Expression of rat transforming growth factor beta 1 gene in the transfected osteoblasts

ABSTRACT

BACKGROUND:

The cells of biomaterial compound cytokine gene or compound transfected cytokine gene that are transplanted into the area of bone defect can promote bone repair.

To investigate the feasibility of gene therapy for bone defect after rat transforming growth factor (TGF)β1 gene is transfected into osteoblasts.

A controlled and observational experiment.

Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS Five newborn Sprague-Dawley rats of either gender were included.

The experiment was conducted in the laboratory of Orthopedic Department, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, from February to September 2000.Rat osteoblasts were transfected with pcDNA3-TGF-β1 plasmid by lipofectamine mediated gene transfer, and the plasmid pcDNA transfected cells were set as control group. The transient expression of TGF-β1 was detected by strept avidin-biotin-peroxidase complex (SABC) method and in situ hybridization detection 24 hours later. The cells transfected by G418 for 2 weeks were detected with SABC to investigate the stable expression of TGF-β1 gene.MAIN OUTCOME

SABC method and in situ hybridization detection were applied to detect gene expression.of pcDNA3-TGF-β1 transfected osteoblasts and in situ hybridization detection After the osteoblasts were transfected for 24 hours, cytoplast was full of brown granules and there were no brown granules in the cytoplast of blank carrier transfected cells, indicating that TGF-β1 mRNA was signifiG418 transfected cells still had high expression of TGF-β1 after 2-week G418 screening.

Osteoblasts can express cytokine immediately with high effect using gene transfection technique. The stable and high expression is presented after TGF-β1 gene is transfected into osteoblasts at the moment of transfection and after 2-week screening, proving the feasibility of gene therapy for bone defect when cytokine gene is transfected into osteoblasts.