Differential regulation of MMP-13 by chemical modified tetracyclines in osteoblasts.

Abstract

Tetracyclines have been shown to regulate matrix metalloproteinase (MMP) expression in numerous cell types with various periodontal disease models. MMP-13, or collagenase-3, has been shown to be induced by a number of osteotropic cytokines and hormones in osteoblastic cells. In this study, we studied MMP-13 gene expression and regulation in osteoblasts by chemically modified tetracyclines (CMTs). Preliminary cytotoxicity studies indicated that 1-10 microg/ml of CMT-8 did not result in statistically significant cell death. Additional fluorescent microscopy experiments indicated that CMT-8 but not CMT-5 had a nuclear distribution within one hour of addition CMTs. Using primary rat calvarial osteoblastic cells obtained from 21-day old neonatal rats, we determined MMP-13 gene expression when stimulated by parathyroid hormone (PTH), interleukin (IL)-1b, or tumour necrosis factor (TNF)-alpha in the presence or absence of CMT-8 or -5. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) using total RNA was used to determine relative expression of MMP-13 compared to glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a constitutively expressed housekeeping gene. Results indicate that all agents were consistently able to induce MMP-13 expression in primary calvarial osteoblastic cells. However, CMT-8 inhibited MMP-13 expression from IL-1b and PTH stimulated cells while having little effect on TNF-alpha stimulated MMP-13 expression. In contrast, CMT-5 only inhibited PTH stimulated MMP-13 expression, with no effect on IL-1b or TNF-alpha stimulated MMP-13 expression. These results suggest that CMT-8 and CMT-5 may differentially affect cell signaling pathways in osteoblastic cells that mediate MMP-13 gene expression.

• Authors: Keith Kirkwood, Thomas Martin, Kimberly Agnello, Young Joon Kim
• Journal: Journal of the International Academy of Periodontology (J Int Acad Periodontol) Vol. 6 Issue 2 Pg. 39-46 (Apr 2004) ISSN: 1466-2094 [Print] England
• PMID: 15125014 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• CMT-5 compound
• Interleukin-1
• Parathyroid Hormone
• Tetracyclines
• Tumor Necrosis Factor-alpha
• tetracycline CMT-8
• Glyceraldehyde-3-Phosphate Dehydrogenases
• Collagenases
• Matrix Metalloproteinase 13
• Mmp13 protein, rat

Topics

• Animals
• Animals, Newborn
• Cell Nucleus (drug effects, metabolism)
• Cell Survival (drug effects)
• Cells, Cultured
• Collagenases (drug effects, genetics)
• Gene Expression Regulation, Enzymologic (drug effects)
• Glyceraldehyde-3-Phosphate Dehydrogenases (drug effects, genetics)
• Interleukin-1 (pharmacology)
• Matrix Metalloproteinase 13
• Microscopy, Fluorescence
• Osteoblasts (drug effects, enzymology)
• Parathyroid Hormone (pharmacology)
• Rats
• Rats, Sprague-Dawley
• Signal Transduction (genetics)
• Tetracyclines (pharmacokinetics, pharmacology)
• Time Factors
• Tumor Necrosis Factor-alpha (pharmacology)