Enhanced surface activation of fibronectin upon adsorption on hydroxyapatite.
In this study the adsorption characteristics and the structure of fibronectin adsorped on hydroxyapatite (Ha) and a reference gold substrate (Au) is examined by quartz crystal microbalance with dissipation (QCM-D) and atomic force microscopy (AFM) at the following concentrations: 20 microg/mL, 30 microg/mL, 40 microg/mL, 100 microg/mL, 200 microg/mL, and 500 microg/mL. The conformational changes of the fibronectin molecules upon surface binding were examined as well with monoclonal antibody directed against the cell binding-domain (CB domain) of fibronectin. The QCM-D and AFM results show that the fibronectin uptake is larger on Au as compared with Ha regardless of the protein bulk concentration used in the experiment, suggesting that the individual fibronectin molecules in general attach to the surfaces in a more unfolded configuration on Ha. Moreover the dissipation values obtained with QCM-D indicate that the individual fibronectin molecules bind in a more compact and rigid configuration on Au compared to the Ha surface. In particular the monoclonal antibody data show that the CB domain on fibronectin is more available on Ha, where such cell-recognizing abilities are more pronounced at low fibronectin surface coverage. The results demonstrate that the detailed molecular structure of fibronectin and its functional activity depend significantly on both the underlying surface chemistry as well as the fibronectin surface coverage.
Interdisciplinary Nanoscience Center (iNANO) and Department of Physics and Astronomy, Aarhus University, DK-8000 Aarhus C, Denmark.
A
A
Dolatshahi-Pirouz
T
T
Jensen
M
M
Foss
J
J
Chevallier
F
F
Besenbacher
D000327:Adsorption; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D017886:Durapatite / Q000737:chemistry*; D004563:Electrochemistry / Q000379:methods; D005353:Fibronectins / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D007700:Kinetics; D018625:Microscopy, Atomic Force; D008968:Molecular Conformation; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D013499:Surface Properties
D000906:Antibodies; D005353:Fibronectins; D011506:Proteins; D006046:Gold; D017886:Durapatite