doi:10.1111/j.1365-2591.2009.01660.
Radio-opaque nanosized bioactive glass for
potential root canal application: evaluation of
radiopacity, bioactivity and alkaline capacity
D. Mohn1, M. Zehnder2, T. Imfeld2 & W. J. Stark1
1
Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, ETH Zurich, Zurich; 2Department of
Preventive Dentistry, Periodontology, and Cariology, University of Zürich Center of Dental Medicine, Zurich, Switzerland
Abstract
Mohn D, Zehnder M, Imfeld T, Stark WJ. Radio-opaque
nanosized bioactive glass for potential root canal application:
evaluation of radiopacity, bioactivity and alkaline capacity.
International Endodontic Journal, 43, 210–217, 2010.
Aim To produce novel nanosized bioactive glass
particles with radio-opaque properties and high
alkaline capacity and to evaluate their performance
as a potential bioactive root canal dressing or filling
material.
Methodology Flame spray-derived bioactive glass
particles in the nanometre range were produced
including bismuth oxide as a radiopacifier. Calcium
hydroxide, barium sulphate and bismuth oxide served
as controls. Corresponding materials were compressed
to obtain dense specimens with increased alkaline
capacity. Radiopacity was evaluated, and in vitro
Introduction
Bioactive glasses of the SiO2–Na2O–CaO–P2O5 system
are of potential interest in dentistry because of their
antimicrobial properties (Stoor et al. 1998) and their
ability to remineralize dentine (Vollenweider et al.
2007). Microorganisms remaining or re-entering the
root canal system are the main cause of post-treatment
disease following root canal treatment (Nair 2006).
Consequently, topical antimicrobials with a long-
Correspondence: Wendelin J. Stark, Department of Chemistry
and Applied Biosciences, Institute for Chemical and Bioengi-
neering, ETH Zurich, Wolfgang-Pauli-Str. 10, HCI E107, 8093
Zurich, Switzerland (Tel.: +41 44 632 09 80; fax: +41 44
633 10 83; e-mail: wendelin.stark@chem.ethz.ch).
210 International Endodontic Journal, 43, 210–217, 2010
bioactivity was monitored using Raman spectroscopy
and scanning electron microscopy. Leaching of
bismuth was controlled using atomic absorption
spectroscopy.
Results Bioactive glass particles with up to 50 wt%
bismuth oxide revealed radiopacity with an equivalent
of 4.94-mm aluminium. The introduction of bismuth
into the bioactive glass altered the alkaline capacity
and the in vitro bioactivity only for high bismuth oxide
quantities. Bismuth oxide leaching out of the glass
matrix was hardly detectable.
Conclusion Bioactive glass can be modified with
bismuth oxide to become radio-opaque.
Keywords: antimicrobial, biomaterial, bismuth,
nanoparticles, radiopacifier, sealer.
Received 25 August 2009; accepted 9 November 2009
lasting effect have been used to treat or prevent apical
periodontitis. Calcium hydroxide (slaked lime), origi-
nally devised as a root canal disinfectant and filling
material (Hermann 1920), remains a popular topical
antiseptic. Calcium hydroxide suspensions release Ca
ions and provide an alkaline environment. As an
untoward long-term effect, they make teeth more prone
to fracture (Andreasen et al. 2002). Similarly, antibac-
terial properties of bioactive glasses are based on their
potential to raise the pH in aqueous suspensions (Allan
et al. 2001), resulting from the sodium and proton
exchange of the glass matrix and the aqueous envi-
ronment (Hench 1991). Bioactive glasses, however, do
not only dissolve or release calcium; they can also
release silica, phosphate and sodium. This ion release
promotes an additional indirect pH-related antibacterial
ª 2010 International Endodontic Journal

effect (Gubler et al. 2008). Furthermore, they change
into inert calcium phosphate in an environment with
constant fluid exchange (Sepulveda et al. 2002) and
can cause calcium phosphate precipitation in their
surroundings (Kangasniemi et al. 1993). Untoward
bioactive glass effects on mechanical dentine properties
appear to be less than those of calcium hydroxide
(Marending et al. 2009).
