Calophyllolide Content in Calophyllum inophyllum at Different Stages of Maturity and Its Osteogenic Activity
Abstract
:1. Introduction
2. Results and Discussion
2.1. Calophyllolide Isolated from C. inophyllum Nuts
2.2. Quantification of the Calophyllolide in Nuts
2.3. Calophyllolide-Induced Osteoblast MC3T3-E1 Cell Differentiation
2.4. Calophyllolide Induced Osteoblastic Gene Expression in MC3T3-E1 Cells
2.5. Calophyllolide Enhanced Mineralization and ALP Expression of MC3T3-E1 Cells
2.6. Discussion
3. Experimental Section
3.1. General Information
3.2. Plant Material
3.3. Chemicals
3.4. Calophyllolide Isolation, Purification and Identification
3.5. Calophyllolide
3.6. Quantitative Analysis of Calophyllolide in the Fruits of C. inophyllum
3.6.1. Preparation of the Calophyllolide Standard Solution
3.6.2. Preparation of the C. inophyllum Nut Sample Solutions
3.6.3. HPLC Analysis
3.7. Cell Culture and Differentiation of MC3T3-E1 Cells
3.8. Alkaline Phosphatase (ALP) Assay
3.9. Quantitative Real-Time RT-PCR
Gene Name | Primer Sequence | Size (bp) | Annealing Temperature (°C) | Accession No. |
---|---|---|---|---|
ALP | F: 5′-TGACCTTCTCTCCTCCATCC-3′ | 126 | 55 | NM_001287176.1 |
R: 5′-CTTCCTGGGAGTCTCATCCT-3′ | ||||
RUNX2 | F: 5′-CCCTGAACTCTGCACCAAGT | 147 | 55 | NM_001145920.2 |
R: 5′-TGGAGTGGATGGATGGGGAT | ||||
OSX | F: 5′-GTCAAGAGTCTTAGCCAAACTC-3′ | 124 | 55 | NM_130458.3 |
R: 5′-AAATGATGTGAGGCCAGATGG-3′ | ||||
OCN | F: 5′-CAATAAGGTAGTGAACAGAC-3′ | 133 | 50 | NM_001305448.1 |
R: 5′-CTTTAAGCCATACTGGTTT-3′ | ||||
GAPDH | F: 5′-GTATGACTCCACTCACGGCAAA-3′ | 101 | 60 | NM_008084 |
R: 5′-GGTCTCGCTCCTGGAAGATG-3′ |
3.10. Mineralized Matrix Formation Assay and ALP Staining
3.11. Statistical Analysis
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Liu, W.-H.; Liu, Y.-W.; Chen, Z.-F.; Chiou, W.-F.; Tsai, Y.-C.; Chen, C.-C. Calophyllolide Content in Calophyllum inophyllum at Different Stages of Maturity and Its Osteogenic Activity. Molecules 2015, 20, 12314-12327. https://doi.org/10.3390/molecules200712314
Liu W-H, Liu Y-W, Chen Z-F, Chiou W-F, Tsai Y-C, Chen C-C. Calophyllolide Content in Calophyllum inophyllum at Different Stages of Maturity and Its Osteogenic Activity. Molecules. 2015; 20(7):12314-12327. https://doi.org/10.3390/molecules200712314
Chicago/Turabian StyleLiu, Wei-Hsien, Yen-Wenn Liu, Zih-Fong Chen, Wen-Fei Chiou, Ying-Chieh Tsai, and Chien-Chih Chen. 2015. "Calophyllolide Content in Calophyllum inophyllum at Different Stages of Maturity and Its Osteogenic Activity" Molecules 20, no. 7: 12314-12327. https://doi.org/10.3390/molecules200712314