Expression of matrix metalloproteinase‐3 and ‐10 is up‐regulated in the periodontal tissues of aged mice

Abstract

Objective The present study was designed to investigate the whole transcriptome of periodontal tissues of both young and aged mice to identify the characteristic up‐regulation of protease genes with aging and to localize their translated protein products in the periodontal tissues. Background The metzincin protease superfamily is composed of matrix metalloproteinases (MMPs), a disintegrin and metalloproteinases, and a disintegrin and metalloproteinases with thrombospondin motifs. Up‐regulation of these extracellular matrix‐degrading proteases has been implicated in senescence of tissues and organs, including the skin. However, few studies have investigated the expression profiles of these proteases and potential involvement in aging of periodontal tissues. Methods Periodontal tissues with the surrounding mandibular bones were collected from 50‐ and 10‐week‐old mice. Total RNA was extracted from the periodontal tissue and analyzed by cap analysis of gene expression (CAGE) to identify differentially expressed genes encoding the metzincin proteases. Furthermore, quantitative real‐time polymerase chain reaction (qRT‐PCR) was performed to validate the CAGE results, and the phenotypic expression of proteases involved in aging was localized via immunohistochemical analysis. Results The CAGE results showed that the expression levels of MMP‐3, ‐10, and ‐12 were up‐regulated at 50 weeks. Subsequent qRT‐PCR analysis showed that the gene expression levels of MMP‐3 and ‐10 were significantly increased with age. MMP‐10 immunoreactivity was localized exclusively in the cementum and alveolar bone adjacent to the periodontal ligament and was stronger and broader in aged mice than young mice. MMP‐3 immunoreactivity was localized in the periodontal ligaments at both 10 and 50 weeks. Conclusion In the present study, we demonstrated that the expression of MMP‐3 and ‐10 increased with aging and identified their characteristic localizations in aged periodontal tissues.

Age-Related Gene and Protein Expression in Mouse Mandibular Condyle Analyzed by Cap Analysis of Gene Expression and Immunohistochemistry

“…In the aged mandibular condyle as well as in the joints of patients with OA and RA, the release of proinflammatory cytokines is increased due to inflammaging and may promote the secretion of MMP-3, which might then facilitate the degradation of the articular cartilage. The gene expression of MMP-3 has been also shown to be elevated in other aged connective tissues, such as human skin dermis, human gingiva, and mouse periodontal tissues [13, 24, 25], which supports the idea that MMP-3 is a crucial age-related factor. In our immunohistochemical investigation, MMP-3 immunoreactivity was detected in the medial and lateral parts of the aged MCC, not in the middle part.…”

“…The total RNA extracted from the mandibular condyle samples of 10- and 50-week-old mice (two biological replicates) was used for CAGE. The quality of the total RNA was examined, and CAGE was performed using DNAFORM (Yokohama, Japan), as described in our previous report [13]. Briefly, a CAGE library was prepared and sequenced using an Illumina NextSeq 500 System (Illumina, San Diego, CA, USA).…”

Impact of Inflammatory Markers and Senescence-Associated Secretory Phenotype in the Gingival Crevicular Fluid on the Outcomes of Periodontal Regeneration

“…Mainly released by neutrophil and macrophage populations [ 39 ], MMPs are regarded as the effectors of the unbalanced matrix degradation during the active phases of periodontitis, being useful diagnostic and prognostic markers for periodontitis [ 40 ]. These zinc-dependent proteases were also detected at higher levels in aged periodontal tissues, being a potential proxy for the local inflammaging status of tissues [ 41 ]. Finally, IL-12 was found to be positively correlated with PPD reduction in our study.…”

Development and Characterization of a Novel FVB-PrkdcR2140C Mouse Model for Adriamycin-Induced Nephropathy

“…The reaction was analyzed using CFX Maestro (Bio-Rad Laboratories Inc., Hercules, CA, USA). The PCR primer sequences used in this study were acquired from previous studies [21][22][23][24][25][26][27] and are listed in Table 1.…”