The optimal dose of recombinant human osteogenic protein-1 enhances differentiation of mouse osteoblast-like cells: an in vitro study
| Autor: | Feng Zhang, Mei-nv Yin, Hai-sheng Lin, Ling-fei Ren, Geng-sheng Shi, Yong-qing Tong |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
medicine.medical_specialty
Time Factors Cellular differentiation Osteocalcin Core Binding Factor Alpha 1 Subunit In Vitro Techniques Real-Time Polymerase Chain Reaction Cell Line Extracellular matrix Mice Internal medicine medicine Animals General Dentistry Cells Cultured Cell Proliferation DNA Primers Analysis of Variance Osteoblasts biology Dose-Response Relationship Drug Staining and Labeling Cell growth Cell Differentiation Cell Biology General Medicine Alkaline Phosphatase Molecular biology In vitro Extracellular Matrix RUNX2 Endocrinology Otorhinolaryngology Cell culture Bone Morphogenetic Proteins biology.protein Alkaline phosphatase Collagen Carrier Proteins |
| Zdroj: | Archives of oral biology. 57(5) |
| ISSN: | 1879-1506 |
| Popis: | Objective There is no certain conclusion on the effect of recombinant human Osteogenic Protein-1 (OP-1, BMP-7) on the proliferation of the osteoblast-like cell line, MC3T3-E1. Furthermore, the optimal dose of rhOP-1 on cell differentiation still needs to be elucidated. This investigation aims to delineate the biofunctional characteristics of rhOP-1 in inducing osteoblastogenesis of MC3T3-E1 through in vitro time-course and dose–response studies. Design MC3T3-E1 cells were cultured for 1, 4, 7 days with the addition of different rhOP-1 concentrations (0, 10, 20, 50, 100, 200, 400 ng/ml), and cell proliferation and cell differentiation were examined. Results MC3T3-E1 cell proliferation was stimulated by rhOP-1 in a dose-dependent manner (0–400 ng/ml) on day 1, whereas on day 4 and 7, it was still stimulated at low concentrations (10, 20, 50 ng/ml) but inhibited at high ones (200, 400 ng/ml). The alkaline phosphatase (ALP) activity, osteocalcin (OC) production, collagen deposition and extracellular matrix mineralization were dramatically elevated by rhOP-1 treatment, as a function of culture time and rhOP-1 concentration, and all of them reached a plateau at the concentration of 200 ng/ml. Real-time quantitative RT-PCR results showed Runx2, AKP-2, OC and Nog mRNA expressions increased in a dose- and time-dependent manner, and their expressions were significantly higher at high rhOP-1 concentrations than that of low ones. No significant differences were found between the effects of 200 ng/ml rhOP-1 and 400 ng/ml rhOP-1 on the differentiation of MC3T3-E1 cells, except the expression of Nog mRNA, whose expression level was much higher at 400 ng/ml than that at 200 ng/ml. Conclusions These results suggest that cell proliferation of MC3T3-E1 is depended on culture time and rhOP-1 concentration, rhOP-1 could stimulate the differentiation of MC3T3-E1 cells and the optimal concentration could be 200 ng/ml. |
| Databáze: | OpenAIRE |
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