Journal Article

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

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

2021

Overview

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-α , Ptgs2 , and Rankl (qPCR). Protein levels of TGF-β 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-α, as well as the protein levels of IL-6 and TNF-α, 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. Clinical relevance The results suggest that site-specific inhibition of MMP-13 may be an interesting strategy to modulate inflammation and reduce bone resorption in osteolytic inflammatory diseases.

• Publisher: Springer Berlin Heidelberg,Springer Nature B.V
• Subject: Animal models / Animals / Bone Resorption / Collagenase 3 / Computed tomography / Dentistry / Enzyme-linked immunosorbent assay / Gum disease / Inflammation / Inflammatory diseases / Interleukin 10 / Interleukin 6 / Kinases / Lipopolysaccharides / MAP kinase / Matrix metalloproteinase / Matrix Metalloproteinase 13 - genetics / Medicine / Metalloproteinase / Mice / Molars / Original Article / Osteolysis / Periodontal Diseases / Periodontitis / Polymerase chain reaction / TRANCE protein / Tumor necrosis factor-α / X-Ray Microtomography