Human T-Cell Responses to Metallic Ion-Doped Bioactive Glasses.

Biomaterials are extensively used as replacements for damaged tissue with bioactive glasses standing out as bone substitutes for their intrinsic osteogenic properties. However, biomaterial implantation has the following risks: the development of implant-associated infections and adverse immune responses. Thus, incorporating metallic ions with known antimicrobial properties can prevent infection, but should also modulate the immune response. Therefore, we selected silver, copper and tellurium as doping for bioactive glasses and evaluated the immunophenotype and cytokine profile of human T-cells cultured on top of these discs. Results showed that silver significantly decreased cell viability, copper increased the T helper (Th)-1 cell percentage while decreasing that of Th17, while tellurium did not affect either cell viability or immune response, as evaluated via multiparametric flow cytometry. Multiplex cytokines assay showed that IL-5 levels were decreased in the copper-doped discs, compared with its undoped control, while IL-10 tended to be lower in the doped glass, compared with the control (plastic) while undoped condition showed lower expression of IL-13 and increased MCP-1 and MIP-1ß secretion. Overall, we hypothesized that the Th1/Th17 shift, and specific cytokine expression indicated that T-cells might cross-activate other cell types, potentially macrophages and eosinophils, in response to the scaffolds.

Design, Stereolithographic 3D Printing, and Characterization of TPMS Scaffolds.

Anatomical and functional tissue loss is one of the most debilitating problems and involves a great cost to the international health-care sector. In the field of bone tissue, the use of scaffolds to promote tissue regeneration is a topic of great interest. In this study, a combination of additive manufacturing and computational methods led to creating porous scaffolds with complex microstructure and mechanical behavior comparable to those of cancellous bone. Specifically, some representative models of triply periodic minimal surfaces (TPMSs) were 3D-printed through a stereolithographic technique using a dental resin. Schwarz primitive and gyroid surfaces were created computationally: they are characterized by a complex geometry and a high pore interconnectivity, which play a key role in the mechanism of cell proliferation. Several design parameters can be varied in these structures that can affect the performance of the scaffold: for example, the larger the wall thickness, the lower the elastic modulus and compressive strength. Morphological and mechanical analyses were performed to experimentally assess the properties of the scaffolds. The relationship between relative density and elastic modulus has been analyzed by applying different models, and a power-law equation was found suitable to describe the trend in both structures.

Micro-CT imaging and finite element models reveal how sintering temperature affects the microstructure and strength of bioactive glass-derived scaffolds.

This study focuses on the finite element simulation and micromechanical characterization of bioactive glass-ceramic scaffolds using Computed micro Tomography ([Formula: see text]CT) imaging. The main purpose of this work is to quantify the effect of sintering temperature on the morphometry and mechanical performance of the scaffolds. In particular, the scaffolds were produced using a novel bioactive glass material (47.5B) through foam replication, applying six different sintering temperatures. Through [Formula: see text]CT imaging, detailed three-dimensional images of the scaffold's internal structure are obtained, enabling the extraction of important geometric features and how these features change with sintering temperature. A finite element model is then developed based on the [Formula: see text]CT images to simulate the fracture process under uniaxial compression loading. The model incorporates scaffold heterogeneity and material properties-also depending on sintering temperature-to capture the mechanical response, including crack initiation, propagation, and failure. Scaffolds sintered at temperatures equal to or higher than 700 [Formula: see text]C exhibit two-scale porosity, with micro and macro pores. Finite element analyses revealed that the dual porosity significantly affects fracture mechanisms, as micro-pores attract cracks and weaken strength. Interestingly, scaffolds sintered at high temperatures, the overall strength of which is higher due to greater intrinsic strength, showed lower normalized strength compared to low-temperature scaffolds. By using a combined strategy of finite element simulation and [Formula: see text]CT-based characterization, bioactive glass-ceramic scaffolds can be optimized for bone tissue engineering applications by learning more about their micromechanical characteristics and fracture response.

Development of Super-Paramagnetic Iron Oxide Nanoparticle-Coated Melt Electrowritten Scaffolds for Biomedical Applications.

