Lysyl oxidase modulates the osteoblast differentiation of primary mouse calvaria cells

Int J Mol Med. 2015 Dec;36(6):1664-70. doi: 10.3892/ijmm.2015.2384. Epub 2015 Oct 21.

Abstract

Lysyl oxidase (LOX) is an extracellular amine oxidase that mediates the formation of collagen fibers. Thus far, five LOX family genes [LOX, lysyl oxidase-like (LOXL)1, LOXL2, LOXL3 and LOXL4] have been identified in humans, each encoding the characteristic C-terminal domains that are required for amine oxidase activity. During osteoblastogenesis, collagen fibers function as a three-dimensional scaffold for organizing mineral deposition. In this study, to assess the functional roles of the LOX family members in osteoblastogenesis, we investigated the temporal expression of these genes as a function of phenotypic development during the osteoblast differentiation of primary cultured mouse calvaria cells. Of the LOX family members, only LOX was prominently expressed during osteoblast differentiation. LOX expression was highest on day 9 of differentiation, as shown by RT-PCR and western blot analysis. The expression pattern of collagen, type I, alpha 2 (COL1A2), which encodes the α2-chain of mouse collagen type I, was similar to that of LOX. The total amine oxidase activity of the differentiating calvaria cells exhibited a temporal pattern that paralleled LOX expression, reaching the highest level on day 9 of differentiation. We also noted that the inhibition of the amine oxidase activity of LOX significantly suppressed both mineral nodule formation and the expression of osteoblast marker genes during the differentiation of primary calvaria cells. Taken together, these findings suggest that the LOX-mediated organization of collagen fibers in the extracellular matrix is an important regulator of osteoblastogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amine Oxidase (Copper-Containing) / genetics
  • Amine Oxidase (Copper-Containing) / metabolism
  • Aminopropionitrile / pharmacology
  • Animals
  • Animals, Newborn
  • Blotting, Western
  • Cell Differentiation / genetics*
  • Cells, Cultured
  • Collagen Type I / genetics
  • Collagen Type I / metabolism
  • Extracellular Matrix / genetics
  • Extracellular Matrix / metabolism
  • Gene Expression Profiling / methods
  • Mice, Inbred ICR
  • Osteoblasts / cytology
  • Osteoblasts / drug effects
  • Osteoblasts / metabolism*
  • Osteogenesis / genetics*
  • Protein-Lysine 6-Oxidase / genetics*
  • Protein-Lysine 6-Oxidase / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Skull / cytology

Substances

  • Collagen Type I
  • Aminopropionitrile
  • Protein-Lysine 6-Oxidase
  • Amine Oxidase (Copper-Containing)