However, in spite of these properties, a recent study
has suggested that a currently commercially available
bioactive glass (S53P4, Abmindent; Abmin Technolo-
gies, Turku, Finland) is still inferior to calcium
hydroxide in disinfecting human root canals infected
with Enterococcus faecalis (Zehnder et al. 2006). An
approach to overcome this limitation is the reduction in
particle size of bioactive glasses to increase the specific
surface area (SSA), and thus, the active exchange
surface area for improved ion release (Sepulveda et al.
2002). Flame spray synthesis, a dry material prepara-
tion method, enables the rapid and flexible production
of nanoparticulate bioactive glasses (Brunner et al.
2006). The reduced particle size (20–50 nm instead of
>5 lm for commercially available products) revealed
improved antibacterial properties against different
E. faecalis strains in a direct exposure model (Waltimo
et al. 2007). Furthermore, nanosized bioactive glass
particles were able to kill an adherent biofilm on
dentine discs (Gubler et al. 2008).
Besides the antibacterial properties in standard
laboratory tests, however, there are further require-
ments for an effective topical antiseptic used for root
canal treatments. As a result of the entombed environ-
ment in a necrotic root canal system, not only alkaline
induction but also alkaline capacity is important to
sustain the antibacterial properties (Waltimo et al.
2009). Furthermore, a certain level of radiopacity is
necessary for a clear distinction between the material
and the surrounding tissues in clinical practice. The
addition of the radio-opaque agent, however, should
not negatively affect a material’s clinical performance.
In the case of commercial calcium hydroxide products,
barium sulphate has commonly been added in an
amount of up to 54 wt% to render the suspension
radio-opaque (Calxyl; OCO Präparate, Dirmstein,
Germany). There are, however, no data on the conse-
quences of the addition of Ba on clinically desired
calcium hydroxide properties such as alkaline capacity.
The current study aimed at producing a nanopartic-
ulate bioactive glass with radiopacity and consistently
high bioactivity and alkaline capacity through the
addition of a bismuth precursor in the flame spray
ª 2010 International Endodontic Journal
Mohn et al. Radio-opaque bioactive glass
process. As shown in preliminary studies, Bi had a
higher radiopacity than comparable radio-opaque
agents such as Ba when directly incorporated via flame
spray synthesis. Nanometric Bi-containing bioactive
glass particles were characterized and tested for radio-
pacity, alkaline capacity, in vitro bioactivity and
bismuth leaching. Conventional mechanical mixtures
of bioactive glass with bismuth oxide and calcium
hydroxide with barium sulphate served as controls.
Materials and methods
Material preparation and characterization
Nanosized bioactive glass particles were produced in a
flame spray setup as described previously (Brunner
et al. 2006). To introduce radiopacity, an organic
bismuth precursor (Grass & Stark 2006) was added to
the starting material and yielded bioactive glass nano-
particles with 20 wt% and 50 wt% of bismuth oxide,
next to nanometric bioactive glass 45S5 (45% SiO2,
24.5% Na2O, 24.5% CaO and 6% P2O5, all in wt%). The
ratios of all constituents of bioactive glass 45S5 in the
radio-opaque bioactive glass were kept constant. Cal-
cium hydroxide [(‡98.5% pure) Riedel-de Haen, Seelze,
Germany] was used as standard material for root canal
disinfection. Bismuth oxide (‡95% pure) and barium
sulphate (‡95% pure) as a commercially available
radiopacifier were procured from Fluka (Buchs,
Switzerland). The SSA of the as prepared nanomaterials
was measured on a Micromeritics Tristar (Gosford,
NSW, Australia) by nitrogen adsorption at 77 K using
the Brunauer–Emmet–Teller (BET) method after out-
gasing for 1 h at 150 C. A transmission electron
microscope (CM30 ST, LaB6 cathode; Philips Electron
Optics, Eindhofen, The Netherlands) was operated at
300 kV with a point resolution of 0.4 nm to study the
morphology and to confirm the particle size.
