Ulinastatin intervention for polymethyl methacrylate-induced MC3T3-E1 mouse preosteoblast apoptosis

doi:10.3969/j.issn.2095-4344.2014.43.010

Abstract

Abstract:

BACKGROUND: Previous studies have indicated that ulinastatin can inhibit RANKL-induced osteoclastogenesis on RAW264.7 cells and also lower matrix metalloproteinase-9 expression and activity. However, it remains be unclear whether ulinastatin has the intervention effect on polymethyl methacrylate (PMMA)-induced MC3T3-E1 mouse preosteoblast apoptosis or not.
OBJECTIVE: To explore the intervention role of ulinastatin on the PMMA-induced MC3T3-E1 mouse preosteoblast apoptosis and its effects on type I collagen, osteocalcin, matrix metalloproteinase-2 mRNA
expression.
METHODS: MC3T3-E1 mouse preosteoblasts at passages 6 and 7 were divided into four groups: blank group (only cultured MC3T3-E1 mouse preosteoblast), PMMA-induced group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension), low dose ulinastatin group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension+500 U/mL ulinastatin) and high dose ulinastatin group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension+5 000 U/mL ulinastatin). MTT method was adopted to detect the proliferation activity of proliferative activity of MC3T3-E1 mouse preosteoblast; alizarin red staining method was used to observe mineralization nodules of MC3T3-E1 mouse preosteoblast among different groups; the change of apoptosis rate for MC3T3-E1 cells was detected by flow cytometry analysis; semi-quantitative RT-PCR was taken to analyze type I collagen, osteocalcin, matrix metalloproteinase-2 mRNA expression level in MC3T3-E1 mouse preosteoblasts among different groups.
RESULTS AND CONCLUSION: Compared with the blank group, PMMA significantly inhibited the proliferation activity of MC3T3-E1 mouse preosteoblast (P < 0.05), and however significantly promoted cells apoptosis (P < 0.05). After addition of different concentrations of ulinastatin (500, 5 000 U/mL), the proliferation activity of MC3T3-E1 mouse preosteoblasts significantly raised (P < 0.05), and cells apoptosis rate significantly decreased (P < 0.05), showing the dose and time-dependent relation. Type I collagen and osteocalcin mRNA expression levels both significantly decreased after co-culture in PMMA group compared with the blank group (P < 0.05), matrix metalloproteinase-2 mRNA expression level, however, significantly increased (P < 0.05). After intervention with 5000 U/mL ulinastatin, type I collagen and osteocalcin mRNA expression levels both significantly increased, while matrix metalloproteinase-2 mRNA expression level significantly decreased (P < 0.05). PMMA group showed no obvious mineralization nodules. Yet, mineralization nodules were formed in the blank group, high and low dose ulinastatin groups. These results indicate that ulinastatin could have the inhibitory effect on the PMMA-induced MC3T3-E1 mouse preosteoblast apoptosis, and it could promote type I collagen and osteocalcin mRNA expression and yet suppress matrix metalloproteinase-2 mRNA expression.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: osteoblasts, osteocalcin, matrix metalloproteinase-2

CLC Number:

R318

Ru Jiang-ying, Cong Yu, Zhao Jian-ning, Guo Ting, Yu Lei, Ding Hao, Jiang Hui . Ulinastatin intervention for polymethyl methacrylate-induced MC3T3-E1 mouse preosteoblast apoptosis[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(43): 6945-6950.

Figures/Tables 3

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