Polycaprolactone (PCL) is usually the material chosen for melt electrowriting (MEW) due to its biocompatibility, mechanical strength, and melt processability. This work first investigates the effect of different processing parameters to obtain optimum PCL-MEW scaffolds. Secondly, to increase PCL`s hydrophilicity and cell affinity, and to enable coating with superparamagnetic iron oxide nanoparticles (SPIONs) and silica-coated-SPIONs (Si-SPIONs), the scaffolds are modified with alkaline surface treatment. Finally, SPIONs and Si-SPIONs are successfully coated on MEW scaffolds. Results show that reproducible scaffolds are fabricated. Additionally, the alkaline treatment does not change the three-dimensional morphology of scaffolds while reducing fiber diameter. Furthermore, SEM images and ATR-FTIR results confirmed that SPIONs and Si-SPIONs-were coated on scaffolds. A cytocompatibility assay showed a non-toxic effect on MG-63 osteoblast-like cells in all scaffolds. Additionally, higher MC3T3-E1 pre-osteoblastic cell adhesion efficiency and proliferation are achieved for the alkaline-treated scaffolds and SPIONs/Si-SPIONs-coated scaffolds. All samples demonstrated the ability to generate heat, useful for hyperthermia-treatment, when subjected to an alternating magnetic field. Overall, the findings suggest that the strategy of coating PCL-MEW scaffolds with SPIONs/Si-SPIONs has the potential to improve scaffold performance for biomedical applications, especially for producing magnetically responsive MEW scaffolds.

3D bioprinting of multifunctional alginate dialdehyde (ADA)-gelatin (GEL) (ADA-GEL) hydrogels incorporating ferulic acid.

The present work explores the 3D extrusion printing of ferulic acid (FA)-containing alginate dialdehyde (ADA)-gelatin (GEL) scaffolds with a wide spectrum of biophysical and pharmacological properties. The tailored addition of FA (≤0.2 %) increases the crosslinking between FA and GEL in the presence of calcium chloride (CaCl2) and microbial transglutaminase, as confirmed using trinitrobenzenesulfonic acid (TNBS) assay. In agreement with an increase in crosslinking density, a higher viscosity of ADA-GEL with FA incorporation was achieved, leading to better printability. Importantly, FA release, enzymatic degradation and swelling were progressively reduced with an increase in FA loading to ADA-GEL, over 28 days. Similar positive impact on antibacterial properties with S. epidermidis strains as well as antioxidant properties were recorded. Intriguingly, FA incorporated ADA-GEL supported murine pre-osteoblast proliferation with reduced osteosarcoma cell proliferation over 7 days in culture, implicating potential anticancer property. Most importantly, FA-incorporated and cell-encapsulated ADA-GEL can be extrusion printed to shape fidelity-compliant multilayer scaffolds, which also support pre-osteoblast cells over 7 days in culture. Taken together, the present study has confirmed the significant potential of 3D bioprinting of ADA-GEL-FA ink to obtain structurally stable scaffolds with a broad spectrum of biophysical and therapeutically significant properties, for bone tissue engineering applications.

Computational models for the simulation of the elastic and fracture properties of highly porous 3D-printed hydroxyapatite scaffolds.

Bone scaffolding is a promising approach for the treatment of critical-size bone defects. Hydroxyapatite can be used to produce highly porous scaffolds as it mimics the mineralized part of bone tissue, but its intrinsic brittleness limits its usage. Among 3D printing techniques, vat photopolymerization allows for the best printing resolution for ceramic materials. In this study, we implemented a Computed micro-Tomography based Finite Element Model of a hydroxyapatite porous scaffold fabricated by vat photopolymerization. We used the model in order to predict the elastic and fracture properties of the scaffold. From the stress-strain diagram of a simulated compression test, we computed the stiffness and the strength of the scaffolds. We found that three morphometric features substantially affect the crack pattern. In particular, the crack propagation is not only dependent on the trabecular thickness but also depends on the slenderness and orientation of the trabeculae with respect to the load. The results found in this study can be used for the design of ceramic scaffolds with heterogeneous pore distribution in order to tailor and predict the compressive strength.

Tannic-acid-mediated synthesis and characterization of magnetite-gold nanoplatforms for photothermal therapy.

