ORIGINAL ARTICLE

Storage media effect on bond strength

of orthodontic brackets
Salman Jaffer,a Larry J. Oesterle,b and Sheldon M. Newmanc
Aurora, Colo

Introduction: In testing bond strengths, various storage media are used. The purpose of this study was to de-
termine the effect of 6 storage media and rinsing on the shear bond strength of bonded orthodontic brackets.
Methods: Bovine teeth were stored in 6 storage media for 7 months before bonding: dry (no medium), filtered
water, 10% formalin, 1% chloramine T, 10% chloramine T, isotonic saline solution, and 70% ethanol. These
teeth were further subdivided into 2 groups; 1 group was rinsed with oil-free air and water spray before bond-
ing orthodontic brackets, and the other group not rinsed. All specimens were tested in the shear-peel mode to
failure. Results: The 10% formalin rinsed sample had statistically significantly higher bond strength (16.9 6
6.56 MPa), and storage in ethanol (rinsed, 9.04 6 5.61 MPa; not rinsed, 9.08 6 3.5 MPa) and dry (8.34 6
3.80 MPa) produced significantly lower bond strengths. No difference was found between the other modes
of storage or rinsing. The adhesive remnant index values showed no statistically significant difference between
any groups. Conclusions: For bond strength studies, storage media can have an effect on bond strength re-
sults. Dry, formalin, and ethanol storage should be avoided. Water, isotonic saline solution, and chloramine T
storage produced comparable bond strengths. Rinsing or not rinsing had no effect on bond strength with
these storage media. (Am J Orthod Dentofacial Orthop 2009;136:83-6)

Retief et al5 described the effect of duration of storage

V
arious storage media have been used in ortho-
dontic bracket shear bond strength studies;
however, the effect of the storage media on
enamel has not been studied. Storing teeth in media
other than water decreases bacterial, viral, and fungal
growth; prevents enamel desiccation; and allows teeth
to be stored before testing. Chloramine T is a disinfec-
tant1; formaldehyde, or formalin, is a disinfectant and
potent germicidal agent against all organisms.2 Ethanol,
pure or denatured, is a solvent in cleaning solutions and
has been used as a surface disinfectant.3
Titley et al4 investigated the effect of 11 storage
methods and media on shear bond strengths of compos-
ite resin restorations applied to bovine dentin. Their
results showed that 4 storage methods or media (irradi-
ation or storage in thymol, methanol, and glutaralde-
hyde) showed lower shear bond strengths to bovine
dentin compared with the controls stored in water.
From the Department of Orthodontics, School of Dentistry, University of
Colorado at Denver and Health Sciences Center, Aurora, Colo.
a
Postdoctoral resident.
b
Professor and chair.
c
Associate professor, Department of Restorative Dentistry.
The authors report no commercial, proprietary, or financial interest in the
products or companies described in this article.
Reprint requests to: Larry Oesterle, Department of Orthodontics, University of
Colorado, School of Dentistry, Mail Stop F849, PO Box 6508, Aurora, CO
80045; e-mail, larry.oesterle@UCDenver.edu.
Submitted, February 2007; revised and accepted, July 2007.
0889-5406/$36.00
Copyright Ó 2009 by the American Association of Orthodontists.
doi:10.1016/j.ajodo.2007.07.028
on extracted human teeth and the effect of storage media
on the shear bond strength of Scotchbond 2/Silux to
dentin. Their results showed that the mean shear bond
strength after 6 months of storage was highest for phys-
iologic saline solution and 1% chloramine T (12.4 6
4.4 MPa) (no significant difference between them),
whereas 0.05% thymol (9.6 6 4.0 MPa) and 70% etha-
nol (9.5 6 2.9 MPa) showed the lowest shear bond
strengths. The duration of storage had no significant ef-
fect on shear bond strength. Goodis et al6 studied the ef-
fect of various storage media on extracted human third-
molar dentin and found that physiologic saline solution
decreased the shear bond strength by decreasing the per-
meability of dentin. These studies represent a body of
evidence describing the effect of different storage media
on the shear bond strengths of various composite restor-
ative materials to bovine and human dentin. However,
no study has determined the effect of various storage
media on the shear bond strength of orthodontic
brackets bonded to bovine or human enamel.
The purpose of this investigation was to determine
the effect of 6 storage media and rinsing on the shear-
peel bond strength of orthodontic brackets.
MATERIAL AND METHODS
Bovine mandibular incisors were acquired and
stored in filtered water at 4 C. All teeth were debrided,
and notches were placed in their roots to provide
83
84 Jaffer, Oesterle, and Newman American Journal of Orthodontics and Dentofacial Orthopedics
July 2009

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Fig. Comparative histogram of the shear mean breaking stress (MPa) and standard deviations.