Radiopacity assessment
According to previous studies (Waltimo et al. 2009),
the amount of bioactive glass per volume plays an
important role in the closed environment of a root
canal. To achieve a denser packing of the naturally
relatively light nanomaterial (low-density powder with
snow-like behaviour), uniaxial pressure of 2.6 MPa
was applied for 30 s. Grinding of the densified material
subsequently resulted in a smooth and dense powder.
The thus improved nanometric filler (with and without
bismuth oxide) was compared to mechanical mixtures
International Endodontic Journal, 43, 210–217, 2010 211
 Radio-opaque bioactive glass Mohn et al.
of nanometric bioactive glass pressed in the same
manner and barium sulphate (50/50 wt%) or bismuth
oxide (80/20 wt%; 50/50 wt%). Calcium hydroxide
with and without additional barium sulphate was used
as commercially available control material. Bovine
dentine slices with a thickness of 1 mm served as
control for the clear distinction between the material
and the surrounding tissue in clinical situations. Prior
to use, the corresponding powder was homogeneously
mixed with unbuffered physiological saline (0.9 wt%
NaCl; Braun, Sempach, Switzerland) to obtain a final
weight to volume ratio of 60% (plastic limit, i.e. the
maximum amount of powder per liquid to yield a paste-
like suspension). A radiograph was taken after prepar-
ing the paste and placing it in a polycarbonate mould
(10 mm diameter, 1 mm thick). Specimens were placed
on a radiographic sensor (Digora, Soredex, Finland)
together with an aluminium step-wedge with a variable
thickness from 0.5 to 6 mm (0.5 and 1 mm incre-
ments). A Trophy Irix (Trophy, Paris, France) X-ray
unit operating at 65 kV, 8 mA and 0.22 s with a focus-
film distance of 25 cm was used. The paste samples
were prepared immediately prior to use in the X-ray
unit, and radiographic analysis was performed in
triplicates. Digital radiographic images were imported
with a Digora system, using Digora software version
1.51 for Windows without grey scale correction.
Optimas imaging analysis software (Meyer Instruments
Inc., Houston, TX, USA) was used to determine the
mean grey value of the samples and converted the
mean grey values to millimetres of aluminium equiv-
alent (AE).
In vitro bioactivity
The bioactivity of the novel derived bioactive glass
nanoparticles incorporating bismuth oxide was tested
using simulated body fluid (SBF) according to Kokubo
et al. (1990). Pressed bioactive glass particles were
placed up to 7 days in SBF at 37 C (Brunner et al.
2006). Raman spectra (EQUINOX 55 and FRA 160/S;
Bruker optics, Ettlingen, Germany) were recorded in
backscattering mode (Notingher et al. 2002). Surface
examination of bioactive glasses before and after
soaking in SBF can show the formation of carbonated
hydroxyapatite. The formation is an indicator for in
vitro bioactivity of biomaterials. To study the carbon-
ated hydroxyapatite, scanning electron microscope
(SEM) analysis was carried out on a LEO 1530 Gemini
(Zeiss, Oberkochen, Germany) after sputtering the
samples with 4 nm of platinum.
212 International Endodontic Journal, 43, 210–217, 2010
Alkaline capacity
The alkaline capacity of dense packed material was
investigated by titration following the procedure as
described by Waltimo et al. (2009). Samples of 1.25 g
of bioactive glass nanoparticles or control materials
were suspended in 10 mL unbuffered physiological
saline to obtain a five times less concentrated suspen-
sion compared to the pastes mentioned earlier. A
calibrated pH electrode (Seven Easy, Mettler-Toledo,
Greifensee, Switzerland) recorded the continuous titra-
tion curve (1.78 mL h)1, tubing pump ecoline VC-MS/
CA8-6; Ismatec Glattbrugg, Switzerland) of 1 mol L)1
hydrochloric acid (Merck, Darmstadt, Germany).
