A comparative in vitro and in vivo analysis of the impact of copper substitution on the cytocompatibility, osteogenic, and angiogenic properties of a borosilicate bioactive glass.

The 0106-B1-bioactive glass (BG) composition (in wt %: 37.5 SiO2, 22.6 CaO, 5.9 Na2O, 4.0 P2O5, 12.0 K2O, 5.5 MgO, and 12.5 B2O3) has demonstrated favorable processing properties and promising bone regeneration potential. The present study aimed to evaluate the biological effects of the incorporation of highly pro-angiogenic copper (Cu) in 0106-B1-BG in vitro using human bone marrow-derived mesenchymal stromal cells (BMSCs) as well as its in vivo potential for bone regeneration. CuO was added to 0106-B1-BG in exchange for CaO, resulting in Cu-doped BG compositions containing 1.0, 2.5 and 5.0 wt % CuO (composition in wt %: 37.5 SiO2, 21.6/ 20.1/17.6 CaO, 5.9 Na2O, 4.0 P2O5, 12.0 K2O, 5.5 MgO, 12.5 B2O3, and 1.0/ 2.5/ 5.0 CuO). In vitro, the BGs' impact on the viability, proliferation, and growth patterns of BMSCs was evaluated. Analyses of protein secretion, matrix formation, and gene expression were used for the assessment of the BGs' influence on BMSCs regarding osteogenic differentiation and angiogenic stimulation. The presence of Cu improved cytocompatibility, osteogenic differentiation, and angiogenic response when compared with unmodified 0106-B1-BG in vitro. In vivo, a critical-size femoral defect in rats was filled with scaffolds made from BGs. Bone regeneration was evaluated by micro-computed tomography. Histological analysis was performed to assess bone maturation and angiogenesis. In vivo effects regarding defect closure, presence of osteoclastic cells or vascular structures in the defect were not significantly changed by the addition of Cu compared with undoped 0106-B1-BG scaffolds. Hence, while the in vitro properties of the 0106-B1-BG were significantly improved by the incorporation of Cu, further evaluation of the BG composition is necessary to transfer these effects to an in vivo setting.

Bio-response of copper-magnesium co-substituted mesoporous bioactive glass for bone tissue regeneration.

Mesoporous bioactive glass (MBG) is widely acknowledged in bone tissue engineering due to its mesoporous structure, large surface area, and bioactivity. Recent research indicates that introduction of metallic ions has beneficial impacts on bone metabolism and angiogenesis. Thus, the features of MBG can be modified by incorporating combinations of ions, such as magnesium (Mg) and copper (Cu), which can play a considerable role in bone formation, influencing angiogenesis, osteogenesis, as well as antibacterial properties. In this study, Mg and Cu were co-doped for the first time (in a ratio of 1 : 1) in 80SiO2-5P2O5-(15 - 2x)CaO-xMgO-xCuO glass composition with x = 0, 0.5, 1, and 2 mol%, synthesized using the sol-gel and evaporation-induced self-assembly method. X-ray diffraction analysis confirmed the amorphous nature of the powders, while inductively coupled plasma-optical emission spectrometry verified the existence of dopant ions in the respective amounts. The nitrogen sorption method indicated the formation of uniform cylindrical mesopores which are open at both ends and a high surface area of the powders. TEM images show fringes, indicating an ordered mesoporous structure in all MgCu co-doped systems. In vitro bioactivity was observed in all MBG powders, confirmed by the formation of an apatite phase when placed in simulated body fluid (SBF). Flake-like microstructure characteristics of HAp crystals found on the surface of MBG powders were visualized using FESEM. Cytotoxicity tests at lower concentrations (0.1 and 1 wt/vol%) of co-doped 2MC MBG (co-doping up to 2 mol%) showed cell proliferation and viability of osteoblast-like MG-63 cells and normal human dermal fibroblast (NHDF) cells similar to the basic glass 80S. Antibacterial study of MBG pellets showed an increment in the zone of inhibition with the sequential addition of doping ions. The turbidity measurement of bacterial cultures revealed that the optimal concentration for effectively inhibiting bacterial growth was 1 wt/vol% (i.e., 10 mg mL-1) concentration of MBG extracts. The result suggested that the incorporation of Mg and Cu ions in MBG in lower concentrations of up to 2 mol% can be useful in bone regeneration owing to bioactivity, cell proliferation, and antibacterial characteristics.

Influence of Copper-Strontium Co-Doping on Bioactivity, Cytotoxicity and Antibacterial Activity of Mesoporous Bioactive Glass.

