Growth characteristics of basic fibroblast growth factor gene-transfected bone marrow mesenchymal stem cells seeded on coral skeleton in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND: Jaw defects are common clinically. It is desirable to find ideal seed cells combined with scaffolds to construct tissue engineered jaws for curing these diseases. OBJECTIVE: To investigate the growth characteristics of bone marrow mesenchymal stem cells (BMSCs) transfected with basic fibroblast growth factor (bFGF) gene after seeded on coral scaffold in vitro. DESIGN, TIME AND SETTING: An experimental study of bone tissue engineering was performed in the Research Institute of Stomatology, Sun Yat-sen University between March 2006 and June 2008. MATERIALS Natural coral from China Hainan bench was made into pieces of 8 mm×8 mm×2 mm. METHODS: BMSCs were isolated from New England rabbits by density gradient centrifugation and then purified by adherent separation. bFGF-pcDNA3 gene was transfected into BMSCs using Lipofectamine TM 2000. bFGF gene-transfected (transfected group) or untransfected (untransfected group)BMSCs were seeded on different coral scaffolds. In addition, bFGF gene-transfected BMSCs were simply cultured but not on the coral scaffold for control (simple culture group). MAIN OUTCOME MEASURES: BMSC proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay and BMSC growth on coral scaffold was observed under the scanning electron microscope. RESULTS: MTT assay showed that the BMSC proliferation rate was significantly higher in the transfected group than in the untransfected group (P < 0.05) and that there was no significant difference in BMSC proliferation between the transfected and simple culture groups (P > 0.05). Scanning electron microscope results displayed that BMSCs adhered to and spread over the coral scaffold, exhibiting various appearances, with some cells had grown into scaffold micropores or spanned micropore surface, and some extracellular matrix secreted by BMSCs were found. CONCLUSION: The transfected group exhibited better growth of BMSCs transfected by bFGF gene than the untransfected group. These findings indicate that coral skeleton does not influence BMSC proliferation and can be used as a scaffold of BMSCs to construct tissue-engineered bone.