Journal List > Restor Dent Endod > v.45(1) > 1148378

Javid, Panahandeh, Torabzadeh, Nazarian, Parhizkar, and Asgary: Bioactivity of endodontic biomaterials on dental pulp stem cells through dentin

Abstract

Objectives

This study investigated the indirect effect of calcium-enriched mixture (CEM) cement and mineral trioxide aggregate (MTA), as 2 calcium silicate-based hydraulic cements, on human dental pulp stem cells (hDPSCs) through different dentin thicknesses.

Materials and Methods

Two-chamber setups were designed to simulate indirect pulp capping (IPC). Human molars were sectioned to obtain 0.1-, 0.3-, and 0.5-mm-thick dentin discs, which were placed between the 2 chambers to simulate an IPC procedure. Then, MTA and CEM were applied on one side of the discs, while hDPSCs were cultured on the other side. After 2 weeks of incubation, the cells were removed, and cell proliferation, morphology, and attachment to the discs were evaluated under scanning electron microscopy (SEM). Energy-dispersive X-ray (EDXA) spectroscopy was performed for elemental analysis. Alkaline phosphatase (ALP) activity was assessed quantitatively. The data were analyzed using the Kruskal-Wallis and Mann-Whitney tests.

Results

SEM micrographs revealed elongated cells, collagen fibers, and calcified nucleations in all samples. EDXA verified that the calcified nucleations consisted of calcium phosphate. The largest calcifications were seen in the 0.1-mm-thick dentin subgroups. There was no significant difference in ALP activity across the CEM subgroups; however, ALP activity was significantly lower in the 0.1-mm-thick dentin subgroup than in the other MTA subgroups (p < 0.05).

Conclusions

The employed capping biomaterials exerted biological activity on hDPSCs, as shown by cell proliferation, morphology, and attachment and calcific precipitations, through 0.1- to 0.5-mm-thick layers of dentin. In IPC, the bioactivity of these endodontic biomaterials is probably beneficial.

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Figure 1.
The setup for the experiment. (A) Separated view; (B) Assembled view. The setup had 1) 2 plexiglass rectangular chambers for the placement of materials/cells and 2) a hole (red arrowhead) for placement of the dentin discs.
rde-45-e3f1.tif
Figure 2.
Scanning electron micrographs of calcium-enriched mixture cement (left column) with (A) 0.1 mm, (C) 0.3 mm, and (E) 0.5 mm dentin thicknesses and mineral trioxide aggregate cement (right column) with (B) 0.1 mm, (D) 0.3 mm, and (F) 0.5 mm dentin thicknesses. White arrowheads indicate elongated cells and blue arrow heads show calcified nodules.
rde-45-e3f2.tif
Figure 3.
Energy-dispersive X-ray spectroscopy of the (A) Calcium-enriched mixture and (B) Mineral trioxide aggregate groups with 0.1-mm-thick dentin. Calcium and phosphorus ions were the predominant ions in the experimental groups.
rde-45-e3f3.tif
Table 1.
Alkaline phosphatase (ALP) activity at various concentrations
Group Activity 30 Activity 40 Activity 50 Activity 60 Activity 80
CEM 0.1 (n = 4) 0.0012 ± 0.0006 (14.88) a 0.0012 ± 0.0004 (13.75) a 0.0012 ± 0.0005 (14.00) a 0.0013 ± 0.0006 (14.88) a,c 0.0012 ± 0.0005 (13.75) a
CEM 0.3 (n = 3) 0.0018 ± 0.0003 (17.50) a 0.0018 ± 0.0003 (18.00) a,b 0.0018 ± 0.0002 (17.33) a,b 0.0018 ± 0.0001 (16.67) a,c 0.0019 ± 0.0003 (17.50) a,b
CEM 0.5 (n = 4) 0.0007 ± 0.0014 (10.25) a,b,c 0.0005 ± 0.0011 (10.50)a,c,d 0.0007 ± 0.0013 (10.25) a,c,d 0.0006 ± 0.0011 (11.25) a,c 0.0007 ± 0.0013 (10.50) a,c,d
MTA 0.1 (n = 5) 0.0000 ± 0.0000 (5.00) b 0.0000 ± 0.0000 (5.00) c,e 0.0000 ± 0.0000 (4.50) c 0.0000 ± 0.0000 (4.00) b 0.0000 ± 0.0000 (4.00) c
MTA 0.3 (n = 4) 0.0002 ± 0.0003 (8.25) b 0.0004 ± 0.0008 (9.50) a,e 0.0005 ± 0.0010 (11.13) a,e 0.0005 ± 0.0009 (10.00) c 0.0006 ± 0.0010 (11.25) a,e
MTA 0.5 (n = 3) 0.0027 ± 0.0006 (21.67) c 0.0024 ± 0.0007 (20.67) b,d 0.0022 ± 0.0006 (20.00) b,d,e 0.0022 ± 0.0005 (20.50) a 0.0025 ± 0.0006 (20.50) b,d,e

The values are presented as mean ± standard deviation, and the numbers in parentheses are the mean ranks in the group. Activity 30, activity 40, activity 50, activity 60, and activity 80 refer to 30%, 40%, 50%, 60%, and 80% levels of ALP activity, respectively.

Different lowercase letters in the same column indicate the presence of statistically significant differences.

CEM, calcium-enriched mixture; MTA, mineral trioxide aggregate.

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