Aim: The design of new hybrid nanoplatforms (HNPs) through the innovative and eco-friendly use of tannic acid (TA) for the synthesis and stabilization of the nanoplatforms. Materials & methods: The size, morphology, composition and magnetic and plasmonic properties of HNPs were investigated together with their ability to generate heat under laser irradiation and the hemotoxicity to explore their potential use for biomedical applications. Results & conclusion: The use of TA allowed the synthesis of the HNPs by adopting a simple and green method. The HNPs preserved the peculiar properties of both magnetic and plasmonic nanoparticles and did not show any hemotoxic effect.

The aim of this research was to prepare new nanoparticles (called nanoplatforms) made from two parts: a magnetic core and the addition of gold particles. These particles can be used for cancer treatment because, when stimulated by light, they are able to release heat, which can kill cancer cells. In particular, in this work, we investigated the preparation of these particles using green methods, without the use of toxic reagents. The obtained nanoparticles were studied to investigate their size, shape, composition, magnetic properties, ability to generate heat and possible toxic effect toward blood cells. The results show that these particles can be produced with green methods, release heat and are not toxic.

UV-Cured Bio-Based Acrylated Soybean Oil Scaffold Reinforced with Bioactive Glasses.

In this study, a bio-based acrylate resin derived from soybean oil was used in combination with a reactive diluent, isobornyl acrylate, to synthetize a composite scaffold reinforced with bioactive glass particles. The formulation contained acrylated epoxidized soybean oil (AESO), isobornyl acrylate (IBOA), a photo-initiator (Irgacure 819) and a bioactive glass particle. The resin showed high reactivity towards radical photopolymerisation, and the presence of the bioactive glass did not significantly affect the photocuring process. The 3D-printed samples showed different properties from the mould-polymerised samples. The glass transition temperature Tg showed an increase of 3D samples with increasing bioactive glass content, attributed to the layer-by-layer curing process that resulted in improved interaction between the bioactive glass and the polymer matrix. Scanning electron microscope analysis revealed an optimal distribution on bioactive glass within the samples. Compression tests indicated that the 3D-printed sample exhibited higher modulus compared to mould-synthetized samples, proving the enhanced mechanical behaviour of 3D-printed scaffolds. The cytocompatibility and biocompatibility of the samples were evaluated using human bone marrow mesenchymal stem cells (bMSCs). The metabolic activity and attachment of cells on the samples' surfaces were analysed, and the results demonstrated higher metabolic activity and increased cell attachment on the surfaces containing higher bioactive glass content. The viability of the cells was further confirmed through live/dead staining and reseeding experiments. Overall, this study presents a novel approach for fabricating bioactive glass reinforced scaffolds using 3D printing technology, offering potential applications in tissue engineering.

Cobalt-Doped Bioactive Glasses for Biomedical Applications: A Review.

Improving angiogenesis is the key to the success of most regenerative medicine approaches. However, how and to which extent this may be performed is still a challenge. In this regard, cobalt (Co)-doped bioactive glasses show promise being able to combine the traditional bioactivity of these materials (especially bone-bonding and osteo-stimulatory properties) with the pro-angiogenic effect associated with the release of cobalt. Although the use and local delivery of Co2+ ions into the body have raised some concerns about the possible toxic effects on living cells and tissues, important biological improvements have been highlighted both in vitro and in vivo. This review aims at providing a comprehensive overview of Co-releasing glasses, which find biomedical applications as various products, including micro- and nanoparticles, composites in combination with biocompatible polymers, fibers and porous scaffolds. Therapeutic applications in the field of bone repair, wound healing and cancer treatment are discussed in the light of existing experimental evidence along with the open issues ahead.

Physical, Mechanical, and Biological Properties of PMMA-Based Composite Bone Cement Containing Silver-Doped Bioactive and Antibacterial Glass Particles with Different Particles Sizes.

In the present work, antibacterial composite bone cement was designed by introducing a bioactive and antibacterial glass into a commercial formulation. The effect of glass particles' addition on the curing parameters of the polymeric matrix was evaluated; moreover, the influence of the glass particle size on the glass dispersion, compressive and bending strength, bioactivity, and antibacterial effect was estimated. The results evidence a delay in the polymerization kinetics of the composite cement, which nevertheless complies with the requirements of the ISO standard. Morphological characterization provides evidence of good dispersion of the glass in the polymeric matrix and its exposition on the cement surface. The different glass grain sizes do not affect the composites' bioactivity and compressive strength, while a slight reduction in bending strength was observed for samples containing glass powders with greater dimensions. The size of the glass particles also appears to have an effect on the antibacterial properties, since the composites containing larger glass particles do not produce an inhibition halo towards the S. aureus strain. The obtained results demonstrate that, by carefully tailoring the glass amount and size, a multifunctional device for artificial joint fixing, temporary prostheses, or spinal surgery can be obtained.

