The recombinant human dentin matrix protein 1-coated titanium and its effect on the attachment, proliferation and ALP activity of MG63 cells.
The aim of the present work was to design a bio-interactive implant surface by coating recombinant human dentin matrix protein 1 (hDMP1) onto titanium and to investigate the biological function of this material. Firstly, the plasmid containing the hDMP1 cDNA was constructed and hDMP1 was expressed, purified and characterized. Then, hDMP1 was coated onto the surface of Ti substrates via a biochemical technique and the procedure was divided into three steps: in the beginning, titanium was treated by regular polishing and denoted as Cp-Ti; then, Cp-Ti received alkaline and water treatment and was nominated as AW-Ti; finally, AW-Ti was coated with hDMP1 and referred to as hDMP1-Ti. The inserts of hDMP1 genes were detected by enzyme digestion as well as gel electrophoresis, and the complete nucleotide sequence of hDMP1 was tested. The purified recombinant hDMP1 was electrophoresed on a 10 % SDS-PAGE gel. Cp-Ti, AW-Ti and hDMP1-Ti were characterized by X-ray photoelectron spectroscope and water contact angles tests. The biological activity of MG63 cells cultured in the three groups was investigated by the cell attachment, proliferation and alkaline phosphatase activity assays. The results show that hDMP1 was successfully constructed and coated onto the titanium surface, and hDMP1-Ti had higher hydrophilicity than Cp-Ti. Compared with Cp-Ti and AW-Ti, hDMP1-Ti showed better in vitro bioactivity.
Department of Prosthodontics, West China College of Stomatology, Sichuan University, Renmin Nanlu, Chengdu, China.
Xibo
X
Pei
Lanlan
L
Pan
Fenglin
F
Cui
Rui
R
He
Hong
H
Bao
Qianbing
Q
Wan
Jian
J
Wang
D000469:Alkaline Phosphatase / Q000378:metabolism*; D001483:Base Sequence; D002448:Cell Adhesion; D002460:Cell Line; D049109:Cell Proliferation; D017931:DNA Primers; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000737:chemistry* / Q000235:genetics; D056951:Photoelectron Spectroscopy; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013499:Surface Properties
C513309:DMP1 protein, human; D017931:DNA Primers; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D010750:Phosphoproteins; D011994:Recombinant Proteins; D000469:Alkaline Phosphatase