Mesoporous bioactive glass (MBG) is an extensively studied biomaterial used for the healing of bone defects. Its biological applications can be tailored by introducing metallic ions, such as strontium (Sr) and copper (Cu), which can enhance its functionalities, including osteogenetic, angiogenetic and antibacterial functionalities. In this study, Cu and Sr ions were co-doped (ratio 1:1) with x = 0.5, 1 and 2 mol% each in glass with an intended nominal composition of 80SiO2-(15-2x)CaO-5P2O5-xCuO-xSrO and synthesized with an evaporation-induced self-assembly (EISA)-based sol-gel technique. XRD confirmed the amorphous nature of the glass, while compositional analysis using ICP-OES confirmed the presence of dopant ions with the required amounts. A TEM study of the MBG powders showed fringes that corresponded to the formation of a highly ordered mesoporous structure. The Cu-Sr-doped MBG showed a positive effect on apatite formation when immersed in SBF, although the release of Cu and Sr ions was relatively slow for 1 mol% of each co-dopant, which signified a stable network structure in the glass. The impact of the Cu and Sr ions on the osteoblast-like cell line MG-63 was assessed. At the particle concentrations of 1 wt./vol.% or lower, the cell viability was above 50%. An antibacterial test was conducted against Gram-negative E. coli and Gram-positive S. aureus bacteria. With a sequential increase in the co-doped ion content in the glass, the zone of inhibition for bacteria increased. The results suggest that the doping of MBG with Cu and Sr ions at up to 2 mol% can result in tailored sustained release of ions to enhance the applicability of the studied glass as a functional biomaterial for bone regeneration applications.

Porous Glass Microspheres from Alkali-Activated Fiber Glass Waste.

Fiber glass waste (FGW) was subjected to alkali activation in an aqueous solution with different concentrations of sodium/potassium hydroxide. The activated materials were fed into a methane-oxygen flame with a temperature of around 1600 °C. X-ray diffraction analysis confirmed the formation of several hydrated compounds, which decomposed upon flame synthesis, leading to porous glass microspheres (PGMs). Pore formation was favored by using highly concentrated activating alkali solutions. The highest homogeneity and yield of PGMs corresponded to the activation with 9 M KOH aqueous solution.

Hydrothermal Corrosion of Double Layer Glass/Ceramic Coatings Obtained from Preceramic Polymers.

Polysilazane-based double layer composite coatings consisting of a polymer-derived ceramic (PDC) bond-coat and a PDC top-coat that contains ceramic passive and glass fillers were developed. To investigate the environmental protection ability of the prepared coatings, quasi-dynamic corrosion tests under hydrothermal conditions were conducted at 200 °C for 48-192 h. The tested PDC coatings exhibited significant mass loss of up to 2.25 mg/cm2 after 192 h of corrosion tests, which was attributed to the leaching of elements from the PDC coatings to the corrosion medium. Analysis of corrosion solutions by inductively coupled plasma optical emission spectrometry (ICP-OES) confirmed the presence of Ba, Al, Si, Y, Zr, and Cr, the main component of the steel substrate, in the corrosion medium. Scanning electron microscopy (SEM) of the corroded surfaces revealed randomly distributed globular crystallites approximately 3.5 µm in diameter. Energy-dispersive X-ray spectroscopy (EDXS) of the precipitates showed the presence of Ba, Al, Si, and O. The predominant phases detected after corrosion tests by X-ray powder diffraction analysis (XRD) were monoclinic and cubic ZrO2, originating from the used passive fillers. In addition, the crystalline phase of BaAl2Si2O8 was also identified, which is in accordance with the results of EDXS analysis of the precipitates formed on the coating surface.

Early-Stage Dissolution Kinetics of Silicate-Based Bioactive Glass under Dynamic Conditions: Critical Evaluation.

This manuscript presents a systematic and detailed study of ion release from 45S5 bioactive glass to develop a methodology to directly monitor dissolved ions in a simulated fluid via inductively coupled plasma optical emission spectrometry (ICP OES). For the kinetic study, two dynamic tests, an inline ICP test and a flow-through test, are performed with the same flow rate, temperature, pH, ionic strength of the solution, and sample surface to leaching solution volume ratio. The flow-through test allows for the measurement of an initial dissolution rate, as well the maximum amount of any species released from the surface of the glass. In addition, the data from the inline ICP test are obtained by immediate and direct monitoring of ions from the first minutes of contact of the glass with aqueous fluids with pH values of 4 and 7.4. The overall dissolution rates of the tested commercial bioactive glass in simulated body fluid (SBF) (pH 7.4) were significantly lower compared to the initial rate acquired. The methodology developed in this study can be applied to monitor the controlled release of ions with additional therapeutic functionalities, where the amount of ions released in the first minutes can be critical for the resulting biological performance.

