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Expression, purification, and characterization of a dentin phosphoprotein produced by Escherichia coli, and its odontoblastic differentiation effects on human dental pulp cells.
- Source :
-
The protein journal [Protein J] 2012 Aug; Vol. 31 (6), pp. 504-10. - Publication Year :
- 2012
-
Abstract
- To investigate the functions of recombinant human dentin phosphoprotein (rhDPP), we examined cell adhesion, viability and the odontoblastic differentiation activity of human dental pulp cells (hDPCs). Firstly, rhDPP was constructed using pBAD-HisA plasmid in Escherichia coli. Cell adhesion and viability of hDPCs by rhDPP was examined using a crystal violet assay and a MTT assay, ALP, mineralization activity and odontoblastic differentiation-related mRNA levels of hDPCs were measured to elucidate the odontoblastic differentiation effect of rhDPP on hDPCs. Initially, rhDPP significantly and dose-dependently increased hDPCs adhesion versus the untreated control (p < 0.05). Cell viability was also significantly increased by rhDPP at 5 days (p < 0.001). Furthermore, the odontoblastic differentiation effect of rhDPP was verified by measuring ALP activity, mineralization activity and the mRNA levels of odontoblastic differentiation markers. Taken together, rhDPP is expected to play an important role on hDPCs, thereby suggesting its potential use for tooth repair and regeneration.
- Subjects :
- Alkaline Phosphatase genetics
Alkaline Phosphatase metabolism
Calcification, Physiologic
Cell Adhesion
Cell Proliferation
Cell Survival drug effects
Cells, Cultured
Collagen Type I genetics
Collagen Type I metabolism
Escherichia coli
Extracellular Matrix Proteins genetics
Extracellular Matrix Proteins isolation & purification
Extracellular Matrix Proteins metabolism
Extracellular Matrix Proteins pharmacology
Gene Expression
Humans
Odontoblasts enzymology
Odontoblasts metabolism
Phosphoproteins genetics
Phosphoproteins isolation & purification
Phosphoproteins metabolism
Phosphoproteins pharmacology
Recombinant Proteins biosynthesis
Recombinant Proteins isolation & purification
Recombinant Proteins pharmacology
Sialoglycoproteins isolation & purification
Sialoglycoproteins pharmacology
Cell Differentiation drug effects
Dental Pulp cytology
Extracellular Matrix Proteins biosynthesis
Odontoblasts physiology
Phosphoproteins biosynthesis
Sialoglycoproteins biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1875-8355
- Volume :
- 31
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- The protein journal
- Publication Type :
- Academic Journal
- Accession number :
- 22752830
- Full Text :
- https://doi.org/10.1007/s10930-012-9430-9