Yeungnam Univ J Med.  2020 Jul;37(3):217-225. 10.12701/yujm.2020.00248.

Mineralization-inducing potentials of calcium silicate-based pulp capping materials in human dental pulp cells

• Kang S

• KMID: 2506521
• DOI: http://doi.org/10.12701/yujm.2020.00248

Abstract

Background
To provide a long-term bacterial seal through the formation of reparative dentin bridge, calcium silicate-based pulp capping materials have been used at sites of pulpal exposure. The aim of this study was to evaluate the mineralization-inducing potentials of calcium silicate-based pulp capping materials (ProRoot MTA [PR], Biodentine [BD], and TheraCal LC [TC]) in human dental pulp cells (HDPCs).
Methods
Specimens of test materials were placed in deionized water for various incubation times to measure the pH variation and the concentration of calcium released. The morphology of HDPCs cultured on the specimens was examined using a confocal laser scanning microscope (CLSM). Alizarin red S staining and alkaline phosphatase assays were used to evaluate mineralization-inducing potentials of the capping materials.
Results
BD showed the highest calcium release in all test periods, followed by PR and TC. (p<0.05). All experimental groups showed high alkalinity after 1 day, except at 14 days. BD showed the highest cell viability compared with PR and TC after 1 and 3 days, while TC showed the lowest value (p<0.05). The CLSM analysis showed that cells were well adhered and expressed actin filaments for all pulp capping materials. Mineralization by PR and BD groups was higher than that by TC group based on alizarin red S staining. BD showed significantly higher alkaline phosphatase activity than PR and TC, while TC showed the lowest value (p<0.05).
Conclusion
Within the limitations of the in vitro study, BD had higher mineralization-inducing potential than PR and TC.

Keyword

Biocompatibility; Biodentine; Calcium silicate material; ProRoot mineral trioxide aggregate

Figure

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  Fig. 1. (A) Stainless-steel frame with five holes, (B) schematic diagram of the specimen, (C) 48-well cell culture plates, having flat bottom which matches the specimen in size.

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  Fig. 2. Images of specimens. (A) ProRoot MTA, (B) Biodentine, (C) TheraCal LC.

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  Fig. 3. The amount of calcium released from ProRoot MTA, Biodentine, and TheraCal LC in deionized water as a function of immersion time. The amount of calcium released from ProRoot MTA and Biodentine increased constantly with the immersion time, whereas the released calcium from TheraCal LC had a tendency to increase until day 4, and almost stopped increasing. For each of the three types of samples, five samples were used at each time of the experiment. Error bars indicate standard errors of the means.

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  Fig. 4. pH values of aqueous medium exposed to the extracts of the capping materials as a function of immersion time. pH values for ProRoot MTA, Biodentine, and TheraCal LC were stable in near 11.0 after 1 day, except at 14 days. Values not sharing a common letter (a, b) are significantly different (p<0.05). For each of the three types of samples, five samples were used at each time of the experiment. Error bars indicate standard errors of the means.

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  Fig. 5. Effects of ProRoot MTA, Biodentine, and TheraCal LC on cell viability measured by MTT assay. On both 1 and 3 days, the cell viability for Biodentine was highest. However, ProRoot MTA and TheraCal LC showed no significant difference (p<0.05). For each of the three types of samples, six samples were used at each time of the experiment. Error bars indicate standard errors of the means. MTT OD value: cell viability absorbance (490 nm).

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  Fig. 6. Confocal laser scanning microscopic images (actin [red], nucleus [blue]) of HDPCs cultured on (A) ProRoot MTA, (B) Biodentine, and (C) TheraCal LC which were incubated in the extract of the materials for 3 days. Cells were well adhered and expressed actin filaments for all pulp capping materials (×200).

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  Fig. 7. Alizarin red S staining images of odontoblasts cultured on (A) ProRoot MTA, (B) Biodentine, and (C) TheraCal LC for 14 days. There was a high increase in mineralization in the ProRoot MTA and Biodentine groups compared with the TheraCal LC group based on alizarin red S staining for calcium.

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  Fig. 8. Alkaline phosphatase (ALP) activity of odontoblasts cultured on pulp capping materials for 14 and 21 days. The ALP activity of odontoblasts in Biodentine group was highest on both observation days. However, the odontblasts in TheraCal LC group showed the lowest ALP activity (p<0.05). For each of the three types of samples, five samples were used at each time of the experiment. Values not sharing a common letter (a, b, c) are significantly different (p<0.05). Error bars indicate standard errors of the means.

Reference

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