Leena Hupa

Antibacterial effects and dissolution behavior of six bioactive glasses

Dissolution behavior of six bioactive glasses was correlated with the antibacterial effects of the same glasses against sixteen clinically important bacterial species. Powdered glasses (<45 microm) were immersed in simulated body fluid (SBF) for 48 h. The pH in the solution inside the glass powder was measured in situ with a microelectrode. After 2, 4, 27, and 48 h, the pH and concentration of ions after removing the particles and mixing the SBF were measured with a normal glass pH electrode and ICP-OES. The bacteria were cultured in broth with the glass powder for up to 4 days, after which the viability of the bacteria was determined. The antibacterial effect of the glasses increased with increasing pH and concentration of alkali ions and thus with increased dissolution tendency of the glasses, but it also depended on the bacterium type. The changes in the concentrations of Si, Ca, Mg, P, and B ions in SBF did not show statistically significant influence on the antibacterial property. Bioactive glasses showed strong antibacterial effects for a wide selection of aerobic bacteria at a high sample concentration (100 mg/mL). The antibacterial effects increased with glass concentration and a concentration of 50 mg/mL (SA/V 185 cm(-1)) was required to generate the bactericidal effects. Understanding the dissolution mechanisms of bioactive glasses is essential when assessing their antibacterial effects.

Copper-releasing, boron-containing bioactive glass-based scaffolds coated with alginate for bone tissue engineering

The aim of this study was to synthesize and characterize new boron-containing bioactive glass-based scaffolds coated with alginate cross-linked with copper ions. A recently developed bioactive glass powder with nominal composition (wt.%) 65 SiO2, 15 CaO, 18.4 Na2O, 0.1 MgO and 1.5 B2O3 was fabricated as porous scaffolds by the foam replica method. Scaffolds were alginate coated by dipping them in alginate solution. Scanning electron microscopy investigations indicated that the alginate effectively attached on the surface of the three-dimensional scaffolds leading to a homogeneous coating. It was confirmed that the scaffold structure remained amorphous after the sintering process and that the alginate coating improved the scaffold bioactivity and mechanical properties. Copper release studies showed that the alginate-coated scaffolds allowed controlled release of copper ions. The novel copper-releasing composite scaffolds represent promising candidates for bone regeneration.

In situ pH within particle beds of bioactive glasses.

The in vitro behavior of three bioactive glasses with seven particle size distributions was studied by measuring the in situ pH inside the particle beds for 48h in simulated body fluid (SBF). After immersion, the surface of the particles was characterized with a field emission scanning electron microscope equipped with an energy-dispersive X-ray analyzer. In addition, the results were compared with the reactions of the same glasses formed as plates. A similar trend in pH as a function of immersion time was observed for all systems. However, the pH inside the particle beds was markedly higher than that in the bulk SBF of the plates. The pH decreased as power functions with increasing particle size, i.e. with decreasing surface area. The in vitro reactivity expressed as layer formation strongly depended on the particle size and glass composition. The average thickness of the total reaction layer decreased with the increase in sample surface area. Well-developed silica and calcium phosphate layers typically observed on glass plates could be detected only on some particles freely exposed to the solution. No distinct reaction layers were observed on the finest particles, possibly because the layers spread out on the large surface area. Differences in the properties of the bulk SBF and the solution inside the particle bed were negligible for particles larger than 800microm. The results enhance our understanding of the in vitro reactions of bioactive glasses in various product forms and sizes.

Fiber glass–bioactive glass composite for bone replacing and bone anchoring implants

OBJECTIVE Although metal implants have successfully been used for decades, devices made out of metals do not meet all clinical requirements, for example, metal objects may interfere with some new medical imaging systems, while their stiffness also differs from natural bone and may cause stress-shielding and over-loading of bone. METHODS Peer-review articles and other scientific literature were reviewed for providing up-dated information how fiber-reinforced composites and bioactive glass can be utilized in implantology. RESULTS There has been a lot of development in the field of composite material research, which has focused to a large extent on biodegradable composites. However, it has become evident that biostable composites may also have several clinical benefits. Fiber reinforced composites containing bioactive glasses are relatively new types of biomaterials in the field of implantology. Biostable glass fibers are responsible for the load-bearing capacity of the implant, while the dissolution of the bioactive glass particles supports bone bonding and provides antimicrobial properties for the implant. These kinds of combination materials have been used clinically in cranioplasty implants and they have been investigated also as oral and orthopedic implants. SIGNIFICANCE The present knowledge suggests that by combining glass fiber-reinforced composite with particles of bioactive glass can be used in cranial implants and that the combination of materials may have potential use also as other types of bone replacing and repairing implants.

