Silencing matrix metalloproteinase-13 (Mmp-13) reduces inflammatory bone resorption associated with LPS-induced periodontal disease in vivo

Author(s)

Guimaraes-Stabili, Morgana R. ; de Medeiros, Marcell Costa ; Rossi, Danuza ; Camilli, Angelo Constantino ; Zanelli, Cleslei Fernando ; Valentini, Sandro Roberto ; Spolidorio, Luis Carlos ; Kirkwood, Keith Lough ; Rossa, Carlos, Jr.

• Document type: Journal article
• Source: CLINICAL ORAL INVESTIGATIONS; v. 25, n. 5, p. 12-pg., 2020-11-02.

Abstract

Objectives The aim of this study was to evaluate the effect of specific inhibition of MMP-13 on inflammation and inflammatory bone resorption in a murine model of lipopolysaccharide (LPS)-induced periodontitis. Materials and methods Periodontitis was induced in mice by micro-injections of LPS into the gingival tissues adjacent to the palatal surfaces of maxillary molars twice a week for 15 days. Matrix metalloproteinase-13 (Mmp-13) shRNA or a specific biochemical inhibitor were also injected into the same sites in alternating days with the LPS injections. Efficacy of shRNA-mediated silencing of Mmp-13 was verified by quantitative real-time polymerase chain reaction (qPCR) and immunoblot. Bone resorption was assessed by microcomputed tomography (uCT). Histological sections stained with hematoxylin/eosin (H/E) were used in the stereometric analysis of the inflammatory infiltrate. Gingival tissues were used to evaluate expression of Mmp-13, Il-6, Tnf-alpha, Ptgs2, and Rankl (qPCR). Protein levels of TGF-beta and IL-10 in the tissues were determined by enzyme-linked immunosorbent assays (ELISA) or by MMP-13 and p38 immunoblot. Results Silencing Mmp-13 expression reduced bone resorption significantly. Expression of Mmp-13, Il-6, and Tnf-alpha, as well as the protein levels of IL-6 and TNF-alpha, was reduced in the animals treated with adenovirus-delivered shRNA; however, these effects were not associated with modulation of p38 MAPK signaling. Interestingly, inhibition Mmp-13 did not affect the severity of inflammatory infiltrate. Conclusions Site-specific inhibition of MMP-13 reduced bone resorption and production of inflammatory mediators associated with periodontal disease. (AU)

• FAPESP's process: 07/05583-3 - Inhibition of MMP-13 and p38 MAPK gene expression by adenoviral-delivered shRNA in a murine model of experimentally induced periodontal disease
• Grantee: Luis Carlos Spolidorio
• Support Opportunities: Regular Research Grants