Electrospun PCL Fiber Mats Incorporating Multi-Targeted B and Co Co-Doped Bioactive Glass Nanoparticles for Angiogenesis.

Vascularization is necessary in tissue engineering to keep adequate blood supply in order to maintain the survival and growth of new tissue. The synergy of biologically active ions with multi-target activity may lead to superior angiogenesis promotion in comparison to single-target approaches but it has been rarely investigated. In this study, polycaprolactone (PCL) fiber mats embedded with B and Co co-doped bioactive glass nanoparticles (BCo.BGNs) were fabricated as a tissue regeneration scaffold designed for promoting angiogenesis. BCo.NBGs were successfully prepared with well-defined spherical shape using a sol-gel method. The PCL fiber mats embedding co-doped bioactive glass nanoparticles were fabricated by electrospinning using benign solvents. The Young's moduli of the nanoparticle containing PCL fiber mats were similar to those of the neat fiber mats and suitable for scaffolds utilized in soft tissue repair approaches. The mats also showed non-cytotoxicity to ST-2 cells. PCL fiber mats containing BCo.BGNs with a relatively high content of B and Co promoted the secretion of vascular endothelial growth factor to a greater extent than PCL fiber mats with a relatively low B and Co contents, which demonstrates the potential of dual ion release (B and Co) from bioactive glasses to enhance angiogenesis in soft tissue engineering.

Multi-targeted B and Co co-doped 45S5 bioactive glasses with angiogenic potential for bone regeneration.

Many elements, such as copper, cobalt (Co), strontium, magnesium and boron (B) have been used for single or multiple doping of bioactive glasses to promote angiogenesis during bone regeneration. However, few works have focused on promoting angiogenesis through stimulating several different signalling pathways which can be called a multi-target approach. In this study, 45S5 bioactive glass (BG) co-doped with B and Co was prepared by conventional melt quenching method. The synergistic effect of the two co-doping elements on angiogenesis promotion was expected. ATR-Fourier transform infrared spectroscopy, energy-dispersive X-ray spectroscopy and inductively coupled plasma-optical emission spectrometry were used to characterize the composition, element distribution and ion release of the samples, respectively. Results showed that the compositions of all samples were consistent with the design compositions and all elements were homogeneously distributed in the bulk glass. Different contents of B and Co led to different release rates of these elements. All samples showed bioactivity after immersion in simulated body fluid, regardless of the amount of doped B and Co. The 1% and 0.1% extracts of all samples did not show any cytotoxicity to MG-63 and ST-2 cells. Compared with single B or Co doping, the B and Co dual doped sample led to a stronger increase of the secretion of vascular endothelial growth factor from ST-2 cells, which supports and confirms the synergistic effect on angiogenesis of B and Co as co-doping elements in 45S5 BG.

Multifunctional zinc ion doped sol - gel derived mesoporous bioactive glass nanoparticles for biomedical applications.

Mesoporous bioactive glasses have been widely investigated for applications in bone tissue regeneration and, more recently, in soft tissue repair and wound healing. In this study we produced mesoporous bioactive glass nanoparticles (MBGNs) based on the SiO2-CaO system. With the intention of adding subsidiary biological function, MBGNs were doped with Zn2+ ions. Zn-MBGNs with 8 mol% ZnO content were synthesized via microemulsion assisted sol-gel method. The synthesized particles were homogeneous in shape and size. They exhibited spherical shape, good dispersity, and a size of 130 ±â€¯10 nm. The addition of zinc precursors did not affect the morphology of particles, while their specific surface area increased in comparison to MBGNs. The presence of Zn2+ ions inhibited the formation of hydroxycarbonate apatite (HCAp) on the particles after immersion in simulated body fluid (SBF). No formation of HCAp crystals on the surface of Zn-MBGNs could be observed after 14 days of immersion. Interestingly, powders containing relatively high amount of zinc released Zn2+ ions in low concentration (0.6-1.2 mg L-1) but in a sustained manner. This releasing feature enables Zn-MBGNs to avoid potentially toxic levels of Zn2+ ions, indeed Zn-MBGNs were seen to improve the differentiation of osteoblast-like cells (MG-63). Additionally, Zn-MBGNs showed higher ability to adsorb proteins in comparison to MBGNs, which could indicate a favourable later attachment of cells. Due to their advantageous morphological and physiochemical properties, Zn-MBGNs show great potential as bioactive fillers or drug delivery systems in a variety of applications including bone regeneration and wound healing.