Detection of Bi in the liquid phase of suspensions
Leaching of bismuth out of mechanical mixtures of
bioactive glass and bismuth oxide was compared to
bismuth oxide-modified bioactive glasses (flame de-
rived). One hundred milligrams of powder was contin-
uously stirred in 10 mL unbuffered physiological
saline. Samples of 0.5 mL were taken at 1, 3 and
51 h and centrifuged at 21 500 g for 10 min. The
concentration of bismuth in the supernatant was
measured using an atomic absorption spectrophotom-
eter (contrAA 300; Analytik Jena, Jena, Germany) with
a wavelength of 223 nm and an acetylene/air flame
with a gas flow of 50 L h)1.
Results
Flame spray synthesis resulted in spherically and
highly agglomerated bioactive glass nanoparticles.
The SSA using BET showed that bioactive glass 45S5
had 71 m2 g)1, whilst modifying the glass with
bismuth oxide reduced it to 60 m2 g)1 and
51 m2 g)1 for glasses with 20 wt% and 50 wt%
bismuth oxide, respectively (Table 1). Transmission
electron microscopy confirmed the morphology and
size of the novel bioactive glass particles to be in the
nanometre range (Fig. 1) and showed bismuth oxide
(2–7 nm) sticking to the surface of the particles.
Pressing the material with uniaxial pressure com-
pacted the powder by a factor of three (Fig. 2), resulting
in a homogenous powder. The radiopacity of the
applied pastes differed strongly from each other
(Fig. 3) depending on the amount of radio-opaque
agent and incorporation of bismuth oxide. Pure
calcium hydroxide had an AE of 0.56 ± 0.14 mm
which was close to that of bovine dentine slices
ª 2010 International Endodontic Journal
 Mohn et al. Radio-opaque bioactive glass

Table 1 Nominal composition of differ-

SSAa Particle
ent nanoparticulate bioactive glasses SiO2 Na2O CaO P2O5 Bi2O3 (m2 g)1) sizeb (nm)
(BG) in weight per cent, specific surface
area (SSA) and particle size BG 45.0 24.5 24.5 6.0 – 71 34
BG 20 wt% Bi2O3 36.0 19.6 19.6 4.8 20.0 60 26
BG 50 wt% Bi2O3 23.4 12.8 12.8 3.1 47.9 51 21
a
Error ± 10%.
b
Brunauer–Emmet–Teller equivalent particle diameter, calculated by 6=SSA
qBG.

(a) (b)

Figure 1 Transmission electron micro-

scopy images of classical nanometric
bioactive glass (a) and bioactive glass
(flame derived) containing 50 wt%
bismuth oxide (b).

Figure 2 Photographic image showing

1 g of as prepared nanometric bioactive
glass (left), pressed bioactive glass (mid-
dle) and pressed bioactive glass (flame
derived) containing 50 wt% bismuth
oxide, respectively.

0.62 ± 0.03 mm AE (Fig. 4). The addition of barium
sulphate at a similar ratio to commercially available
products (50 wt%) resulted in 2.47 ± 0.09 mm AE.
Nanometric bioactive glass 45S5 showed an AE of
0.14 ± 0.10 mm; the mechanical mixture with barium
sulphate (50 wt%) increased this value to
2.24 ± 0.22 mm. Modifying the bioactive glass with
20 wt% bismuth oxide in the flame spray process
increased the radiopacity to an AE of 1.84 ± 0.07 mm,
whereas the mechanical mixture with 20 wt% bismuth
oxide equalled 1.22 ± 0.36 mm. An increased bismuth
oxide quantity (modified by the flame) up to 50 wt%
resulted in 4.94 ± 0.17 mm AEs whilst adding
mechanically 50 wt% bismuth oxide corresponded to
4.66 ± 0.68 mm (Fig. 4).
