Bioactive Glass Microparticles Research Articles

Spray-dried ternary bioactive glass microspheres: Direct and indirect structural effects of copper-doping on acellular degradation behavior

Silicate-based bioactive glass nano/microspheres hold significant promise for bone substitution by facilitating osteointegration through the release of biologically active ions and the formation of a biomimetic apatite layer. Cu-doping enhances properties such as pro-angiogenic and antibacterial behavior. While sol-gel methods usually yield homogeneous spherical particles for pure silica or binary glasses, synthesizing poorly aggregated Cu-doped ternary glass nano/microparticles without a secondary CuO crystalline phase remains challenging. This article introduces an alternative method for fabricating Cu-doped ternary microparticles using sol-gel chemistry combined with spray-drying. The resulting microspheres exhibit well-defined, poorly aggregated particles with spherical shapes and diameters of a few microns. Copper primarily integrates into the microspheres as Cu0 nanoparticles and as Cu2+ within the amorphous network. This doping affects silica network connectivity, as calcium and phosphorus are preferentially distributed in the glass network (respectively as network modifiers and formers) or involved in amorphous calcium phosphate nano-domains depending on the doping rate. These differences affect the interaction with simulated body fluid. Network depolymerization, ion release (SiO44−, Ca2+, PO43−, Cu2+), and apatite nanocrystal layer formation are impacted, as well as copper release. The latter is mainly provided by the copper involved in the silica network and not from metal nanoparticles, most of which remain in the microspheres after interaction. This understanding holds promising implications for potential therapeutic applications, offering possibilities for both short-term and long-term delivery of a tunable copper dose. Statement of significanceA novel methodology, scalable to industrial levels, enables the synthesis of copper-doped ternary bioactive glass microparticles by combining spray-drying and sol-gel chemistry. It provides precise control over the copper percentage in microspheres. This study explores the influence of synthesis conditions on the copper environment, notably Cu0 and Cu2+ ratios, characterized by EPR spectroscopy, an aspect poorly described for copper-doped bioactive glass. Additionally, copper indirectly affects silica network connectivity and calcium/phosphorus distribution, as revealed by SSNMR. Multiscale characterization illustrates how these features impact acellular degradation in simulated body fluid, highlighting the therapeutic potential for customizable copper dosing to address short- and long-term needs.

Synthesis and Characterization of Bioactive Glass and Zinc Oxide Nanoparticles with Enamel Remineralization and Antimicrobial Capabilities.

We investigated the effect of bioactive glass and zinc oxide nanoparticles on enamel remineralization, as well as their antimicrobial effect on cariogenic microbes. This is the first study that investigated the properties of bioactive glass and zinc oxide nanoparticles with mixed materials. Fluoride gel (F), bioactive glass microparticles (µB), bioactive glass nanoparticles (nB), zinc oxide nanoparticles (Z), and a mixed suspension of nB and Z (nBZ) were prepared and characterized by scanning and transmission electron microscopy, zeta potential measurement, X-ray diffraction, and acid buffering capacity testing. Further, we performed a remineralization cycle test of 28 days, and nanoindentation testing was carried out during the immersion period, and then the enamel surfaces were examined using scanning electron microscopy. Additionally, the antimicrobial effects of the sample suspensions were evaluated by measuring their minimum microbicidal concentrations against various cariogenic microbes. Our results revealed that nB had a near-circular shape with an amorphous structure and a considerably large specific surface area due to nanoparticulation. Additionally, nB possessed a rapid acid buffering capacity that was comparable to that of μB. In the remineralization test, faster recovery of mechanical properties was observed on the enamel surface immersed in samples containing bioactive glass nanoparticles (nB and nBZ). After remineralization, demineralized enamel immersed in any of the samples showed a rough and porous surface structure covered with mineralized structures. Furthermore, nBZ exhibited a broad antimicrobial spectrum. These results demonstrated that bioactive glass and zinc oxide nanoparticles have superior demineralization-suppressing and remineralization-promoting effects.

A 3D-Printed Wound-Healing Material Composed of Alginate Dialdehyde-Gelatin Incorporating Astaxanthin and Borate Bioactive Glass Microparticles.