Poly(ε-caprolactone)/bioactive glass composite electrospun fibers for tissue engineering applications.

In this work, composite electrospun fibers containing innovative bioactive glass nanoparticles were produced and characterized. Poly(ε-caprolactone), benign solvents, and sol-gel B- and Cu-doped bioactive glass powders were used to fabricate fibrous scaffolds. The retention of bioactive glass nanoparticles in the polymer matrix, the electrospinnability of this novel solution and the obtained electrospun composites were extensively characterized. As a result, composite electrospun fibers characterized by biocompatibility, bioactivity, and exhibiting overall properties adequate for both hard and soft tissue engineering applications, have been produced. The addition of these bioactive glass nanoparticles was, indeed, able to impart bioactive properties to the fibers. Cell culture studies show promising results, demonstrating proliferation and growth of cells on the composite fibers. Wettability, degradation rate, and mechanical performance were also tested and are in line with previous results.

Mechanical characterization of miniaturized 3D-printed hydroxyapatite parts obtained through vat photopolymerization: an experimental study.

Hydroxyapatite is one of the materials of choice for tissue engineering bone scaffolds manufacturing. Vat photopolymerization (VPP) is a promising Additive Manufacturing (AM) technology capable of producing scaffolds with high resolution micro-architecture and complex shapes. However, mechanical reliability of ceramic scaffolds can be achieved if a high fidelity printing process is obtained and if knowledge of the intrinsic mechanical properties of the constituent material is available. As the hydroxyapatite (HAP) obtained from VPP is subjected to a sintering process, the mechanical properties of the material should be assessed with specific reference to the process parameters (e.g. sintering temperature) and to the specific characteristic size of the microscopic features in the scaffolds. In order to tackle this challenge the HAP solid matrix of the scaffold was mimicked in the form of miniaturized samples suitable for ad hoc mechanical characterization, which is an unprecedented approach. To this purpose small scale HAP samples, having a simple geometry and size similar to that of the scaffolds, were produced through VPP. The samples were subjected to geometric characterization and to mechanical laboratory tests. Confocal laser scanning and Computed micro-Tomography (micro-CT) were used for geometric characterization; while, micro-bending and nanoindentation were used for mechanical testing. Micro-CT analyses have shown a highly dense material with negligible intrinsic micro-porosity. The imaging process allowed quantifying the variation of geometry with respect to the nominal size showing high accuracy of the printing process and identifying printing defects on one specific sample type, depending on the printing direction. The mechanical tests have shown that the VPP produces HAP with an elastic modulus as high as approximately 100GPa and flexural strength of approximately 100MPa. The results of this study have shown that vat photopolymerization is a promising technology capable of producing high quality HAP with reliable geometric fidelity.

Iron Oxide-Au Magneto-Plasmonic Heterostructures: Advances in Their Eco-Friendly Synthesis.

In recent years, nanotechnologies have attracted considerable interest, especially in the biomedical field. Among the most investigated particles, magnetic based on iron oxides and Au nanoparticles gained huge interest for their magnetic and plasmonic properties, respectively. These nanoparticles are usually produced starting from processes and reagents that can be the cause of potential human health and environmental concerns. For this reason, there is a need to develop simple, green, low-cost, and non-toxic synthesis methods and reagents. This review aims at providing an overview of the most recently developed processes to produce iron oxide magnetic nanoparticles, Au nanoparticles, and their magneto-plasmonic heterostructures using eco-friendly approaches, focusing the attention on the microorganisms and plant-assisted syntheses and showing the first results of the development of magneto-plasmonic heterostructures.

Mechanical Properties of Robocast Glass Scaffolds Assessed through Micro-CT-Based Finite Element Models.

