Additional bone and cartilage are formed if transforming growth factor-beta is injected into the periosteum of calvariae or long bones. To investigate this further, transforming growth factor-beta 2 was injected into the periosteum of the tibia of 3-day-old, 3-month-old and 2-year-old rabbits. In all instances, there was an increase in proliferation of the cells of the cambial layer of the periosteum, that is, the osteoprogenitor cells, and breakdown of the fibrous layer. Oedema was induced in the surrounding connective tissues. Over the experimental period the normal neonatal tibia is undergoing rapid growth; there is periosteal bone formation and endosteal resorption. In the experimental neonatal tibiae, an increase in periosteal bone formation is seen after three injections of 20 ng of transforming growth factor-beta 2, which is accompanied by cartilage after five injections; the amounts of induced bone and cartilage increase with the number of injections. The chondrocytes hypertrophy after 4 days and the cartilage is replaced by bone endochondrally. In contrast, after seven injections of 20 ng transforming growth factor-beta 2, there is only a small amount of new bone on the 3-month-old tibia and none on the 2-year-old tibia. One day after seven injections of 200 ng transforming growth factor-beta 2, there is a small amount of bone formation, while seven days after cartilage is found as small discrete nodules on the 3-month-old tibia, but as small areas within the bone on the 2-year-old tibia. It is concluded that the primary effect of transforming growth factor-beta 2 in this experimental model is to increase the proliferative rate of the osteoprogenitor cells in the periosteum. It is argued that transforming growth factor-beta 2 does not initiate osteoblastic or chondrocytic differentiation of osteoprogenitor cells. It is suggested that their differentiation is controlled by the local environment, in particular, the vascularity and locally circulating growth factors.

REFERENCES

Ashhurst
D. E.
(
1986
).
The influence of mechanical conditions on the healing of experimental fractures in the rabbit: a microscopical study.
Phil. Trans. R. Soc. Lond
313
,
271
302
Ballock
R.T.
,
Heydemann
A.
,
Wakefield
L.M.
,
Flanders
K.C.
,
Roberts
A.B.
,
Sporn
M.B.
) (
1993
).
TGF-1 prevents hypertrophy of epiphyseal chondrocytes: regulation of gene expression for cartilage matrix proteins and metalloproteases.
Dev. Biol
158
,
414
429
Bassett
C. A. L.
,
Herrmann
I.
(
1961
).
Influence of oxygen concentration and mechanical factors on differentiation of connective tissues in vitro.
Nature
190
,
460
461
Ber
R.
,
Kubota
T.
,
Sodek
J.
,
Aubin
J. E.
(
1991
).
Effects of transforming growth factor-beta on normal clonal bone cell populations.
Biochem. Cell Biol
9
,
132
140
Bonewald
L. F.
,
Mundy
G. R.
(
1990
).
Role of transforming growth factor-beta in bone remodelling.
Clin. Orthop
250
,
261
276
Caplan
A. I.
,
Koutroupas
S.
(
1973
).
The control of muscle and cartilage development in the chick limb: the role of differential vascularization.
J. Embryol. Exp. Morph
29
,
571
583
Centrella
M.
,
Canalis
E.
(
1985
).
Transforming and nontransforming growth factors are present in medium conditioned by fetal rat calvariae.
Proc. Nat. Acad. Sci. USA
82
,
7335
7339
Centrella
M.
,
Massague
J.
,
Canalis
E.
(
1986
).
Human platelet-derived transforming growth factor-stimulates parameters of bone growth in fetal rat calvariae.
Endocrinology
119
,
2306
2312
D'Angelo
M.
,
Greene
R. M.
(
1991
).
Transforming growth factor-modulation of glycosaminoglycan production by mesenchymal cells of the developing murine secondary palate.
Dev. Biol
145
,
374
378
Farnum
C. E.
,
Wilsman
N. J.
(
1993
).
Determination of proliferative characteristics of growth plate chondrocytes by labeling with bromodeoxyuridine.
Calc. Tiss. Int
52
,
110
119
Flanders
K. C.
,
Thompson
N. L.
,
Cissel
D. S.
,
Van Obberghen-Schilling
E.
,
Baker
C. C.
,
Kass
M. E.
,
Ellingsworth
L. R.
,
Roberts
A. B.
,
Sporn
M. B.
(
1989
).
Transforming growth factor-1: histochemical localization with antibodies to different epitopes.
J. Cell Biol
108
,
653
660
Gatherer
D.
,
Ten Dijke
P.
,
Baird
D. T.
,
Akhurst
R. J.
(
1990
).
Expression of TGF-isoforms during first trimester human embryogenesis.
Development
110
,
445
460
Hallman
R.
,
Feinberg
R. N.
,
Latker
C. H.
,
Sassa
J.
,
Risau
W.
(
1987
).
Regression of blood vessels precedes cartilage differentiation during chick limb development.