appropriate retention for subsequent mounting in
acrylic cylinders. After debriding, the enamel surfaces
were smoothed and flattened by sequentially using
320, 400, and 600 grit sandpaper. After preparing the
surface, the teeth, in groups of 20, were stored in 1 of
the following storage media for 7 months at 4 C: filtered
water, 10% formalin, 1% chloramine T, 10% chlora-
mine T, isotonic saline solution, and 70% ethanol. Dur-
ing storage, the containers were observed for visible
signs of bacterial growth, but no solutions required re-
placement. There was an additional storage condition
in which 20 specimens were stored dry for 7 months be-
fore bracket bonding. After 7 months in the media, the
incisors were mounted in acrylic cylinders and divided
into 2 groups: 1 group was rinsed with oil-free air and
water spray before bonding, and the other not rinsed.
The teeth were mounted in the acrylic testing cylinders
after storage to eliminate any possible effect of the
acrylic on the storage media. The original intent of the
study was to test after a 6-month storage period, but
a breakdown of the testing machine required extending
the testing time to 7 months.
After mounting in acrylic cylinders and either
rinsing or not rinsing, the teeth were bonded by using
a conventional bracket bonding technique. The adhesive
for all specimens, Transbond II APC (Adhesive Pre-
Coated) (3M Unitek, Monrovia, Calif), was supplied
by the manufacturer preapplied to the bracket bases to
standardize the amount of adhesive. A maxillary right
central incisor mini-twin .018-in bracket (number
3017-201, 3M Unitek) with a bonding surface area of
9.604 mm2 was used for all specimens. All teeth were
acid etched for 40 seconds with an etchant gel contain-
ing 35% phosphoric acid, washed, and dried. The
enamel surface was inspected for the frosted appearance
that indicates an adequate etch before the adhesive
primer (Transbond XT, 3M Unitek) was applied. The
preadhesive-coated bracket was pushed firmly to place,
excess adhesive removed, and the specimen was cured
with a light-emitting diode curing light (Ortholux
LED, 3M Unitek) for 40 seconds, 20 seconds on the me-
sial and 20 seconds on the distal surfaces. By using the
light guide diameter (7.2 mm 5 0.72 cm) and a power
meter (Ophir Optronics, Littleton, Colo), the power out-
put was determined (radiance, 420 mW), and the power
density of 1000 mW/cm2 was calculated.
All specimens were tested in the shear-peel mode to
failure after 24 hours of storage in water at 37 C. A
stainless steel wire was placed under the bracket wings
to apply a shear-peel force parallel to the bracket base.
The shear force was applied until bond failure at an ac-
tuator speed of 1 mm per minute.
A reflecting microscope was used to examine the
enamel surface after testing and determine the adhesive
remnant index (ARI) scores.7
Descriptive statistics were performed, and signifi-
cant differences (P #0.05) in bracket bond strength or
ARI values were tested by using the Kruskal-Wallis
(nonparametric) 1-way analysis of variance (ANOVA)
on ranks followed by the Newman-Keuls test.
RESULTS
The results are summarized in the Figure and
Tables I and II. The 10% formalin rinsed sample had
statistically significant greater bond strength than any
American Journal of Orthodontics and Dentofacial Orthopedics Jaffer, Oesterle, and Newman 85
Volume 136, Number 1

Table I. Mean shear breaking stress, standard deviations, Table II. ARI score distribution and statistical signifi-
and statistical significance with the Kruskal-Wallis cance (no statistically significant difference was found
(nonparametric) 1-way ANOVA on ranks between the samples)
Mean 6 SD Significance ARI score
Sample n (MPa) (P #0.05) Significance
Sample 0 1 2 3 (P #0.05)
Dry 20 8.34 6 3.80 d
Filtered water-rinse 20 14.5 6 5.84 b Dry 2 18 0 0 a
Filtered water-no rinse 20 10.5 6 4.22 b Filtered water-rinse 1 19 0 0 a
Isotonic saline solution-rinse 20 12.7 6 5.26 b Filtered water-no rinse 2 18 0 0 a
Isotonic saline solution-no 20 11.8 6 5.39 b Isotonic saline solution-rinse 0 20 0 0 a
rinse Isotonic saline solution-no rinse 0 20 0 0 a
1% chloramine T-rinse 20 14.4 6 5.39 b 1% chloramine T-rinse 1 19 0 0 a
1% chloramine T-no rinse 20 13.2 6 4.77 b 1% chloramine T-no rinse 0 20 0 0 a
10% chloramine T-rinse 20 15.6 6 8.65 b 10% chloramine T-rinse 0 20 0 0 a
10% chloramine T-no rinse 20 14.1 6 5.27 b 10% chloramine T-no rinse 1 19 0 0 a
10% formalin-rinse 20 16.9 6 6.56 a 10% formalin-rinse 1 19 0 0 a
10% formalin-no rinse 20 14.2 6 6.62 b 10% formalin-no rinse 1 19 0 0 a
70% ethanol-rinse 20 9.04 6 5.61 c 70% ethanol-rinse 5 15 0 0 a
70% ethanol-no rinse 20 9.08 6 3.50 c 70% ethanol-no rinse 2 18 0 0 a