Bioactive glass ions as strong enhancers of osteogenic differentiation in human adipose stem cells

Bioactive glasses are known for their ability to induce osteogenic differentiation of stem cells. To elucidate the mechanism of the osteoinductivity in more detail, we studied whether ionic extracts prepared from a commercial glass S53P4 and from three experimental glasses (2-06, 1-06 and 3-06) are alone sufficient to induce osteogenic differentiation of human adipose stem cells. Cells were cultured using basic medium or osteogenic medium as extract basis. Our results indicate that cells stay viable in all the glass extracts for the whole culturing period, 14 days. At 14 days the mineralization in osteogenic medium extracts was excessive compared to the control. Parallel to the increased mineralization we observed a decrease in the cell amount. Raman and Laser Induced Breakdown Spectroscopy analyses confirmed that the mineral consisted of calcium phosphates. Consistently, the osteogenic medium extracts also increased osteocalcin production and collagen Type-I accumulation in the extracellular matrix at 13 days. Of the four osteogenic medium extracts, 2-06 and 3-06 induced the best responses of osteogenesis. However, regardless of the enhanced mineral formation, alkaline phosphatase activity was not promoted by the extracts. The osteogenic medium extracts could potentially provide a fast and effective way to differentiate human adipose stem cells in vitro.

Dissolution patterns of biocompatible glasses in 2-amino-2-hydroxymethyl-propane-1,3-diol (Tris) buffer

A continuous flow measurement system with sensitive on-line ion analysis has been applied to study the initial dissolution behaviour of biocompatible glasses in Tris. Altogether 16 glasses with widely varying compositions were studied. The measurement system allowed for quantitative determination of the time-dependent rates of dissolution of sodium, potassium, calcium, magnesium, silicon and phosphorus during the first 10-15 min in contact with Tris solution. The dissolution rates of the different ions showed significant glass to glass variations, but all glasses studied showed one of four distinct dissolution patterns. The ion dissolution rates after an exposure of 1000 s, expressed as the normalized surface-specific mass loss rates, were compared with the in vitro and in vivo reactivity of the glasses as predicted by models in the literature. The results showed a clear correlation between the dissolution rates of the glasses in Tris and their reactivity as measured by other different methods. Consequently, the measured short-term dissolution patterns could be used to determine which glasses are suitable as bioactive, biodegradable, or inert biomaterials for medical devices.

Mechanical verification of soft-tissue attachment on bioactive glasses and titanium implants

Soft-tissue attachment is a desired feature of many clinical biomaterials. The aim of the current study was to design a suitable experimental method for tensile testing of implant incorporation with soft-tissues. Conical implants were made of three compositions of bioactive glass (SiO(2)-P(2)O(5)-B(2)O(3)-Na(2)O-K(2)O-CaO-MgO) or titanium fiber mesh (porosity 84.7%). The implants were surgically inserted into the dorsal subcutaneous soft-tissue or back muscles in the rat. Soft-tissue attachment was evaluated by pull-out testing using a custom-made jig 8 weeks after implantation. Titanium fiber mesh implants had developed a relatively high pull-out force in subcutaneous tissue (12.33+/-5.29 N, mean+/-SD) and also measurable attachment with muscle tissue (2.46+/-1.33 N). The bioactive glass implants failed to show mechanically relevant soft-tissue bonding. The experimental set-up of mechanical testing seems to be feasible for verification studies of soft-tissue attachment. The inexpensive small animal model is beneficial for large-scale in vivo screening of new biomaterials.