The formation of a carbonated hydroxyapatite layer
on the surface of bioactive glass nanoparticles after
immersion in SBF for 7 days was confirmed using
Raman spectroscopy (Fig. 5). It was observed that the
bismuth oxide-modified bioactive glasses formed a
hydroxyapatite layer also. This was corroborated by
SEM images showing the typical cauliflower shape of
this layer on both, the pure and the Bi-modified
bioactive glass (Fig. 6).
The alkaline capacity of compacted bioactive glasses
was lower than that of calcium hydroxide, although the
initial pH value was in the same range. Adding barium
sulphate to calcium hydroxide to render the powder
radio-opaque reduced the alkaline capacity by half.
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)
Figure 4 Relative radiopacity of tested materials in compar-
ison with an aluminium step wedge showing the mean and
the standard deviation (n = 3). Dashed bars indicate mechan-
ical mixtures and empty bars are mere materials.
derived) showed the same titration curve as the pure
counterpart. The glass containing 50 wt% bismuth oxide
had a faster pH drop in comparison with the counterpart
with 20 wt% bismuth oxide (both flame derived) (Fig. 7).
However, the incorporation of 50 wt% bismuth oxide
using the flame spray synthesis into the bioactive glass
reduced alkaline capacity less than mechanically adding
50 wt% barium sulphate or bismuth oxide.
The assessment of bismuth in the liquid phase of the
suspensions from bismuth oxide-modified bioactive
glasses and mechanical mixtures gave bismuth con-
centrations between 0 and 1.3 ppm in the nondiluted
214 International Endodontic Journal, 43, 210–217, 2010
Figure 3 Radiographic image of bio-
active glass (flame derived), reference
material and an aluminium step wedge;
(a) calcium hydroxide, (b) nanometric
bioactive glass 45S5, (c) bioactive glass
with 20 wt% bismuth oxide, (d) bioactive
glass with 50 wt% bismuth oxide.
Figure 5 Raman spectroscopy of as prepared bioactive glass
(flame derived) and after immersion in simulated body fluid
(SBF) up to 7 days showing the formation of carbonated
hydroxyapatite.
liquid phase which was in the error range of the
measurement.
Discussion
This study showed that bioactive glasses with bismuth
oxide still have high in vitro bioactivity and alkaline
capacity. This could be a step towards a clinical
application of radio-opaque bioactive glass as a topical
root canal disinfectant. Furthermore, the particles
were packed more densely to increase the alkaline
capacity and thus the antibacterial properties in the
ª 2010 International Endodontic Journal

(a) (b)
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).
Figure 7 Continuous titration of calcium hydroxide and
bioactive glass (BG) suspensions with 1M HCl and a constant
flow rate of 1.78 mL per hour.
closed environment of a root canal, which is a key
factor for a successful root canal dressing (Waltimo
et al. 2009).
Flame spray synthesis is a one-step method to
prepare not only complex nanomaterials (Stark &
Pratsinis 2002) but also modified glasses (Brunner
et al. 2006) to further extend material properties. In
this study, an increasing amount of bismuth oxide was
added to bioactive glass nanoparticles whilst reducing
simultaneously the SSA but also decreasing the
primary particle size (Table 1). This finding can be
correlated with the increasing density of the entire
bioactive glass composition and the increased incorpo-
ration of small bismuth oxide particles not only in the
glass matrix but also on the surface (Fig. 1). However,
the characteristics of the as prepared materials were
not further investigated.
ª 2010 International Endodontic Journal
Mohn et al. Radio-opaque bioactive glass
(c)
It has recently been shown that as-prepared bioac-
tive glass nanoparticles exhibited a dramatically strong
pH loss during titration and thus only a low alkaline
capacity. This further correlated with the reduced
antibacterial properties of the material when exposed to
E. faecalis in a root canal (Waltimo et al. 2009). As the
amount of material plays a relevant role for alkalinity,
the nanoparticles were uniaxially pressed in this study
to further compact the relatively light nanopowder
(Fig. 2). An increase up to 60 wt/vol% of particulate
bioactive glass in saline was achieved in the suspension
described here compared to a ratio of 20 wt/vol% with
the unpressed material (Waltimo et al. 2009). There-
fore, a more successful antibacterial behaviour as a
result of higher alkaline capacity (see later) can be
expected, because more material per volume can be
filled into an entombed environment.