Differentiation
34
,
98
105
Harashima
Y.
,
Tanaka
T.
,
Gotoh
K.
,
Satoh
R.
,
Inazu
M.
(
1992
).
Difference in transforming growth factor1-induced osteogenesis between neonatal and adult rats.
Calc. Tiss. Int
50
,
1
–.
Heine
U. I.
,
Munoz
E. F.
,
Flanders
K. C.
,
Ellingsworth
L. R.
,
Lam
H.-Y. P.
,
Thompson
N. L.
,
Roberts
A. B.
,
Sporn
M. B.
(
1987
).
Role of transforming growth factor-in the development of the mouse embryo.
J. Cell Biol
105
,
2861
2876
Hock
J. M.
,
Canalis
E.
,
Centrella
M.
(
1990
).
Transforming growth factor-stimulates bone matrix apposition and bone cell replication in cultured fetal rat calvariae.
Endocrinology
126
,
421
426
Hoffman
F. M.
(
1991
).
Transforming growth factor--related genes in Drosophila and vertebrate development.
Curr. Opin. Cell Biol
3
,
947
952
Izumi
T.
,
Scully
S. P.
,
Heydemann
A.
,
Bolander
M. E.
(
1992
).
Transforming growth factor1 stimulates type II collagen expression in cultured periosteum-derived cells.
J. Bone Miner. Res
7
,
115
121
Joyce
M. E.
,
Roberts
A. B.
,
Sporn
M. B.
,
Bolander
M. E.
(
1990
).
Transforming growth factor-and the initiation of chondrogenesis and osteogenesis in the rat femur.
J. Cell Biol
110
,
2195
2207
Mackie
E. J.
,
Trechsel
U.
(
1990
).
Stimulation of bone formation in vivo by transforming growth factor-: remodeling of woven bone and lack of inhibition by indomethacin.
Bone
11
,
295
300
Marcelli
C.
,
Yates
A. J.
,
Mundy
G. R.
(
1990
).
In vivo effects of human recombinant transforming growth factoron bone turnover in normal mice.
J. Bone Miner. Res
5
,
1087
1096
Morales
T. I.
(
1991
).
Transforming growth factor-beta 1 stimulates synthesis of proteoglycan aggregates in calf articular cartilage organ cultures.
Arch. Biochem. Biophys
286
,
99
106
Nakahara
H.
,
Bruder
S. P.
,
Haynesworth
S. E.
,
Holecek
J. J.
,
Baber
M. A.
,
Goldberg
V. M.
,
Caplan
A. I.
(
1990
).
Bone and cartilage formation in diffusion chambers by subcultured cells derived from the periosteum.
Bone
11
,
181
188
Nakahara
H.
,
Dennis
J. E.
,
Bruder
S. P.
,
Haynesworth
S. E.
,
Lennon
D. P.
,
Caplan
A. I.
(
1991
).
In vitro differentiation of bone and hypertrophic cartilage from periosteal-derived cells.
Exp. Cell Res
195
,
492
503
Noda
M.
,
Rodan
G. A.
(
1986
).
Type-transforming growth factor inhibits proliferation and expression of alkaline phosphatase in murine osteoblast-like cells.
Biochem. Biophys. Res. Commun
140
,
56
65
Noda
M.
,
Camilliere
J. J.
(
1989
).
In vivo stimulation of bone formation by transforming growth factor-.
Endocrinology
124
,
2991
2994
Owen
M.
(
1963
).
Cell population kinetics of an osteogenic tissue- I.
J. Cell Biol
19
,
19
32
Owen
M.
,
Macpherson
S.
(
1963
).
Cell population kinetics of an osteogenic tissue- II.
J. Cell Biol
19
,
33
44
Pawelek
J. M.
(
1969
).
Effects of thyroxine and low oxygen tension on chondrogenic expression in cell culture.
Dev. Biol
19
,
52
72
Pfeilschifter
J.
,
D'Souza
S. M.
,
Mundy
G. R.
(
1987
).
Effects of transforming growth factor-on osteoblastic osteosarcoma cells.
Endocrinology
121
,
212
218
Pfeilschifter
J.
,
Diel
I.
,
Pilz
U.
,
Brunotte
K.
,
Naumann
A.
,
Ziegler
R.
(
1993
).
Mitogenic responsiveness of human bone cells in vitro to hormones and growth factors decreases with age.
J. Bone Miner. Res
8
,
707
717
Reddi
A. H.
(
1981
).
Cell biology and biochemistry of endochondral bone development.
Coll. Rel. Res
1
,
209
226
Redini
F.
,
Daireaux
M.
,
Mauviel
A.
,
Galera
P.
,
Loyau
G.
,
Pujol
J.-P.
(
1991
).
Characterization of proteoglycans synthesized by rabbit articular chondrocytes in response to transforming growth factor-(TGF-).
