Tobias J. Brunner

Comparison of nanoscale and microscale bioactive glass on the properties of P(3HB)/Bioglass composites.

This study compares the effects of introducing micro (m-BG) and nanoscale (n-BG) bioactive glass particles on the various properties (thermal, mechanical and microstructural) of poly(3hydroxybutyrate) (P(3HB))/bioactive glass composite systems. P(3HB)/bioactive glass composite films with three different concentrations of m-BG and n-BG (10, 20 and 30 wt%, respectively) were prepared by a solvent casting technique. The addition of n-BG particles had a significant stiffening effect on the composites, modulus when compared with m-BG. However, there were no significant differences in the thermal properties of the composites due to the addition of n-BG and m-BG particles. The systematic addition of n-BG particles induced a nanostructured topography on the surface of the composites, which was not visible by SEM in m-BG composites. This surface effect induced by n-BG particles considerably improved the total protein adsorption on the n-BG composites compared to the unfilled polymer and the m-BG composites. A short term in vitro degradation (30 days) study in simulated body fluid (SBF) showed a high level of bioactivity as well as higher water absorption for the P(3HB)/n-BG composites. Furthermore, a cell proliferation study using MG-63 cells demonstrated the good biocompatibility of both types of P(3HB)/bioactive glass composite systems. The results of this investigation confirm that the addition of nanosized bioactive glass particles had a more significant effect on the mechanical and structural properties of a composite system in comparison with microparticles, as well as enhancing protein adsorption, two desirable effects for the application of the composites in tissue engineering.

Effect of nanoparticulate bioactive glass particles on bioactivity and cytocompatibility of poly(3-hydroxybutyrate) composites

This work investigated the effect of adding nanoparticulate (29 nm) bioactive glass particles on the bioactivity, degradation and in vitro cytocompatibility of poly(3-hydroxybutyrate) (P(3HB)) composites/nano-sized bioactive glass (n-BG). Two different concentrations (10 and 20 wt %) of nanoscale bioactive glass particles of 45S5 Bioglass composition were used to prepare composite films. Several techniques (Raman spectroscopy, scanning electron microscopy, atomic force microscopy, energy dispersive X-ray) were used to monitor their surface and bioreactivity over a 45-day period of immersion in simulated body fluid (SBF). All results suggested the P(3HB)/n-BG composites to be highly bioactive, confirmed by the formation of hydroxyapatite on material surfaces upon immersion in SBF. The weight loss and water uptake were found to increase on increasing bioactive glass content. Cytocompatibility study (cell proliferation, cell attachment, alkaline phosphatase activity and osteocalcin production) using human MG-63 osteoblast-like cells in osteogenic and non-osteogenic medium showed that the composite substrates are suitable for cell attachment, proliferation and differentiation.

IN VIVO AND IN VITRO EVALUATION OF FLEXIBLE, COTTONWOOL-LIKE NANOCOMPOSITES AS BONE SUBSTITUTE MATERIAL FOR COMPLEX DEFECTS

The easy clinical handling and applicability of biomaterials has become a focus of materials research due to rapidly increasing time and cost pressures in the public health sector. The present study assesses the in vitro and in vivo performance of a flexible, mouldable, cottonwool-like nanocomposite based on poly(lactide-co-glycolide) and amorphous tricalcium phosphate nanoparticles (PLGA/TCP 60:40). Immersion in simulated body fluid showed exceptional in vitro bioactivity for TCP-containing fibres (mass gain: 18%, 2 days, HAp deposition). Bone regeneration was quantitatively investigated by creating four circular non-critical-size calvarial defects in New Zealand White rabbits. The defects were filled with the easy applicable cottonwool-like PLGA/TCP fibres or PLGA alone. Porous bovine-derived mineral (Bio-Oss) was used as a positive control and cavities left empty served as a negative control. The area fraction of newly formed bone (4 weeks implantation) was significantly increased for TCP-containing fibres compared to pure PLGA (histological and micro-computed tomographic analysis). A spongiosa-like structure of the newly formed bone tissue was observed for PLGA/TCP nanocomposites, whereas Bio-Oss-treated defects afforded a solid cortical bone.

