Effect of zoladronate on the proliferation and osteogenic differentiation of rat bone mesenchymal stem cells

doi:10.7518/hxkq.2019.03.003

Abstract

Abstract:

Objective To evaluate the effect of zoledronate acid (ZA) on the proliferation and osteogenic differentiation of rat mesenchymal stem cells (BMSCs). Methods The BMSCs isolated from the SD rats were cultured with different con-centrations of ZA (1, 5, 10, and 20 μmol·L ^-1), and the contro1 group received the same volume of culture medium but without ZA. Cell counting kit-8 was used to detect proliferation activity in each group. Alkaline phosphatase (ALP) staining and alizarin red staining were used to detect the osteogenic differentiation ability in each group. The gene expression levels of ALP, bone morphogenetic protein-2 (BMP-2), typeⅠcollagenase (COL-Ⅰ), runt-related transcription factor-2 (Runx-2), zinc finger structure transcription factor (Osx), osteocalcin (OCN), and osteopontin (OPN) were evaluated by real-time quantitative polymerase chain reaction (qRT-PCR). Results Zoledronate at 1 μmol·L ^-1concentration had no effect on the proliferation and osteogenic differentiation of BMSCs. No significant difference was observed between this group and the control group (P>0.05). When the ZA concentration was more than 1 μmol·L ^-1, ZA inhibited the proliferation and osteogenic differentiation of BMSCs, and the effect was concentration dependent. The difference between each group and the control group was statistically significant (P<0.05). At ZA concentration of 5 μmol·L ^-1, ZA enhanced the expression of ALP, BMP-2, COL-Ⅰ, Runx-2, Osx, OCN, and OPN (P<0.05). However, at ZA concentration of more than 5 μmol·L ^-1, the expression levels of osteogenicrelated genes in each group was lower than those of the control group (P<0.05). Conclusion Low ZA concen-tration has no effect on the proliferation and osteogenic differentiation of BMSCs. ZA at 5 μmol·L ^-1 concentration inhibits the proliferation but promotes the osteogenic differentiation of BMSCs. High ZA concentration inhibits the proliferation and osteogenic differentiation of BMSCs.

Key words: bisphosphonates, bone mesenchymal stem cells, proliferation, osteogenic differentiation

CLC Number:

Q254

Jiehang Li,Zhifei Su,Xuan Bai,He Yuan,Jiyao Li. Effect of zoladronate on the proliferation and osteogenic differentiation of rat bone mesenchymal stem cells[J]. West China Journal of Stomatology, 2019, 37(3): 242-247.

Figures/Tables 5

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基因名称	引物序列
GAPDH	F:5’-GCCTTCCGTGTTCCTACC-3’
	R:5’-AGAGTGGGAGTTGCTGTTG-3’
ALP	F:5’-CCAACTCTTTTGTGCCAGAGA-3’
	R:5’-GGCTACATTGGTGTTGAGCTTTT-3’
BMP-2	F:5’-ACATGCAGCGTGAAATCCTG-3’
	R:5’-CGTGGTATAGGTCCAACATGAAG-3’
COL-Ⅰ	F:5’-ACGTCCTGGTGAAGTTG-3’
	R:5’-CAGGGAAGCCTCTTTCTCCT-3’
Runx-2	F:5’-GACTGTGGTTACCGTCATGGC-3’
	R:5’-ACTTGGTTTTTCATAACAGCGGA-3’
Osx	F:5’-CCACCAAGAATTACTCAGAACCC-3’
	R:5’-AAGGACCGACTCACTCAGTCT-3’
OCN	F:5’-CTGACCTCACAGATCCCAAGC-3’
	R:5’-TGGTCTGATAGCTCGTCACAAG-3’
OPN	F:5’-GGGGACTATGCACCTGAGC-3’
	R:5’-GACTGTCGAAATGGGCTACCT-3’