Efficacy for Bone Deterioration in Mice with Osteoporosis and Pharmaceutical Properties of a Novel Compound Calcium Carbonate Granule

Figure 5

CCCG promoted osteogenic differentiation of HOS cell in vitro: (a) the effect of CCCG on HOS cells activity was determined by CCK-8 compared to control groups (n = 5). (b) ALP staining and activity quantitative in HOS cells after drug administration (n = 3). Under the catalysis of alkaline phosphatase, BCIP is hydrolyzed to produce a highly reactive product, which reacted with NBT to form an insoluble dark blue to blue-purple NBT-Formazan. The red arrow indirectly represented the enzyme activity of ALP. (c) Mineralized nodule staining and quantitative analysis of mineralized nodules in HOS cells. Red arrows represented calcium nodules (orange-red deposits). (d) Representative western blots showing β-catenin and Runx-2 protein expression of HOS cells, treated with or without CCCG for 48 h (n = 3). (e) RT-qPCR was used to quantify the mRNA expression of osteogenic marker genes ALP, RUNX2, COL-I, OTC, OPN, BMP2, and OPG, treated with or without CCCG for 48 h. The mRNA expression levels were normalized to the mRNA expression of GAPDH (n = 4). Control: HOS cells were cultured in osteogenic induction medium; CCCG-L: HOS cells were cultured in osteogenic induction medium with low-dose CCCG; CCCG-M: HOS cells were cultured in osteogenic induction medium with middle-dose CCCG; CCCG-H: HOS cells were cultured in osteogenic induction medium with high-dose CCCG; CCD: HOS cells were cultured in osteogenic induction medium with CCD. , and .

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