1HNMR metabolomics of MC3T3-E mouse osteoblast proliferation and alkaline phosphatase content by deer antler peptide amine

Author(s):  
Bian Xuefeng ◽  
Li Jingfeng ◽  
Sun Jiaming ◽  
Zhang Hui
2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Xinyu Shao ◽  
Xiaojun Cao ◽  
Ge Song ◽  
Yuan Zhao ◽  
Bimin Shi

Aims. To study the proliferation of osteoblasts and genes expression under normal glucose, high glucose, and metformin (Met).Methods. MG63 osteoblast-like cells were cultured in osteogenic medium supplemented with normal glucose (glucose 5.5 mmol/L) or high glucose (glucose 16.7 mmol/L) and metformin + high glucose (Met 300 μmol/L + glucose 16.7 mmol/L). Proliferation was detected with CCK-8 assay at days 1, 3, and 7. Real-time PCR and Western blot were performed to compare the expression of collagen I (Col I), osteocalcin (OCN), osteoprotegerin (OPG), receptor activator for NF-κB ligand (RANKL), and metal matrix proteinases 1 and 2 (MMP1, MMP2). Alkaline phosphatase (ALP) activity was also detected at days 6, 12, and 18.Results. Exposure to high glucose inhibited the proliferation of osteoblasts (P<0.05), with suppressed OCN and OPG. Meanwhile, Col I, RANKL, MMP1, and MMP2 were unaffected. Metformin attenuated the suppression on proliferation with increased expression of Col I, OCN, and OPG, meanwhile suppressing MMP1 and MMP2. High glucose lowered the intracellular ALP, while metformin raised it. Metformin attenuated the downregulation of ALP completely at day 6, partly at day 12, but not at day 18.Conclusions. Metformin attenuated the suppression effect of high glucose to the osteoblast proliferation and gene expression, more prominently in earlier stage.


2008 ◽  
Vol 3 (1) ◽  
pp. 31-37 ◽  
Author(s):  
Elena Oprita ◽  
Lucia Moldovan ◽  
Oana Craciunescu ◽  
Otilia Zarnescu

AbstractThe purpose of the present study was to investigate the effect of a collagen/β-tricalcium phosphate (COL/β-TCP) composite on osteoblast growth and proliferation. The COL/β-TCP composite was prepared by mixing COL type I with β-TCP, in 1:1 (w/w) ratio and conditioned as sponge by freeze-drying. The osteoblast culture was obtained from rat calvaria bones by enzymatic digestion and cells were seeded in the COL/β-TCP composite. The cell morphology and viability, alkaline phosphatase and osteocalcin, as markers of osteoblast proliferation were evaluated at 3, 7 and 25 days of culture. Histological sections revealed that cell colonization progressively increased inside the COL/β-TCP scaffold, and osteoblasts had a random distribution throughout the scaffold. Cells cultured into the COL/β-TCP scaffold presented osteoblast phenotype, intense staining of alkaline phosphatase and increased production of osteocalcin. Transmission electron micrographs revealed intimate contacts between osteoblasts and the scaffold. MTT test indicated that the viability of the cells cultivated in the presence of COL/β-TCP scaffold was similar to that of the control. All these results show that our COL/β-TCP composite act as a good substrate for rat osteoblast proliferation and migration and could be a promising substitute for bone repair.


2010 ◽  
Vol 30 (7) ◽  
pp. 501-510 ◽  
Author(s):  
Young-Eun Cho ◽  
Ethel Alcantara ◽  
Santhy Kumaran ◽  
Kun-Ho Son ◽  
Ho-Yong Sohn ◽  
...  

2008 ◽  
Vol 46 (6) ◽  
pp. 2135-2142 ◽  
Author(s):  
Un-Ho Jin ◽  
Seok-Jong Suh ◽  
Sang-Dong Park ◽  
Kap-Sung Kim ◽  
Dae Young Kwon ◽  
...  

1994 ◽  
Vol 143 (2) ◽  
pp. R9-R16 ◽  
Author(s):  
Joanna S. Price ◽  
Babatunde O. Oyajobi ◽  
Richard O. C. Oreffo ◽  
R. Graham G. Russell

ABSTRACT Deer antler growth provides a unique natural model of rapid and complete bone regeneration. In this study, the distal antler tips of male red deer (Cervus elaphus) were collected post-mortem during the annual growth period (April-August), and an in vitro system established for the culture of cells from three regions; the inner layer of the perichondrium, the reserve mesenchyme and the cartilage zone. Alkaline phosphatase (ALP) expression by cultured cells, as demonstrated by enzyme histochemistry and biochemical assay, reflected the stage of cellular differentiation. ALP activity was highest in cells cultured from the hypertrophic cartilage region (3.6 ± 0.2 μmol/μg cell protein/minute), and lowest in undifferentiated mesenchymal cells (0.3 ± 0.01 μmol/μg cell protein/minute). ALP expression was lost with passage in culture. Levels of ALP activity in cultured cells correlated with the pattern and extent of enzyme expression in tissue sections as demonstrated by histochemical staining. Insulin-like growth factor (IGF)-I (10−9M-10−7M) was found to be mitogenic for cultured cells from all three zones as shown by increased incorporation of [3H]thymidine into DNA. These results demonstrate that cells from three different regions of the antler tip can be maintained in culture, and that antler cells share certain phenotypic characteristics of growth plate chondrocytes. These data provide further evidence of a role for IGF-1 in the regulation of antler growth. Antler regrowth is a potentially useful model for the study of the factors that regulate bone formation.


2021 ◽  

Abstract Marine organisms have attracted considerable attention in recent years. In this study, peptides with osteogenic activity from Pinctada martensii were isolated and identified. Additionally, the effects of the hydrolysates on MC3T3-E1 cell proliferation and differentiation were evaluated using the MTT and alkaline phosphatase (ALP) assays, respectively. First, trypsin, pancreatin, and neutral protease were used to hydrolyse the intact shellfish. The hydrolysates with the greatest effects on osteoblast proliferation and ALP activity were separated and purified. Second, fraction WP2 was isolated and purified using a Sephadex G-25 column. WP2, which had the highest osteogenic activity, increased cell growth by 48.57 ± 0.05% and ALP activity by 6.27 ± 0.07 mU. Finally, four novel peptides were identified in WP2 (FDNEGKGKLPEEY, IVLDSGDGVTH, IVLDSGDGVSH, and SSENSDLQRQ) by Orbitrap Fusion Lumos Tribrid orbital liquid chromatography-mass spectrometry. Our findings revealed that P. martensii contains peptides with potential osteogenic activity.


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