Dental pulp cells cocultured with macrophages aggravate the inflammatory conditions stimulated by LPS

Min-Ching Wang; Kuo-Wei Chang; Shu-Chun Lin; Ling-Hsin Hsu; Pei-Shih Hung

doi:10.1186/s12903-023-03625-4

Dental pulp cells cocultured with macrophages aggravate the inflammatory conditions stimulated by LPS

BMC Oral Health. 2023 Dec 9;23(1):991. doi: 10.1186/s12903-023-03625-4.

Authors

Min-Ching Wang^{1

2

3}, Kuo-Wei Chang^{2

4}, Shu-Chun Lin^{2

4}, Ling-Hsin Hsu⁵, Pei-Shih Hung^{6

7}

Affiliations

¹ Division of Pediatric Dentistry, Department of Dentistry, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan.
² Department of Dentistry, National Yang Ming Chiao Tung University, Taipei, Taiwan.
³ Department of Stomatology, Taipei Veterans General Hospital, Taipei, Taiwan.
⁴ Institute of Oral Biology, School of Dentistry, National Yang Ming Chiao Tung University, Taipei, Taiwan.
⁵ Department of Dentistry, Taipei City Hospital, Taipei, Taiwan.
⁶ Institute of Oral Biology, School of Dentistry, National Yang Ming Chiao Tung University, Taipei, Taiwan. g39017005@gmail.com.
⁷ Department of Medical Research, National Yang Ming Chiao Tung University Hospital, Yilan, Taiwan. g39017005@gmail.com.

Abstract

Background: Pulp inflammation is complex interactions between different types of cells and cytokines. To mimic the interactions of different types of cells in inflamed dental pulp tissues, dental pulp cells (DPCs) were cocultured with different ratios of macrophages (THP-1) or LPS treatment.

Methods: DPCs were cocultured with various ratios of THP-1, then photographed cell morphology and determined cell viability by MTT assay at preset times. Total RNA was also extracted to measure the inflammation marker-IL-6 and IL-8 expressions by RT-Q-PCR. The DPCs and THP-1 were treated with 0.01 - 1μg/ml lipopolysaccharide (LPS) and extract RNA at preset times, and detected IL-6 and IL-8 expression. DPCs were cocultured with various ratios of THP-1 with 0.1 μg/mL LPS, and detected IL-6 and IL-8 expression after 24 and 48 h. The data were analyzed by unpaired t-test or Mann-Whitney test. Differences were considered statistically significant when p < 0.05.

Results: THP-1 and DPCs coculture models did not suppress the viability of DPCs and THP-1. Cocultured with various ratios of THP-1 could increase IL-6 and IL-8 expressions of DPCs (p = 0.0056 - p < 0.0001). The expressions of IL-6 and IL-8 were stronger in higher ratio groups (p = 0.0062 - p < 0.0001). LPS treatment also induced IL-6 and IL-8 expressions of DPCs and THP-1 (p = 0.0179 - p < 0.0001 and p = 0.0189 - p < 0.0001, separately). Under the presence of 0.1 μg/mL LPS, DPCs cocultured with THP-1 for 24 h also enhanced IL-6 and IL-8 expression (p = 0.0022). After cocultured with a higher ratio of THP-1 for 48 h, IL-6 and IL-8 expressions were even stronger in the presence of LPS (p = 0.0260).

Conclusions: Coculturing dental pulp cells and macrophages under LPS treatment aggravate the inflammatory process. The responses of our models were more severe than traditional inflamed dental models and better represented what happened in the real dental pulp. Utilizing our models to explore the repair and regeneration in endodontics will be future goals.

Keywords: Coculture; Dental pulp cells (DPCs); Inflammation; LPS; Macrophage.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Coculture Techniques
Dental Pulp*
Humans
Inflammation
Interleukin-6 / metabolism
Interleukin-8 / metabolism
Lipopolysaccharides* / metabolism
Lipopolysaccharides* / pharmacology
Macrophages
RNA / metabolism

Substances

Lipopolysaccharides
Interleukin-6
Interleukin-8
RNA