Bioactivity and antibacterial activity against E-coli of calcium-phosphate-based glasses: Effect of silver content and crystallinity

In this work we developed improved bioactive glasses and glass-ceramics for biomedical applications, investigating their in vitro bioactivity, biocompatibility and antibacterial properties against E-Coli. A melt-quenched bioactive glass of the SiO₂-CaO-P₂O₅-MgO system was modified with the addition of 1 and 2 mol% Ag₂O and the 1 mol% Ag₂O-containing glasses were then heat treated to produce glass-ceramics. Surface modifications after soaking in SBF and ionic concentration changes showed that addition of silver and crystallization did not affect bioactivity although crystalline phases promoted a decrease in the degradation rate.Biocompatibility of all Ag-containing glasses and glass-ceramics was confirmed for certain samples concentrations. The antibacterial activity of the glasses against E-Coli was generally improved with decreasing particle size or increasing Ag₂O. The Ag-containing glass-ceramics with higher content of crystalline phase appears as a promising biocompatible biocidal material with potential applications in bone-related diseases.     

Apatite formation on melt-derived bioactive glass powder based on SiO2-CaO-Na2O-P2O5 system

The higher melting temperature and longer soaking time during conventional glass melting route promoted the search for alternative in developing new bioactive glass (BG) composition with improved in fabrication temperature and melting time. The current project involved fabrication of new BG compositions based on SiO₂-CaO-Na₂O-P₂O₅ system via melt derived route. It was confirmed that all bioactive glass composition can be melted at temperature lower than 1400 °C. Formation of Si-O-Si (tetrahedral) functional group highlighted that silicate based glass was established as detected by Fourier transform infrared spectroscope (FTIR). BG bioactivity was performed by incubating the BG powder in Tris-buffer solution (pH 8) for 7, 14 and 21 days. In vitro test confirmed the apatite formation on the bioactive glass surface upon soaking in Tris-buffer solution with characteristic of carbonate group (C-O) and P-O band noticed from FTIR and present of crystalline peak observed in X-ray diffraction (XRD). Morphology of apatite formation on BG surface was observed using scanning electron microscope (SEM).     

Sodium is not essential for high bioactivity of glasses

This study aims to demonstrate that excellent bioactivity of glass can be achieved without the presence of an alkali metal component in glass composition. In vitro bioactivity of two sodium-free glasses based on the quaternary system SiO₂-P₂O₅-CaO-CaF₂ with 0 and 4.5 mol% CaF₂ content was investigated and compared with the sodium-containing glasses with equivalent amount of CaF₂. The formation of apatite after immersion in Tris buffer was followed by Fourier-transform infrared spectroscopy, X-ray diffraction, ³¹P and ¹⁹F solid-state MAS-NMR. The dissolution study was completed by ion release measurements in Tris buffer. The results show that sodium-free bioactive glasses formed apatite at 3 h of immersion in Tris buffer, which is as fast as the corresponding sodium-containing composition. This signifies that sodium is not an essential component in bioactive glasses and it is possible to make equally degradable bioactive glasses with or without sodium. The results presented here also emphasize the central role of the glass compositions design which is based on understanding of structural role of components and/or predicting the network connectivity of glasses.     

Effect of ZrO2 addition on in-vitro bioactivity and mechanical properties of SiO2–Na2O–CaO–P2O5 bioactive glass-ceramic

Herein, a multicomponent bioactive glass (0Z, 46SiO₂–30CaO–18Na₂O–6P₂O₅, wt.%) is prepared via melting. ZrO₂ is introduced into the bioglass using two different methods, and then three types of glass-ceramic bulks are manufactured using low-cost pressureless sintering. The effect of ZrO₂ addition on the bioactivity and mechanical properties of the bioactive glass-ceramic is assessed. The results indicate that the main crystalline precipitate from the bioactive glass-ceramic is Na₂Ca(Si₂O₆). The crystallisation ability of the 5Z glass-ceramic, bioactive glass-ceramic with ZrO₂ added during melting at high temperature, is reduced because ZrO₂ participated in the reconstruction of the glass network. Further, the ZrO₂ addition led to a low rate of cation release when soaked in simulated body fluid, indicating a decreased bioactivity. At the same time, the 5Y bioactive glass-ceramic, prepared by mixing YSZ particles with 0Z using ball-milling, possesses not only the highest mechanical strength (about twice the strength of 0Z) but also a high bioactivity. This study presents a promising method for the production of excellent bioactive glass-ceramic and a candidate (5Y) for the clinical applications where load bearing is required.     

