Improving the affinity of fibroblasts for bacterial cellulose using carbohydrate-binding modules fused to RGD.
The attachment of cells to biomedical materials can be improved by using adhesion sequences, such as Arg-Gly-Asp (RGD), found in several extracellular matrix proteins. In this work, bifunctional recombinant proteins, with a Cellulose-Binding Module (CBM), from the cellulosome of Clostridium thermocellum and cell binding sequences-RGD, GRGDY-were cloned and expressed in E.coli. These RGD-containing cellulose binding proteins were purified and used to coat bacterial cellulose fibres. Its effect on the cell adhesion/biocompatibility properties was tested using a mouse embryo fibroblasts culture. Bacterial cellulose (BC) secreted by Gluconacetobacter xylinus (=Acetobacter xylinum) is a material with unique properties and promising biomedical applications. CBMs adsorbs specifically and tightly on cellulose. Thus, they are a useful tool to address the fused RGD sequence (or other bioactive peptides) to the cellulose surface, in a specific and simple way. Indeed, fibroblasts exhibit improved ability to interact with bacterial cellulose sheets coated with RGD-CBM proteins, as compared with cellulose treated with the CBM, that is, without the adhesion peptide. The effect of the several fusion proteins produced was analyzed.
Andrade
F K
FK
IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal.
Moreira
S M G
SM
Domingues
L
L
Gama
F M P
FM
D016475:3T3 Cells; D000818:Animals; D002448:Cell Adhesion; D002470:Cell Survival / Q000187:drug effects; D002482:Cellulose / Q000378:metabolism*; D048013:Acetivibrio thermocellus; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000166:cytology*; D041822:Gluconacetobacter xylinus / Q000737:chemistry; D051379:Mice; D008856:Microscopy, Fluorescence; D009842:Oligopeptides / Q000378:metabolism*; D011137:Polystyrenes / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D044967:Serum
D003470:Culture Media; D009842:Oligopeptides; D011137:Polystyrenes; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C055213:saccharide-binding proteins; C047981:arginyl-glycyl-aspartic acid; D002482:Cellulose