J Korean Acad Periodontol.  2000 Jun;30(2):347-358. 10.5051/jkape.2000.30.2.347.

The Effects of Platelet-Derived Growth Factor-BB on the Expression of Bone Matrix Protein in the MC3T3-E1 Cells

• Kim MS
• Lee JM
• Suh JY

• KMID: 2191816
• DOI: http://doi.org/10.5051/jkape.2000.30.2.347

Abstract

Bone remodeling results from the combined process of bone resorption and new bone formation which is regulated in part by some of the polypeptide growth factors such as platelet derived growth factor(PDGF), which has been known to be an important local regulator of bone cell activity and participate in normal bone remodeling. This process includes strictly regulated gene expression of several bone matrix proteins such as type I collagen and osteopontin, a 44 kDa phosphorylated glycoprotein, which has important roles in bone formation. The purpose of this study is to evaluate the effecs of PDGF-BB on the mRNA expression of bone matrix protein, type I collagen and osteopontin, in MC3T3-E1 cell culture. Cells were seeded at 5x10(5) cells in 10 ml of minimum essential medium alpha(alpha-MEM) containig 10% fetal bovine serum, 10 mM beta glycerophosphate. 0.1, 1, 10 ng/ml PDGF-BB were added to the cells for the day 3, 7, 14, 21, 28 and cultured for 24 hours. Type I collagen cDNA, Hf677, and osteopontin cDNA were used as probes for northern blot analysis. Total cellular RNA was purified at indicated day and northern blot analysis was performed. The results were as follows: Type I collagen mRNA expressions were higher at the day 3 and 7, and lower in the day 14, 21 in the control groups. In the experimental groups, mRNA expressions were increased when 0.1 ng/ml PDGF-BB were added on the day 3, 7, 21, and decreased in dose-dependent manner on the day 14, decreased at all added dose on the day 28. Osteopontin mRNA expressions were highest in the day 21 groups and lowest in the day 14 groups in the control groups. Interesting results were shown in the day 14 and 21 groups. We found that osteopontin mRNA level was increased in dose dependent manner in the day 14 groups, and decreased dose dependent manner in the day 21 groups. In conclusion, PDGF-BB may have various control effects on type I mRNA expression in the growth and differentiation process of MC3T3-E1 cells and may have contrary regulatory effects on osteopontin mRNA expression. For examples, when the baseline level of osteopontin mRNA was low, as in the day 14, PDGF-BB up-regulated osteopontin mRNA expression in dose dependent manner, and when the baseline level was high as in the day 21, PDGF-BB down-regulated dose dependent manner. Thus, it may be useful for clinical application in periodontal regeneration procedure if further study were performed.

MeSH Terms

Blood Platelets
Blotting, Northern
Bone Matrix*
Bone Remodeling
Bone Resorption
Cell Culture Techniques
Collagen Type I
DNA, Complementary
Gene Expression
Glycoproteins
Intercellular Signaling Peptides and Proteins
Osteogenesis
Osteopontin
Regeneration
RNA
RNA, Messenger
Collagen Type I
DNA, Complementary
Glycoproteins
Intercellular Signaling Peptides and Proteins
Osteopontin
RNA
RNA, Messenger