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出生后生长及机械应激期间大鼠膜内成骨中基质金属蛋白酶基因的表达

Expression of matrix metalloproteinase genes in the rat intramembranous bone during postnatal growth and upon mechanical stresses.

作者信息

Collins John M, Ramamoorthy Kamalanathan, Da Silveira Adriana, Patston Philip, Mao Jeremy J

机构信息

Tissue Engineering Laboratory Rm 237, University of Illinois at Chicago, MC 841, 801 South Paulina Street, Chicago, IL 60612-7211, USA.

出版信息

J Biomech. 2005 Mar;38(3):485-92. doi: 10.1016/j.jbiomech.2004.04.018.

Abstract

A cranial suture consists of neural-crest derived cells and matrices between mineralized skull bones. Little is known regarding the involvement of matrix metalloproteinases (MMPs) in the degradation of extracellular matrix of cranial sutures. In the postnatal rat model, the posterior frontal suture (PFS) undergoes complete ossification between P12-P22, whereas the sagittal suture (SS) remains patent. The present study utilized reverse transcriptase-polymerase chain reaction (RT-PCR) to explore the expression of MMP-1 and MMP-2 genes in the PFS and SS in P8 and P32 rats, and also to determine whether these MMP genes are modulated by exogenous mechanical forces. RNA was isolated from P8 and P32 normal PFS and SS each by pooling sutural specimens from 14 to 20 rats. RT-PCR analysis and semi-quantitative luminosity demonstrated the expression of MMP-1 and MMP-2 genes in the patent P8 PFS, P8 SS, and P32 SS, but no apparent MMP-2 expression in the physiologically ossified P32 PFS. Exogenous cyclic forces applied to the maxilla at 1000 mN and 4 Hz elicited corresponding cyclic bone strain waveforms with peak strain of 134.14+/-38.15 muepsilon (mean+/-S.D.) for the PFS, and 28.35+/-10.86 muepsilon for the SS in P32 rats. These cyclic forces delivered for 20 min/d over 2 consecutive days induced the expression of MMP-2 gene in the physiologically fused P32 PFS that was not expressed without mechanical stresses. Taken together, these data suggest potentially important roles of MMP genes in the postnatal development of cranial sutures, and their susceptibility to mechanical stresses.

摘要

颅缝由神经嵴衍生细胞和矿化颅骨之间的基质组成。关于基质金属蛋白酶(MMPs)参与颅缝细胞外基质降解的情况,人们了解甚少。在出生后的大鼠模型中,额后缝(PFS)在出生后第12至22天之间完全骨化,而矢状缝(SS)则保持开放。本研究利用逆转录聚合酶链反应(RT-PCR)来探究出生后第8天和第32天大鼠的PFS和SS中MMP-1和MMP-2基因的表达情况,并确定这些MMP基因是否受外源性机械力的调节。通过汇集14至20只大鼠的缝合标本,分别从出生后第8天和第32天的正常PFS和SS中分离RNA。RT-PCR分析和半定量光度法显示,在开放的出生后第8天的PFS、出生后第8天的SS和出生后第32天的SS中存在MMP-1和MMP-2基因的表达,但在生理上已骨化的出生后第32天的PFS中未观察到明显的MMP-2表达。以1,000 mN和4 Hz的频率对上颌施加外源性循环力,在出生后第32天的大鼠中,PFS产生了相应的循环骨应变波形,峰值应变为134.14±38.15 μɛ(平均值±标准差),SS的峰值应变为28.35±10.86 μɛ。连续2天每天施加这些循环力20分钟,可诱导生理上已融合的出生后第32天的PFS中MMP-2基因的表达,而在没有机械应力的情况下该基因不表达。综上所述,这些数据表明MMP基因在颅缝出生后发育中可能具有重要作用,并且它们对机械应力敏感。

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