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.
Highlights
Calcitonin gene-related peptide (CGRP) is one of the most abundant neuropeptides
Calcitonin gene-related peptide (CGRP) has been confirmed with induction osteoblastic differentiation, but if it can make the three-dimensional culture of adiposederived stem cells (ADSCs) to the osteoblastic differentiation, constructing tissue-engineered bone rare reports
Cells were mixed with sodium alginate to prepare calcium alginate gel, and the cells were assigned into two-group cultivates in 24 orifice plates
Summary
Calcitonin gene-related peptide (CGRP) is one of the most abundant neuropeptides. So far CGRP research has concentrated on its role on the nervous and the cardiovascular systems [1,2,3], but not on bone defect repair. Hukkanen et al (1993) found that CGRP-positive nerve fibers initially show degradation of periosteum and quickly proliferate around the bone fracture in rats [4] It is followed by the progress of the callus formation and bone remodeling along with the change in distribution and density. Aoki et al (2004) confirmed that a large number of CGRP-positive nerve fibers were emerged in fiber granulation tissue, periosteum, and bone tissue during healing process of fracture in rats in an orderly manner with the degree of callus formation and hyperplasia of bone remodeling [5]. We examined the effect of CGRP on the rabbit adipose-derived stem cells (ADSCs) to the proliferation and bone cell differentiation using alginate three-dimensional culture method in order to provide experimental basis for repair of bone defect with tissue engineering bone
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call