The novel albumin–chitosan core–shell nanoparticles for gene delivery: preparation, optimization and cell uptake investigation

Natural polymers and proteins such as chitosan (CS) and albumin (Alb) have recently attracted much attention both in drug delivery and gene delivery. The underlying rationale is their unique properties such as biodegradability, biocompatibility and controlled release. This study aimed to prepare novel albumin–chitosan–DNA (Alb-CS-DNA) core–shell nanoparticles as a plasmid delivery system and find the best conditions for their preparation. Phase separation method and ionic interaction were used for preparation of Alb nanoparticles and Alb-CS-DNA core–shell nanoparticles, respectively. The effects of three important independent variables (1) CS/Alb mass ratio, (2) the ratios of moles of the amine groups of cationic polymers to those of the phosphate groups of DNA (N/P ratio), and (3) Alb concentration, on the nanoparticle size and loading efficiency of the plasmid were investigated and optimized through Box–Behnken design of response surface methodology (RSM). The optimum conditions were found to be CS/Alb mass ratio = 3, N/P ratio = 8.24 and Alb concentration = 0.1 mg/mL. The most critical factors for the size of nanoparticles and loading efficiency were Alb concentration and N/P ratio. The optimized nanoparticles had an average size of 176 ± 3.4 nm and loading efficiency of 80 ± 3.9 %. Cytotoxicity experiments demonstrated that the prepared nanoparticles were not toxic. The high cellular uptake of nanoparticles (~85 %) was shown by flow cytometry and fluorescent microscopy.Tarbiat Modares UniversityUnited States. National Institutes of Health (Grant R01A1050875

A textile platform using mechanically reinforced hydrogel fibres towards engineering tendon niche

INTRODUCTION: Tendon injuries can result from tendon overuse or trauma, resulting in substantial pain and disability. Given that natural or surgical repair of tendons lead to a poor outcome in terms of mechanical properties and functionality, there is a great need for tissue engineering strategies. Textile platforms enable the generation of biomimetic constructs [1]. Therefore, the main goal of this study is the development of cell-laden hybrid hydrogel fibers reinforced with a mechanically robust core fiber and their assembly into braided constructs towards replicating tendon mechanical properties and architecture. METHODS: To fabricate mechanically reinforced hydrogel fibres, a commercially available suture was coated using a cell-hydrogel mixture of methacryloyl gelatine (GelMA) and alginate. Composite fibres (CFs) were obtained by ionic crosslinking of alginate followed by photocrosslinking of GelMA. CFs were assembled using braiding technique and the mechanical properties of single fibres and braided constructs were evaluated. Different cells were encapsulated in the hydrogel layer, including MC-3T3, mesenchymal stem cells (MSCs) and human tendon-derived cells (TDCs). Cell viability and metabolic activity were evaluated by LIVE/DEAD staining and presto blue assay of metabolic activity. The expression of tendon-related markers and matrix deposition were also investigated. RESULTS: CFs were fabricated with a GelMA:alginate hydrogel layer and using multifilament twisted cotton or biodegradable suturing threads. The biocompatibility of this system was evaluated on encapsulated cells (Fig.1a). Cells (MC-3T3, MSCs and TDCs) were homogeneously distributed along the hydrogel layer, being viable up to 14 days in culture. In addition, TDCs were spreading inside the hydrogel after less than 48 h. Moreover, to further improve the mechanical properties of CFs, braided constructs were generated (Fig. 1b). Braiding CFs together enhanced their tensile strength and the process did not affect the viability of encapsulated cells.DISCUSSION & CONCLUSIONS: CFs were generated with a load bearing core and a hydrogel layer towards mimicking both mechanical properties and the matrix-rich microenvironment of tendon tissue. Accordingly, cell behaviour can be further modulated by modifying the hydrogel composition or, ultimately, through the addition of bioactive cues. Finally, braiding CFs together allows tuning the mechanical properties of developed constructs to match those of native tendon tissues.Fundação para a Ciência e a Tecnologia in the framework of FCT-POPH-FSE, the PhD grant SFRH/BD/96593/2013 of R.C-

