Fibroblast growth factor-18 is a trophic factor for mature chondrocytes and their progenitors.
OBJECTIVE
The aim of this study was to examine the effects of recombinant human Fgf18 on chondrocyte proliferation and matrix production in vivo and in vitro. In addition, the expressions of Fgf18 and Fgf receptors (Fgfr) in adult human articular cartilage were examined.


METHODS
Adenovirus-mediated transfer of Fgf18 into murine pinnae and addition of FGF18 to primary cultures of adult articular chondrocytes were used to assess the effects of FGF18 on chondrocytes. In situ hybridization was used to examine the expression of Fgf18 and Fgfr s in adult human articular cartilage.


RESULTS
Expression of Fgf18 by adenovirus-mediated gene transfer in murine pinnae resulted in a significant increase in chondrocyte number. Chondrocytes were identified by staining with toluidine blue and a monoclonal antibody directed against type II collagen. Fgf18, Fgfr 2-(IIIc), Fgfr 3-(IIIc), and Fgfr 4 mRNAs were detected within these cells by in situ hybridization. The nuclei of the chondrocytes stained with antibodies to PCNA and FGF receptor (FGFR) 2. Addition of FGF18 to the culture media of primary articular chondrocytes increased the proliferation of these cells and increased their production of extracellular matrix. To assess the receptor selectivity of FGF18, BaF3 cells stably expressing the genes for the major splice variants of Fgfr1-3 were used. Proliferation of cells expressing Fgfr 3-(IIIc) or Fgfr 2-(IIIc) was increased by incubation with FGF18. Using FGFR-Fc fusion proteins and BaF3 cells expressing Fgfr 3-(IIIc), only FGFR 3-(IIIc)-Fc, FGFR 2-(IIIc)-Fc or FGFR 4-Fc reduced FGF18-mediated cell proliferation. Expression of Fgf18, Fgfr 3-(IIIc) and Fgfr 2-(IIIc) mRNAs was localized to chondrocytes of human articular cartilage by in situ hybridization.


CONCLUSIONS
These data demonstrate that Fgf18 can act as a trophic factor for elastic chondrocytes and their progenitors in vivo and articular chondrocytes cultured in vitro. Expression of Fgf18 and the genes for two of its receptors in chondrocytes suggests that Fgf18 may play an autocrine role in the biology of normal articular cartilage.
Ellsworth
J L
JL
ZymoGenetics, Inc., 1201 Eastlake Avenue East, Seattle, Washington 98102, USA. ellsworj@zgi.com
Berry
J
J
Bukowski
T
T
Claus
J
J
Feldhaus
A
A
Holderman
S
S
Holdren
M S
MS
Lum
K D
KD
Moore
E E
EE
Raymond
F
F
Ren
HongPing
H
Shea
P
P
Sprecher
C
C
Storey
H
H
Thompson
D L
DL
Waggie
K
K
Yao
L
L
Fernandes
R J
RJ
Eyre
D R
DR
Hughes
S D
SD
D000328:Adult; D000818:Animals; D002358:Cartilage, Articular / Q000737:chemistry; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D024043:Collagen Type II / Q000378:metabolism; D004431:Ear, External; D005260:Female; D005346:Fibroblast Growth Factors / Q000032:analysis / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization / Q000379:methods; D051379:Mice; D008819:Mice, Nude; D011509:Proteoglycans / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D017468:Receptors, Fibroblast Growth Factor / Q000032:analysis / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D013552:Swine
D024043:Collagen Type II; D011509:Proteoglycans; D012333:RNA, Messenger; D017468:Receptors, Fibroblast Growth Factor; D011994:Recombinant Proteins; C113569:fibroblast growth factor 18; D005346:Fibroblast Growth Factors