Samples with the same letter were not statistically different, and those ARI scores: 0, no adhesive left on the specimen; 1, less than 50% of
with different letters were different: a, highest statistically significant adhesive left on the specimen; 2, more than 50% of adhesive left on
bracket bond strength; d, lowest statistically significant bracket bond the specimen; 3, 100% of adhesive left on the specimen.
strength.

other test sample, with a mean breaking stress value of
16.9 6 6.56 MPa. This was approximately 10% higher
than the mean stress values for the following samples
that were statistically the same: 1% chloramine T
rinsed and not rinsed, 10% chloramine T rinsed and
not rinsed, filtered water rinsed and not rinsed, 10%
formalin not rinsed, and isotonic saline solution rinsed
and not rinsed. The 10% formalin rinsed sample had
approximately 15% greater bond strength than the
70% alcohol rinsed and nonrinsed samples. The 70%
alcohol rinsed sample (9.04 6 5.61 MPa) and the
70% alcohol nonrinsed sample (9.08 6 3.50 MPa)
were not statistically different, but, along with the
dry sample (8.34 6 3.80 MPa), resulted in statistically
lower bond strengths than the other storage media. The
ARI values showed no statistically significant differ-
ence between the samples.
DISCUSSION
Laboratory testing of bond strengths of orthodontic
adhesive materials is a valuable source of information
for their clinical usefulness. However, storage media
might affect the laboratory values obtained. Storage me-
dia effects on bovine enamel provide predictive values
for the effects of storage media on bonding to human
enamel. According to Fonseca et al,8 human and bovine
enamel have similar radiodensities—ie, both are 95% to
96% inorganic. Although the radiodensities were simi-
lar, the actual enamel structures might still have signif-
icant differences. Furthermore, Oesterle et al9 found
that ‘‘bovine enamel has been shown to provide an ac-
ceptable comparable substrate to test bonding, although
actual bond strengths were lower than those found with
human enamel.’’ Equally important, Fowler et al10 and
Nakamichi et al11 found that human and bovine enamel
have comparable bond strength measurements.
Since enamel is 95% to 96% inorganic, we and
probably other researchers assumed that, because of
its high inorganic component and low organic compo-
nent, various storage media would have little effect on
bonding properties. Although dentin with a high organic
content was affected by storage media, no previous
study had been done on enamel.4-6 Our results, however,
show an effect on enamel also. Storage of the enamel
dry or in ethanol had a negative effect on bond strength,
possibly due to the desiccating effects of these storage
variables. This is similar to the findings of Retief
et al5 when testing the effect of ethanol on dentin bond-
ing. Whether a specimen was rinsed or not rinsed ap-
peared to have little effect on most samples. The only
exception was formalin. When the formalin was rinsed
off before bonding, it produced a higher bond strength
than no rinsing. The reason for this is not clear and could
have been a testing aberration, since the actual mean
difference was only 2.7 MPa. Storage in the other me-
dia, whether rinsed or not rinsed before bonding, pro-
duced statistically equivalent results. Therefore,
storage in 1% or 10% chloramine T, filtered water,
10% formalin nonrinsed, or isotonic saline solution
86 Jaffer, Oesterle, and Newman
had no or a similar effect on bond strength. The shear
bond strength was slightly greater for all samples rinsed
with oil-free air and water spray (13.9 6 6.69 MPa)
when compared with all nonrinsed samples (12.1 6
5.30 MPa), but this difference did not reach statistical
significance. This small increase might be because rins-
ing with oil-free air and water spray before etching
aided in removing the biofilm consisting of any micro-
organisms or other organic or chemical matter that built
up when the samples were stored.
Another explanation for the differences found is that
the storage media might affect the surface tension on the
enamel and alter the effectiveness of etching. A further
undocumented mechanism that could result in lower
bonding strength values is residual media in the enamel
porosities that survived etching and affected primer po-
lymerization. These and any other mechanisms should
be investigated further to determine their validity. Rins-
ing the enamel after storage is probably a recommended
procedure to minimize any of these effects.
Most specimens had ARI scores of 1 or 0. Most of
the composite resin was left on the bracket when the
brackets were debonded. This means that the bond fail-
ure occurred purely or primarily at the adhesive-enamel
interface under all storage conditions; this has been
found previously.12 The various storage media did not
affect where the failure occurred.
CONCLUSIONS
Proper selection of storage media for enamel is im-
portant. For bond strength studies, the storage media can
affect the results. Dry, formalin, and ethanol storage me-
dia should be avoided. Water, isotonic saline solution,
and chloramine T storage produce comparable bond
strengths. Although rinsing the teeth before bonding is
preferred, rinsing or not rinsing had no effect on bond
strength with these storage media.
American Journal of Orthodontics and Dentofacial Orthopedics
July 2009
We thank 3M Unitek for generously donating pre-
coated brackets, adhesive primer, etchant gel, and a cur-
ing light for this study.
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