Effects of Kangguzengsheng Capsules-containing serum on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Our previous studies have found that the Kangguzengsheng Capsules can promote fracture healing.OBJECTIVE: To explore the influence of Kangguzengsheng Capsules-containing serum on the proliferation andosteogenic differentiation of bone marrow mesenchymal stem cells.METHODS: Forty Sprague-Dawley rats were randomly divided into four groups. The capsule powder was resolved into1 mL natural saline and intragastrically administered into rats according to 1.16 g/100 g, 3.48 g/100 g, 10.44 g/100 g in low,medium and high dose groups. Rats in control group were given equal volume of natural saline. After consecutiveadministration for 12 days, blood samples from the abdominal aorta were collected and serum samples were isolated andpreserved until use. Bone marrow mesenchymal stem cells were isolated and purified by the whole marrow adhesion method,and identified by flow cytometry. Harvested cells were divided into four groups and cultured in the osteogenic culture mediumcontaining different kinds of serum samples as described above. MTT method was adopted to test the cell proliferation at 24,48 and 72 hours of culture. Alkaline phosphatase activity in cells was detected at 7 and 14 days of culture. Alkalinephosphatase staining was performed at 7 days of culture, and alizarin red staining performed at 14 days of culture.RESULTS AND CONCLUSION: Compared with the blank control group, bone marrow mesenchymal stem cells showedno significant changes in the proliferative ability and alkaline phosphatase activity in the low dose group, but these twoindices were significantly increased in the high and medium dose groups at 48 and 72 hours of culture or at 7 and 14days of culture, respectively (P < 0.05). Cells positive for alkaline phosphatase and alizarin red staining were observed inthe low, medium and high dose groups, and the cell staining was most remarkable in the high dose group. To conclude,Kangguzengsheng Capsules-containing serum can promote the proliferation and osteogenic differentiation of bonemarrow mesenchymal stem cells.