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Radio-Opaque Nanosized Bioactive Glass For Potential Root Canal Application Evaluation of Radio Pa City Bio Activity and Alkaline Capacity-Endoexpert
Radio-Opaque Nanosized Bioactive Glass For Potential Root Canal Application Evaluation of Radio Pa City Bio Activity and Alkaline Capacity-Endoexpert
210 International Endodontic Journal, 43, 210–217, 2010 ª 2010 International Endodontic Journal
Mohn et al. Radio-opaque bioactive glass
effect (Gubler et al. 2008). Furthermore, they change process. As shown in preliminary studies, Bi had a
into inert calcium phosphate in an environment with higher radiopacity than comparable radio-opaque
constant fluid exchange (Sepulveda et al. 2002) and agents such as Ba when directly incorporated via flame
can cause calcium phosphate precipitation in their spray synthesis. Nanometric Bi-containing bioactive
surroundings (Kangasniemi et al. 1993). Untoward glass particles were characterized and tested for radio-
bioactive glass effects on mechanical dentine properties pacity, alkaline capacity, in vitro bioactivity and
appear to be less than those of calcium hydroxide bismuth leaching. Conventional mechanical mixtures
(Marending et al. 2009). of bioactive glass with bismuth oxide and calcium
However, in spite of these properties, a recent study hydroxide with barium sulphate served as controls.
has suggested that a currently commercially available
bioactive glass (S53P4, Abmindent; Abmin Technolo-
Materials and methods
gies, Turku, Finland) is still inferior to calcium
hydroxide in disinfecting human root canals infected
Material preparation and characterization
with Enterococcus faecalis (Zehnder et al. 2006). An
approach to overcome this limitation is the reduction in Nanosized bioactive glass particles were produced in a
particle size of bioactive glasses to increase the specific flame spray setup as described previously (Brunner
surface area (SSA), and thus, the active exchange et al. 2006). To introduce radiopacity, an organic
surface area for improved ion release (Sepulveda et al. bismuth precursor (Grass & Stark 2006) was added to
2002). Flame spray synthesis, a dry material prepara- the starting material and yielded bioactive glass nano-
tion method, enables the rapid and flexible production particles with 20 wt% and 50 wt% of bismuth oxide,
of nanoparticulate bioactive glasses (Brunner et al. next to nanometric bioactive glass 45S5 (45% SiO2,
2006). The reduced particle size (20–50 nm instead of 24.5% Na2O, 24.5% CaO and 6% P2O5, all in wt%). The
>5 lm for commercially available products) revealed ratios of all constituents of bioactive glass 45S5 in the
improved antibacterial properties against different radio-opaque bioactive glass were kept constant. Cal-
E. faecalis strains in a direct exposure model (Waltimo cium hydroxide [(‡98.5% pure) Riedel-de Haen, Seelze,
et al. 2007). Furthermore, nanosized bioactive glass Germany] was used as standard material for root canal
particles were able to kill an adherent biofilm on disinfection. Bismuth oxide (‡95% pure) and barium
dentine discs (Gubler et al. 2008). sulphate (‡95% pure) as a commercially available
Besides the antibacterial properties in standard radiopacifier were procured from Fluka (Buchs,
laboratory tests, however, there are further require- Switzerland). The SSA of the as prepared nanomaterials
ments for an effective topical antiseptic used for root was measured on a Micromeritics Tristar (Gosford,
canal treatments. As a result of the entombed environ- NSW, Australia) by nitrogen adsorption at 77 K using
ment in a necrotic root canal system, not only alkaline the Brunauer–Emmet–Teller (BET) method after out-
induction but also alkaline capacity is important to gasing for 1 h at 150 C. A transmission electron
sustain the antibacterial properties (Waltimo et al. microscope (CM30 ST, LaB6 cathode; Philips Electron
2009). Furthermore, a certain level of radiopacity is Optics, Eindhofen, The Netherlands) was operated at
necessary for a clear distinction between the material 300 kV with a point resolution of 0.4 nm to study the
and the surrounding tissues in clinical practice. The morphology and to confirm the particle size.
addition of the radio-opaque agent, however, should
not negatively affect a material’s clinical performance.
