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Copyright of Chinese Journal of Osteoporosis is the property of Chinese Journal of Osteoporosis and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Osteoporosis is the property of Chinese Journal of Osteoporosis and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

To study the effects of Zuoguiwan ( ZGW) on nourishing kidney essence, strengthening bone and marrow, and promotion of the proliferation of bone marrow mesenchymal stem cells ( BMSCs) and differentiation into osteoblasts. Methods The ZGW containing serum was prepared. 10% PBS DMEM group, osteogenic liquid group, 2. 5% serum of health rats with osteogenic liquid group, and 2. 5% serum containing ZGW osteogenic liquid group were set up. MTT method was used to observe the proliferation of BMSCs in each group. The differentiation of BMSCs into osteoblasts was tested using alkaline phosphatase calcium cobalt staining method . Alkaline phosphatase ( AKP) method was used to detect the content of AKP in cell culture medium. The expression of runt related transcription factor 2 ( Runx2 ), osteocalcin ( OPN), and alkaline phosphatase (ALP) protein in BMSCs were detected using Western blotting. Results Rapid proliferation of BMSCs was showed in 2. 5% serum containing ZGW osteogenic liquid group. The plateau stage appeared earlier. It promoted the secretion of AKP in BMSCs, increased the content of ALP, and up-regulated the protein expression levels of Runx2, ALP, and OPN in BMSCs cells. Conclusion The molecular mechanism of ZGW against osteoporosis may be related to the regulation of Runx2, activation of ALP and OPN protein expression, promotion of BMSCs proliferation and differentiation into osteoblasts. [ABSTRACT FROM AUTHOR]

Objective To explore the optimal conditions of cell counting kit-8 assay (CCK-8) and MTT assay on the detection of the proliferation of bone marrow mesenchymal stem cells (BMSCs) intervened by serum containing Zuoguiwan (ZGW). The advantages and disadvantages of the two method were compared. Methods BMSCs were intervened by serum containing ZGW. The optimal wavelength, culture time, sensitivity, and cell proliferation of BMSCs were detected by using CCK-8 and MTT method, respectively. Results Optimal detection wavelengths of CCK-8 and MTT reagents were 450 nm and 550 nm, respectively. The best incubation time of CCK-8 and MTT assay were 3 h and 4 h, respectively. Cell inoculation density should be respectively controlled by 0. 1x 10/ hole and 0. 25x 10/ hole, respectively. When the inoculation density of BMSCs was higher than 0.25x10*/well and 0. 5x10'/well, the OD values detected by CCK-8 assay and MTT assay entered the plateau stage earlier. It was easy to appear the false S-shaped growth curve. However, when the cell density was lower than 0.05x 10/well and 0. 1x 10/ well, it was difficult to form S-shaped growth curve because of slow growing, which could not reflect the proliferation characteristics of BMSCs. Compared with CCK-8 assay, there was no significant change in response to the increase of OD value of BMSCs in MTT assay, when BMSCs density was lower than 0. 1x 10/well or higher than 2x 10/well.. When BMSCs density was between 0.25x 10/well and 1x 10/well, the response of MTT assay to the elevation of OD value of BMSCs increased significantly (P< 0.05). Conclusion MTT assay is more suitable for the study of serum containing ZGW on the proliferation and differentiation of BMSCs. [ABSTRACT FROM AUTHOR]