Lysyl oxidase enhances elastin synthesis and matrix formation by vascular smooth muscle cells.
Lysyl oxidase (LOX) is a copper-dependent enzyme that initiates covalent crosslinking of elastin precursors by oxidizing peptidyl lysine to aminoadipic semi-aldehydes. Previous studies have shown LOX deficiency to affect crosslinking of elastin and collagen in vivo, resulting in disorganized connective tissue formation. In this study, we investigated the utility of exogenously supplemented LOX peptides (50-100 microl/well) to elastin synthesis, crosslinking efficiency and matrix deposition in adult rat aortic smooth muscle cell (RASMC) cultures. Additionally, we also examined the role of LOX peptides on SMC proliferation and matrix metalloproteinase (MMP) synthesis in these cultures. Highly purified bovine aorta LOX peptide was found to increase matrix elastin synthesis by 40-80% to that in control cultures in a dose-dependent manner, while the crosslinking efficiency significantly (as measured by the ratio of matrix elastin protein to the total elastin protein synthesized) improved to 45-55% of total elastin synthesized under these conditions. However, LOX peptides affected neither SMC proliferation relative to controls, nor elastin precursor (tropoelastin) synthesis, nor the total elastin synthesis on a per-cell basis. In general, LOX peptides also did not affect MMP-2 and MMP-9 activities relative to control cultures, except for MMP-9 activity suppression at a higher LOX dose, suggesting that these LOX peptide cues could be safely used to enhance tropoelastin crosslinking into matrix structures and elastin matrix yield, within tissue-engineered constructs, a major challenge in the field.
Kothapalli
Chandrasekhar R
CR
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Ramamurthi
Anand
A
D000447:Aldehydes / Q000737:chemistry; D000818:Animals; D001011:Aorta / Q000166:cytology; D002417:Cattle; D049109:Cell Proliferation; D003432:Cross-Linking Reagents / Q000737:chemistry; D004549:Elastin / Q000096:biosynthesis*; D008239:Lysine / Q000737:chemistry; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D008249:Protein-Lysine 6-Oxidase / Q000502:physiology*; D051381:Rats; D023822:Tissue Engineering / Q000379:methods; D014333:Tropoelastin / Q000737:chemistry
D000447:Aldehydes; D003432:Cross-Linking Reagents; D014333:Tropoelastin; D004549:Elastin; D008249:Protein-Lysine 6-Oxidase; D020780:Matrix Metalloproteinase 9; D008239:Lysine