Effect of a novel recombinant protein of fibronectinIII7-10/cadherin 11 EC1-2 on osteoblastic adhesion and differentiation.
The limitations of specific adhesion and osteoblastic differentiation are current problems in bone tissue engineering. The aim of this study was to investigate the effect of a novel recombinant protein of fibronectin module III7-10/cadherin 11 EC1-2 (rFN/CDH) on cell adhesion and differentiation. Gene coding rFN/CDH was engineered by a homology modeling strategy, and an expression plasmid was constructed by standard DNA techniques. The rFN/CDH protein was expressed in Rosetta-gami (DE3), an improved Escherichia coli system. MC3T3-E1 cell centrifugal adhesive assay indicated that the adhesive capacity of rFN/CDH was significantly improved. Quantitative analysis of two osteogenic markers, osteocalcin mRNA expression and alkaline phosphatase activity, indicated that they were further up-regulated when human mesenchymal stem cells were cultured for 7-10 d on rFN/CDH pre-coated surfaces. These results suggest that rFN/CDH possesses an enhanced dual biofunctionality in osteoblastic adhesion and differentiation, and a promising application can be expected in biomimetic strategies and biomaterial development.
Zhang
Yuan
Y
Department of Orthopedics, Xinqiao Hospital, Third Military Medical University, Xinqiao Street, Shapingba District, Chongqing 400037, PR China.
Zhou
Yue
Y
Zhu
Jie
J
Dong
Shiwu
S
Li
Changqing
C
Xiang
Qiang
Q
D016475:3T3 Cells; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D015820:Cadherins / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects*; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005353:Fibronectins / Q000494:pharmacology*; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D015675:Osteocalcin / Q000235:genetics; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D023822:Tissue Engineering
D015820:Cadherins; D017931:DNA Primers; D005353:Fibronectins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D015675:Osteocalcin; C086960:osteoblast cadherin; D000469:Alkaline Phosphatase