In this study, the mechanical properties of two classes of robocast glass scaffolds are obtained through Computed micro-Tomography (micro-CT) based Finite Element Modeling (FEM) with the specific purpose to explicitly account for the geometrical defects introduced during manufacturing. Both classes demonstrate a fiber distribution along two perpendicular directions on parallel layers with a 90∘ tilting between two adjacent layers. The crack pattern identified upon compression loading is consistent with that found in experimental studies available in literature. The finite element models have demonstrated that the effect of imperfections on elastic and strength properties may be substantial, depending on the specific type of defect identified in the scaffolds. In particular, micro-porosity, fiber length interruption and fiber detaching were found as key factors. The micro-pores act as stress concentrators promoting fracture initiation and propagation, while fiber detachment reduces the scaffold properties substantially along the direction perpendicular to the fiber plane.

In Vivo Evaluation of 3D-Printed Silica-Based Bioactive Glass Scaffolds for Bone Regeneration.

Bioactive glasses are often designed as porous implantable templates in which newly-formed bone can grow in three dimensions (3D). This research work aims to investigate the bone regenerative capability of silicate bioactive glass scaffolds produced by robocasting in comparison with powder and granule-like materials (oxide system: 47.5SiO2-10Na2O-10K2O-10MgO-20CaO-2.5P2O5, mol.%). Morphological and compositional analyses performed by scanning electron microscopy (SEM), combined with energy dispersive spectroscopy (EDS) after the bioactivity studies in a simulated body fluid (SBF) confirmed the apatite-forming ability of the scaffolds, which is key to allowing bone-bonding in vivo. The scaffolds exhibited a clear osteogenic effect upon implantation in rabbit femur and underwent gradual resorption followed by ossification. Full resorption in favor of new bone growth was achieved within 6 months. Osseous defect healing was accompanied by the formation of mature bone with abundant osteocytes and bone marrow cells. These in vivo results support the scaffold's suitability for application in bone tissue engineering and show promise for potential translation to clinical assessment.

In situ reduction of Ag on magnetic nanoparticles with gallic acid: effect of the synthesis parameters on morphology.

Aim: Synthesis of Fe3O4-Ag composite nanoparticles (NPs) by a new in situ reduction of Ag NPs on the surface of Fe3O4 NPs using gallic acid as a reducing agent. Materials & methods: The influence of process parameters on NP morphology and functionalization was evaluated by means of field-emission scanning/scanning transmission electron microscopy and Fourier-transform IR spectroscopy. Results & conclusion: The synthesis conditions affected the morphology of the obtained NPs, evidence of the formation of polydispersed aggregates, nanoflower-like or nanodumbbell nanocomposites. In particular, well-defined nanodumbbells were obtained in aqueous media, with an NP/gallic acid ratio of 10:1, while the presence of a silica shell did not improve the morphology of Ag NPs nucleated on the Fe3O4 core.

Nanoparticles (NPs) have been extensively investigated in the biomedical field for their use in diagnosing and treating tumors. The aim of this work was to develop a simple and green method to produce NPs formed from iron oxides (which are magnetic) and silver. Magnetic NPs can be moved utilizing an external magnetic field, which can be used to localize NPs in a particular location, such as in tumors. Silver NPs have antibacterial properties and can be used to generate heat to kill tumor cells when exposed to light. This study showed that, by varying factors when synthesizing these NPs (e.g., amount of reagents, presence of coatings on the particles), it is possible to obtain multifunctional NPs with different shapes and sizes.

Surface Modification of Bioresorbable Phosphate Glasses for Controlled Protein Adsorption.

The traditional silicate bioactive glasses exhibit poor thermal processability, which inhibits fiber drawing or sintering into scaffolds. The composition of the silicate glasses has been modified to enable hot processing. However, the hot forming ability is generally at the expense of bioactivity. Metaphosphate glasses, on the other hand, possess excellent thermal processability, congruent dissolution, and a tailorable degradation rate. However, due to the layer-by-layer dissolution mechanism, cells do not attach to the material surface. Furthermore, the congruent dissolution leads to a low density of OH groups forming on the glass surface, limiting the adsorption of proteins. It is well regarded that the initial step of protein adsorption is critical as the cells interact with this protein layer, rather than the biomaterial itself. In this paper, we explore the possibility of improving protein adsorption on the surface of phosphate glasses through a variety of surface treatments, such as washing the glass surface in acidic (pH 5), neutral, and basic (pH 9) buffer solutions followed or not by a treatment with (3-aminopropyl)triethoxysilane (APTS). The impact of these surface treatments on the surface chemistry (contact angle, ζ-potential) and glass structure (FTIR) was assessed. In this manuscript, we demonstrate that understanding of the material surface chemistry enables to selectively improve the adsorption of albumin and fibronectin (used as model proteins). Furthermore, in this study, well-known silicate bioactive glasses (i.e., S53P4 and 13-93) were used as controls. While surface treatments clearly improved proteins adsorption on the surface of both silicate and phosphate glasses, it is of interest to note that protein adsorption on phosphate glasses was drastically improved to reach similar protein grafting ability to the silicate bioactive glasses. Overall, this study demonstrates that the limited cell/phosphate glass biological response can easily be overcome through deep understanding and control of the glass surface chemistry.

