also real-time analyses for process optimization. In this review, we focus on recent advances and developments in the use of microfluidic devices for synthesis of lipid nanoparticles. We present different designs based on hydrodynamic flow focusing, droplet-based methods and controlled microvortices, and discuss integration of microfluidic platforms with synchrotron small-angle X ray scattering (SAXS) for in situ structural characterization of lipid nano-self-assemblies under continuous flow conditions, along with major challenges and future directions in this research area.

An alginate-poly(acrylamide) hydrogel with TGF-ß3 loaded nanoparticles for cartilage repair: Biodegradability, biocompatibility and protein adsorption.

Current implantable materials are limited in terms of function as native tissue, and there is still no effective clinical treatment to restore articular impairments. Hereby, a functionalized polyacrylamide (PAAm)-alginate (Alg) Double Network (DN) hydrogel acting as an articular-like tissue is developed. These hydrogels sustain their mechanical stability under different temperature (+4 °C, 25 °C, 40 °C) and humidity conditions (60% and 75%) over 3 months. As for the functionalization, transforming growth factor beta-3 (TGF-ß3) encapsulated (NPTGF-ß3) and empty poly(lactide-co-glycolide) (PLGA) nanoparticles (PLGA NPs) are synthesized by using microfluidic platform, wherein the mean particle sizes are determined as 81.44 ± 9.2 nm and 126 ± 4.52 nm with very low polydispersity indexes (PDI) of 0.194 and 0.137, respectively. Functionalization process of PAAm-Alg hydrogels with ester-end PLGA NPs is confirmed by FTIR analysis, and higher viscoelasticity is obtained for functionalized hydrogels. Moreover, cartilage regeneration capability of these hydrogels is evaluated with in vitro and in vivo experiments. Compared with the PAAm-Alg hydrogels, functionalized formulations exhibit a better cell viability. Histological staining, and score distribution confirmed that proposed hydrogels significantly enhance regeneration of cartilage in rats due to stable hydrogel matrix and controlled release of TGF-ß3. These findings demonstrated that PAAm-Alg hydrogels showed potential for cartilage repair and clinical application.

One-Step Microfluidic Coating of Phospholipid Microbubbles with Natural Alginate Polymer as a Delivery System for Human Epithelial Lung Adenocarcinoma.

In this study, the neoplastic drug frequently used in the treatment of lung cancer, carboplatin is loaded to microbubbles via a microfluidic platform. In order to increase the drug loading capacity of microbubbles, carboplatin is encapsulated into alginate polymer layer. The phospholipid microbubbles (MBs) are synthesized by MicroSphere Creator, which is connected with T-junction and micromixer for the treatment with CaCl2 solution to provide gelation of the alginate coated phospholipid microbubbles (AMBs). The carboplatin loaded alginate coated phospholipid microbubbles (CAMBs) result in 12.2 ± 0.21 µm mean size, obtained by mixing with 0.05% CaCl2 using T-junction. The cytotoxic activities of the synthesized MBs, AMBs, and CAMBs are also investigated with the 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) (MTT) and live/dead fluorescent dying assays in the A549 and BEAS-2B cell lines. The one-step microfluidic coating of lipid microbubbles with natural alginate polymer appears to be a promising strategy for enhanced drug reservoir properties.

Nano-vesicular formulation of propolis and cytotoxic effects in a 3D spheroid model of lung cancer.

