Immobilisation orientée du facteur de croissance de l'épiderme à des fins d'application en ingénierie tissulaire
Year: 2010
Type: dissertation
Abstract: RESUME
Les implants generes par bio-ingenierie offrent des perspectives prometteuses en
medecine regeneratrice grâce, entre autre, au developpement de supports mieux adaptes a la
promotion de la regeneration tissulaire. Plusieurs groupes de recherche ont d’ailleurs developpe
des techniques permettant d’immobiliser de maniere non-orientee des proteines afin de
promouvoir l’adhesion et la croissance cellulaire. Bien que des reponses cellulaires aient pu etre
observees, ces strategies de couplage ne sont pas optimales en raison de la creation de surfaces
heterogenes et de l’alteration, dans de nombreux cas, de l’activite biologique des proteines ainsi
greffees. Ces methodes de fixation ont donc rapidement ete remplacees par des strategies
favorisant le greffage oriente de proteines. Pour ce faire, il est necessaire de leur adjoindre une
etiquette (Fc ou polyhistidine), puis de faire interagir ces chimeres avec un support adequatement
traite (proteines A/G ou ions metalliques).
Recemment, une nouvelle approche basee sur l’interaction existant entre deux peptides,
denommes E coil et K coil, a ete rapportee. Ces peptides synthetiques possedent une forte affinite
et une haute specificite l’un pour l’autre et ont deja ete utilises dans diverses applications en
genie biomedical ou en biotechnologie (purification ou capture de proteines a la surface de
biocapteurs). Ce systeme peptidique pourrait egalement etre utilise afin de permettre
l’immobilisation orientee de proteines telles que des facteurs de croissance. Les proteines devront
alors etre etiquetees avec un peptide coil tandis que le support choisi devra presenter le peptide
coil complementaire prealablement greffe. Apres une simple incubation, les proteines devraient etre capables de se fixer de maniere orientee a la surface du materiau via l’interaction coiled-coil
ayant lieu entre les deux peptides.----------ABSTRACT
Bioengineered implants are becoming extremely promising with respect to the
development of biocompatible scaffolds to promote and sustain tissue regeneration. Several
research groups have developed methods for the non-oriented immobilization of growth factors
(GF) with the ultimate goal of modulating cell growth and phenotype. Although unambiguous
effects have been reported with randomly grafted ligands, these protocols suffer from several
caveats such as the creation of heterogeneous surfaces on which protein bioactivity may be
suboptimal. Consequently, new methods based on the interactions between affinity tags such as
Fc or polyhistidine and solid supports, allowing for oriented protein immobilization are currently
developed.
Recently, a heterodimeric coiled-coil system formed by two de novo designed peptides
(the E and K coils) has been characterized and demonstrated to be a promising tool for various
biotechnology and biomedical applications such as protein capture and delivery due to the high
affinity and the strong specificity that these two peptides have for each others. In this work, we
have decided to take advantage of this coiled-coil system to specifically capture a coil-tagged
growth factor, EGF, in an oriented fashion through its interaction with a surface on which the coil
partner will be covalently immobilized. We selected EGF since this growth factor is known to
have a major impact on cellular proliferation during wound healing mechanisms, which make it
an excellent candidate for tissue engineering applications.
Author: Cyril Boucher
Language: French
Subfield: Biomaterials
Field: Materials Science
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W162145425