Osteogenesis by Transfer of Bone Morphogenetic Protein-2 cDNA in Ligamentum Flavum Cells: Approach Toward Tissue Engineering.
10.4184/jkss.2002.9.4.263
- Author:
Seong Hwan MOON
1
;
Hyang KIM
;
Un Hye KWON
;
Jung Hoon WON
;
Hak Sun KIM
;
Soo Bong HAHN
;
Hwan Mo LEE
Author Information
1. Department of Orthopedic Surgery, Yonsei University, College of Medicine, Seoul, Korea. hwanlee@yumc.yonsei.ac.kr
- Publication Type:Original Article
- Keywords:
BMP-2;
adenovirus;
osteogenesis;
Tissue engineering
- MeSH:
Adenoviridae;
Alkaline Phosphatase;
Animals;
Cell Culture Techniques;
CHO Cells;
Cricetinae;
DNA, Complementary*;
Gene Expression;
Humans;
Immunohistochemistry;
Ligamentum Flavum*;
Osteoblasts;
Osteocalcin;
Osteogenesis*;
Phenotype;
Spinal Stenosis;
Tissue Engineering*
- From:Journal of Korean Society of Spine Surgery
2002;9(4):263-269
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
STUDY DESIGN: In-vitro experiment. 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 pcDNA3.1/Hygro/BMP-2 into CHO cells using Lipofectamine 2000. Adenovirus-lacZ (Ad/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 Ad/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. Also, cell culture with BMP-2 showed strong positivity with osteocalcin by immunocytochemistry. CONCLUSION: BMP-2 more strongly induced the osteogenesis of ligamentum flavum, and also its gene transfer to ligamentum flavum was found to be feasible. These results may open a new era of ligamentum flavum tissue engineering.
