Magnetic Surfactants and Polymers with Gadolinium Counterions for Protein Separations

New magnetic surfactants, (cationic hexadecyltrimethlyammonium bromotrichlorogadolinate (CTAG), decyltrimethylammonium bromotrichlorogadolinate (DTAG), and a magnetic polymer (poly­(3-acrylamidopropyl)­trimethylammonium tetrachlorogadolinate (APTAG)) have been synthesized by the simple mixing of the corresponding surfactants and polymer with gadolinium metal ions. A magnetic anionic surfactant, gadolinium tri­(1,4-bis­(2-ethylhexoxy)-1,4-dioxobutane-2-sulfonate) (Gd­(AOT)₃), was synthesized via metathesis. Both routes enable facile preparation of magnetically responsive magnetic polymers and surfactants without the need to rely on nanocomposites or organic frameworks with polyradicals. Electrical conductivity, surface tensiometry, SQUID magnetometry, and small-angle neutron scattering (SANS) demonstrate surface activity and self-aggregation behavior of the magnetic surfactants similar to their magnetically inert parent analogues but with added magnetic properties. The binding of the magnetic surfactants to proteins enables efficient separations under low-strength (0.33 T) magnetic fields in a new, nanoparticle-free approach to magnetophoretic protein separations and extractions. Importantly, the toxicity of the magnetic surfactants and polymers is, in some cases, lower than that of their halide analogues

Antiviral Properties of Polymeric Aziridine- and Biguanide-Modified Core–Shell Magnetic Nanoparticles

Polycationic superparamagnetic nanoparticles (∼150–250 nm) were evaluated as virucidal agents. The particles possess a core–shell structure, with cores consisting of magnetite clusters and shells of functional silica covalently bound to poly­(hexamethylene biguanide) (PHMBG), polyethyleneimine (PEI), or PEI terminated with aziridine moieties. Aziridine was conjugated to the PEI shell through cationic ring-opening polymerization. The nanometric core–shell particles functionalized with biguanide or aziridine moieties are able to bind and inactivate bacteriophage MS2, herpes simplex virus HSV-1, nonenveloped infectious pancreatic necrosis virus (IPNV), and enveloped viral hemorrhagic septicaemia virus (VHSV). The virus–particle complexes can be efficiently removed from the aqueous milieu by simple magnetocollection

Mobility of Water and Polymer Species and Rheological Properties of Supramolecular Polypseudorotaxane Gels Suitable for Bone Regeneration

The aim of this work was to prepare polypseudorotaxane-based supramolecular gels combining αCD with two temperature-responsive copolymers of different architecture (i.e., linear poloxamer P and X-shaped poloxamine T), at the lowest concentration as possible to form syringeable depots, and to shed light on the self-diffusion and spatial closeness of all components (including water) which in turn may determine the cooperative self-assembly phenomena and the performance of the gels at the macroscopic level. The exchange rate between bound water and bulk water was measured with a novel NMR experiment Water Diffusion Exchange-Diffusion Optimized Spectroscopy (WDE-DOSY). Polypseudorotaxane formation caused opposite effects on the mobility of αCD species (decreased) and internal water (increased) but did not affect PPO-water interaction. Consequently, designed ternary P/T/αCD supramolecular gels exhibited in situ gelling at body temperature could host large amounts of PLA/PLGA microspheres and behaved as porous 3D-scaffolds for mesenchymal stem cells (MSCs) supporting their osteogenic differentiation. Interestingly, the gels withstood freeze-drying and reconstitution with minor changes in inner structure and rheological properties. The gathered information may help to understand better the supramolecular gels and provide tools for the rational design of syringeable bone scaffolds that can simultaneously accommodate cells and drug microcarriers for efficient tissue regeneration

Superhydrophobic Chips for Cell Spheroids High-Throughput Generation and Drug Screening

We suggest the use of biomimetic superhydrophobic patterned chips produced by a benchtop methodology as low-cost and waste-free platforms for the production of arrays of cell spheroids/microtissues by the hanging drop methodology. Cell spheroids have a wide range of applications in biotechnology fields. For drug screening, they allow studying 3D models in structures resembling real living tissues/tumors. In tissue engineering, they are suggested as building blocks of bottom-up fabricated tissues. We used the wettability contrast of the chips to fix cell suspension droplets in the wettable regions and evaluated on-chip drug screening in 3D environment. Cell suspensions were patterned in the wettable spots by three distinct methods: (1) by pipetting the cell suspension directly in each individual spot, (2) by the continuous dragging of a cell suspension on the chip, and (3) by dipping the whole chip in a cell suspension. These methods allowed working with distinct throughputs and degrees of precision. The platforms were robust, and we were able to have static or dynamic environments in each droplet. The access to cell culture media for exchange or addition/removal of components was versatile and opened the possibility of using each spot of the chip as a mini-bioreactor. The platforms’ design allowed for samples visualization and high-content image-based analysis on-chip. The combinatorial analysis capability of this technology was validated by following the effect of doxorubicin at different concentrations on spheroids formed using L929 and SaOs-2 cells

