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Marr"*
MS""
School of Dentistry
Medical College of Georgia
Augusts, Georgia
I o Pro5tliodnrtii
122
), Number?, 1995
123
ining the interactions. Data were converted to percent control to facilitate comparison. Since tbe
buffer itself produced a slight inhibition of growtb at
all pHs, the cells in medium containing appropriate
buffer but no eluate were used as controls.
duced was twice the concentration of that from previous, comparable studies,"-''' The final pH for the
media containing eluates in the pH 4,0, 5,0, and
6,8 buffers was pH 7,2, 7.6, and 7,7, respectively.
Cell Culture
Results
Metabolic Assays
Three thousand oral epithelial cells, suspended in
50 |JL of eluate plus 50 [jL of fresh medium, were
added to each well of 96 well culture plates. This
allowed eight samples per material, per test, and
treatment. This final 1:2 dilution in medium
brought the buffer to a final concentration of 0,125
M, and the ratio of disk and eluate:culture medium
was comparable to that of the authors' prior
studies.""'" Plates were placed on a rocking platform for 30 minutes and returned to the incubator
for 24 hours, Weils containing cells in medium
alone and medium containing suitably diluted
buffer in which no resin had been soaked were
used as controls.
After this initial 24-hour period, cellular
metabolism, as reflected by RNA synthesis, was
monitored. Cells were labeled by adding 15
pCi/mL' H-Uridine (Specific Activity equal to 35,8
Ci/mol; New England Nuclear, Boston, MA, After
24 hours of incubation the isotope-containing
media were removed, the cells washed with Trisbuffered saline, pH 7,4, then 100 [jL of 10% (V/V)
trichloroacetic acid (TCA) was put into each well.
After washing an additional three times with TCA,
the samples were solubilized in Laemmelis' solubilization buffer-'' (0,0625moi/L Tris-hydrochloride,
pH 6.8, 2% sodium dodecyl sulfate, 5% 2-mercaptoethanol, 10% glycerol), and duplicate aliquots
were counted in a liquid scintillation counter
(model 3801, Beckman Instruments, Fullerton,
CA), All studies were run twice and results were
based on counts per minute (CPM)/weII,
Statistical analysis o tbe data was performed
using a multifactoriai ANOVA (alpha = 0.05] to
determine differences in cytotoxicity based on material, pH, and time. If significant differences were
found, levels of tbe interaction term were used to
define groups for a one-way ANOVA and Duncan's
multiple range test as a post hoc method for exam-
The Irternalfoiia
I of Prosthodontii
124
Volumes.
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Day 1
Day 2
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Day 3
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Discussion
3, Number 2, 1995
^ H
I 1
125
Pe
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1
-
25 -
Dayi
TB.H
1
1
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^ H
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pH 4.0-Lot 2
1 pH 6.8-Lot 2
Day 2
126
i. Number 2, 1995
Conclusion
This in vitro study examined the effects of environmental pH on elution of potentially toxic substances from heat-, light-, and dual- (chemical
plus light) polymerized denture base resins. Three
reins were evaluated at pH levels of 4.0, 5.0, and
6.8. Within the design of this study it can be concluded that:
1. Components that leach out of denture base
resins can have an adverse effect on oral
epithelial cells,
2. Leaching of such components can occur in different amounts and at different rates, depending
on environmental pH and the material.
3. Since the pH in the oral cavity varies over a
wide range, the pH-related elution profiles can
affect the clinical response to a material.
References
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iitersture Abstract -
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Volume 8, N