Resolvin D1 and periodontal ligament fibroblast function
Date
2013
DOI
Authors
Zarrough, Ahmed Elmagtouf
Version
OA Version
Citation
Abstract
BACKGROUND AND OBJECTIVES: Periodontitis is an irreversible destruction of the tooth supporting tissues initiated by infection and mediated by the host's immune system. Periodontal ligament fibroblasts are capable of producing and releasing cytokines that participate in the local inflammatory response triggered by bacteria. Resolution of inflammation is currently considered an active process that is mediated by specialized lipid mediators (lipoxins and resolvins). The purpose of the study is to test the actions of resolvin D1 (RvD1) on human periodontal ligament fibroblasts (hPDLFs).
MATERIALS AND METHODS: The expression of RvD1 receptors (GPR32 and ALX/FPR2) on hPDLFs was assessed using QPCR, immunofluorescence, and FACS. hPDLFs were stimulated with IL-1[beta] (500 pg/mL) in the presence or absence of RvDl (100 nM). Cell migration and proliferation were measured using scratch assay and BrdU proliferation assay, respectively. The production of IL-6, IL-8, MCP-1, MMP-1, MMP-2, MMP-3, TIMP-1, and TIMP-2 were measured using multiplex immunoassay. The production of PGE2 and OPG were measured using ELISA. The mRNA levels of IL-6, IL-8, MCP-1, Cox-2, MMP-1, MMP-2, MMP-3, TIMP-1, TIMP-2, and OPG were examined using real time QPCR. The intracellular protein levels of IL-6, IL-8, MCP-1, COX-2, and OPG were assessed using western blotting, whereas the amounts of intracellular MMP-1, MMP-2, MMP-2, TIMP-1, and TIMP-2 proteins were determined by multiplex immunoassays.
RESULTS: hPDLF cells expressed GPR32 and ALX/FPR2 genes and proteins. RvDl counteracted the inhibitory actions of IL-l[beta] on hPDLF cell migration and proliferation. RvDl reversed the IL-l[beta]-induced increase in the production of IL-6, IL-8, MCP-1, PGE2, MMP-1, and MMP-3 by hPDLFs. RvDl reduced MMP-2 production only when cells were challenged with IL-l[beta]. TIMP-1 production was increased when RvDl was added to IL-l[beta] stimulated hPDLFs. RvDl reversed the IL-l[beta]-induced inhibition in TIMP-2 and OPG production by hPDLFs. While treating with IL-l[beta] upregulated the genes for IL-6, IL-8, MCP- 1, COX-2, MMP-1, and MMP-3, addition of RvDl did not alter this IL-l[beta]-induced upregulation; however, neither IL-l[beta] nor RvDl affected the gene expression profiles of MMP-2, TIMP-1, TIMP-2, and OPG. Interestingly, RvDl rescued the IL-l[beta]-induced stimulation in the intracellular protein levels of IL-6, IL-8, MCP-1, MMP-1, and MMP-3; however, RvDl did not alter the IL-l[beta]-induced stimulation in intracellular production of COX-2. RvDl also rescued the IL-l[beta]-induced inhibition in the intracellular protein levels of TIMP-2 and OPG, but neither IL-l[beta] nor RvDl altered those of MMP-2 and TIMP-1.
CONCLUSION AND CLINICAL SIGNIFICANCE: Our data suggest that RvDl plays key pro resolution and anti-inflammatory roles promoting wound healing and regeneration of periodontal ligament tissues during periodontal ligament disease by regulating hPDLF functions. These data may contribute to the development of novel treatment modalities of periodontal disease by exploiting the anti-inflammatory and pro-resolution properties of RvDl.
Description
Dissertation (DScD) --Boston University, Henry M. Goldman School of Dental Medicine, 2013 (Department of Oral Biology and Periodontology).
Includes bibliographic references: leaves 112-131.
Includes bibliographic references: leaves 112-131.
License
This work is being made available in OpenBU by permission of its author, and is available for research purposes only. All rights are reserved to the author.