Biochim. Biophys. Acta
1093
,
196
206
Roberts
A. B.
,
Sporn
M. B.
,
Assoian
R. K.
,
Smith
J. M.
,
Roche
N. S.
,
Wakefield
L. M.
,
Heine
U. I.
,
Liotta
L. A.
,
Falanga
V.
,
Kehrl
J. H.
,
Fauci
A. S.
(
1986
).
Transforming growth factor type: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro.
Proc. Nat. Acad. Sci. USA
83
,
4167
4171
Robey
P. G.
,
Young
M. F.
,
Flanders
K. C.
,
Roche
N. S.
,
Kondiah
P.
,
Reddi
A. H.
,
Termine
J. D.
,
Sporn
M. B.
,
Roberts
A. B.
(
1987
).
Osteoblasts synthesize and respond to transforming growth factor-type(TGF-) in vitro.
J. Cell Biol
105
,
457
463
Rosier
R. N.
,
O'Keefe
R. J.
,
Crabb
I. D.
,
Puzas
J. E.
(
1989
).
Transforming growth factor beta: an autocrine regulator of chondrocytes.
Conn. Tiss. Res
20
,
295
301
Rubin
C. T.
,
Bain
S. D.
,
McLeod
K. J.
(
1992
).
Suppression of the osteogenic response in the aging skeleton.
Calc. Tiss. Int
50
,
306
313
Shull
M. M.
,
Ormsby
I.
,
Kier
A. B.
,
Pawlowski
S.
,
Diebold
R. J.
,
Yin
M.
,
Allen
R.
,
Sidman
C.
,
Proetzel
G.
,
Calvin
D.
,
Annunziata
N.
,
Doetschman
T.
(
1992
).
Targeted disruption of the mouse transforming growth factor-1 gene results in multifocal inflammatory disease.
Nature
359
,
693
699
Sprugel
K. H.
,
McPherson
J. M.
,
Clowes
A. W.
,
Ross
R.
(
1987
).
Effects of growth factors in vivo. I. Cell ingrowth into porous subcutaneous chambers.
Amer. J. Path
129
,
601
613
Strong
D. D.
,
Beachler
A. L.
,
Wergedal
J. E.
,
Linkhart
T. A.
(
1991
).
Insulin like growth factor II and transforming growth factorregulate collagen expression in human osteoblast like cells in vitro.
J. Bone Miner. Res
6
,
15
23
Ten Dijke
P.
,
Iwata
K. K.
,
Goddard
C.
,
Pieler
C.
,
Canalis
E.
,
McCarthy
T. L.
,
Centrella
M.
(
1990
).
Recombinant transforming growth factor type3: biological activities and receptor-binding properties in isolated bone cells.
Mol. Cell. Biol
10
,
4473
4479
Thompson
N. L.
,
Flanders
K. C.
,
Smith
J. M.
,
Ellingsworth
L. R.
,
Roberts
A. B.
,
Sporn
M. B.
(
1989
).
Expression of transforming growth factor-1 in specific cells and tissues of adult and neonatal mice.
J. Cell Biol
108
,
661
669
Tonna
E. A.
,
Cronkite
E. P.
(
1962
).
An autoradiographic study of periosteal cell proliferation with tritiated thymidine.
Lab. Invest
11
,
455
462
Tschan
T.
,
Höerler
I.
,
Houze
Y.
,
Winterhalter
K. H.
,
Richter
C.
,
Bruckner
P.
(
1990
).
Resting chondrocytes in culture survive without growth factors, but are sensitive to toxic oxygen metabolites.
J. Cell Biol
111
,
257
260
Tschan
T.
,
Böhme
K.
,
Conscience-Egli
M.
,
Zenke
G.
,
Winterhalter
K. H.
,
Bruckner
P.
(
1993
).
Autocrine or paracrine transforming growth factor-modulates the phenotype of chick sternal chondrocytes in serum-free agarose culture.
J. Biol. Chem
268
,
5156
5161
Turner
R. T.
,
Spelsberg
T. C.
(
1991
).
Correlation between mRNA levelsfor bone cell proteins and bone formation in long bones of maturing rats.
Amer. J. Physiol
261
,
348
–.
Wilson
D. J.
(
1986
).
Development of avascularity during cartilage differentiation in the embryonic limb.
Differentiation
30
,
183
187
Wrana
J. L.
,
Kubota
T.
,
Zhang
Q.
,
Overall
C. M.
,
Aubin
J. E.
,
Butler
W. T.
,
Sodek
J.
(
1991
).
Regulation of transformation-sensitive secreted phosphoprotein (SPPI osteopontin) expression by transforming growth factor-. Comparisons with expression of SPARC (secreted acidic cysteine-rich protein).
Biochem. J
273
,
523
531
Young
R. W.
(
1962
).
Regional differences in cell generation time in growing rat tibiae.
Exp. Cell Res
26
,
562
567
This content is only available via PDF.