Moving from a Product-Based Economy to a Service-Based Economy for a More Sustainable Future

Traditionally, economic growth and prosperity have been linked with the availability, production and distribution of tangible goods as well as the ability of consumers to acquire such goods. Early evidence regarding this connection dates back to Adam Smith's Wealth of Nations (1776), in which any activity not resulting in the production of a tangible good is characterized as unproductive of any value." Since then, this coupling of economic value and material production has been prevalent in both developed and developing economies throughout the world. One unintended consequence of this coupling has been the exponential increase in the amount of solid waste being generated. The reason is that any production and consumption of material goods eventually generates the equivalent amount of (or even more) waste. Exacerbating this problem is the fact that, with today's manufacturing and supply chain management technologies, it has become cheaper to dispose and replace most products rather than to repair and reuse them. This has given rise to what some call a disposable society." To put things in perspective: In 2012 households in the U.K. generated approximately 22 thousand tons of waste, which amounted to 411 kg of waste generated per person (Department for Environment, Food & Rural Affairs, 2015). During the same time period, households in the U.S. generated 251 million tons of waste, which is equivalent to a person generating approximately 2 kg of waste every day (U.S. Environmental Protection Agency, 2012). Out of these 251 million tons of total waste generated, approximately 20% of the discarded items were categorized as durable goods. The disposal of durable goods is particularly worrisome because they are typically produced using material from non- renewable resources such as iron, minerals, and petroleum-based raw materials

Selection of the best proper DC-SQUIDs in a multi-SQUID configuration

We have carried out experimental investigation of multi-DC-SQUID magnetometer configuration fabricated on YBa2Cu30 7-δ thin films onto 24 degree SrTiO3 bicrystal substrates by directly coupling the pick-up loop to DC-SQUIDs. The layout of the magnetometer pick-up loop was chosen as a square washer configuration by maximizing loop effective area and minimizing loop inductance. We have used De-Magnetron Sputtering technique for deposition of the films and chemical etching process for patterning the Josephson junctions having 4 μm widths. The use of multi-SQUID configuration is related to the selection of the best proper junctions for SQUID to improve the chip sensitivity with selectivity option of choosing the squid junctions rather than multichannel operation. Selection of the best junctions compared to each other depending on the junction critical currents and noise levels caused by the fabrication process and placements of the junctions on the grain boundary enable having an increased output signal of the DC-SQUID. © 2007 IEEE

Selection of aptamers against triple negative breast cancer cells using high throughput sequencing

Triple-negative breast cancer is the most aggressive subtype of invasive breast cancer with a poor prognosis and no approved targeted therapy. Hence, the identification of new and specific ligands is essential to develop novel targeted therapies. In this study, we aimed to identify new aptamers that bind to highly metastatic breast cancer MDA-MB-231 cells using the cell-SELEX technology aided by high throughput sequencing. After 8 cycles of selection, the aptamer pool was sequenced and the 25 most frequent sequences were aligned for homology within their variable core region, plotted according to their free energy and the key nucleotides possibly involved in the target binding site were analyzed. Two aptamer candidates, Apt1 and Apt2, binding specifically to the target cells with Kd values of 44.3 ± 13.3 nM and 17.7 ± 2.7 nM, respectively, were further validated. The binding analysis clearly showed their specificity to MDA-MB-231 cells and suggested the targeting of cell surface receptors. Additionally, Apt2 revealed no toxicity in vitro and showed potential translational application due to its affinity to breast cancer tissue sections. Overall, the results suggest that Apt2 is a promising candidate to be used in triple-negative breast cancer treatment and/or diagnosis. © 2021, The Author(s).Tis study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020—Programa Operacional Regional do Norte. Débora Ferreira (DF) is the recipient of a fellowship supported by a doctoral advanced training (call NORTE-69-2015-15) funded by the European Social Fund under the scope of Norte2020—Programa Operacional Regional do Norte. Joaquim Barbosa (JB) and Diana A. Sousa (DAS) acknowledge FCT for the Grants SFRH/BD/51109/2010 and PD/BD/139083/2018, respectively.info:eu-repo/semantics/publishedVersio