Targeted Combinatorial Therapy Using Gold Nanostars as Theranostic Platforms

This paper reports the development of a multimodal therapy nanoplatform based on gold nanostars (Au NS) as core particles. These NS were functionalized with the chemotherapeutic drug doxorubicin (DOXO), which was conjugated to the NS surface by means of a cleavable heterobifunctional cross-linker (sulfo-LC-SPDP) to allow its release under the action of reducing enzymes. To ensure a specific delivery of the chemotherapeutic drug, the nanoplatform was additionally functionalized with folic acid (FA) as targeting ligand and cellular uptake adjuvant. By synthetically modifying the plasmon band of Au NS to the near-infrared (NIR) region of the electromagnetic spectrum, the present nanoplatform was able to simultaneously combine the capability of photothermal therapy (PTT) through the conversion of absorbed light energy into localized heat and chemotherapy, enabling their monitoring by means of optical fluorescence imaging thanks to DOXO’s autofluorescence. Cellular uptake was observed to be enhanced when the Au NPs were decorated with the targeting ligand. In addition, the therapeutic efficiency of the nanoplatform tested in HeLa cells demonstrated the larger cytotoxicity efficiency of the combined therapy if compared to individual ones

Materials with Fungi-Bioinspired Surface for Efficient Binding and Fungi-Sensitive Release of Antifungal Agents

Materials with fungi-bioinspired surface have been designed to host ergosterol-binding polyene antibiotics and to release them via a competitive mechanism only when fungi are present in the medium. Silicone rubber (SR) surfaces were endowed with selective loading and fungi-triggered release of polyene antifungal agents by means of a two-step functionalization that involved the grafting of glycidyl methacrylate (GMA) via a γ-ray preirradiation method (9–21.3% wt grafting) and the subsequent immobilization of ergosterol (3.9–116.8 mg/g) to the epoxy groups of polyGMA. The functionalized materials were characterized using FTIR and Raman spectroscopy, thermogravimetric analysis (TGA), and fluorescence, scanning electron microscopy (SEM), and atomic force microscopy (AFM) image analyses. Specific interactions between natamycin or nystatin and ergosterol endowed SR with ability to take up these polyene drugs, while immobilization of ergosterol did not modify the loading of antifungal drugs that did not interact in vivo with ergosterol (e.g., miconazole). In a buffer medium, polyene-loaded ergosterol-immobilized slabs efficiently retained the drug (<10% released at day 14), while in the presence of ergosterol-containing liposomes that mimic fungi membranes the release rate was 10-to-15-fold enhanced due to a competitive displacement of the drug from the ergosterol-immobilized slab to the ergosterol-containing liposomes. Release in the presence of cholesterol liposomes was slower due to a weaker interaction with polyene agents. The fungi-responsive release was demonstrated for both polyene drugs tested and for slabs prepared with a wide range of amounts of immobilized GMA and ergosterol, demonstrating the robustness of the approach. Nystatin-loaded functionalized slabs were challenged with Candida albicans and showed improved capability to inhibit biofilm formation compared to nystatin-soaked pristine SR, confirming the performance of the bioinspired materials

Spermidine Cross-Linked Hydrogels as a Controlled Release Biomimetic Approach for Cloxacillin

The intrinsic ability of albumin to bind active substances in the physiological fluids has been explored to endow hydrogels with improved capability to regulate drug release. To develop such biomimetic-functional hydrogels, it is critical that albumin conformation is not altered and that the protein remains retained inside the hydrogel keeping its conformational freedom, i.e., it should be not chemically cross-linked. Thus, the hydrogels were prepared with various proportions of albumin by physical cross-linking of anionic polysaccharides (gellan gum and chondroitin sulfate) with the cationic endogen polyamine spermidine under mild conditions in order to prevent albumin denaturation. Texture and swelling properties of hydrogels with various compositions were recorded, and the effect of the preparation variables was evaluated applying neurofuzzy logic tools for hydrogels prepared with and without albumin and associating the antibiotic cloxacillin. Developed hydrogel systems were extensively analyzed by means of nuclear magnetic resonance (NMR) to determine weak-to-medium and strong binding modes and the equilibrium constants of the albumin–cloxacillin association. NMR techniques were also employed to demonstrate the successful modulation of the cloxacillin release from the albumin-containing hydrogels. <i>In vitro</i> microbiological tests carried out with <i>Staphylococcus aureus</i> and <i>Staphylococcus epidermidis</i> confirmed the interest of the albumin-containing hydrogels as efficient platforms for cloxacillin release in its bioactive form

Antimicrobial Properties and Osteogenicity of Vancomycin-Loaded Synthetic Scaffolds Obtained by Supercritical Foaming

Advanced porous synthetic scaffolds are particularly suitable for regeneration of damaged tissues, but there is the risk of infections due to the colonization of microorganisms, forming biofilms. Supercritical foaming is an attractive processing method to prepare bone scaffolds, regulating simultaneously the porosity and loading of bioactive compounds without loss of activity. In this work, scaffolds made of poly-ε-caprolactone (50 kDa), containing chitosan and an antimicrobial agent (vancomycin), were processed by supercritical CO₂ foaming for bone regeneration purposes. The obtained scaffolds showed a suitable combination of morphological (porosity, pore size distribution, and interconnectivity), time-dependent in vitro vancomycin release behavior and biological properties (cell viability and proliferation, osteodifferentiation, and tissue-scaffold integration). The scaffolds sustained vancomycin release for more than 2 weeks. Finally, the antimicrobial activity of the scaffolds was tested against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria after 24 h of incubation with full growth inhibition for S. aureus