BACKGROUND: Propolis exhibits therapeutic properties due to the presence of phenolic acids, esters, and flavonoids. The scope of this study was to develop a nano-vesicular formulation and establish a three-dimensional (3D) spheroid model in which lung cancer is recapitulated. RESULTS: Niosome vesicles doped with galangin-rich propolis extract were synthesized by the ether injection method using a cholesterol : surfactant mass ratio of 1 : 3 at 40 °C for 1 h. Formulated niosomes were administered to 3D lung cancer spheroid model and the cytotoxicity was compared with that of a two-dimensional (2D) setting. The galangin content was determined as 86 µg mg-1 propolis extract by ultra-performance liquid chromatography (UPLC). The particle size of loaded niosome was 151 ± 2.84 nm with a polydispersity index (PDI) of about 0.232, and an encapsulation efficiency of 70% was achieved. CONCLUSION: The decrease in cell viability and the scattering in the 3D spheroids of A549 lung cancer cells treated with propolis-loaded niosomes were notable, indicating a profound cytotoxic effect and suggesting that they can be utilized as an effective nano-vesicle. © 2020 Society of Chemical Industry.

Quantitative determination of H2O2 for detection of alanine aminotransferase using thin film electrodes.

The abnormal concentrations or absence of biomolecules (e.g., proteins) in blood can further be used in diagnosis of a particular pathology at an early stage. Current studies are intensely focusing on the analysis of interaction and detection of biomolecules via point-of-care systems (POCs), allowing miniaturized and parallelized reactions, simultaneously. Recent developments have shown that the collaboration of electrochemical sensing techniques and POCs to overcome challenging problems in health-care settings provides new approaches in diagnosis and treatment of diseases. The aim of this study was to adapt the alanine aminotransferase (ALT) enzyme to the platinum (Pt) thin film electrode system and quantitatively determine the enzyme levels via enzymatically generated H2O2 with differential pulse voltammetry (DPV). A simple potentiostat architecture with expanded sweep range utilizing dual LMP91000 devices was developed and adapted to the needs of the biosensor. In order to calibrate the system, known concentrations of H2O2 were also tested. Moreover, signals associated with the other electroactive species coming from the ALT reaction were eliminated. Resulted potential range has been achieved between +500 mV and +900 mV and the linear range was found to be 0.05 M-0.5 M for H2O2, whereas 5 UL-1 to 120 UL-1 for ALT enzyme.

Effects of hydrostatic pressure and supercritical carbon dioxide on the viability of Botryococcus braunii algae cells.

In bio-based industries, Botryococcus braunii is identified as a potential resource for production of hydrocarbons having a wide range of applications in chemical and biopolymer industries. For a sustainable production platform, the algae cultivation should be integrated with downstream processes. Ideally the algae are not harvested, but the product is isolated while cultivation and growth is continued especially if the doubling time is slow. Consequently, hydrocarbons can be extracted while keeping the algae viable. In this study, the effects of pressure on the viability of B. braunii cells were tested hydrostatically and under supercritical CO2 conditions. Viability was determined by light microscopy, methylene blue uptake and by re-cultivation of the algae after treatments to follow the growth. It was concluded that supercritical CO2 was lethal to the algae, whereas hydrostatic pressure treatments up to 150 bar have not affected cell viability and recultivation was successful.

Silica-based organic-inorganic hybrid nanoparticles and nanoconjugates for improved anticancer drug delivery.

After the introduction of first generation MSNs for drug delivery with some challenges such as large particle sizes, irregular morphologies and aggregations, second generation provided uniform spherical morphologies, tunable pore/particle sizes and compositions. Henceforth, organic-inorganic hybrid mesoporous silica nanosystems have grown rapidly and utilized for active and passive targeting of tumorigenic cells especially conjugated with organic polymers followed by third generation counterparts with improved functionalities for cancer therapy. The aim of this review article is to focus on the advancements in mesoporous silica based organic-inorganic hybrid nanoparticles developed as drug carriers targeting cancer cells. Brief introduction to the state-of-the-art in passive and active targeting methods is presented. Specifically, therapeutic, diagnostic and theranostic applications are discussed with emphases on triggered and ligand conjugated organic-inorganic hybrid mesoporous silica nanomaterials. Although mesoporous silica nanoparticles perform well in preclinical tests, clinical translation progresses slowly as appropriate doses needs to be evaluated for human use along with biocompatibility and efficiency depending on surface modifications.