Radiopacity assessment
In the case of commercial calcium hydroxide products,
barium sulphate has commonly been added in an According to previous studies (Waltimo et al. 2009),
amount of up to 54 wt% to render the suspension the amount of bioactive glass per volume plays an
radio-opaque (Calxyl; OCO Präparate, Dirmstein, important role in the closed environment of a root
Germany). There are, however, no data on the conse- canal. To achieve a denser packing of the naturally
quences of the addition of Ba on clinically desired relatively light nanomaterial (low-density powder with
calcium hydroxide properties such as alkaline capacity. snow-like behaviour), uniaxial pressure of 2.6 MPa
The current study aimed at producing a nanopartic- was applied for 30 s. Grinding of the densified material
ulate bioactive glass with radiopacity and consistently subsequently resulted in a smooth and dense powder.
high bioactivity and alkaline capacity through the The thus improved nanometric filler (with and without
addition of a bismuth precursor in the flame spray bismuth oxide) was compared to mechanical mixtures
ª 2010 International Endodontic Journal International Endodontic Journal, 43, 210–217, 2010 211
Radio-opaque bioactive glass Mohn et al.
212 International Endodontic Journal, 43, 210–217, 2010 ª 2010 International Endodontic Journal
Mohn et al. Radio-opaque bioactive glass
(a) (b)
0.62 ± 0.03 mm AE (Fig. 4). The addition of barium The formation of a carbonated hydroxyapatite layer
sulphate at a similar ratio to commercially available on the surface of bioactive glass nanoparticles after
products (50 wt%) resulted in 2.47 ± 0.09 mm AE. immersion in SBF for 7 days was confirmed using
Nanometric bioactive glass 45S5 showed an AE of Raman spectroscopy (Fig. 5). It was observed that the
0.14 ± 0.10 mm; the mechanical mixture with barium bismuth oxide-modified bioactive glasses formed a
sulphate (50 wt%) increased this value to hydroxyapatite layer also. This was corroborated by
2.24 ± 0.22 mm. Modifying the bioactive glass with SEM images showing the typical cauliflower shape of
20 wt% bismuth oxide in the flame spray process this layer on both, the pure and the Bi-modified
increased the radiopacity to an AE of 1.84 ± 0.07 mm, bioactive glass (Fig. 6).
whereas the mechanical mixture with 20 wt% bismuth The alkaline capacity of compacted bioactive glasses
oxide equalled 1.22 ± 0.36 mm. An increased bismuth was lower than that of calcium hydroxide, although the
oxide quantity (modified by the flame) up to 50 wt% initial pH value was in the same range. Adding barium
resulted in 4.94 ± 0.17 mm AEs whilst adding sulphate to calcium hydroxide to render the powder
mechanically 50 wt% bismuth oxide corresponded to radio-opaque reduced the alkaline capacity by half.
4.66 ± 0.68 mm (Fig. 4). Bioactive glass with 20 wt% bismuth oxide (flame
ª 2010 International Endodontic Journal International Endodontic Journal, 43, 210–217, 2010 213
Radio-opaque bioactive glass Mohn et al.
(a) (b)
(c) (d)
derived) showed the same titration curve as the pure liquid phase which was in the error range of the
counterpart. The glass containing 50 wt% bismuth oxide measurement.
had a faster pH drop in comparison with the counterpart
with 20 wt% bismuth oxide (both flame derived) (Fig. 7).
Discussion
However, the incorporation of 50 wt% bismuth oxide
using the flame spray synthesis into the bioactive glass This study showed that bioactive glasses with bismuth
reduced alkaline capacity less than mechanically adding oxide still have high in vitro bioactivity and alkaline
50 wt% barium sulphate or bismuth oxide. capacity. This could be a step towards a clinical
The assessment of bismuth in the liquid phase of the application of radio-opaque bioactive glass as a topical
suspensions from bismuth oxide-modified bioactive root canal disinfectant. Furthermore, the particles
glasses and mechanical mixtures gave bismuth con- were packed more densely to increase the alkaline
centrations between 0 and 1.3 ppm in the nondiluted capacity and thus the antibacterial properties in the
214 International Endodontic Journal, 43, 210–217, 2010 ª 2010 International Endodontic Journal
Mohn et al. Radio-opaque bioactive glass
Figure 6 Scanning electron microscopy images showing (a) nanometric bioactive glass with 45S5, (b) bioactive glass with 20 wt%
bismuth oxide and (c) bioactive glass with 50 wt% bismuth oxide after immersion in simulated body fluid for 7 days (all flame-
derived).
