Interaction of specifically chemically modified dextrans with transforming growth factor beta1: potentiation of its biological activity.
Transforming growth factor beta (TGFbeta), a potent multifunctional cytokine, is well known to demonstrate heparin binding ability. This study investigated the binding capacity of heparin-like family of chemically modified dextrans to TGFbeta1. Dextran derivatives with various substitution contents in carboxymethyl, benzylamide and sulfate groups were evaluated using a gel mobility shift assay. This structure-function study indicated that a synergistic role of benzylamide and sulfate substituents resulted in an optimal interaction with the growth factor. The effect of these polymers on the biological response of TGFbeta1 was assessed using mink lung epithelial cells transfected with a plasminogen activator inhibitor-1 promoter-luciferase construct (PAI/Luc). When the growth factor was mixed with 250 microg/mL of carboxymethyl-benzylamide-dextran (DCMB) or carboxymethyl-benzylamide-sulfate-dextran (DCMBSu), the luciferase gene expression was enhanced. Only polymers exhibiting TGFbeta1 binding demonstrated a biological potentiating effect. However, this effect was strongly amplified as the cell plating time increased (35-fold increase with a 2 days plating time versus 1.1-fold increase with a 4 hr plating time at a 0.25 ng/mL concentration of TGFbeta1). TGFbeta1 induced the PAI/Luc construct in a dose-dependent fashion but its effect diminished when added to cells previously cultured for 24 and 48 hr. The results indicated that the potentiating effect required a complex formation between TGFbeta1 and polymers, the action of which seeming to locally maintain TGFbeta1 in an active form. TGFbeta isoforms playing a key role in the process of bone repair, specifically designed functionalized dextrans could potentiate the in vivo TGFbeta1 biological effect and be used in the field of wound healing applications.
Logeart-Avramoglou
Delphine
D
Laboratoire de Recherches Orthopediques, CNRS UMR 7052, Universite Paris 7 Denis Diderot, Paris, France. logeart@ext.jussieu.fr
Huynh
Remi
R
Chaubet
Frederic
F
Sedel
Laurent
L
Meunier
Alain
A
D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D003911:Dextrans / Q000737:chemistry* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D017930:Genes, Reporter; D008156:Luciferases / Q000378:metabolism; D008907:Mink; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000737:chemistry / Q000494:pharmacology*; D053773:Transforming Growth Factor beta1
D003911:Dextrans; D017395:Plasminogen Activator Inhibitor 1; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D008156:Luciferases