Stimuli-responsive materials have the potential to enable the generation of new bioinspired devices with unique physicochemical properties and cell-instructive ability. Enhancing biocompatibility while simplifying the production methodologies, as well as enabling the creation of complex constructs, i.e., via 3D (bio)printing technologies, remains key challenge in the field. Here, a novel method is presented to biofabricate cellularized anisotropic hybrid hydrogel through a mild and biocompatible process driven by multiple external stimuli: magnetic field, temperature, and light. A low-intensity magnetic field is used to align mosaic iron oxide nanoparticles (IOPs) into filaments with tunable size within a gelatin methacryloyl matrix. Cells seeded on top or embedded within the hydrogel align to the same axes of the IOPs filaments. Furthermore, in 3D, C2C12 skeletal myoblasts differentiate toward myotubes even in the absence of differentiation media. 3D printing of the nanocomposite hydrogel is achieved and creation of complex heterogeneous structures that respond to magnetic field is demonstrated. By combining the advanced, stimuli-responsive hydrogel with the architectural control provided by bioprinting technologies, 3D constructs can also be created that, although inspired by nature, express functionalities beyond those of native tissue, which have important application in soft robotics, bioactuators, and bionic devices.
Crosslinking and denaturation were two variables that deeply affected the performance of collagen-based scaffolds designed for tissue regeneration. If crosslinking enhances the mechanical properties and the enzymatic resistance of collagen, while masking or reducing the available cell binding sites, denaturation has very opposite effects, as it impairs the mechanical and the enzymatic stability of collagen, but increases the number of exposed cell adhesive domains. The quantification of both crosslinking and denaturation was thus fundamental to the design of collagen-based scaffolds for selected applications. The aim of this work was to investigate the extents of crosslinking and denaturation of collagen-based films upon dehydrothermal (DHT) treatment, that is, one of the most commonly employed methods for zero-length crosslinking that shows the unique ability to induce partial denaturation. Swelling measurements, differential scanning calorimetry, Fourier transform infrared spectroscopy, colorimetric assays for the quantification of primary amines, and mechanical tests were performed to analyze the effect of the DHT temperature on crosslinking and denaturation. In particular, chemically effective and elastically effective crosslink densities were evaluated. Both crosslinking and denaturation were found to increase with the DHT temperature, although according to different trends. The results also showed that DHT treatments performed at temperatures up to 120°C maintained the extent of denaturation under 25%. Coupling a mild DHT treatment with further crosslinking may thus be very useful not only to modulate the crosslink density, but also to induce a limited amount of denaturation, which shows potential to partially compensate the loss of cell binding sites caused by crosslinking.
A porous collagen-based hydrogel scaffold was prepared in the presence of iron oxide nanoparticles (NPs) and was characterized by means of infrared spectroscopy and scanning electron microscopy. The hybrid scaffold was then loaded with fluorescein sodium salt as a model compound. The release of the hydrosoluble species was triggered and accurately controlled by the application of an external magnetic field, as monitored by fluorescence spectroscopy. The biocompatibility of the proposed matrix was also tested by the MTT assay performed on 3T3 cells. Cell viability was only slightly reduced when the cells were incubated in the presence of the collagen-NP hydrogel, compared to controls. The economicity of the chemical protocol used to obtain the paramagnetic scaffolds as well as their biocompatibility and the safety of the external trigger needed to induce the drug release suggest the proposed collagen paramagnetic matrices for a number of applications including tissue engeneering and drug delivery.
In this work, tunable nonwoven mats based on poly(3-hydroxybutyrate) (PHB) and type I collagen (Coll) were successfully produced by electrospinning. The PHB/Coll weight ratio (fixed at 100/0, 70/30, and 50/50, resp.) was found to control the morphological, thermal, mechanical, and degradation properties of the mats. Increasing collagen amounts led to larger diameters of the fibers (in the approximate range 600–900 nm), while delaying their thermal decomposition (from 245°C to 262°C). Collagen also accelerated the hydrolytic degradation of the mats upon incubation in aqueous medium at 37°C for 23 days (with final weight losses of 1%, 15%, and 23% for 100/0, 70/30, and 50/50 samples, resp.), as a result of increased mat wettability and reduced PHB crystallinity. Interestingly, 70/30 meshes were the ones displaying the lowest stiffness (~116 MPa; p < 0.05 versus 100/0 and 50/50 meshes), while 50/50 samples had an elastic modulus comparable to that of 100/0 ones (~250 MPa), likely due to enhanced physical crosslinking of the collagen chains, at least at high protein amounts. All substrates were also found to allow for good viability and proliferation of murine fibroblasts, up to 6 days of culture. Collectively, the results evidenced the potential of as-spun PHB/Coll meshes for tissue engineering applications.
A supramolecular adduct formed by the interaction among octadecylamine (ODA) and zinc oxide nanostructures was promoted. A stable dispersion of the ZnO@ODA adduct was obtained and characterized by means of thermogravimetric analysis, infrared and Raman spectroscopy. Then, the supramolecular adduct was spread at the air/water interface of a Langmuir trough. The presence of octadecylamine gave amphiphilic features to the ZnO@ODA adduct that was transferred from the air/water interface to solid substrates by Langmuir-Schaefer (LS) method. The transferred film was characterized by tunnel electron microscopy that highlighted rectangular well-shaped structures assembled by nanostructure of ZnO arranged in an ODA matrix. Piezoelectric feature of large LS film (1 cm 2 ) was tested and a very promising response was observed as a consequence of the application of a pressure of 1 kPa.
Folic Acid decorated SPIONs selective internalization was monitored by an innovative Ellipsometry imaging approach.
This work demonstrates that lipid-detergent mixed micelles can be employed successfully in order to achieve and modulate the transfer of bio-active hydrophobic compounds into lipid carriers by means of a simple and bio-safe procedure. In our specific investigation, liposome preparations incorporating mixtures of natural carotenoids with high lycopene content were developed and characterized, aiming to obtain formulations of potential nutraceutical and pharmaceutical interest. The starting material was a solvent-free high-quality lycopene rich oleoresin (LRO) obtained by extracting a freeze-dried tomato matrix with supercritical carbon dioxide (SC-CO 2 ). Mixed micelles containing 1-palmitoyl-2-oleoyl-snglycero-3-phosphocholine (POPC) and cholate were loaded with LRO antioxidants by means of two slightly different procedures, which surprisingly resulted in significant differences in both quality and quantity of incorporated carotenoids. In particular, the selective incorporation of (all-E)-lycopene was achieved by extracting the oleoresin with a pre-formed cholate/POPC micelle suspension whilst (Z)-isomers were preferentially integrated when treating a POPC/LRO mixed film with cholate. The micelle to vesicle transition (MVT) method was employed in order to produce vesicles of well-defined lamellarity and size. Visible and infrared (IR) spectroscopy as well as Dynamic Light Scattering (DLS) andTransmission Electron Microscopy (TEM) measurements allowed the extensive characterization of LRO-loaded micelles and liposomes. The antioxidant potential of preparations was assessed by measuring the radical scavenging activity towards the coloured radical cation of 2,2 0 -azinobis(3-ethylbenzothiazoline-6-sulphonate) (ABTS). Important information about the reliability of different approaches for antioxidant capacity evaluation of micelle and liposome preparations was gained and the successful incorporation of LRO antioxidant power in a bio-deliverable water-dispersed form was demonstrated.
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