ª 2010 International Endodontic Journal International Endodontic Journal, 43, 210–217, 2010 215
Radio-opaque bioactive glass Mohn et al.
glass to 4.94 mm AE (Fig. 4) compared to a conven- hand, did not show a difference in alkaline capacity
tional mechanical mixture of bioactive glass and compared to original nanometric bioactive glass. The
bismuth oxide (50/50 wt%) with 4.66 mm. Regarding addition of 50 wt% bismuth oxide (flame derived)
the International standard 6876 for root canal sealers, resulted in a lower alkaline capacity, which neverthe-
this novel material exceeded the requested value of less was still higher than that of a 50/50 mechanical
3 mm. In contrast, the mechanical mixture of calcium mixture of bioactive glass with barium sulphate or
hydroxide and barium sulphate had an AE of merely bismuth oxide. Comparing this finding with a historic
2.47 mm. Bovine dentine had 0.62 mm AE, which is control from a previous study under identical condi-
lower than 1.74 mm reported in a recent study tions (Waltimo et al. 2009) shows that the compressed
(Húngaro Duarte et al. 2009). Consequently, depending material revealed a higher alkaline capacity than the
on the measurement conditions, the radiopacity of the unpressed counterpart, although a radio-opaque agent
novel flame-derived bioactive glass nanoparticles might was included.
even be higher than that reported here. Bismuth oxide is known to be only poorly soluble or
Although bioactive glasses were modified with insoluble in water depending on the acidity of the
bismuth oxide to a content of up to 50 wt%, they still corresponding medium (Slikkerveer & Dewolff 1989).
formed an apatite layer on their surface upon immer- The bismuth concentration in unbuffered saline varied
sion in SBF. Raman spectra revealed the formation of between 0 and 1.3 ppm. This means that less than
carbonated hydroxyapatite confirming the in vitro 0.05% of the bismuth in the powders was present in the
bioactivity (Fig. 5). This suggests that bismuth oxide solution. Hence, the rest is probably still incorporated
incorporation did not have a significant influence on in the silica network, sticking to the silica network or
the ability to form hydroxyapatite. On the other hand, precipitated as insoluble bismuth oxide next to the
the alteration of other properties of the glass cannot be flame-derived bioactive glass.
excluded, i.e. a decelerated formation of hydroxyapatite Further studies should assess the antimicrobial effect
which can be speculated as a result of the smaller of the glasses under investigation in root canals and
Raman spectroscopy peak at 960 cm)1 (Fig. 5) that is compare the cyto- and biocompatibility of these mate-
characteristic for P–O symmetric vibration of PO34 rials to that of calcium hydroxide.
groups of hydroxyapatite (Penel et al. 1998). The peak
at 1070 cm)1 corresponds to the m1 vibrational mode of
CO23 indicating a carbonated hydroxyapatite. Never- Conclusion
theless, it has been shown that modification of bioactive Bismuth oxide-modified bioactive glass nanoparticles
glass does not necessarily have a negative influence produced by flame spray synthesis and then pressed to
on bioactive properties (Bellantone et al. 2000). In form a dense powder showed radiopacity, alkaline
the present study, SEM images further validated the capacity and bioactivity that would encourage their
formation of apatite on the surface (Fig. 6). The application in further pre-clinical tests.
bismuth oxide-modified bioactive glass showed smaller
and less apparent hydroxyapatite structures compared
to pure bioactive glass after immersion in SBF, which Acknowledgements
can be an indicator for the slower formation. The Funding by the Department of Preventive Dentistry and
bioactive glass containing 50 wt% bismuth oxide Oral Epidemiology, University of Zurich is kindly
showed considerably less apatite, suggesting that this acknowledged. We also thank Beatrice Sener and
content is probably the limit for radiopacifier addition Henning Scriba for their help with the digital radio-
without loosing the favoured properties. graphic analysis, Fabian Koehler for SEM images, Frank
The mechanical addition of 50 wt% barium sul- Krumeich for TEM images and Evagelos Athanassiou
phate to calcium hydroxide reduced alkaline capacity for help with the Raman spectroscopy.
by half (Fig. 7) compared to pure calcium hydroxide.
Hence, the potential of this topical disinfectant could
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ª 2010 International Endodontic Journal International Endodontic Journal, 43, 210–217, 2010 217