Collagen cross-linking influences osteoblastic differentiation
Collagen cross-linking influences osteoblastic differentiation
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DOI：
10.1007/s00223-008-9136-3
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发表时间：
2008-05-01
影响因子：
4.2
通讯作者：
Varga, F.
中科院分区：
文献类型：
--
作者：
Turecek, C.;Fratzl-Zelman, N.;Varga, F.
成骨细胞合成胶原基质，胶原基质自身调节前体细胞向成熟成骨细胞的分化。它们表达赖氨酸氧化酶（LOX），它参与胶原交联过程。甲状腺素，如-氨基丙腈（- APN），抑制稳定基质的形成。本研究旨在探讨交联对成骨细胞分化的影响。分别给MC3T3-E1细胞接种400 μ M β APN或不加APN处理1周。然后，取出活细胞，在细胞外基质上播种新的MC3T3-E1细胞，并在无β APN的情况下培养1周。分离RNA，用定量逆转录-聚合酶链反应测定特异性标记基因的表达。用傅里叶变换红外光谱法测定β - APN处理后特异性交联的变化。形成的胶原基质显示，与对照培养物相比，骨胶原的两种主要交联deH-DHLNL和pyr显著减少。基因表达研究显示胶原α 1 (I) （COL1A1）增加150%。LOX和骨钙素（OCN） mRNA的表达显著下调至75%左右。当新鲜的MC3T3-E1细胞接种在没有β - APN的改变基质上时，COL1A1 mRNA表达上调（140%），OCN下调（60%），LOX mRNA表达不受影响。这些结果表明，β - APN处理不仅破坏胶原交联的形成，而且影响成骨细胞的活性和表达。综上所述，在有甲状腺激素存在的情况下产生的被破坏的基质会影响成骨细胞基因的表达，即使在它不存在的情况下也是如此。
Osteoblasts synthesize collagen matrix, which itself regulates the differentiation of precursor cells into mature osteoblasts. They express lysyl oxidase (LOX), which is involved in the collagen cross-linking process. Lathyrogens, like beta-aminopropionitrile (beta APN), inhibit the formation of a stable matrix. The aim of the present study was to investigate the influence of cross-linking on osteoblastic differentiation. MC3T3-E1 cells were seeded and treated with or without 400 mu M beta APN for 1 week. Thereafter, living cells were removed and, on this extracellular matrix, new MC3T3-E1 cells were seeded and cultured for 1 week without beta APN. RNA was isolated, and expression of specific marker genes was determined by quantitative reverse transcription-polymerase chain reaction. Changes in specific cross-links after beta APN treatment were measured with Fourier-transform infrared spectroscopy. The collagen matrix that formed showed a significant reduction of two major cross-links of bone collagen, deH-DHLNL and pyr, compared to control cultures. Gene expression studies showed an increase of collagen alpha 1 (I) (COL1A1) to 150%. Expression of LOX and osteocalcin (OCN) mRNA was significantly downregulated to about 75%. When fresh MC3T3-E1 cells were seeded on this altered matrix without beta APN, COL1A1 mRNA expression was upregulated (140%), OCN was downregulated (60%), and LOX mRNA expression remained unaffected. These results indicate that beta APN treatment not only disrupts collagen cross-link formation but also affects osteoblastic activity and expression. In conclusion, the disrupted matrix produced in the presence of lathyrogen influences, even in its absence, the expression of osteoblastic genes.