Effect of transforming growth factor-beta3 on the proliferation and osteogenic capability of osteoblasts

doi:10.3969/j.issn.2095-4344.3156

Abstract

Abstract: BACKGROUND: Less is reported on the effect of transforming growth factor-β3 (TGF-β3) on the proliferation and osteogenic capability of osteoblasts till now.
OBJECTIVE: To investigate the ability of TGF-β3 in different doses on the proliferation and osteogenic capability of osteoblasts.
METHODS: We isolated osteoblasts from the calvarium of neonatal New Zealand white rabbit according to our previous method, purified and identified the cells using differential attachment method. The osteoblasts were divided into a control group and an experimental group. The osteoblasts in the control group were cultured with conventional culture medium, and those in the experimental group were cultured in the medium containing separately 0.1, 1, 10, and 100 μg/L TGF-β3. The morphology and growth of osteoblasts were observed daily. Cell proliferation was detected by MTT assay and the growth curve was drawn at 1, 3, 5, 7, 9, 11, and 13 days of culture. Alkaline phosphatase activity was measured at 1, 3, 5, 7, 9, 11, 13, and 15 days of culture. The expression levels of collagen 1A1, Runt related transcription factor 2 (Runx-2) and osteocalcin genes were measured by RT-qPCR at 7 and 14 days of culture. Alizarin red S staining was carried out to testify mineralized matrix productivity of each group at 21 days of culture. The study was approved by the Animal Experimental Ethics Committee of the First Affiliated Hospital of Xinjiang Medical University.
RESULTS AND CONCLUSION: The osteoblasts were isolated, purified and identified successfully. MTT assay results showed that the cell proliferation activity of 10 μg/L TGF-β3 group was higher than that in the control group (P < 0.05). The alkaline phosphatase activity of 10 and 100 μg/L TGF-β3 groups was higher than that in the control group (P < 0.05). RT-qPCR results revealed that the mRNA expressions of collagen 1A1, Runx-2 and osteocalcin in the 10 μg/L TGF-β3 group were always higher than those in the control group (P < 0.05). The mRNA expressions of collagen 1A1, Runx-2 and osteocalcin in the 100 μg/L TGF-β3 group were higher than those in the control group at 7 days of culture (P < 0.05). Alizarin red S staining showed that the mineralized nodules of the 10 and 100 μg/L TGF-β3 groups, especially in the 10 μg/L TGF-β3 group (P < 0.05), were bigger and more than those in the control group. To conclude, 10-100 μg/L TGF-β3 can improve the proliferation and osteogenic capability of osteoblasts and these effects are more obvious when the concentration of TGF-β3 is 10 μg/L.

Key words: bone, growth factor, osteogenesis, osteoblast, cell proliferation, alkaline phosphatase, gene

CLC Number:

Ailimaierdan·Ainiwaer, Wang Ling, Gu Li, Dilidaer•Taxifulati, Wang Shan, Yin Hongbin. Effect of transforming growth factor-beta3 on the proliferation and osteogenic capability of osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(17): 2664-2669.

Figures/Tables 5

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