In this study, a wound dressing composed of an alginate dialdehyde-gelatin (ADA-GEL) hydrogel incorporated by astaxanthin (ASX) and 70B (70:30 B2O3/CaO in mol %) borate bioactive glass (BBG) microparticles was developed through 3D printing. ASX and BBG particles stiffened the composite hydrogel construct and delayed its in vitro degradation compared to the pristine hydrogel construct, mainly due to their cross-linking role, likely arising from hydrogen bonding between the ASX/BBG particles and ADA-GEL chains. Additionally, the composite hydrogel construct could hold and deliver ASX steadily. The composite hydrogel constructs codelivered biologically active ions (Ca and B) and ASX, which should lead to a faster, more effective wound-healing process. As shown through in vitro tests, the ASX-containing composite hydrogel promoted fibroblast (NIH 3T3) cell adhesion, proliferation, and vascular endothelial growth factor expression, as well as keratinocyte (HaCaT) migration, thanks to the antioxidant activity of ASX, the release of cell-supportive Ca2+ and B3+ ions, and the biocompatibility of ADA-GEL. Taken together, the results show that the ADA-GEL/BBG/ASX composite is an attractive biomaterial to develop multipurposed wound-healing constructs through 3D printing.

Coatings of hydroxyapatite-bioactive glass microparticles for adhesion to biological tissues.

Adsorption of particles across interfaces has been proposed as a way to create adhesion between hydrogels and biological tissues. Here, we explore how this particle bridging approach can be applied to attach a soft polymer substrate to biological tissues, using bioresorbable and nanostructured hydroxyapatite–bioactive glass microparticles. For this, microparticles of aggregated flower-like hydroxyapatite and bioactive glass (HA–BG) were synthesized via a bioinspired route. A deposition technique using suspension spreading was developed to tune the coverage of HA–BG coatings at the surface of weakly cross-linked poly(beta-thioester) films. By varying the concentration of the deposited suspensions, we produced coatings having surface coverages ranging from 4% to 100% and coating densities ranging from 0.02 to 1.0 mg cm−2. The progressive dissolution of these coatings within 21 days in phosphate-buffered saline was followed by SEM. Ex vivo peeling experiments on pig liver capsules demonstrated that HA–BG coatings produce an up-to-two-fold increase in adhesion energy (9.8 ± 1.5 J m−2) as compared to the uncoated film (4.6 ± 0.8 J m−2). Adhesion energy was found to increase with increasing coating density until a maximum at 0.2 mg cm−2, well below full surface coverage, and then it decreased for larger coating densities. Using microscopy observations during and after peeling, we show that this maximum in adhesion corresponds to the appearance of particle stacks, which are easily separated and transferred onto the tissue. Such bioresorbable HA–BG coatings give the possibility of combining particle bridging with the storage and release of active compounds, therefore offering opportunities to design functional bioadhesive surfaces.

Degradable magnesium implants: improving bioactive and antibacterial performance by designed hybrid coatings

Magnesium and its alloys are promising material candidates for degradable fracture fixation devices due to their suitable mechanical properties and biocompatibility; however, their fast corrosion in aqueous media causes pain and swelling. In this study, a hybrid coating system composed by a sol–gel silica-based matrix with bioactive glass microparticles and silica–gentamicin nanoparticles was deposited by spray technology on magnesium AZ91D alloy. The coating was homogeneously distributed on the surface and protected the degradation of AZ91D alloy in simulated body fluid for at least 28 days, preventing the pH increase of the solution and accelerating the formation of calcium phosphate-related compounds on the surface. Moreover, inhibition of bacteria growth was proved for Staphylococcus aureus and Escherichia coli and increased cell adhesion and proliferation of ST-2 and differentiated MC3T3-E1 cells was shown. The generated coating is a promising surface treatment for providing bioactive and antibacterial proprieties to new degradable implants.

Bioactive glass-containing hydrogel delivery system for osteogenic differentiation of human dental pulp stem cells.

Osteoconductive hydrogels can be fabricated by incorporating necessary growth factors and bioactive particles or simply utilizing the ability of the hydrogel itself to induce bone regeneration. The osteogenic inductive potential of the bioactive glass microparticles (BG MPs) has been well-studied. However, the role of the hydrogel embedding the BG MPs on the osteogenic differentiation of the encapsulated stem cells has not been well established. Moreover, it has been reported that the dental pulp stem cell (DPSC) has the capability of regenerating the craniofacial bone tissue when receiving the necessary osteogenic signals from the microenvironment. In the current study, we have fabricated a composite hydrogel based on alginate and Matrigel containing BG MPs and evaluated the role of the BG MPs and Matrigel on osteogenic differentiation of the encapsulated DPSCs. Our results confirmed that presence of Matrigel enhances the osteogenic differentiation of the DPSCs regardless of the decrease in elasticity of the hydrogel in presence of the BG MPs. This strategy of modifying the microenvironment can be a promising approach for craniofacial bone tissue engineering.