Synthesis,Characterization, and In Vitro Bioactivity of Sol‐Gel‐Derived Zn,Mg, and Zn‐Mg Co‐Doped Bioactive Glasses

Bioactive glasses in the systems CaO‐SiO₂‐P₂O₅‐ZnO, CaO‐SiO₂‐P₂O₅‐MgO, and CaO‐SiO₂‐P₂O₅‐MgO‐ZnO were prepared and characterized. Bioactive glass powders were produced by the sol‐gel method. The prepared bioactive glass powders were immersed in a simulated body fluid (SBF) for periods of up to 28 days at 310 K to investigate the bioactivity of the produced samples. Inductively coupled plasma (ICP) and ultraviolet (UV) spectroscopic techniques were used to detect changes in the SBF composition. X‐Ray diffraction (XRD) was utilized to recognize and confirm the formation of a hydroxyapatite (HA) layer on the bioactive glass powders. Microstructural characterizations of the bioactive glass samples were investigated by scanning electron microscopy (SEM) techniques. Density, porosity, and surface area values of bioactive glass powders were also determined in order to characterize the textural properties of the samples. The results revealed the growth of an HA layer on the surface of the bioactive glass samples. MgO in the glass sample increases the rate of formation of an HA layer while ZnO in the glass slows it down.     

Bioactive nanoglasses and xerogels (SiO2–CaO and SiO2–CaO–P2O5) as promising candidates for biomedical applications

A binary and ternary system of highly bioactive nanoparticles of xerogel (88SiO₂–12CaO (wt.%), solvXG88, 78SiO₂–16CaO–6P₂O₅, solvXG78P6) and glass (69SiO₂–25CaO–6P₂O_5, solvBG69P6) were prepared following an unconventional solvothermal path of the synthesis. These nanosized (<50 nm), non-spherical in shape and mesoporous materials with different compositions greatly enhanced the formation process of hydroxyapatite (HA) in simulated physiological fluids compared to reference 45S5 glass, commonly used in orthopedics and dentistry. The deposition of a HA layer on the glasses was analyzed by various techniques, namely XRD, IR-ATR, ³¹P CP-MAS NMR, EDS analyses, SEM, and HR-TEM imaging. For both types of nanoparticles (glass and obtained at lower temperature xerogels) superior apatite-mineralization ability in time as short as 4 h in the physiological-like buffer was achieved thus, exceeded the bioactivity of the 45S5 glass. This unique bioactivity was complemented by biocompatibility with human dermal fibroblasts and MC3T3 mouse osteoblast precursors, verified in a wide range of concentrations, as well as hemocompatibility studies. The most promising candidate which can compete with clinically used Bioglass is solvXG78P6.     

Hydroxyapatite forming ability,ion release and antibacterial activity of the melt-derived SiO2–P2O5–Na2O–CaO–F glasses modified by replacing CaO with SrO and ZnO

Since the discovery of 1970s, bioactive glass has been a hot topic of research because of its excellent biological activity, which makes it a material that can repair and replace human bone tissue organs. In this work, the bioactive glasses in the system SiO₂–P₂O₅–Na₂O–CaO–F with different amounts of strontium oxide (SrO) and zinc oxide (ZnO) were prepared by the conventional melt quenching technology. The hydroxyapatite (HA) forming ability, ion release and antibacterial activity of these prepared glasses were investigated and the obtained results illustrated that SrO-doped samples had a better ability to form HA in modified simulated body fluid (MSBF) than ZnO-doped samples. As the immersion time of the sample in MSBF increased, the content of HA phase gradually increased. In the same immersion time, the formation ability of HA and the variation of SrO substitution amount showed a non-linear trend, which is mainly related to the influence of SrO content on the glass network structure. The results of ion concentration showed that the formation of HA was the result of the comprehensive action of various ions in the solution, especially the release rate of Si⁴⁺ ions, which had a direct impact on the formation ability of HA. The antibacterial test illustrated that the difference in antibacterial activity of bacteria solution at different sample concentrations may be related to the high pH environment and the osmotic effects caused by the non-physiological concentration of ions in the solution. The glass sample contained 4 wt% SrO showed the minimum bactericidal concentration at 64 mg/mL. The glass samples prepared in this experiment had good biological activity and antibacterial effect, making them suitable for using in dentistry and orthopedic applications, as well as providing a valuable composition reference for the preparation of bioactive glass with excellent comprehensive properties.     

