OP-1 (BMP-7) Affects mRNA Expression of Type I, II, X Collagen, and Matrix Gla Protein in Ossifying Long Bones In Vitro

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  • A. Haaijman
    Department of Oral Cell Biology ACTA, Vrije Universiteit, Amsterdam, The Netherlands
  • R. N. D'Souza
    Department of Basic Sciences, University of Texas at Houston, Health Science Center, Houston, Texas, U.S.A.
  • A. L. J. J. Bronckers
    Department of Oral Cell Biology ACTA, Vrije Universiteit, Amsterdam, The Netherlands
  • S. W. Goei
    Department of Oral Cell Biology ACTA, Vrije Universiteit, Amsterdam, The Netherlands
  • E. H. Burger
    Department of Oral Cell Biology ACTA, Vrije Universiteit, Amsterdam, The Netherlands

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<jats:title>Abstract</jats:title> <jats:p>In long bone development, a regulating role of OP-1 is suggested by the local correlated expression of both OP-1 ligand and OP-1 binding receptors in developing mouse hind limbs. OP-1 is expressed in the interdigital mesenchyme, whereas OP-1 binding receptors are found in the bordering perichondrium, and both OP-1 ligand and receptors are present in the zone of (pre)hypertrophic chondrocytes. We investigated the role of OP-1 in long bone development experimentally by treating organ cultures of embryonic mouse metatarsals with rhOP-1. The mRNA expression patterns of type I, II, X collagen, and matrix Gla protein (MGP) were studied using in situ hybridization and cell proliferation using [3H]thymidine and BrdU labeling. In the epiphyseal perichondrium, treatment with 40 ng/ml OP-1 enhanced cell proliferation after day 2, while 6-day treatment caused a shift in expression from type I collagen to type II collagen mRNA. This supports previous histochemical findings that OP-1 induced the transition of perichondrium into cartilage. In the center of the rudiment, OP-1 inhibited the expression of type X collagen mRNA, indicating inhibition of chondrocyte hypertrophy. An arrest of differentiation at the prehypertrophic chondrocyte stage was also indicated by the large area of cells expressing MGP mRNA in the OP-1–treated rudiments. We conclude that OP-1 affected the expression of marker genes of chondrocyte differentiation by acting on two steps in endochondral ossification. First, cell proliferation was enhanced, particularly so in the perichondrium where cells started to express the chondrocyte phenotype. Second, the terminal differentiation of mature chondrocytes into hypertrophic chondrocytes was inhibited. These results, combined with the known pattern of OP-1 ligand and BMP receptor expression in the embryo, suggest that OP-1 plays a local role in the cascade of events during endochondral ossification.</jats:p>

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