Novel injectable gellan gum hydrogel composites incorporating Zn- and Sr-enriched bioactive glass microparticles: High-resolution X-ray microcomputed tomography, antibacterial and in vitro testing.

Mineralization of hydrogel biomaterials is desirable to improve their suitability as materials for bone regeneration. In this study, gellan gum (GG) hydrogels were formed by simple mixing of GG solution with bioactive glass microparticles of 45S5 composition, leading to hydrogel formation by ion release from the amorphous bioactive glass microparticles. This resulted in novel injectable, self-gelling composites of GG hydrogels containing 20% bioactive glass. Gelation occurred within 20min. Composites containing the standard 45S5 bioactive glass preparation were markedly less stiff. X-ray microcomputed tomography proved to be a highly sensitive technique capable of detecting microparticles of diameter approximately 8μm, that is, individual microparticles, and accurately visualizing the size distribution of bioactive glass microparticles and their aggregates, and their distribution in GG hydrogels. The widely used melt-derived 45S5 preparation served as a standard and was compared with a calcium-rich, sol-gel derived preparation (A2), as well as A2 enriched with zinc (A2Zn5) and strontium (A2Sr5). A2, A2Zn, and A2Sr bioactive glass particles were more homogeneously dispersed in GG hydrogels than 45S5. Composites containing all four bioactive glass preparations exhibited antibacterial activity against methicillin-resistant Staphylococcus aureus. Composites containing A2Zn5 and A2Sr5 bioactive glasses supported the adhesion and growth of osteoblast-like cells and were considerably more cytocompatible than 45S5. All composites underwent mineralization with calcium-deficient hydroxyapatite upon incubation in simulated body fluid. The extent of mineralization appeared to be greatest for composites containing A2Zn5 and 45S5. The results underline the importance of the choice of bioactive glass when preparing injectable, self-gelling composites.

Comparison of the effects of 45S5 and 1393 bioactive glass microparticles on hMSC behavior.

Bioactive glasses (BAGs) are highly interesting materials for bone regeneration applications in orthopedic and dental defects. It is quite well known that ionic release from BAGs influences cell behavior and function. Mindful of the clinical scenario, we hypothesized that local cell populations might additionally physically interact with the implanted BAG particles and respond differently than to just the ionic stimuli. We therefore studied the biological effect of two BAG types (45S5 and 1393) applied to human mesenchymal stromal cells (hMSCs) in three distinct presentation modes: (a) direct contact; and to dissolution products in (b) 2D, and (c) 3D culture. We furthermore investigated how the dose‐dependence of these BAG particles, in concentrations ranging from 0.1 to 2.5 w/v %, influenced hMSC metabolic activity, proliferation, and cell spreading. These cellular functions were significantly hampered when hMSCs were exposed to high concentrations of either glasses, but the effects were more pronounced in the 45S5 groups and when the cells were in direct contact with the BAGs. Furthermore the biological effect of 1393 BAG outperformed that of 45S5 BAG in all tested presentation modes. These outcomes highlight the importance of investigating cell–BAG interactions in experimental set‐ups that recapitulate host cell interactions with BAG particles. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2772–2782, 2017.

Modification of electrophoretic deposition of chitosan–bioactive glass–hydroxyapatite nanocomposite coatings for orthopedic applications by changing voltage and deposition time

This article presents electrophoretic depositions of chitosan (CS), bioactive glass (BG), and hydroxyapatite (HA) applied on titanium (Ti) substrate from ethanol-based suspension. The strategy was utilized to modify the deposition of particles with different values of sizes and Zeta potentials, and consequently obtain desirable nanocomposite coating. SEM studies showed that uniform distribution of particles was obtained on isolated substrate at pH 4.5. Moreover, modified deposition of BG microparticles (<37µm) and HA nanoparticles (<150nm) in chitosan matrix was reached by changing voltage and deposition time. The results of X-ray diffraction (XRD) revealed a composite with high content of HA was fabricated at pH 4.5, V=20V, and t=15min. The deposition mechanism at the initial and final times was significantly related to self-assembly of HA nanoparticles that chelated by cationic macromolecular chains and located on BG microparticles, and CS-coated particulates of HA nanoparticles, respectively. The results of potentiodynamic polarization and electrochemical impedance spectroscopy studies demonstrated that the CS–BG–HA composite coating acts as a barrier layer in corrected simulated body fluid (C-SBF).