Characterization,in vitro bioactivity and biological studies of sol-gel synthesized SrO substituted 58S bioactive glass

Bioactive glasses (BGs) are considered as a high potential candidate in bone repair and replacement. In the present study, sol–gel derived BGs based on 60% SiO₂-(36%-x) CaO-4%P₂O₅-x SrO (where x = 0, 5 and 10 mol%) quaternary system were synthesized and characterized. The effect of Sr substitutions on bioactivity, proliferation, alkaline phosphatase activity of osteoblast cell line MC3T3-E1 and antibacterial activity were investigated. Dried gels were stabilized at 700 °C to eliminate the nitrates and prevent the crystallization of bioactive glasses. X-ray diffraction, Fourier transform infrared spectroscopy and scanning electron microscopy results confirmed the formation of hydroxycarbonate apatite on the BG surfaces. The 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and alkaline phosphate activity results showed that 5% SrO increased both differentiation and proliferation of MC3T3-E1 cells, while 10% SrO resulted in a decrease in bioactivity. Live/Dead and DAPI/Actin staining exhibited viable cell and the morphology of actin fibers and nuclei of MC3T3 cells treated with BG-0 and BG-5. The result of antibacterial test showed that strontium substituted 58S BG exhibited antibacterial effect against methicillin-resistant Staphylococcus aureus bacteria. Taken together, results suggest that 58S BG with 5 mol% SrO is a good candidate for bone tissue engineering with maximum cell proliferation and ALP activity, good bioactivity and high antibacterial efficiency.     

Carbon sphere influence on textural properties and bioactivity of mesoporous bioactive glass/hydroxyapatite nanocomposite

Mesoporous bioactive glass/hydroxyapatite nanocomposite (MBG-HA) synthesis was conducted through evaporation-induced self-assembly (EISA) method followed by in situ carbonization, with non-ionic block co-polymer as mesoporous template and glucose-derived carbon sphere as co-template. The mixture of different carbon sphere contents into a glass network in the CaO–SiO₂–P₂O₅ system tailored the structural, morphological, and textural properties of MBG–HAs. Based on the preliminary results, the carbon sphere content and textural and structural parameters of as synthesized MBG–HAs showed a negative trend. The MBG–HA 0.5 with additional low carbon sphere content, showed a high surface area and large pore volume. The inclusion of HA nanoparticles inside the channels strongly influenced the high carbon sphere content of the MBG–HA8 mesoporous structure. For in vitro bioactivity tests, MBG–HAs with higher textural parameters possessed a faster apatite phase formation kinetics following the sequence MBG–HA0.5>MBG–HA2>MBG–HA5>MBG–HA8.     

Tailoring bioactive and mechanical properties in polycrystalline CaO–SiO2–P2O5 glass-ceramics

Biological functions and mechanical properties are vital factors for artificial bone materials, with great clinic demand for bone injuries and defects. This study highlights mechanical strengths, in vitro and in vivo biological behaviors of the bioactive CaO–SiO₂–P₂O₅ glass-ceramics tailored by the nucleating agent P₂O₅. A high mechanical flexural strength of ～170 MPa and hardness of ～720 HV were achieved, owing to strengthened Si–O bonds in the network. In vitro cell tests demonstrate remarkable viability (using L929 cells) and bioactivity (using bone marrow-derived mesenchymal stromal cells), associated with the release of Ca²⁺ ions in the solution due to weakened Ca–O bonds in the glass-ceramic network. The assay in the simulated body ?uid revealed a formation of the hydroxyapatite-phase layer, which may act as a bridge to facilitate bioactivity on the CaO–SiO₂–P₂O₅ glass-ceramics. In vivo assay shows a significant bone-ingrowth capability on the CaO–SiO₂–P₂O₅ glass-ceramic. This work paves a promising route to utilize P₂O₅–nucleated CaO–SiO₂–P₂O₅ glass-ceramics for load-bearing bone replacement.     

