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The osteogenic differentiation stimulating activity of Sea cucumber methanolic crude extraction on rat bone marrow mesenchymal stem cells.
Baharara J
,
Amini E
,
Kerachian MA
,
Soltani M
.
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OBJECTIVES: Sea cucumber derived bioactive compound is considered efficient in treatment of bone disorders. This study was performed to evaluate the effect of this extract on differentiation of rat bone marrow mesenchymal stem cells (rBMMSc) into osteogenic lineage.
MATERIALS AND METHODS: Isolated rBMMSc were grown in DMEM supplemented with 10% FBS. The cells were exposed to different concentration of extract. After 21 days, Alizarin red staining, alkaline phosphatase assay and RT-PCR were performed. The results were analyzed by ANOVA software and P value <0.05 was considered significant.
RESULTS: Morphological methods revealed that appropriate concentrations of extract increased osteogenic differentiation in a dose-dependent manner. RT-PCR revealed that extract without or with osteogenic medium due to osteopontin expression had a potential role in osteogenesis.
CONCLUSION: Based on our data it concluded that S. cucumber extract stimulated Bone marrow mesenchymal cells to differentiate into osteogenic lineage without existence of osteogenic medium.
Figure 1. Part 1- Obtained results from flowcytometry. A) Isotype CD44:IgG2a-PE. B) 87. 13% Expression of CD44 marker in BMSCs. C)
Isotype CD29:IgG2a-PE. D) Approximately 55.41% BMSCs expressed CD29 marker. Part 2- The dose dependent effect of Sea cucumber
methanol extraction the presence of osteogenic mediumon BMSCs: the morphology of BMSCs 21 days after treatment period were assessed
by Alizarin red staining, As shown in the Figures, suitable concentrations of Sea cucumber alcoholic extract (6.25, 12.5, 25 µg/ml) formed
red mineral amorphous deposits that it was indicating one of the prominent marker differentiation to osteoblasts. A) Control. B - E) (3.5,
6.25, 12.5, 25 µg/ml). Magnification 200 µm
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