2. Academic Validation
  3. Targeting the Nod-like receptor protein 3 Inflammasome with inhibitor MCC950 rescues lipopolysaccharide-induced inhibition of osteogenesis in Human periodontal ligament cells

Targeting the Nod-like receptor protein 3 Inflammasome with inhibitor MCC950 rescues lipopolysaccharide-induced inhibition of osteogenesis in Human periodontal ligament cells

  • Arch Oral Biol. 2021 Nov;131:105269. doi: 10.1016/j.archoralbio.2021.105269.
Wei Peng 1,  Bo Zhang 2,  Zhengfan Sun 3,  Meifeng Zhang 4,  Ling Guo 5
Affiliations

Affiliations

  • 1 Department of prosthodontics, Hospital of Stomatology Affiliated to Southwest Medical University, Luzhou, Sichuan, China. Electronic address: [email protected].
  • 2 Department of prosthodontics, Hospital of Stomatology Affiliated to Southwest Medical University, Luzhou, Sichuan, China. Electronic address: [email protected].
  • 3 Department of prosthodontics, Hospital of Stomatology Affiliated to Southwest Medical University, Luzhou, Sichuan, China. Electronic address: [email protected].
  • 4 Department of prosthodontics, Hospital of Stomatology Affiliated to Southwest Medical University, Luzhou, Sichuan, China. Electronic address: [email protected].
  • 5 Department of prosthodontics, Hospital of Stomatology Affiliated to Southwest Medical University, Luzhou, Sichuan, China. Electronic address: [email protected].
PMID: 34601319 DOI: 10.1016/j.archoralbio.2021.105269
Abstract

Objective: We aim to investigate whether lipopolysaccharide-stimulated activition of Nod-like receptor protein 3 (NLRP3) Inflammasome inhibits osteogenesis in Human periodontal ligament cells (HPDLCs). Futhermore, to study whether MCC950 (a inhibitor of NLRP3 Inflammasome) rescues lipopolysaccharide-induced inhibition of osteogenesis in HPDLCs as well as the underlying mechanisms.

Methods: HPDLCs were isolated from periodontal ligament of healthy orthodontic teeth from teenagers, and cells surface marker protein were detected by flow cytometry. Cells viability were determined by Cell Counting kit 8 assay. Enzyme-linked immunosorbent assay was used to analyze the secretion of proinflammatory factors. Western blot and real-time quantitative polymerase chain reaction (RT-qPCR) were measured assessing the expression of NLRP3 and Caspase-1. RT-qPCR, Alizarin red staining and Alkaline Phosphatase staining were tested to determine the osteogenic differentiation capacity of HPDLCs.

Results: It was found that lipopolysaccharide in the range of concentrations from 10 to 100 μg/ml significantly inhibited HPDLCs viability at 24 h and significantly improved proinflammatory cytokine expressions at 8 h and 24 h. MCC950 reversed lipopolysaccharide-stimulated proinflammatory cytokine expressions including interleukin-1β and interleukin-18, but not tumor necrosis factor-α. In addition, MCC950 rescued the lipopolysaccharide-inhibited osteogenic gene (Alkaline Phosphatase, Runt-related transcription factor 2, and Osteocalcin). Moreover, MCC950 downregulated lipopolysaccharide-induced relative protein of NLRP3 Inflammasome signaling pathway, such as NLRP3 and Caspase-1.

Conclusion: MCC950 rescues lipopolysaccharide-induced inhibition of osteogenesis in HPDLCs via blocking NLRP3 Inflammasome signaling pathway, and it may be used as a promising therapeutic agent for periodontitis or periondontal regenerative related disease.

Keywords

Human periodontal ligament cells MCC950; Lipopolysaccharide; Nod-like receptor protein 3; Osteogenesis; Periodontitis.

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