Effects of Sr and Mg dopants on biological and mechanical properties of SiO2–CaO–P2O5 bioactive glass

In the present study, the effects of Sr and Mg were investigated on mechanical and biological properties of 58S bioactive glass (BG). SiO₂-P₂O₅-CaO BG with different contents of Sr and Mg were synthesized via the sol-gel method and immersed in simulated body fluid (SBF) for several days to explore their biocompatibility. Precise analyses of the BG using X-ray powder diffraction, scanning electron microscopy, and transmission electron microscopy showed that the Mg-doped BG containing 8 wt % MgO possessed better biocompatibility. It was also found that mechanical properties of the BG could be improved by increasing the amounts of MgO and SrO. Both 5Sr-BG and 8Mg-BG samples did not exhibit any cytotoxicity while showing high alkaline phosphatase activity in comparison with control specimens. However, the Sr-doped BG sample including 5 wt % SrO demonstrated enhanced bioactivity and biocompatibility.     

Modification of mesoporous structure of silver-doped bioactive glass with antibacterial properties for bone tissue applications

Silver-containing mesoporous bioglasses powders with SiO₂–CaO–P₂O₅–Ag₂O composition have been successfully synthesized by sol-gel and evaporation-induced self-assembly (EISA) methods in presence of various amounts of surfactant (Pluronic-F127). The morphology and crystal structure of the powders were characterized by scanning electron microscopy (SEM) and X-ray diffraction spectroscopy (XRD). Also, the textural properties of samples have been evaluated by adsorption-desorption, Langmuir and BET methods. Accordingly, powders had a smooth surface morphology with cubic mesoporous structure and a desired surface area and pore volume. The in vitro bioactivity was assessed by SEM, XRD and Fourier transform infrared spectroscopy (FTIR) analyses. All samples enhance the formation of HA after soaking the material in simulated body fluid (SBF) solution. The antibacterial property of samples was evaluated to investigate the effect of silver content in chemical composition. The results showed an adequate antibacterial activity of the samples against Escherichia coli, Salmonella and Listeria monocytogenes.     

Two-step sinter-crystallization of K2O–CaO–P2O5–SiO2 (45S5-K) bioactive glass

Bioactive glasses and glass-ceramics (GCs) effectively regenerate bone tissue, however most GCs show improved mechanical properties. In this work, we developed and tested a rarely studied bioactive glass composition (24.4K₂O-26.9CaO-46.1SiO₂-2.6P₂O₅ mol%, identified as 45S5-K) with different particle sizes and heating rates to obtain a sintered GC that combines good fracture strength, low elastic modulus, and bioactivity. We analyzed the influence of the sintering processing conditions in the elastic modulus, Vickers microhardness, density, and crystal phase formation in the GC. The best GC shows improved properties compared with its parent glass. This glass achieves a good densification degree with a two-step viscous flow sintering approach and the resulting GC shows as high bioactivity as that of the standard 45S5 Bioglass®. Furthermore, the GC elastic modulus (56 GPa) is relatively low, minimizing stress shielding. Therefore, we unveiled the glass sintering behavior with concurrent crystallization of this complex bioactive glass composition and developed a potential GC for bone regeneration.     

Sol‐gel preparation and properties of Ag‐containing bioactive glass films on titanium

Ag‐containing bioactive glass films (Ag/Ca atomic ratios of 0, 5% and 10%) were sol‐gel prepared for bioactive and antibacterial modification of titanium. The gel powders calcined at 610°C are mainly amorphous confirmed by x‐ray diffraction, but small diffraction peaks of Ca₃SiO₅ and silver are detected. The film surface is porous with the pore size of ~200 nm. Silver‐rich sub‐micro particles with sizes of 100‐480 nm are present at the surface of Ag‐glass films. CaP phase, metallic silver and silica are detected by x‐ray photoelectron spectroscopy. The mean apparent bonding strength of the films is as high as 21±1 MPa measured by the pull‐off test. The potentiodynamic polarization test shows that the coated samples have better corrosion resistance than the polished sample. The Ag‐glass coatings and their wafer samples exhibit antibacterial activity against S. aureus. The coated samples are covered by apatite layer after soaked in the simulated body fluid for 2 weeks, demonstrating their bioactivity.     