A distinct differentiation of filling material and the
surrounding tissue is mandatory for clinical practice;
hence a positive X-ray contrast medium (radio-opaque
medium) which absorbs X-rays stronger than the
surrounding tissue should be added. Various radiopa-
cifiers such as barium sulphate in calcium hydroxide
(Alacam et al. 1990) or bismuth subcarbonate, bismuth
oxide and barium sulphate in different sealing materials
(Tanomaru et al. 2008) have been reported. Further-
more, the manufacturer of a commercially available
product (Sealapex; Sybron Kerr, Romulus, MI, USA)
reported the use of bismuth oxide, because it increases
the sealer’s shelf life and radiopacity (Tanomaru et al.
2008). Most recent studies reported the successful use
of bismuth oxide as radiopacifier for Portland cements
used as root-end filling materials (Kim et al. 2008,
Hwang et al. 2009, Saliba et al. 2009). Adding bismuth
oxide up to 50 wt% by the flame spray process in the
current study increased the radiopacity of bioactive
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-
tional mechanical mixture of bioactive glass and
bismuth oxide (50/50 wt%) with 4.66 mm. Regarding
the International standard 6876 for root canal sealers,
this novel material exceeded the requested value of
3 mm. In contrast, the mechanical mixture of calcium
hydroxide and barium sulphate had an AE of merely
2.47 mm. Bovine dentine had 0.62 mm AE, which is
lower than 1.74 mm reported in a recent study
(Húngaro Duarte et al. 2009). Consequently, depending
on the measurement conditions, the radiopacity of the
novel flame-derived bioactive glass nanoparticles might
even be higher than that reported here.
Although bioactive glasses were modified with
bismuth oxide to a content of up to 50 wt%, they still
formed an apatite layer on their surface upon immer-
sion in SBF. Raman spectra revealed the formation of
carbonated hydroxyapatite confirming the in vitro
bioactivity (Fig. 5). This suggests that bismuth oxide
incorporation did not have a significant influence on
the ability to form hydroxyapatite. On the other hand,
the alteration of other properties of the glass cannot be
excluded, i.e. a decelerated formation of hydroxyapatite
which can be speculated as a result of the smaller
Raman spectroscopy peak at 960 cm)1 (Fig. 5) that is
characteristic for P–O symmetric vibration of PO34
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-
theless, it has been shown that modification of bioactive
glass does not necessarily have a negative influence
on bioactive properties (Bellantone et al. 2000). In
the present study, SEM images further validated the
formation of apatite on the surface (Fig. 6). The
bismuth oxide-modified bioactive glass showed smaller
and less apparent hydroxyapatite structures compared
to pure bioactive glass after immersion in SBF, which
can be an indicator for the slower formation. The
bioactive glass containing 50 wt% bismuth oxide
showed considerably less apatite, suggesting that this
content is probably the limit for radiopacifier addition
without loosing the favoured properties.
The mechanical addition of 50 wt% barium sul-
phate to calcium hydroxide reduced alkaline capacity
by half (Fig. 7) compared to pure calcium hydroxide.
Hence, the potential of this topical disinfectant could
be reduced also. However, further studies are required
to explore this. Adding barium sulphate or bismuth
oxide to classical bioactive glass revealed the same
finding. Modifying the bioactive glass by flame spray
synthesis with 20 wt% bismuth oxide, on the other
216 International Endodontic Journal, 43, 210–217, 2010
hand, did not show a difference in alkaline capacity
compared to original nanometric bioactive glass. The
addition of 50 wt% bismuth oxide (flame derived)
resulted in a lower alkaline capacity, which neverthe-
less was still higher than that of a 50/50 mechanical
mixture of bioactive glass with barium sulphate or
bismuth oxide. Comparing this finding with a historic
control from a previous study under identical condi-
tions (Waltimo et al. 2009) shows that the compressed
material revealed a higher alkaline capacity than the
unpressed counterpart, although a radio-opaque agent
was included.
