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

The Author(s) 2015. This article is published with open access at Springerlink.com 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 differen- tiation, thus constructing tissue-engineered bone rare re- ports. 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 re- gionof NewZealandrabbits.At thethird 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 b-g- lycerophosphate sodium, 10 -7 mol/L dexamethasone, 50 mg/L ascorbic acid, 0.1 % volume fraction of fetal bovine serum. ADSCs in the experimental group were in- cubated with the same medium as above, and in addition 1.5 lg/L CGRP was added. The cells proliferation and the mRNA expressions of collagen I and osteocalcin were de- tected by MTT and RT-PCR assays, respectively and alka- line phosphatase(ALP)and calcium concentration at differentinductiontimeweredetected.Thecellproliferation 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.