Bioactivity of endodontic biomaterials on dental pulp stem cells through dentin

Bahar Javid; Narges Panahandeh; Hassan Torabzadeh; Hamid Nazarian; Ardavan Parhizkar; Saeed Asgary

doi:10.5395/rde.2018.45.e3

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

Research Article

Restor Dent Endod 2020;45(1):e3.

Published online: 4 February 2020

DOI: https://doi.org/10.5395/rde.2018.45.e3

Bioactivity of endodontic biomaterials on dental pulp stem cells through dentin

Bahar Javid¹, Narges Panahandeh^1,^2,^*, Hassan Torabzadeh², Hamid Nazarian³, Ardavan Parhizkar⁴, Saeed Asgary⁴

¹Department of Restorative Dentistry, School of Dentistry, Shahid Beheshti University of Medical Sciences, Tehran, Iran

²Dental Research Center, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran

³Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

⁴Iranian Center for Endodontic Research, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran

^*Correspondence to Narges Panahandeh, DDS, MS Assistant Professor, Dental Research Center, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Daneshjoo Blvd., Velenjak, Tehran 1983963113, Iran. E-mail: nargespanahandeh@yahoo.com

Received 17 June 2019 Revised 14 August 2019 Accepted 14 September 2019

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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.

Keywords: Calcium-enriched mixture cement, Dental pulp, Dental pulp capping, Endodontics, Mineral trioxide aggregate, Stem cells

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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.

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.

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.

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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