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KMID : 1160620200250010071
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Preventive Nutrition and Food Science
2020 Volume.25 No. 1 p.71 ~ p.77
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Effects of the Whey Protein Hydrolysates of Various Protein Enzymes on the Proliferation and Differentiation of 3T3-E1 Osteoblasts
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Jo Kyung-Ae
Hong Ki-Bae
Suh Hyung-Joo
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Abstract
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In this study, we used various proteinases to investigate the effect of whey protein hydrolysates on proliferation and differentiation of MC3T3-E1 osteoblasts. To confirm hydrolysis of the whey protein hydrolysates, the yield and α-amino acid content were determined. Since osteogenic cell activity is an important factor in osteogenesis, we evaluated the proliferation of osteogenic cells by measuring 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and alkaline phosphatase (ALP) activity. To analyze bone matrix formation, we identified calcium deposition by staining with Alizaline red-S. The free amino acid content was significantly higher in the whey protein hydrolysates prepared using Protamex, Flavourzyme, and Alcalase than in the control. When cells were treated with 500 μg/mL of whey protein hydrolysates prepared using Protamex and Alcalase, cell proliferation increased by 120% and 130%, respectively, compared with the control group. In addition, ALP activity was significantly higher following treatment with 500 μg/mL of whey protein hydrolysates prepared using Protamex and Alcalase (142.61% and 135.06%, respectively; p<0.05). Furthermore, when treated with 125 ?g/mL of the same hydrolysates, the rate of calcium deposition increased significantly to 157.56% compared with the control group (p<0.05). Therefore, our results suggest that whey protein hydrolysates prepared using Protamex and Alcalase may have more beneficial effects on osteoblast proliferation and bone health than those prepared using other proteolytic enzymes.
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KEYWORD
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alkaline phosphatase, calcium deposition, MC3T3-E1, proliferation, whey protein hydrolysates
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