International Dentistry - Vol. 12, No. 2 - 31
SCIENTIFIC
Table 1: Life/Dead Assay 1 hour and 24 hours after
thermal treatment.
Temperature
Vitality %
°C
1h
24 h
37
0
0
39
0
0
42
0
0
45
0
0
46
0,5
2
47
10
17
48.5
18
29
50
17
27
55
24
59
56.5
48
54
58
100
100
60
100
100
65
100
100
Figure 1: Vitality test of thermally treated DPSC.
In order to perform examinations with scanning electron
microscope, the cells were processed as follows:
- Washing of the cells in cacodylate buffer (0.1 M)
- Fixation with 2.5 % Glutaraldehyde in cacodylate buffer
for 20'
- Washing with cacodylate buffer for two times, followed
by two washings with Aqua dest
- Dehydration with increasing alcohol concentration: 20 %,
30 %, 50 %, 70 %, 90 %, 2x in 100 %EtOH for 10' each
- Further processing of the samples at the Centre for Electron
Microscopy (Critical Point Drying and sputtering with gold;
SCD 005, BAL TEC AG, Liechtenstein)
- Microscope: Zeiss EM 902 A.
Examinations with the transmission electron microscope were
conducted:
- Washing of the cells with cacodylate buffer (0.1 M) with
6.8 % Sucrose
- Fixation of 30' with 1 % glutaraldehyde
- Washing with cacodylate buffer
- Contrasting with 1 % Osmiumtetroxyde and 1 %potassium
ferrocyanide for two hours
- Dehydration with increasing alcohol concentration: 20 %,
30 %, 50 %, 70 %, 90 %, 2x in 100 %EtOH for 10' each
- Embedding in Epon (epoxy resin), polymerisation for four
days at 60 °C
- Ultramicrotomy, ultra-thin sections (70 nm; Leica Ultracut S,
Leica Mikrosysteme GmbH, Bensheim, Germany)
- Dyeing of the sections with 1 % Uranyl acetate in methanol
and 1 drop of acetic acid for 10'
- Microscope: Zeiss EM 906.
Results
Light microscopy and vitality test
The cells received thermal treatment at temperatures ranging
from 37 °C to 60 °C and varying inter-mediate temperature
levels. Light microscopy examinations showed significant
morphological changes at temperatures from 46.5 °C ±
0.5 °C.
At temperatures from 37 °C to 45 °C, all cells exhibited
a green calcein fluorescence. At temperatures of 46 °C and
above, lethal results were detected in some of the cells that
had undergone thermal treatment. The number of lethal cells
increased in correspondence to a rise in temperature.
At temperatures of 46 °C to 56.5 °C, the number of lethal
cells had almost doubled 24 h after thermal treatment in
comparison to the number of lethal cells one hour after
VOL. 12, NO. 2 INTERNATIONAL DENTISTRY - AUSTRALASIAN EDITION 31
Table of Contents for the Digital Edition of International Dentistry - Vol. 12, No. 2
Contents
International Dentistry - Vol. 12, No. 2 - Cover1
International Dentistry - Vol. 12, No. 2 - Cover2
International Dentistry - Vol. 12, No. 2 - Contents
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