The interaction of cellular fibronectin with collagen during fibroblast-mediated contraction of collagen gels.
In the first instance highly hydrated collagen gels contract to dense and compact gels when populated by fibroblasts. We previously reported the involvement of fibronectin (FN) in an early process of the collagen gel contraction, utilizing a specific monoclonal antibody dubbed A3A5 (MoAb-A3A5) that inhibits the gel contraction. This study was performed to further characterize the role of the epitope for MoAb-A3A5 in the interaction between fibroblasts and collagen fibrils. Although both cellular FN (cFN) and plasma FN (pFN) were reactive with MoAb-A3A5, the FN that actually participates in a process of the gel contraction was shown to be cFN. The gel contraction was significantly accelerated when fibroblasts were pretreated with excess amounts of cFN and was significantly inhibited when the collagen molecules were pretreated with excess cFN. Such effects of the pretreatments were not observed for pFN. The involvement of cFN, but not pFN, in the interaction of fibroblasts with collagen fibrils was additionally shown by the similar inhibitory action of cFN, but not pFN, on the spreading and elongation of fibroblasts on collagen fibrils. The epitope for MoAb-A3A5 was strongly suggested to be a new functional domain responsible for the interactions between fibroblasts and native collagen molecules. This was not the case for those with denatured one, because fibroblasts on collagen fibrils were not stainable with MoAb-A3A5, whereas the interactions on gelatin were stainable. The lack of the reactivity of fibroblasts on collagen fibrils toward MoAb-A3A5 was not a result of the absence of FN on the cell membrane, but seemed to be a steric hindrance to the access of the antibody.
Yoshizato
K
K
Tissue Regeneration Project of Regional Intensive Program, Hiroshima Institute of Industrial Science and Technology, Higashihiroshima, Japan. kyoshiz@ipcl.hiroshima-u.ac.jp
Tsukahara
M
M
Oki
T
T
Hayashi
M
M
Obara
M
M
Morpho
Y
Y
D000911:Antibodies, Monoclonal / Q000378:metabolism; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology*; D005353:Fibronectins / Q000378:metabolism*; D005782:Gels; D006801:Humans; D007150:Immunohistochemistry
D000911:Antibodies, Monoclonal; D005353:Fibronectins; D005782:Gels; D003094:Collagen