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Calcitonin Gene-Related Peptide-Induced Calcium Alginate Gel Combined with Adipose-Derived Stem Cells Differentiating to Osteoblasts

View Article: PubMed Central

ABSTRACT

Calcitonin gene-related peptide (CGRP) has been confirmed with induction osteoblastic differentiation, but if it can make the three-dimensional culture of adipose-derived stem cells (ADSCs) to the osteoblastic differentiation, thus constructing tissue-engineered bone rare reports. To investigate the feasibility of exogenous CGRP-induced calcium alginate gel combined with ADSCs from rabbits in three-dimensional condition to construct tissue-engineered bone. ADSCs were obtained by collagenase I digestion of the subcutaneous adipose tissue of inguinal region of New Zealand rabbits. At the third passage, cells were mixed with sodium alginate to prepare calcium alginate gel, and the cells were assigned into two-group cultivates in 24 orifice plates. ADSCs in the control group were treated with DMEM/F-12 medium supplemented with 10−2 mol/L β-glycerophosphate sodium, 10−7mol/L dexamethasone, 50 mg/L ascorbic acid, 0.1 % volume fraction of fetal bovine serum. ADSCs in the experimental group were incubated with the same medium as above, and in addition 1.5 µg/L CGRP was added. The cells proliferation and the mRNA expressions of collagen I and osteocalcin were detected by MTT and RT-PCR assays, respectively and alkaline phosphatase(ALP)and calcium concentration at different induction time were detected. The cell proliferation curves were S shaped. The OD values of experimental group were higher than those of control group at 1, 3, 5, 7, 14, and 21 days after osteogenic induction (P < 0.05). ALP and alizarin red stains of ADSCs were all positive, but golden round nodes became bigger and more in the experimental group compared with the control group after 2 weeks. At 7 and 14 days, collagen I and osteocalcin mRNA expression were greater in the experimental group than the control group. ALP and calcium concentration of experimental group were higher than that of control group at 1, 2, 3, 4 weeks after osteogenic induction (P < 0.05). Thus, these results show that the CGRP-induced ADSCs combined with calcium alginate gel to osteoblasts differentiation.

No MeSH data available.


The cells proliferation of both groups (*Compared with control group,P < 0.05,n = 6)
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Fig4: The cells proliferation of both groups (*Compared with control group,P < 0.05,n = 6)

Mentions: MTT detection shows that OD values were increased with the prolongation of time of cells in both groups. But except for the first day, the differences in the different time points were statistically significant in the CGRP-induced group and the control group (P < 0.05). And change in different induced phases was statistically significant in CGRP-induced group (P < 0.05), showing that the CGRP promotes osteogenic differentiation of ADSCs (Fig. 4).Fig. 4


Calcitonin Gene-Related Peptide-Induced Calcium Alginate Gel Combined with Adipose-Derived Stem Cells Differentiating to Osteoblasts
The cells proliferation of both groups (*Compared with control group,P < 0.05,n = 6)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4628090&req=5

Fig4: The cells proliferation of both groups (*Compared with control group,P < 0.05,n = 6)
Mentions: MTT detection shows that OD values were increased with the prolongation of time of cells in both groups. But except for the first day, the differences in the different time points were statistically significant in the CGRP-induced group and the control group (P < 0.05). And change in different induced phases was statistically significant in CGRP-induced group (P < 0.05), showing that the CGRP promotes osteogenic differentiation of ADSCs (Fig. 4).Fig. 4

View Article: PubMed Central

ABSTRACT

Calcitonin gene-related peptide (CGRP) has been confirmed with induction osteoblastic differentiation, but if it can make the three-dimensional culture of adipose-derived stem cells (ADSCs) to the osteoblastic differentiation, thus constructing tissue-engineered bone rare reports. To investigate the feasibility of exogenous CGRP-induced calcium alginate gel combined with ADSCs from rabbits in three-dimensional condition to construct tissue-engineered bone. ADSCs were obtained by collagenase I digestion of the subcutaneous adipose tissue of inguinal region of New Zealand rabbits. At the third passage, cells were mixed with sodium alginate to prepare calcium alginate gel, and the cells were assigned into two-group cultivates in 24 orifice plates. ADSCs in the control group were treated with DMEM/F-12 medium supplemented with 10&minus;2 mol/L &beta;-glycerophosphate sodium, 10&minus;7mol/L dexamethasone, 50&nbsp;mg/L ascorbic acid, 0.1&nbsp;% volume fraction of fetal bovine serum. ADSCs in the experimental group were incubated with the same medium as above, and in addition 1.5&nbsp;&micro;g/L CGRP was added. The cells proliferation and the mRNA expressions of collagen I and osteocalcin were detected by MTT and RT-PCR assays, respectively and alkaline phosphatase(ALP)and calcium concentration at different induction time were detected. The cell proliferation curves were S shaped. The OD values of experimental group were higher than those of control group at 1, 3, 5, 7, 14, and 21&nbsp;days after osteogenic induction (P&nbsp;&lt;&nbsp;0.05). ALP and alizarin red stains of ADSCs were all positive, but golden round nodes became bigger and more in the experimental group compared with the control group after 2&nbsp;weeks. At 7 and 14&nbsp;days, collagen I and osteocalcin mRNA expression were greater in the experimental group than the control group. ALP and calcium concentration of experimental group were higher than that of control group at 1, 2, 3, 4&nbsp;weeks after osteogenic induction (P&nbsp;&lt;&nbsp;0.05). Thus, these results show that the CGRP-induced ADSCs combined with calcium alginate gel to osteoblasts differentiation.

No MeSH data available.