A novel strategy to synthesize bioactive glass based on the eutectic reaction of B2O3–K2O

Melt-derived route was used to prepare modified bioactive glass-ceramic based on the 45S5 composition with the same network connectivity. Their phase composition, sinterability, and bioactivity were studied. A modified composition was proposed using potassium tetraborate (K₂B₄O₇) to reduce the melting temperature during manufacture. The phase composition and the bioactivity was determined by X-ray diffraction and Fourier transform infrared spectroscopy. Furthermore, the antibacterial properties were evaluated against Enterococcus faecalis. The result shows that glass-ceramics already had P–O and C–O bond functional groups on day 2. These bonds are responsible for the creation of the HCA layer. Scanning electron microscopy (SEM) pictures and Energy Dispersive X-ray Spectroscopy (EDX) investigations showed that, after being immersed in SBF solution, a layer of hydroxyapatite (HA) formed on both BG surfaces on day 2 and that by day 21, HCA cluster crystals had developed. Inductively coupled plasma-optical emission spectroscopy metrics of ionic release from the prepared glass-ceramic, mainly calcium and phosphorus ions in SBF solution, revealed that HCA formation occurred on both BG surfaces, which correlated to the increasing pH within 2 days of incubation; furthermore, it exhibited good antibacterial behavior against the Enterococcus faecalis.     

Structural,mechanical, and in vitro characterization of freeze-cast scaffolds prepared using a sol-gel-derived bioactive glass from the SiO2–CaO–Na2O–P2O5–K2O–MgO system

This work deals with the preparation of freeze-cast scaffolds using a bioactive glass from the SiO₂–CaO–Na₂O–P₂O₅–K₂O–MgO system. This material could be sintered at lower temperatures (650 °C) than other variations of bioactive glasses, which is an important advantage in terms of energy and cost savings. This behavior represents a great advantage in terms of energy and cost savings. The freeze-casting step was conducted using water as a solvent and liquid nitrogen as a coolant. The prepared samples were examined according to their pore structure, thermal behavior, mechanical stability, and bioactivity. The glass transition temperature (T_g), crystallization onset temperature (T_x), and maximum crystallization temperature (T_c) evaluated for this bioactive glass were about 660 °C, 690 °C, and 705 °C. Consequently, the freeze-cast scaffolds could be sintered at 650 °C for 2–8 h, which favored viscous flow sintering without crystallization. Bioactivity assays were conducted by soaking the scaffolds in simulated body fluid for up to 21 days, showing that these materials present a bioactive behavior, inducing hydroxyapatite formation. These materials' mechanical properties and biocompatibility make them promising candidates for use in trabecular bone repair.     

Bioactivity of Fe2O3-CaO-SiO2 glass ceramics modified through the addition of P2O5 and TiO2

The bioactivity of ferromagnetic glass ceramics is one key factor affecting the success of hyperthermia treatments. In order to investigate the effect of the addition of a nucleating agent on the bioactivity of the resulting glass ceramics, two different nucleating agents were used for the fabrication of Fe₂O₃-CaO-SiO₂ glass ceramics, and their short-term and long-term effects on the cytotoxicity of human fetal normal colonic mucosa cells were evaluated. The results show that P₂O₅ promotes the formation of an apatite-like phase on the surface of Fe₂O₃-CaO-SiO₂ glass ceramics immersed in simulated body fluid. While TiO₂ had some adverse effect on the cell viability, P₂O₅ had little effect on the cell viability over a certain concentration range. In this paper, the prepared Fe₂O₃-CaO-SiO₂ glass ceramics with P₂O₅ added as nucleating agent are demonstrated to exhibit a good bioactivity and low cytotoxicity, which suggests good application prospects in the field of hyperthermia.     