Bismuth oxide is known to be only poorly soluble or
insoluble in water depending on the acidity of the
corresponding medium (Slikkerveer & Dewolff 1989).
The bismuth concentration in unbuffered saline varied
between 0 and 1.3 ppm. This means that less than
0.05% of the bismuth in the powders was present in the
solution. Hence, the rest is probably still incorporated
in the silica network, sticking to the silica network or
precipitated as insoluble bismuth oxide next to the
flame-derived bioactive glass.
Further studies should assess the antimicrobial effect
of the glasses under investigation in root canals and
compare the cyto- and biocompatibility of these mate-
rials to that of calcium hydroxide.
Conclusion
Bismuth oxide-modified bioactive glass nanoparticles
produced by flame spray synthesis and then pressed to
form a dense powder showed radiopacity, alkaline
capacity and bioactivity that would encourage their
application in further pre-clinical tests.
Acknowledgements
Funding by the Department of Preventive Dentistry and
Oral Epidemiology, University of Zurich is kindly
acknowledged. We also thank Beatrice Sener and
Henning Scriba for their help with the digital radio-
graphic analysis, Fabian Koehler for SEM images, Frank
Krumeich for TEM images and Evagelos Athanassiou
for help with the Raman spectroscopy.
References
Alacam T, Gorgul G, Omurlu H (1990) Evaluation of
diagnostic radiopaque contrast materials used with calcium
hydroxide. Journal of Endodontics 16, 365–8.
ª 2010 International Endodontic Journal

Allan I, Newman H, Wilson M (2001) Antibacterial activity of
particulate Bioglass (R) against supra- and subgingival
bacteria. Biomaterials 22, 1683–7.
Andreasen JO, Farik B, Munksgaard C (2002) Long-term
calcium hydroxide as a root canal dressing may increase
risk of root fracture. Dental Traumatology 18, 134–7.
Bellantone M, Coleman NJ, Hench LL (2000) Bacteriostatic
action of a novel four-component bioactive glass. Journal of
Biomedical Materials Research 51, 484–90.
Brunner TJ, Grass RN, Stark WJ (2006) Glass and bioglass
nanopowders by flame synthesis. Chemical Communications
13, 1384–6.
Grass RN, Stark WJ (2006) Flame spray synthesis under a
non-oxidizing atmosphere: preparation of metallic bismuth
nanoparticles and nanocrystalline bulk bismuth metal.
Journal of Nanoparticle Research 8, 729–36.
Gubler M, Brunner TJ, Zehnder M, Waltimo T, Sener B, Stark
WJ (2008) Do bioactive glasses convey a disinfecting
mechanism beyond a mere increase in pH? International
Endodontic Journal 41, 670–8.
Hench LL (1991) Bioceramics: from concept to clinic. Journal
of the American Ceramic Society 74, 1487–510.
Hermann BW (1920) Calciumhydroxid als Mittel zum Behandeln
und Füllen von Zahnwurzelkanälen. PhD Thesis. Würzburg, D.
Húngaro Duarte MA, de Oliveira El Kadre GD, Vivan RR,
Guerreiro Tanomaru JM, Filho MT, de Moraes IG (2009)
Radiopacity of Portland cement associated with different
radiopacifying agents. Journal of Endodontics 35, 737–40.
Hwang YC, Lee SH, Hwang IN et al. (2009) Chemical
composition, radiopacity, and biocompatibility of Portland
cement with bismuth oxide. Oral Surgery, Oral Medicine,
Oral Pathology, Oral Radiology, and Endodontology 107,
E96–102.
