Wang Zhenyu, Leng Jiali, Zhao Yuguang, Yu Dehai, Xu Feng, Song Qingxu, Qu Zhigang, Zhuang Xinming, Liu Yi
Department of Spinal Surgery, the First Hospital of Jilin University, Changchun, China.
Nursing Platform of Bone, Joint and Sports Medicine, the First Hospital of Jilin University, Changchun, China.
Cell Physiol Biochem. 2017;43(6):2327-2337. doi: 10.1159/000484385. Epub 2017 Oct 27.
BACKGROUND/AIMS: Mechanical load can regulate disc nucleus pulposus (NP) biology in terms of cell viability, matrix homeostasis and cell phenotype. N-cadherin (N-CDH) is a molecular marker of NP cells. This study investigated the role of N-CDH in maintaining NP cell phenotype, NP matrix synthesis and NP cell viability under high-magnitude compression.
Rat NP cells seeded on scaffolds were perfusion-cultured using a self-developed perfusion bioreactor for 5 days. NP cell biology in terms of cell apoptosis, matrix biosynthesis and cell phenotype was studied after the cells were subjected to different compressive magnitudes (low- and high-magnitudes: 2% and 20% compressive deformation, respectively). Non-loaded NP cells were used as controls. Lentivirus-mediated N-CDH overexpression was used to further investigate the role of N-CDH under high-magnitude compression.
The 20% deformation compression condition significantly decreased N-CDH expression compared with the 2% deformation compression and control conditions. Meanwhile, 20% deformation compression increased the number of apoptotic NP cells, up-regulated the expression of Bax and cleaved-caspase-3 and down-regulated the expression of Bcl-2, matrix macromolecules (aggrecan and collagen II) and NP cell markers (glypican-3, CAXII and keratin-19) compared with 2% deformation compression. Additionally, N-CDH overexpression attenuated the effects of 20% deformation compression on NP cell biology in relation to the designated parameters.
N-CDH helps to restore the cell viability, matrix biosynthesis and cellular phenotype of NP cells under high-magnitude compression.
背景/目的:机械负荷可在细胞活力、基质稳态和细胞表型方面调节椎间盘髓核(NP)生物学特性。N-钙黏蛋白(N-CDH)是NP细胞的分子标志物。本研究探讨了N-CDH在高幅度压缩下维持NP细胞表型、NP基质合成和NP细胞活力中的作用。
将接种在支架上的大鼠NP细胞使用自行研发的灌注生物反应器进行灌注培养5天。在细胞经受不同压缩幅度(低幅度和高幅度:分别为2%和20%压缩变形)后,研究NP细胞生物学特性,包括细胞凋亡、基质生物合成和细胞表型。未加载的NP细胞用作对照。采用慢病毒介导的N-CDH过表达进一步研究N-CDH在高幅度压缩下的作用。
与2%变形压缩和对照条件相比,20%变形压缩条件下N-CDH表达显著降低。同时,与2%变形压缩相比,20%变形压缩增加了凋亡NP细胞数量,上调了Bax和裂解型半胱天冬酶-3的表达,下调了Bcl-2、基质大分子(聚集蛋白聚糖和胶原蛋白II)以及NP细胞标志物(磷脂酰肌醇蛋白聚糖-3、CAXII和角蛋白-19)的表达。此外,N-CDH过表达减弱了20%变形压缩对NP细胞生物学特性相关指定参数的影响。
N-CDH有助于在高幅度压缩下恢复NP细胞的细胞活力、基质生物合成和细胞表型。