Calcitonin gene-related peptide induces the osteogenic differentiation of adipose derived stem cells combined with calcium alginate gel

doi:10.3969/j.issn.2095-4344.2013.42.003

Abstract

Abstract:

BACKGROUND: Calcitonin gene-related peptide has been confirmed to induce osteogenic differentiation, but whether it can induce the osteogenic differentiation of adipose-derived stem cells under three-dimensional culture to construct tissue-engineered bone is rarely reported.
OBJECTIVE: To investigate the feasibility of exogenous calcitonin gene-related peptide to induce osteogenic differentiation of adipose-derived stem cells combined with calcium alginate gel in three-dimensional condition.
METHODS: Adipose-derived stem cells were gained by collagenase I digestion of the subcutaneous adipose tissue of both inguinal regions of New Zealand rabbits. Passage 3 cells were mixed with sodium alginate to prepare calcium alginate gel, and then the cells were assigned into two groups and cultured in 24-well plates. Adipose-derived stem cells in the control group were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 supplemented with 10^-2 mol/L β-glycerophosphate sodium, 10^-7 mol/L dexamethasone, 50 mg/L ascorbic acid, 10% fetal bovine serum. While, adipose-derived stem cells in the experimental group were incubated with the same medium as above, but 1.5 µg/L calcitonin gene-related peptide was added. The cells proliferation and the mRNA expressions of collagen I and osteocalcin were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and reverse transcription-PCR respectively, and alkaline phosphatase and calcium concentration were detected at different induction time.
RESULTS AND CONCLUSION: The cell proliferation curves were S shaped. The absorbance values of the experimental group were higher than those of the control group at1, 3, 5, 7, 14, 21 days after osteogenic induction (P < 0.05). Alkaline phosphatase and alizarin red staining of adipose-derived stem cells was all positive, but golden round nodes became more and bigger in the experimental group compared with the control group after 2 weeks. At 7 and 14 days, collagen I and osteocalcin mRNA expressions were higher in the experimental group than in the control group. Alkaline phosphatase activity and calcium concentration of the experimental group were higher than those of the control group at 1, 2, 3, 4 weeks after osteogenic induction (P < 0.05). Results showed that the calcitonin gene-related peptide can induce the osteogenic differentiation of adipose-derived stem cells combined with calcium alginate gel.

Key words: biocompatible materials, calcitonin gene-related peptide, gels, stem cells

CLC Number:

R318

Huang Chang-zhi, Yang Xiao-ning, Liu Da-cheng, Sun Yi-gong, Dai Xing-ming . Calcitonin gene-related peptide induces the osteogenic differentiation of adipose derived stem cells combined with calcium alginate gel[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(42): 7356-7362.

Figures/Tables 6

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