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Comparative Study
. 2014 Jul 11:14:87.
doi: 10.1186/1472-6831-14-87.

Physical properties and biological/odontogenic effects of an experimentally developed fast-setting α-tricalcium phosphate-based pulp capping material

Jun-Bong Lee, Su-Jung Park, Hyun-Ha Kim, Young-Sun Kwon, Kwang-Won Lee, Kyung-San Min  1
Affiliations
Comparative Study

Physical properties and biological/odontogenic effects of an experimentally developed fast-setting α-tricalcium phosphate-based pulp capping material

Jun-Bong Lee et al. BMC Oral Health. .

Abstract

Background: Recently, fast-setting α-tricalcium-phosphate (TCP) cement was developed for use in the pulp capping process. The aim of this study was to investigate the physical properties and biological effects of α-TCP cement in comparison with mineral trioxide aggregate (MTA).

Methods: We measured the setting time, pH values, compressive strength, and solubility of the two materials. We evaluated biocompatibility on the basis of cell morphology and a viability test using human dental pulp cells (hDPCs). Chemical composition of each material was analyzed by energy dispersive x-ray spectroscopic (EDS) analysis. The expression of odontogenic-related genes was evaluated by Western blotting and immunofluorescence. The calcified nodule formation was measured by Alizarin red staining. We performed the pulp capping procedure on rat teeth for histological investigation. The data were analyzed by an independent t-test for physical properties, one-way ANOVA for biological effects, and the Mann-Whitney U test for tertiary dentin formation. A P value of less than 0.05 was considered statistically significant for all tests.

Results: The setting time, pH values, and compressive strength of α-TCP was lower than that of MTA (P < 0.05); however, the solubility of α-TCP was higher than that of MTA (P < 0.05). The resultant cell viability observed with the two materials was similar (P > 0.05). Scanning electron microscopy (SEM) revealed that cells attached to both materials were flat and had cytoplasmic extensions. The expression of odontogenic-related markers and mineralized nodule formation were higher in the two experimental groups compared to the control group (P < 0.05). Continuous tertiary dentin was formed underneath the capping materials in all samples of the tested groups.

Conclusions: Our study demonstrated that the α-TCP exhibited biocompatibility and odontogenicity comparable to MTA, whereas it had a quicker setting time.

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Figures

Figure 1
Figure 1
The physical properties and cell viability of the tested materials. The pH values (A), solubility (B), and compressive strength (C) of MTA and TCP. Note that the setting time, pH values, and compressive strength of α-TCP was lower than that of MTA whereas the solubility of α-TCP was higher than that of MTA. (D) Effects of MTA and TCP on cell viability measured by MTT assay. The cell viability of α-TCP-treated samples was higher than those of MTA at day 14. *Significant difference between each group; P < 0.05.
Figure 2
Figure 2
Investigation of cell morphology and chemical composition of the materials. SEM observation of cells incubated for 3 days on (A) MTA (×1000) and (B) TCP (×1000). Both groups showed flattened cells in close proximity to one another, and these were seen to be spreading across the substrate. EDS analysis of the samples: (C) MTA and (D) TCP.
Figure 3
Figure 3
Effects of the tested materials on odontoblastic differentiation of hDPCs. (A) Effects of MTA and TCP on DSPP, DMP1, and ON protein in hDPCs. (B) The graph shows the quantification of protein expression by densitometry and is presented as fold increases compared with control cells. (C) Effects of MTA and TCP on the formation of calcification nodules in hDPCs. *Significant difference between each group; P < 0.05.
Figure 4
Figure 4
Immunofluorescence analysis of hDPCs treated with medium only (A, D, and G), MTA (B, E, and H), or TCP (C, F, and I). Fluorescence images showing anti-DSPP (A-C), anti-DMP1 (D-F), and anti-ON (G-I) signals (green) of cells after 3 days of culture (×400). Note that the protein signals in the cells of the experimental groups were stronger than those in the cells of the control group.
Figure 5
Figure 5
Histological observation. Capped pulps stained with hematoxylin-eosin 4 weeks after treatment with MTA (A) and TCP (B) (×50). (C) A specimen in the control group capped only with glass ionomer cement. (D and E) Higher magnification of boxed areas shown in A and B (×400), respectively. Odontoblasts (arrowheads) are polarized and appear to be arranged in a palisade pattern. *Reparative tertiary dentin formed underneath the capping materials.
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References

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