Do bioactive glasses convey a disinfecting mechanism beyond a mere increase in pH

AIM To test whether bioactive glasses kill microbiota via mineralization or the release of ions other than sodium. METHODOLOGY Flame-spray synthesis was applied to produce nanometric glasses of different sodium content and constant Ca/P ratio: 28S5, 45S5 and 77S. Calcium hydroxide and nanometric tricalcium phosphate (TCP) were used as controls. Apatite induction was monitored by Raman spectroscopy. Bovine dentine disks with adherent Enterococcus faecalis cells were exposed to test and control suspensions or buffered solutions for 1 h, 1 day and 1 week. Colony-forming units were counted and disks were inspected using scanning electron microscopy. Suspension supernatants and solutions were analysed for their pH, osmolarity, calcium and silicon content. RESULTS Sodium containing glasses induced pH levels above 12, compared with less than pH 9 with sodium-free 77S. Calcium hydroxide, 45S5 and 28S5 killed all bacteria after 1 day and lysed them after 1 week. TCP caused the highest apatite induction and substantial calcification on bacteria adhering to dentine, but did not reduce viable counts. 77S achieved disinfection after 1 week without visible apatite formation, whilst the buffer solution at pH 9 caused only minimal reduction in counts. CONCLUSION Bioactive glasses have a directly and an indirectly pH-related antibacterial effect. The effect not directly linked to pH is because of ion release rather than mineralization.

Improved degradation and bioactivity of amorphous aerosol derived tricalcium phosphate nanoparticles in poly(lactide-co-glycolide)

The industrially used flame synthesis of silica polymer fillers was extended to amorphous tricalcium phosphate (a-TCP) nanoparticles and resulted in a similar morphology as the traditionally used polymer fillers. Doping of poly(lactide-co-glycolide) (PLGA) with such highly agglomerated a-TCP was investigated for mechanical properties, increased in vitro biodegradation and the formation of a hydroxyapatite layer on the surface of the nanocomposite. PLGA films with particle loadings ranging from 0 to 30 wt% were prepared by solvent casting. Degradation in simulated body fluid (SBF) at 37 °C under sterile conditions for up to 42 days was followed by Raman spectroscopy, scanning electron microscopy (SEM), thermal analysis and tensile tests. The presence of nanoparticles in the PLGA matrix slightly increased the Youngs modulus up to 30% compared to pure polymer reference materials. The nanoparticle doped films showed a significantly increased loss of polymer mass during degradation. Scanning electron microscopy images of doped films showed that the SBF degraded the PLGA by corrosion as facilitated by the incorporation of nanoparticulate calcium phosphate. Raman spectroscopy revealed that the deposition of about 10 nm sized hydroxyapatite crystallites on the surface of doped PLGA films was strongly increased by the addition of tricalcium phosphate fillers. The combination of increased hydroxyapatite formation and enhanced polymer degradation may suggest the use of such amorphous, aerosol derived a-TCP fillers for applications in non-load-bearing implant sites.

Flexible, silver containing nanocomposites for the repair of bone defects: antimicrobial effect against E. coli infection and comparison to tetracycline containing scaffolds

Bone regeneration in infected tissue or areas with high bacteria concentrations such as the oral cavity requires combining disinfection with biomaterial properties. Classical antibiotics typically provide excellent short term protection against re-infection of a defect but are typically washed out of an operation site within days. The present work investigates the use of silver on amorphous tricalcium phosphate (TCP) nanoparticles for electrospun, highly porous poly(lactide-co-glyclolide) (PLGA) fibrous composites. In vitro bioactivity tests of the wool-like composite PLGA/Ag-TCP (80 : 20) containing 0.5 wt% silver showed rapid hydroxyapatite deposition on the nanocomposite within 2 days. Antibacterial tests using E. coli demonstrated a strongly prolonged antibacterial effect of the scaffolds containing finely dispersed silver on TCP if compared to current clinically used methods based on soaking the scaffolds with a tetracycline solution prior to implantation.