Anisotropic Composites of Desaminotyrosine and Desaminotyrosyl Tyrosine Functionalized Gelatin and Bioactive Glass Microparticles

ABSTRACTFunctionalization of gelatin with desaminotyrosine (DAT) and desamino tyrosyl tyrosine (DATT) has been demonstrated to allow network formation based on non-covalent interactions of the aromatic moieties. Based on the observation that the DAT(T) groups furthermore could interact with hydroxyapatite fillers, here it was investigated whether such interactions of DAT(T) could also be employed to stabilize composites formed by functionalized gelatins and bioactive glass (BG) particles. Because of sedimentation of the BG microparticles during the gelification, anisotropic composites with two distinct layers were formed. The characterization of mechanical properties by tensile tests and rheology showed that all composites of non-functionalized and DAT(T) functionalized gelatins with BG microparticles showed an increased Young’s modulus (E) up to 3 MPa, an increased storage modulus (G’) up to 100 kPa, increased tensile strength (σmax) up to 3.4 MPa, and increased loss modulus (G’’) compared to the pure matrices. As the observed effects were more pronounced in the DAT(T) functionalized gelatins compared to non-functionalized gelatins, and a much increased thermal stability of these composites was found, it is likely that there are binding interactions between the aromatic moieties and the BG microparticles. This effect open opportunities for the further development of this type of gelatin-based composites for bone regeneration applications.

Synthesis and Bioactivity of Sol–Gel Derived Porous, Bioactive Glass Microspheres Using Chitosan as Novel Biomolecular Template

This article reports a novel, simple route for synthesis of sol–gel derived porous, bioactive glass (BG) microspheres using chitosan (CTS) as a biomolecular template in the acid‐catalyzed sol–gel process. The use of CTS, which is a natural biocompatible polymer with a unique molecular structure, allowed the synthesis of BG microparticles with a spherical shape, 5–10 μm in size. Furthermore, pores with a size of 5–40 nm were created entirely throughout BG microspheres, which were formed by removing the network of CTS present initially in the CTS‐BG composite microparticles via heat‐treatment at 600°C for 2 h. These porous BG microspheres induced the vigorous precipitation of apatite crystals on their surface when immersed in simulated body fluid for 24 h, thus suggesting their excellent bioactivity .

Effect of particle size on the in vitro bioactivity, hydrophilicity and mechanical properties of bioactive glass-reinforced polycaprolactone composites

Polycaprolactone (PCL) composite films containing 5 wt.% bioactive glass (BG) particles of different sizes (6 μm, 250 nm, < 100 nm) were prepared by solvent casting methods. The ultra-fine BG particles were prepared by high-energy mechanical milling of commercial 45S5 Bioglass® particles. The characteristics of bioactive glass particles were studied by field-emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), dynamic light scattering (DLS), and X-ray diffraction (XRD) methods. In vitro bioactivity of the PCL/BG composite films was evaluated through immersion in the simulated body fluid (SBF). The films were analyzed by FE-SEM, energy dispersive spectrometry (EDS), XRD, and atomic force microscopy (AFM). The mechanical properties of highly-porous PCL/BG composites were examined on cylindrical specimens under quasi-static compression load. It was found that partial crystallization of amorphous BG particles during a prolonged mechanical milling occurred and calcium silicate (CaSiO 3) and sodium calcium silicate (Na 2CaSiO 4) phases were formed. The introduction of submicron BG particles (250 nm) was shown to improve the bioactivity of PCL films. In contrast to BG microparticles, the submicron BG particles were distributed on the film surfaces, providing a high surface exposure to SBF with an improved nanotopography. A notable increase in the stiffness and elastic modulus of the composite was also obtained. As compared to submicron BG particles, lower bioactivity and elastic modulus were acquired for PCL/BG nanoparticles. It was also shown that in spite of high specific surface area of the nanoparticles, partial crystallization during mechanical milling and agglomeration of the nanoparticles during processing decrease the bioactivity, hydrophilicity and mechanical response of the BG-reinforced PCL composites.

Fabrication of porous bioactive glass particles by one step sintering

In this study, we reported a facile method to prepare porous bioactive glass microparticles. Porous particles were synthesized by sintering hollow bioactive glass microspheres obtained using a sol–gel co-template technology. The results showed that porous bioactive glass particles possessed a narrow particle size distribution, a relatively porous surface morphology and a hollow structure. It is worth to say that the resulting microparticles present an amorphous structure although the sintering temperature was improved compared to hollow microspheres. The presence of macropore on the shell may provide an efficient method to carry drugs in the hollow cores. Considering the high deposit rate of nanoscale apatite for bioactive glass materials, the porous microparticles should have potential applications in drug and bioactive molecules delivery, in addition to bone tissue regeneration.