Abstract
Purpose: To investigate the effect of epigallocatechin-3-gallate (EGCG) on bone metabolism and osteoblastic activity. Methods: MG-63 human osteoblast-like cells were treated with varied concentrations of EGCG. Alkaline phosphatase (ALP) activity and matrix mineralization assays were carried out on the treated and untreated MG-63 human osteoblast-like cells. Beta-catenin mRNA level was determined by quantitative real-time polymerase chain reaction (q-PCR). Results: The results showed that EGCG treatment significantly increased ALP and mineralization activities at concentrations of 15 and 30 μM, in a dose-dependent manner. Furthermore, EGCG treatment significantly increased beta-catenin mRNA expression by 70.7 ± 11.0 and 126.7 ± 35.1 %, respectively, at EGCG concentration of 15 and 30 μM. In addition, the stimulating effect of EGCG treatment on ALP activity was abolished by co-treatment with ICI 182,780, an antagonist of estrogen receptor (ER). Again, the increase in beta-catenin MRNA, when treated with EGCG, was inhibited by co-treatment with ICI 182,780. Conclusion: EGCG promotes osteoblastic activity in human osteoblast-like cells, by Wnt signaling through estrogen receptor (ER) pathway. Keywords: Epigallocatechin-3-gallate, Osteoporosis, Osteoclast, Proliferation, Estrogen receptor
Highlights
Osteoporosis is a musculoskeletal disease characterized by low bone mineral density (BMD) and high risk of fragility fractures
We studied the effects of EGCG on bone metabolism regarding osteoblastic activity in MG-63 human osteoblastlike cells, and evaluated the possible underlying mechanisms
In order to determine whether EGCG could increase osteoblastic cell differentiation, its effect on Alkaline phosphatase (ALP) activity as investigated
Summary
Osteoporosis is a musculoskeletal disease characterized by low bone mineral density (BMD) and high risk of fragility fractures. Osteoblasts, known as bone-forming cells, are the major osteoprogenitor cells, whose proliferation and differentiation will eventually result in the formation of the mineralized extracellular matrix [4]. The degree of matrix mineralization of MG-63 cells was determined by Alizarin Red S staining method after EGCG treatment for 6 days [13]. After removing the unbonded stain by rinsing three times with distilled water, the amount of stain on the tested cells was extracted by shaking the cells with 10 mM sodium phosphate containing 10 % cetylpyridinium chloride (Sigma, USA) for 15 min. The data were expressed as percentage relative to the vehicle control group, after the relative matrix mineralization values were normalized by the relative cell viability. P < 0.05 was considered statistically significant
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