산 처리된 한천의 인접부위에서 배양된 조골세포에서의 알칼리성 인산분해효소 발현
Expression of alkaline Phosphatase in Osteoblasts Cultured Adjacent to Acidified Agar
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Purpose : Many studies working on the influence of pH change on osteoclast and osteoblast have been reported but most of them were about experiments of culture medium being directly acidified and basified. In this study, experimental model with medium which can be cultured with temporary acidic stimulation on osteoblast was designed. When consistent releasing of acidified material from medium was occurred, we were to figure out activation of osteoblast in adjacent area with the expression of ALP. Materials and Methods : For experimental groups, we dilute agar with distilled water to 1% concentration and put phosphoric acid. We made solution of pH 7.2, 6.5, 6.0 and 5.0. The solution was poured into 6 well plates and they were solidified for 24 hours then half of gelled agar was removed from each of plates. Experimental groups were medium with MC3T3-E1 cell cultured in half of the well and control groups were medium cultured with no treatment. Each of them was cultured for 4 days and 7days. Results : When analyzing color density of ALP using Image Pro plus program, we detected lower level in experimental groups than in control groups, but there was no significant difference. In experimental groups of 4-day, osteoblast adjacent to acidified agar showed stronger ALP expression than that of distant part. Conclusion : As an observation with ALP expression we found temporary releasing of acid in initial culture could increase ALP expression of osteoblast cell in adjacent part but could not induce consistent activation.
Abstract
Purpose : Many studies working on the influence of pH change on osteoclast and osteoblast have been reported but most of them were about experiments of culture medium being directly acidified and basified. In this study, experimental model with medium which can be cultured with temporary acidic stimulation on osteoblast was designed. When consistent releasing of acidified material from medium was occurred, we were to figure out activation of osteoblast in adjacent area with the expression of ALP. Materials and Methods : For experimental groups, we dilute agar with distilled water to 1% concentration and put phosphoric acid. We made solution of pH 7.2, 6.5, 6.0 and 5.0. The solution was poured into 6 well plates and they were solidified for 24 hours then half of gelled agar was removed from each of plates. Experimental groups were medium with MC3T3-E1 cell cultured in half of the well and control groups were medium cultured with no treatment. Each of them was cultured for 4 days and 7days. Results : When analyzing color density of ALP using Image Pro plus program, we detected lower level in experimental groups than in control groups, but there was no significant difference. In experimental groups of 4-day, osteoblast adjacent to acidified agar showed stronger ALP expression than that of distant part. Conclusion : As an observation with ALP expression we found temporary releasing of acid in initial culture could increase ALP expression of osteoblast cell in adjacent part but could not induce consistent activation.
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