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J. Biol. Chem., Vol. 261, Issue 27, 12665-12674, 09, 1986
PV Hauschka, AE Mavrakos, MD Iafrati, SE Doleman and M Klagsbrun
The mineralized matrix of osseous tissue harbors abundant mitogenic
activity which is extractable by demineralizing solvents. In bovine bone
powder free of blood and cartilage contamination, the volume concentration
of mitogens is up to 20 times greater than in serum. Growth factor activity
in bone extracts was quantitated on quiescent mouse BALB/c/3T3 fibroblasts,
where [3H]thymidine incorporation for 48 h was stimulated up to 200-fold in
a linear, dose-dependent manner. Six distinct bone-derived growth factors
(BDGFs) have been resolved and partially purified (up to 44,000-fold) on
heparin-Sepharose using NaCl gradient elution. Provisionally named by the
NaCl molarity at which they elute, these BDGFs include BDGF-0.45 (25% of
total activity). This factor is heat-stable and sensitive to
dithiothreitol, and displaces 125I-labelled bovine platelet-derived growth
factor in a radioreceptor assay. BDGF-0.45 (approximately 50 ng/g of bone)
is closely related or identical to bovine platelet-derived growth factor.
BDGF-1.1 (10%) has a pI of 5.2 and shows a 16,600-dalton doublet on sodium
dodecyl sulfate- polyacrylamide gel electrophoresis Western blots stained
with antiserum to bovine anionic fibroblast growth factor. Two activities
with high heparin affinity resemble cationic forms of fibroblast growth
factor. BDGF-1.5 is the dominant factor in fetal membranous bone (50%), but
is less abundant in adult bone (20%). BDGF-1.7, a 17,500-18,400-dalton
triplet, is virtually absent in fetal bone (7%) but abundant (30%) in adult
bone and may be related to cartilage derived growth factor. Two minor
activities, BDGF-0.1 (10%) and BDGF-2.0 (7%) have not been characterized.
Proliferation of bovine capillary endothelial cells was strongly supported
by BDGFs 1.1, 1.5, and 1.7, but not by 0.45. These four purified BDGFs and
the crude bone extract were also strongly mitogenic for rat osteoblasts
while depressing alkaline phosphatase specific activity by 2-3-fold. Bone
exhibits the most complex spectrum of growth factor activities of any
tissue yet described. Bone cells and other indigenous cell types must be
considered as possible sources of the BDGFs, in addition to sequestration
from blood. Mechanisms for unmasking or release of BDGFs from the
mineralized matrix resulting in local action on target cells are
undoubtedly important for the development and maintenance of bone tissue.
Growth factors in bone matrix. Isolation of multiple types by affinity chromatography on heparin-Sepharose
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