Mechanisms of magnesium-stimulated adhesion of osteoblastic cells to commonly used orthopaedic implants.
Poor cell adhesion to orthopaedic and dental implants may result in implant failure. Cellular adhesion to biomaterial surfaces primarily is mediated by integrins, which act as signal transduction and adhesion proteins. Because integrin function depends on divalent cations, we investigated the effect of magnesium ions modified bioceramic substrata (Al(2)O(3)-Mg(2+)) on human bone-derived cell (HBDC) adhesion, integrin expression, and activation of intracellular signalling molecules. Immunohistochemistry, flow cytometry, cell adhesion, cell adhesion blocking, and Western blotting assays were used. Our findings demonstrated that adhesion of HBDC to Al(2)O(3)-Mg(2+) was increased compared to on the Mg(2+)-free Al(2)O(3). Furthermore, HBDC adhesion decreased significantly when the fibronectin receptor alpha5beta1- and beta1-integrins were blocked by functional blocking antibodies. HBDC grown on the Mg(2+)-modified bioceramic expressed significantly enhanced levels of beta1-, alpha5beta1-, and alpha3beta1-integrins receptors compared to those grown on the native unmodified Al(2)O(3). Tyrosine phosphorylation of intracellular integrin-dependent signalling proteins as well as the expression of key signalling protein Shc isoforms (p46, p52, p66), focal adhesion kinase, and extracellular matrix protein collagen type I were significantly enhanced when HBDC were grown on Al(2)O(3)-Mg(2+) compared to the native Al(2)O(3). We conclude that cell adhesion to biomaterial surfaces is probably mediated by alpha5beta1- and beta1-integrin. Cation-promoted cell adhesion depends on 5beta1- and beta1-integrins associated signal transduction pathways involving the key signalling protein Shc and results also in enhanced gene expression of extracellular matrix proteins. Therefore, Mg(2+) supplementation of bioceramic substrata may be a promising way to improve integration of implants in orthopaedic and dental surgery.
Zreiqat
H
H
School of Pathology, UNSW, Sydney 2052, Australia. H.Zreiqat@unsw.edu.au
Howlett
C R
CR
Zannettino
A
A
Evans
P
P
Schulze-Tanzil
G
G
Knabe
C
C
Shakibaei
M
M
D000537:Aluminum Oxide; D000906:Antibodies / Q000276:immunology; D001672:Biocompatible Materials; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D016023:Integrins / Q000096:biosynthesis; D008274:Magnesium / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D010006:Osteoblasts / Q000187:drug effects*; D010590:Phalloidine; D010766:Phosphorylation; D019736:Prostheses and Implants; D015398:Signal Transduction / Q000187:drug effects; D013268:Stimulation, Chemical; D013499:Surface Properties
D000906:Antibodies; D001672:Biocompatible Materials; D016023:Integrins; D010590:Phalloidine; D008274:Magnesium; D000537:Aluminum Oxide