The migration of chondrocytes from healthy to injured tissues is one of the most important challenges during cartilage repair. Additionally, maintenance of the chondrogenic phenotype remains another limitation, especially during monolayer culture in vitro. Using both the differentiated and undifferentiated chondrogenic ATDC5 cell line, we showed that extracellular nucleotides are able to increase the migration rate of chondrocytes without affecting their chondrogenic phenotype. We checked the potency of natural nucleotides (ATP, ADP, UTP, and UDP) as well as their stable phosphorothioate analogs, containing a sulfur atom in the place of one nonbridging oxygen atom in a phosphate group. We also detected P2y1, P2y2, P2y4, P2y6, P2y12, P2y13, and P2y14 mRNA transcripts for nucleotide receptors, demonstrating that P2y1 and P2y13 are highly upregulated in differentiated ATDC5 cells. We showed that ADPβS, UDPβS, and ADP are the best stimulators of migration of differentiated chondrocytes. Additionally, ADP and ADPβS positively affected the expression of type II collagen, a structural component of the cartilage matrix.

译文

软骨细胞从健康组织向受伤组织的迁移是软骨修复过程中最重要的挑战之一。此外,软骨形成表型的维持仍然是另一个限制,尤其是在体外单层培养期间。使用分化和未分化的软骨源性ATDC5细胞系,我们表明细胞外核苷酸能够增加软骨细胞的迁移率,而不会影响其软骨形成表型。我们检查了天然核苷酸 (ATP,ADP,UTP和UDP) 及其稳定的硫代磷酸酯类似物的效力,其中含有硫原子代替磷酸基团中的一个非桥接氧原子。我们还检测到核苷酸受体的P2y1,P2y2,P2y4,P2y6,P2y12,P2y13和P2y14 mRNA转录本,表明P2y1和P2y13在分化的ATDC5细胞中高度上调。我们发现ADP β s,udp β s和ADP是分化软骨细胞迁移的最佳刺激因子。此外,ADP和ADP β 对软骨基质的结构成分II型胶原的表达产生积极影响。

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