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 This is unlike conventional nanoparticle synthesis methods, where a final solution or withdrawn aliquots are separately analysed.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.INP grant from the Danish Agency for Science and Higher Education [8073-00022B]AY and OYC acknowledge the INP grant (Ref. no.: 8073-00022B) from the Danish Agency for Science and Higher Education for initiating and exploring new networking and collaboration opportunities between Danish and Turkish research institutions

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

Anticancer activities of bioactive peptides derived from rice husk both in free and encapsulated form in chitosan

Cereal grain-derived protein hydrolysates exert beneficial effects for human health, thus utilization of by-products with high protein content has drawn attention. In this study, hot water extraction of rice husk has been optimized to obtain protein hydrolysate with the highest anticancer activity. The optimum protein value was obtained as 2.40 g/L corresponding to 43.2 g protein/kg dried rice husk under the conditions of 60 degrees C, 2.0 mL/min flow rate and pH 10.0. The protein hydrolysate was encapsulated with chitosan, where the mean particle size of protein hydrolysate (0.3 %) loaded chitosan nanoparticles was 256.4 +/- 33. 4 nm with 89 % encapsulation efficiency and a 65% release at the end of 6 days. Cytotoxicity assays showed that the lowest cell viabilities have been achieved with A549 and MCF7 cells with IC50 values of 1.98 and 3.58 mu g/mL, respectively, whereas nuclear fragmentation and apoptotic bodies were observed through Hoechst 33342 staining. The cytotoxic effect might be associated with the wide variety of peptide/protein subunits ranging from 10 kDa to more than 180 kDa in the protein hydrolysate of rice husk. (C) 2021 The Korean Society of Industrial and Engineering Chemistry. Published by Elsevier B.V. All rights reserved.Kocaeli University [KOU-BAP-2016-081, KOU-BAP-2018-149]The research funds provided by Kocaeli University (KOU-BAP-2016-081 and KOU-BAP-2018-149) are highly appreciated. Special thanks to Mr. Ahmet Yilmaz who provided the rice husk from Ozserhat Gida Sanayi Tic. Ltd. Sti, Edirne, Turkey. We are grateful to Dilan Yerli for helping with SDS-PAGE, Prof. Dr. Kemal Sami Korkmaz for providing access to his lab at Ege University and Pelin Saglam-Metiner for her contributions to fluorescent staining assays

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

Yesil-Celiktas, Ozlem/0000-0003-4509-2212WOS: 000529144400001PubMed: 32346989In 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 mu m 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.Tide Microfluidics B.V. (The Netherlands)Netherlands Government; Erasmus+ Traineeship Mobility GrantThe authors gratefully acknowledge funding for all consumables and the apparatus needed for MBs provided from the Tide Microfluidics B.V. (The Netherlands) and Erasmus+ Traineeship Mobility Grant for EIA

An alginate-poly(acrylamide) hydrogel with TGF-beta 3 loaded nanoparticles for cartilage repair: Biodegradability, biocompatibility and protein adsorption

Saygili, Ecem/0000-0002-8389-9079; Ilhan Ayisigi, Esra/0000-0003-1880-4261WOS:000619184100039PubMed: 33476613Current 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 degrees C, 25 degrees C, 40 degrees C) and humidity conditions (60% and 75%) over 3 months. As for the functionalization, transforming growth factor beta-3 (TGF-beta 3) encapsulated (NPTGF-beta 3) and empty poly(lactide-co-glycolide) (PLGA) nanopartides (PLGA NPs) are synthesized by using microfluidic plat-form, wherein the mean particle sizes are determined as 81.94 +/- 92 nm and 126 +/- 4.52 nm with very low poly-dispersity indexes (PDI) of 0.194 and 0.137, respectively. Functionaliza lion process of PAAm-Alg hydrogels with ester-end PLGA NPs is confirmed by MR analysis, and higher viscoelastidty 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-beta 3. These findings demonstrated that PAAm-Alg hydrogels showed potential for cartilage repair and clinical application. (C) 2021 Elsevier B.V. All rights reserved.Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [117M843]; National Research Founda-tion of KoreaNational Research Foundation of Korea [2017K2A9A1A06037807]This work was supported by The Scientific and Technological Research Council of Turkey (TUBITAK) under grant number 117M843. Y.-W.KimandJ.-Y.Sun were supported under the framework of international cooperation program managed by the National Research Founda-tion of Korea (2017K2A9A1A06037807, FY2017)

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

Ulucan, Fulden/0000-0001-5567-0261; Yesil-Celiktas, Ozlem/0000-0003-4509-2212; saglam metiner, pelin/0000-0002-5726-1928WOS: 000526841800001PubMed: 32239766BACKGROUND 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 degrees 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 mu g 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. (c) 2020 Society of Chemical Industr

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