Fine-tuning of Bioactive Glass for Root Canal Disinfection

An ideal preparation of 45S5 bioactive glass suspensions/slurries for root canal disinfection should combine high pH induction with capacity for continuing release of alkaline species. The hypothesis of this study was that more material per volume of bioactive glass slurry is obtained with a micrometric material (< 5 μm particle size) or a micrometric/ nanometric hybrid, rather than a solely nanometric counterpart. This should correlate with alkaline capacity and antimicrobial effectiveness. Slurries at the plastic limit were prepared with test and reference materials in physiological saline. Total mass and specific surface area of glass material per volume were determined. Continuous titration with hydrochloric acid was performed, and antimicrobial effectiveness was tested in extracted human premolars mono-infected with E. faecalis ATTC 29212 (N = 12 per material). While the nanometric slurry had a 12-fold higher specific surface area than the micrometric counterpart, the latter had a considerably higher alkaline capacity and disinfected significantly better (Fisher’s exact test, P < 0.05). The hybrid slurry behaved similarly to the micrometric preparation.

Controlling the reactivity of calcium phosphate cements

The effect of composition on the reactivity of a calcium phosphate cement (CPC) consisting of tricalcium phosphate (TCP)–water mixtures was investigated by isothermal calorimetry at 37 °C. The parameters of interest were the mean particle size of the powder, the use of small amounts of hydroxyapatite nanoparticles as nucleating agent, the phosphate concentration and the pH of the aqueous solution. Interactions of various parameters were also investigated. The results show that the main parameter controlling CPC reactivity was TCP particle size, because in contrast to other parameters it markedly reduced the total reaction time.

Comparative assessment of time-related bioactive glass and calcium hydroxide effects on mechanical properties of human root dentin

Suspensions of micro- or nanoparticulate SiO(2)-Na(2)O-CaO-P(2)O(5) bioactive glasses could potentially be used as dressings in traumatized front teeth with open apices as an alternative to Ca(OH)(2). These materials have a disinfecting capacity similar to Ca(OH)(2), but bear the advantage of bioactivity. However, because bioactive glasses initially act as alkaline biocides just as Ca(OH)(2) does, they may also negatively affect mechanical dentin properties over time. This was assessed in the current study using standardized human root dentin bars. Specimens were immersed in 1:20 (wt vol(-1)) suspensions of nanometric bioactive glass 45S5 or calcium hydroxide for 1, 10, or 30 days. Control specimens were immersed in pure saline for 30 days (n = 20 per group). Subsequently, modulus of elasticity (E) and flexural strength (FS) of the specimens were determined. Results were compared between groups using one-way anova and Scheffés post-hoc test. Ca(OH)(2) caused a significant (P < 0.001) 35% drop in mean flexural strength values compared to the control treatment after 10 days. No further change was observed between 10 days and 30 days. Bioactive glass caused a 20% drop in mean flexural strength as compared to the control after 10 days. However, this difference did not reach statistical significance (P > 0.05). No effects of either material on dentin modulus of elasticity values were observed. It was concluded that the calcium hydroxide suspension affected the dentin more than the bioactive glass counterpart; however, the effect was self-limiting and probably restricted to superficial dentin layers, as suggested by the mere decrease in flexural strength but not in modulus of elasticity values.

Effect of thermal treatments on the reactivity of nanosized tricalcium phosphate powders

This study had two aims: (i) to determine the effects of calcination temperature on the hydraulic reactivity of three nanosized X-ray amorphous tricalcium phosphate (ATCP) powders, and (ii) to try to bring a better understanding of the poor reactivity of α-tricalcium phosphate (α-TCP) nanoparticles obtained by calcining ATCP powders. For those purposes, five calcination temperatures were selected in the range of 500 to 800 °C. Calcination led to the formation of powders consisting of ATCP (500 °C), α-TCP (600, 650 and 700 °C) and β-tricalcium phosphate (β-TCP; 700 and 800 °C). The powder reactivity was assessed by mixing 0.2 g of powder and 1 mL Na2HPO4 0.2 M solution in an isothermal calorimeter. The total reaction time was a function of the calcination temperature: from a few minutes for ATCP powders and ATCP powder calcined at 500 °C, it increased to several hours for ATCP powder calcined at 600, 650 and 700 °C. Interestingly, this change of reaction time was mostly due to an increase of the so-called induction time, i.e. time to start the reaction. It increased from a few seconds to a few hours. Strikingly, the induction time and the total reaction time were reduced by an increase of the calcination temperature from 650 to 700 °C. The results were attributed to the formation of a defect-free surface upon calcination and the concomitant difficulty in creating surface defects to allow dissolution and hence reaction.