Journal of Peking University(Health Sciences) ›› 2019, Vol. 51 ›› Issue (6): 1108-1114. doi: 10.19723/j.issn.1671-167X.2019.06.023

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Effects of mineral trioxide aggregate and ethanolic extracts of Shandong propolis on the biological properties of human dental pulp fibroblasts

Bing-qing SHI,Xiao-jing YUAN,Yu-ming ZHAO()   

  1. Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
  • Received:2017-12-26 Online:2019-12-18 Published:2019-12-19
  • Contact: Yu-ming ZHAO E-mail:yumingzhao70@sina.com
  • Supported by:
    Supported by the Capital Featured Clinical Application Research Project of Beijing Municipal Science & Technology Commission(Zl51100004015093)

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Abstract:

Objective: To evaluate the effect of mineral trioxide aggregate (MTA) and propolis from Shangdong province on the cell viability, mineralization and migration and anti-inflammatory ability of dental pulp fibroblasts.Methods: The human dental pulp fibroblasts were cultured and subjected to 10 mg/L of propolis and 1 ∶8 dilution of MTA extraction. The cell viability was evaluated with cell counting kit-8 (CCK-8) after 1, 5, 7 and 9 days. The cells in the upper inserts and the test culture media on the bottoms of 24-well plates interacted for 15 hours. Then the numbers of cells migrated through the per-meable membranes were compared. The cells seeded in the 24-well plates were incubated in osteogenic medium with different materials for 21 days and stained with alizarin red S, then photographed. To evaluate the deposition of calcified matrix, the wells were destained with 100 mmol/L cetylpyridinium chloride. Finally, the cells were exposed to 1 mg/L lipopolysaccharide (LPS) to induce an inflammatory response, in the presence of propolis, MTA extraction. The cells were collected after 3 h, and the expressions of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were determined using real-time polymerase chain reaction (real-time PCR). Statistical analysis was performed by one-way ANOVA and nonparametric tests (P<0.05).Results: The cell viability of propolis group was significantly lower than those of MTA and control groups on days 5, 7 and 9, while MTA significantly increased the numbers of the viable cells on days 7 and 9. The migration cells of propolis group (26.67±2.52) were fewer than control group (61.33±4.93), and the cells of MTA group (80.00±2.65) were statistically more than those of the other two groups. The propolis group significantly induced more calcified matrix deposition than MTA group after 21 days of culture. Propolis significantly suppressed the expressions of IL-1β and IL-6 after LPS exposure compared with MTA and control groups.Conclusion: The propolis from Shandong compared with MTA showed a certain degree of cytotoxicity, and had no significant effect on cell migration. On the other hand, propolis exhibited significant anti-inflammatory and mineralization promotion effect, suggesting that the active ingredients of propolis could be introduced as a supplement of pulp capping materials, or used as an irrigant or intracanal medicament due to its excellent anti-inflammatory effect. Propolis may have potential in vital pulp treatment of young permanent tooth suffering pulp inflammation.

Key words: Propolis, Dental pulp, Fibroblasts, Anti-inflammatory effect, Mineral trioxide aggregate

CLC Number: 

  • R788.2

Figure 1

The first passage human dental pulp cells (×4)"

Table 1

Primer sequences of three inflammatory cytokines"

Gene Primer sequences
TNF-α F: CGAGTGACAAGCCTGTAGCC R: TGAAGAGGACCTGGGAGTAGAT
IL-1β F: AGCTCGCCAGTGAAATGATG R: GCCCTTGCTGTAGTGGTGGT
IL-6 F: GAAAGCAGCAAAGAGGCACT R: TTTCACCAGGCAAGTCTCCT

Figure 2

The effect of various concentrations of Shandong propolis (A) and MTA extraction (B) on cell viability in primary culture of dental pulp cells by CCK-8 after 1 or 3 days; the effect of different materials on cell viability in primary culture of dental pulp cells by CCK-8 after 1, 5, 7, 9 days (C) Control, Dulbecco’s modified eagle medium (DMEM) group; sd, Shandong propolis group; MTA, mineral trioxide aggregate (MTA) extraction group. *P<0.05, ** P<0.01, compared with control; #P<0.05, ## P<0.01, compared with MTA."

Figure 2

The effect of various concentrations of Shandong propolis (A) and MTA extraction (B) on cell viability in primary culture of dental pulp cells by CCK-8 after 1 or 3 days; the effect of different materials on cell viability in primary culture of dental pulp cells by CCK-8 after 1, 5, 7, 9 days (C) Control, Dulbecco’s modified eagle medium (DMEM) group; sd, Shandong propolis group; MTA, mineral trioxide aggregate (MTA) extraction group. *P<0.05, ** P<0.01, compared with control; #P<0.05, ## P<0.01, compared with MTA."

Figure 3

The number of cells migrate through permeable membranes in different groups after seeded for 15 hours Abbreviation as in Figure 2. ** P<0.01, compared with control, ## P<0.01, compared with MTA."

Figure 4

Alizarin red staining of cells cultured with different materials for 21 days A, staining of the different groups; B, quantification of alizarin red stain-ing (1 :10 dilution). ** P<0.01, compared with control, ## P<0.01, compared with MTA. DMEM, ordinary DMEM medium group; om, osteogenic medium group; sd, Shandong propolis + osteogenic medium group; MTA, MTA extraction + osteogenic medium group."

Figure 5

The expression of TNF-α, IL-1β and IL-6 in different groups after LPS stimulation for 3 hours Control, LPS group; sd, Shandong propolis + LPS group; MTA, MTA extraction + LPS group. *P<0.05, compared with control, # P<0.05, compared with MTA."

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