Kangasniemi IMO, Vedel E, Deblickhogerworst J, Yliurpo AU,
Degroot K (1993) Dissolution and scanning electron
microscopic studies of Ca,P particle-containing bioac-
tive glasses. Journal of Biomedical Materials Research 27,
1225–33.
Kim EC, Lee BC, Chang HS, Lee W, Hong CU, Min KS (2008)
Evaluation of the radiopacity and cytotoxicity of Portland
cements containing bismuth oxide. Oral Surgery, Oral
Medicine, Oral Pathology, Oral Radiology, and Endodontology
105, E54–7.
Kokubo T, Kushitani H, Sakka S, Kitsugi T, Yamamuro T
(1990) Solutions able to reproduce in vivo surface-structure
changes in bioactive glass-ceramic A-W3. Journal of Bio-
medical Materials Research 24, 721–34.
ª 2010 International Endodontic Journal
Mohn et al. Radio-opaque bioactive glass
Marending M, Stark WJ, Brunner TJ, Fischer J, Zehnder M
(2009) Comparative assessment of time-related bioactive
glass and calcium hydroxide effects on mechanical proper-
ties of human root dentin. Dental Traumatology 25, 126–9.
Nair PNR (2006) On the causes of persistent apical periodon-
titis: a review. International Endodontic Journal 39, 249–81.
Notingher I, Boccaccini AR, Jones J, Maquet V, Hench LL
(2002) Application of Raman microspectroscopy to the
characterisation of bioactive materials. Materials Character-
ization 49, 255–60.
Penel G, Leroy G, Rey C, Bres E (1998) MicroRaman spectral
study of the PO4 and CO3 vibrational modes in synthetic
and biological apatites. Calcified Tissue International 63,
475–81.
Saliba E, Abbassi-Ghadi S, Vowles R, Camilleri J, Hooper S,
Camilleri J (2009) Evaluation of the strength and radiopac-
ity of Portland cement with varying additions of bismuth
oxide. International Endodontic Journal 42, 322–8.
Sepulveda P, Jones JR, Hench LL (2002) In vitro dissolution of
melt-derived 45S5 and sol-gel derived 58S bioactive glasses.
Journal of Biomedical Materials Research 61, 301–11.
Slikkerveer A, Dewolff FA (1989) Pharmacokinetics and
toxicity of bismuth compounds. Medical Toxicology and
Adverse Drug Experience 4, 303–23.
Stark WJ, Pratsinis SE (2002) Aerosol flame reactors for man-
ufacture of nanoparticles. Powder Technology 126, 103–8.
Stoor P, Soderling E, Salonen JI (1998) Antibacterial effects of
a bioactive glass paste on oral microorganisms. Acta
Odontologica Scandinavica 56, 161–5.
Tanomaru M, Jorge EG, Tanomaru JMG, Goncalves M (2008)
Evaluation of the radiopacity of calcium hydroxide- and
glass-ionomer-based root canal sealers. International End-
odontic Journal 41, 50–3.
Vollenweider M, Brunner TJ, Knecht S et al. (2007) Remin-
eralization of human dentin using ultrafine bioactive glass
particles. Acta Biomaterialia 3, 936–43.
Waltimo T, Brunner TJ, Vollenweider M, Stark WJ, Zehnder M
(2007) Antimicrobial effect of nanometric bioactive glass
45S5. Journal of Dental Research 86, 754–7.
Waltimo T, Mohn D, Paqué F et al. (2009) Fine-tuning of
bioactive glass for root canal disinfection. Journal of Dental
Research 88, 235–8.
Zehnder M, Luder HU, Schatzle M, Kerosuo E, Waltimo T
(2006) A comparative study on the disinfection potentials of
bioactive glass S53P4 and calcium hydroxide in contra-
lateral human premolars ex vivo. International Endodontic
Journal 39, 952–8.
International Endodontic Journal, 43, 210–217, 2010 217