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Abstract
Objectives
To determine the effect of bone morphogenetic protein- 2 in the osteogenesis of human ligamentum flavum cells and test the feasibility of gene transfer to these cells.
Summary of literature review
Bone morphogenetic protein- 2 (BMP- 2) is known to be an important factor in the differentiation and maintenance of the osteoblastic phenotype. Tissue engineering for osteogenesis in ligamentum flavum by BMP- 2 and gene transfer has not been previously studied.
Materials and Methods
Ligmentum flavum cells were harvested and cultured from surgical patients with spinal stenosis. BMP- 2 was produced by transfecting pcDNA 3.1/Hygro/BMP- 2 into CHO cells using Lipofectamine 2000. A denovirus- lacZ (A d/lacZ) was also produced, and administered with BMP- 2 to cell culture. The expression of lacZ was analyzed by X - gal staining. Bone formation was assessed by alkaline phosphatase, von Kossa, and alizarin Red- S staining, and the expression of osteocalcin was determined immunocytochemically.
Results
Ligamentum flavum cell cultures with A d/lacZ showed marker gene expression. BMP- 2 induced osteogenesis in ligamentum flavum cells as evidenced by alkaline phosphatase, von Kosa, and alizarin Red- S staining. A lso, cell culture with BMP- 2 showed strong positivity with osteocalcin by immunocytochemistry.
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