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. 2022 Mar 15;15(6):2170.
doi: 10.3390/ma15062170.

The Effects of 3-Dimensional Bioprinting Calcium Silicate Cement/Methacrylated Gelatin Scaffold on the Proliferation and Differentiation of Human Dental Pulp Stem Cells

Affiliations

The Effects of 3-Dimensional Bioprinting Calcium Silicate Cement/Methacrylated Gelatin Scaffold on the Proliferation and Differentiation of Human Dental Pulp Stem Cells

Dakyung Choi et al. Materials (Basel). .

Abstract

A calcium silicate cement/methacrylated gelatin (GelMa) scaffold has been applied in tissue engineering; however, the research on its applications in dental tissue regeneration remains lacking. We investigate the effect of this scaffold on human dental pulp stem cells (hDPSCs). hDPSCs were cultured in 3D-printed GelMa and MTA-GelMa scaffolds. Cell adhesion was evaluated using scanning electron microscopy images. Cells were cultured in an osteogenic differentiation medium, which contained a complete medium or α-MEM containing aqueous extracts of the 3D-printd GelMa or MTA-GelMa scaffold with 2% FBS, 10 mM β-glycerophosphate, 50 μg/mL ascorbic acid, and 10 nM dexamethasone; cell viability and differentiation were shown by WST-1 assay, Alizarin Red S staining, and alkaline phosphatase staining. Quantitative real-time PCR was used to measure the mRNA expression of DSPP and DMP-1. One-way analysis of variance followed by Tukey’s post hoc test was used to determine statistically significant differences, identified at p < 0.05. hDPSCs adhered to both the 3D-printed GelMa and MTA-GelMa scaffolds. There was no statistically significant difference between the GelMa and MTA-GelMa groups and the control group in the cell viability test. Compared with the control group, the 3D-printed MTA-GelMa scaffold promoted the odontogenic differentiation of hDPSCs. The 3D-printed MTA-GelMa scaffold is suitable for the growth of hDPSCs, and the scaffold extracts can better promote odontoblastic differentiation.

Keywords: bioprinting; gelatin; mineral trioxide aggregates (MTA); scaffold.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
(A,B) Design of the 3D scaffold using the Organ Regenerator Program. (C) A photo image of the 3D scaffolds is a figure.
Figure 2
Figure 2
(AC) SEM images of the 3D-printed GelMa scaffolds. (DF) SEM images of the 3D-printed GelMa/ProRoot MTA scaffolds. (GI) SEM images of the 3D-printed GelMa/Endocem Zr scaffolds. (A,D,G) ×50 magnification, (B,E,H) ×1000 magnification, (C,F,I) ×3000 magnification.
Figure 3
Figure 3
(AC) Adhesion of hDPSCs on the 3D-printed GelMa scaffolds. (DF) Adhesion of hDPSCs on the 3D-printed GelMa/ProRoot MTA scaffolds. (GI) Adhesion of hDPSCs on the 3D-printed GelMa/Endocem Zr scaffolds. (A,D,G) ×500 magnification, (B,E,H) ×1000 magnification, (C,F,I) ×3000 magnification.
Figure 4
Figure 4
The cell viability of hDPSCs exposed for 24 h to extracts of the test materials as measured by the WST-1 assay. There was no significant difference in the cell viability of the GelMa and MTA-GelMa scaffold groups compared to the control (p > 0.05).
Figure 5
Figure 5
The mRNA expression level of odontogenic markers was determined by quantitative real-time PCR. The expression of (A) DSPP and (B) DMP-1 in hDPSCs stimulated with the extraction of scaffolds was determined. Different lower-case letters represent statistically significant differences (p < 0.05).
Figure 6
Figure 6
(A) ALP activity of hDPSCs cultured on the extracts of scaffolds for 7 days. (B) ALP staining of hDPSCs cultured on the extracts of scaffolds for 7 days. * Statistically significant difference (p < 0.05) compared to the control group.
Figure 7
Figure 7
(A) Quantification of calcium mineral deposits of hDPSCs cultured on the extracts of scaffolds for 14 days. (B) Alizarin Red S staining of hDPSCs cultured on the extracts of scaffolds for 14 days. There was no significant difference in the calcium deposition of the GelMa and MTA-GelMa scaffold groups compared to the control (p > 0.05).

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