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Abstract
Polypeptide growth factors belong to a class of potent biologic mediators which regulate cell differentiation, proliferation, migration and metabolism. IGF-I is polypeptides secreted by skeletal cells and is considered as regulators of bone formation. The purpose of this study is to evaluate the effects of IGF-I on bone nodule formation and alkaline phosphatase activity of MC3T3-E1 cells.
MC3T3-E1 cells were seeded at 1×104 cells/well, 1×105 cells/well in alpha-modified Eagle medium containing 10% fetal bovine serum, 10 mM β-glycerophosphate and 50µg/ml of ascorbic acid. Before 48 hours of indicated time, medium were changed with serum free medium. After 24 hours, 0.1, 1, 10 ng/ml IGF-I were added to the cells and cultured for 3, 7, 14, 21, 28 days. And histochemical analysis was done and ALP activity was measured and was expressed as nmol/min/mg of protein.
The bone nodule formation in MC3T3-E1 cells of IGF-I was seen at 21, 28 days, but there were no difference between control group and experimental groups.
The ALP activity decreased when it is compare to control 2 group except for 1 ng/ml, 10 ng/ml IGF-I of 21-day-groups and 1 ng/ml IGF-I of 28-day-groups.
Dose response effects of IGF-I of ALP activity in MC3T3-E1 cells were seen the highest ALP activity at 1ng/ml until 21days and the highest ALP activity at 10 ng/ml of 28 day-groups.
The peak times were seen at 7-day group, 14-day group on control group and experimental group respectively, and 1 ng/ml group was the highest ALP activity.
From the above results, IGF-I was not seen notable effect on bone nodule formation and decreased ALP activity of MC3T3-E1 cells but the use of IGF-I to mediate biological stimulation of MC3T3-E1 cells shows promise for future therapeutic application.
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