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Augmented osteogenesis of mesenchymal stem cells using a fragmented Runx2 mixed with cell-penetrating, dimeric a-helical peptide.
European Journal of Pharmaceutical Sciences ( IF 4.3 ) Pub Date : 2019-12-30 , DOI: 10.1016/j.ejps.2019.105210
So Hee Nam 1 , Yan Lee 1 , Joon Hyung Ahn 1 , Chun Kee Chung 2 , Hee-Jin Yang 3 , Sung Bae Park 3 , Sangmok Jang 4
Affiliation  

The intracellular delivery of transcription factor/cofactor using cell penetrating peptide (CPP) can lead to selective osteogenesis. The present work investigates the cell-penetrating potential of the a cyclic, α-helical cell-penetrating peptide based on leucine and lysine residues (cLK) for intracellular delivery in MC3T3 cells and the osteogenic effects of a C-terminal proline‑serine‑threonine-rich (PST) domain of Runx2 using cLK in rat mesenchymal stem cells (MSCs). We confirmed that the combination of cLK and fluorescein 5-isothiocyanate (FITC)-fragmented-Runx2 (fRunx2) showed an enhanced cell-penetrating activity of FITC-fRunx2 compared with FITC-fRunx2 alone. In addition, the fRunx2-cLK group showed strong staining with alizarin red compared with other groups and the degree of alizarin red staining in the fRunx2-cLK group was also 1.2-fold higher than that in the fRunx2-Tat group. The ALP and osteocalcin gene expression levels in the fRunx2-cLK group were higher than those in the other groups. The fRunx2 transferred effectively into the cytoplasm aided by the cLK peptide and augmented the osteogenic differentiation of MSCs.

中文翻译:

使用片段Runx2与穿透细胞的二聚a-螺旋肽混合,增强间充质干细胞的成骨作用。

使用细胞穿透肽(CPP)在细胞内传递转录因子/辅因子可以导致选择性成骨。本工作研究了基于亮氨酸和赖氨酸残基(cLK)的环状α螺旋细胞穿透肽在MC3T3细胞中向细胞内传递的细胞穿透潜力以及C末端脯氨酸-丝氨酸-苏氨酸的成骨作用大鼠间充质干细胞(MSCs)中使用cLK实现Runx2的富含(PST)域。我们确认,与单独的FITC-fRunx2相比,cLK和5异硫氰酸荧光素(FITC)片段化Runx2(fRunx2)的组合显示出增强的FITC-fRunx2细胞穿透活性。另外,与其他组相比,fRunx2-cLK组的茜素红染色强烈,fRunx2-cLK组的茜素红染色程度也为1。比fRunx2-Tat组高2倍。fRunx2-cLK组的ALP和骨钙素基因表达水平高于其他组。fRunx2在cLK肽的辅助下有效转移到细胞质中,并增强了MSC的成骨分化。
更新日期:2019-12-31
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