Phosphate content affects structure and bioactivity of sol-gel silicate bioactive glasses

Bioactive glasses can heal bone defects and bond with bone through formation of hydroxyl carbonate apatite (HCA) surface layer. Sol-gel derived bioactive glasses are thought to have potential for improving bone regeneration rates over melt-derived compositions. The 58S sol-gel composition (60 mol% SiO₂, 36 mol% CaO, and 4 mol% P₂O₅) has appeared in commercial products. Here, hydroxyapatite (HA) was found to form within the 58S glass during sol-gel synthesis after thermal stabilization. The preformed HA may lead to rapid release of calcium orthophosphate, or nanocrystals of HA, on exposure to body fluid, rather than the release of separate the calcium and phosphate species. Increasing the P₂O₅ to CaO ratio in the glass composition reduced preformed HA formation, as observed by XRD and solid-state NMR. Instead, above 12 mol% phosphate, a phosphate glass network (polyphosphate) formed, creating co-networks of phosphate and silica. Nanopore diameter of the glass and rate of HCA layer formation in simulated body fluid (SBF) decreased when the phosphate content increased.     

The impact of Nb2O5 on in-vitro bioactivity and antibacterial activity of CaF2–CaO–B2O3–P2O5–SrO glass system

This work consists of in vitro bioactivity (in SBF) and antibacterial studies (against S. aureus and E. coli bacteria) of Nb₂O₅ doped bioactive glasses. X-ray diffraction and scanning electron microscopy investigations indicated deposition of Nb-HAp (hydroxyapatite) crystalline layer on the samples after exposing to SBF. The spectroscopy investigations also indicated the deposition of HAp layer on these samples. The magnitude of HAp deposited on the glasses found to be relying on concentration of Nb₂O₅ dopant; this conclusion was drawn by determining weight loss of the glasses due to exposure to SBF and also by assessing the variation of pH of the remnant fluid as functions of Nb₂O₅content. The studies further indicated the maximal content of hydroxyapatite was deposited on the surface the glasses doped with 4.0 mol% of Nb₂O₅. The antibacterial studies (against E. coli and S. aureus bacteria) of these glasses indicated the maximal killing effect of bacteria of the samples admixed with 4.0 mol% of Nb₂O₅. This result is attributed to the occupancy of maximal fraction of Nb ions in NbO₆ structural units (confirmed by IR and Raman spectroscopic results) in this sample that paved the way for easy disintegration of the glass and to act on the bacteria. Overall, the results of bioactivity studies of Nb₂O₅ doped bioglasses indicated that the Nb₂O₅ not only enhanced bioactivity potential but also exhibited antimicrobial activity.     

The effect of magnesium content on in vitro bioactivity,biological behavior and antibacterial activity of sol–gel derived 58S bioactive glass

Bioactive glasses (BGs) have a great potential for bone replacement and regeneration in bone tissue engineering applications. In this research, first, sol–gel derived magnesium substituted 58?S BGs (MBGs) series composed of 60SiO₂–4P₂O₅-(36-x) CaO- xMgO, (x = 0; 1; 3; 5; 8 and 10?mol.%) were synthesized and stabilized at 700?°C to eliminate the nitrates and prevent the crystallization of MBGs. MgO was substituted for CaO in the BG formula up to 10?mol% and the effect of Mg concentration on in vitro bioactivity and cellular properties of the MBGs were investigated by immersing them in simulated body fluid (SBF) followed by structural characterization using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) techniques. The effects Mg on proliferation and differentiation of osteoblastic MC3T3-E1 cells were also evaluated by 3-(4,5dimethylthiazol-2-yl)??2,5-diphenyltetrazolium bromide (MTT) and alkaline phosphate (ALP) activity.Results revealed that magnesium-substituted 58?S BG with 5?mol% MgO (BG-5) had the highest formation rate of hydroxyapatite (HA) while substitution of 8?mol% and10 mol% MgO (BG-8 and BG-10) lowered the bioactivity. MTT and ALP results confirmed that the substitution of the MgO up to 5?mol% increased both proliferation and differentiation of MC3T3-E1 cells, while more substitution had a negative effect and resulted in a decrease of proliferation and differentiation in BG-8 and BG-10. The result of antibacterial test showed that MBGs exhibited antibacterial effect against methicillin-resistant Staphylococcus aureus (MRSA) bacteria. Taken together, results suggest that, among all the synthesized MBGs, sample BG-5 is a promising candidate as multifunctional biomaterial for bone tissue engineering with maximum cell proliferation and ALP activity, good bioactivity and high antibacterial efficiency against MRSA bacteria. Eventually, the BG-5 is suggested to be used in segmental defects in rat model in vivo.