Biological Evaluation of a New Sodium-Potassium Silico-Phosphate Glass for Bone Regeneration: In Vitro and In Vivo Studies.

In vitro and in vivo studies are fundamental steps in the characterization of new implantable materials to preliminarily assess their biological response. The present study reports the in vitro and in vivo characterizations of a novel experimental silicate bioactive glass (BG) (47.5B, 47.5SiO2-10Na2O-10K2O-10MgO-20CaO-2.5P2O5 mol.%). Cytocompatibility tests were performed using human mature osteoblasts (U2OS), human mesenchymal stem cells (hMSCs) and human endothelial cells (EA.hy926). The release of the early osteogenic alkaline phosphatase (ALP) marker suggested strong pro-osteogenic properties, as the amount was comparable between hMSCs cultivated onto BG surface and cells cultivated onto polystyrene control. Similarly, real-time PCR revealed that the osteogenic collagen I gene was overexpressed in cells cultivated onto BG surface without biochemical induction. Acute toxicity tests for the determination of the median lethal dose (LD50) allowed classifying the analyzed material as a slightly toxic substance with LD50 = 4522 ± 248 mg/kg. A statistically significant difference in bone formation was observed in vivo through comparing the control (untreated) group and the experimental one, proving a clear osteogenic effect induced by the implantation at the defect site. Complete resorption of 47.5B powder was observed after only 3 months in favor of newly formed tissue, thus confirming the high osteostimulatory potential of 47.5B glass.

Foam Replica Method in the Manufacturing of Bioactive Glass Scaffolds: Out-of-Date Technology or Still Underexploited Potential?

Since 2006, the foam replica method has been commonly recognized as a valuable technology for the production of highly porous bioactive glass scaffolds showing three-dimensional, open-cell structures closely mimicking that of natural trabecular bone. Despite this, there are important drawbacks making the usage of foam-replicated glass scaffolds a difficult achievement in clinical practice; among these, certainly the high operator-dependency of the overall manufacturing process is one of the most crucial, limiting the scalability to industrial production and, thus, the spread of foam-replicated synthetic bone substitutes for effective use in routine management of bone defect. The present review opens a window on the versatile world of the foam replica technique, focusing the dissertation on scaffold properties analyzed in relation to various processing parameters, in order to better understand which are the real issues behind the bottleneck that still puts this technology on the Olympus of the most used techniques in laboratory practice, without moving, unfortunately, to a more concrete application. Specifically, scaffold morphology, mechanical and mass transport properties will be reviewed in detail, considering the various templates proposed till now by several research groups all over the world. In the end, a comprehensive overview of in vivo studies on bioactive glass foams will be provided, in order to put an emphasis on scaffold performances in a complex three-dimensional environment.

Antioxidant Activity of Silica-Based Bioactive Glasses.

Bioactive glasses are the materials of choice in the field of bone regeneration. Antioxidant properties of interest to limit inflammation and foreign body reactions have been conferred to bioactive glasses by the addition of appropriate ions (such as Ce or Sr). On the other hand, the antioxidant activity of bioactive glasses without specific ion/molecular doping has been occasionally cited in the literature but never investigated in depth. In the present study, three silica-based bioactive glasses have been developed and characterized for their surface properties (wettability, zeta potential, chemical composition, and reactivity) and radical scavenging activity in the presence/absence of cells. For the first time, the antioxidant activity of simple silica-based (SiO2-CaO-Na2O) bioactive glasses has been demonstrated.