Multi-layer porous fiber-reinforced composites for implants: In vitro calcium phosphate formation in the presence of bioactive glass

OBJECTIVES Glass-fiber-reinforced composites (FRCs), based on bifunctional methacrylate resin, have recently shown their potential for use as durable cranioplasty, orthopedic and oral implants. In this study we suggest a multi-component sandwich implant structure with (i) outer layers out of porous FRC, which interface the cortical bone, and (ii) inner layers encompassing bioactive glass granules, which interface with the cancellous bone. METHODS The capability of Bioglass(®) 45S5 granules (100-250μm) to induce calcium phosphate formation on the surface of the FRC was explored by immersing the porous FRC-Bioglass laminates in simulated body fluid (SBF) for up to 28d. RESULTS In both static (agitated) and dynamic conditions, bioactive glass granules induced precipitation of calcium phosphate at the laminate surfaces as confirmed by scanning electron microscopy. SIGNIFICANCE The proposed dynamic flow system is useful for the in vitro simulation of bone-like apatite formation on various new porous implant designs containing bioactive glass and implant material degradation.

Controlling the ion release from mixed alkali bioactive glasses by varying modifier ionic radii and molar volume

Partially substituting one alkali oxide for another reduces the crystallisation tendency and improves the processing of bioactive glasses. Here, we investigate how we can use alkali ions of varying ionic radii to control glass degradation and ion release from Bioglass 45S5. Partially replacing sodium by lithium reduced ion release in static and dynamic dissolution studies in Tris buffer, while ion release increased with increasing potassium for sodium substitution. While the mixed alkali effect is known to reduce ion release from conventional silicate glasses (compared to compositions containing one alkali oxide only), in the glasses studied here ion release was controlled by the packing of the silicate network, described by glass molar volume and oxygen density. Incorporating an alkali ion of smaller ionic radius (Li for Na or Na for K) resulted in a more compact network of higher oxygen density, which reduced ion release. On the other hand, an alkali ion of larger ionic radius (K for Na or Na for Li) expanded the silicate network, allowing for faster ion release. This can be explained by water molecules penetrating an expanded silicate network more easily than a more compact one, thereby directly influencing the ion exchange between modifier ions and protons from the dissolution medium. This shows that the use of modifier ions of varying ionic radii allows for tailoring bioactive glass ion release and degradation while maintaining silicate network polymerisation and network connectivity. And, indeed, recent literature suggests that this concept can be extended to other modifiers besides alkali metal ions, making it possible to design bioactive glasses of tailored solubility.

A glass fiber-reinforced composite - bioactive glass cranioplasty implant: A case study of an early development stage implant removed due to a late infection.

This case study describes the properties of an early development stage bioactive glass containing fiber-reinforced composite calvarial implant with histology that has been in function for two years and three months. The patient is a 33-year old woman with a history of substance abuse, who sustained a severe traumatic brain injury later unsuccessfully treated with an autologous bone flap and a custom-made porous polyethylene implant. She was thereafter treated with developmental stage glass fiber-reinforced composite - bioactive glass implant. After two years and three months, the implant was removed due to an implant site infection. The implant was analyzed histologically, mechanically, and in terms of chemistry and dissolution of bioactive glass. Mechanical integrity of the load bearing fiber-reinforced composite part of the implant was not affected by the in vivo period. Bioactive glass particles demonstrated surface layers of hydroxyapatite like mineral and dissolution, and related increase of pH was considerably less after two and three months period than that for fresh bioactive glass. There was a difference in the histology of the tissues inside the implant areas near to the margin of the implant that absorbed blood during implant installation surgery, showed fibrous tissue with blood vessels, osteoblasts, collagenous fibers with osteoid formation, and tiny clusters of more mature hard tissue. In the center of the implant, where there was less absorbed blood, only fibrous tissue was observed. This finding is in line with the combined positron emission tomography - computed tomography examination with (18F)-fluoride marker, which demonstrated activity of the mineralizing bone by osteoblasts especially at the area near to the margin of the implant 10 months after implantation. Based on these promising reactions found in the bioactive glass containing fiber-reinforced composite implant that has been implanted for two years and three months, calvarial reconstruction with the presented material appears to be a feasible method.