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Effects of bone morphogenetic protein‐7 stimulation on osteoblasts cultured on different biomaterials
Author(s) -
Açil Yahya,
Springer Ingo N.G.,
Broek Vanessa,
Terheyden Hendrik,
Jepsen Søren
Publication year - 2002
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.10197
Subject(s) - bone morphogenetic protein , chemistry , bone morphogenetic protein 2 , type i collagen , in vitro , cell culture , cell , polyacrylamide gel electrophoresis , gel electrophoresis , biomedical engineering , biochemistry , biology , endocrinology , medicine , genetics , gene , enzyme
The objective of the present study was to investigate the effects of an in vitro stimulation of human osteoblasts by recombinant human bone morphogenetic protein‐7 (rhBMP‐7) on the collagen types and the quantity of the collagen cross‐links synthesized in a three‐dimensional culture on various biomaterials for bone replacement. Trabecular bone chips were harvested from human iliac crests, and cell cultures were established at standard conditions. One hundred and fifty nanograms per milliliter of rhBMP‐7 was added. For the second passage a cell scraper was used to bring the cells into suspension, and 100 μl osteoblasts (at a density of 3.3 × 10 5 ) were transferred onto nine blocks of either Bio‐Oss ® , Tutoplast ® , or PepGen p‐15™. Blocks incubated with cells that were not treated with rhBMP‐7 served as controls. Cell colonization of the biomaterials was observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) after a period of 2, 4, and 6 weeks. Throughout the experiment medium, supernatants were collected and collagen was characterized by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). Finally, the collagen cross‐link residues hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) were quantified by HPLC. Within 4 weeks the cells became confluent on all of the studied biomaterials. All samples synthesized bone specific LP and collagen type I. However, in rhBMP‐7‐stimulated samples, the amount of HP and LP found was increased by 45% compared to non‐stimulated samples. Cell proliferation and collagen synthesis was similar on the different biomaterials, but was consistently reduced in specimen not stimulated with rhBMP‐7. In vitro stimulation of osteoblasts on Bio‐Oss, Tutoplast, or PepGen p‐15 with rhBMP‐7 and subsequent transplantation of the constructs might lead to an enhanced osseointegration of the biomaterials in vivo. J. Cell. Biochem. 86: 90–98, 2002. © 2002 Wiley‐Liss, Inc.