siRNA incorporated in slow-release injectable hydrogel continuously silences DDIT4 and regulates nucleus pulposus cell pyroptosis through the ROS/TXNIP/NLRP3 axis to alleviate intervertebral disc degeneration.

IF 4.7 2区 医学 Q2 CELL & TISSUE ENGINEERING
Miao Ma, Chongjing Zhang, Zeyuan Zhong, Yajun Wang, Xuegang He, Daxue Zhu, Zhi Qian, Baoqing Yu, Xuewen Kang
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Abstract

Aims: In this investigation, we administered oxidative stress to nucleus pulposus cells (NPCs), recognized DNA-damage-inducible transcript 4 (DDIT4) as a component in intervertebral disc degeneration (IVDD), and devised a hydrogel capable of conveying small interfering RNA (siRNA) to IVDD.

Methods: An in vitro model for oxidative stress-induced injury in NPCs was developed to elucidate the mechanisms underlying the upregulation of DDIT4 expression, activation of the reactive oxygen species (ROS)-thioredoxin-interacting protein (TXNIP)-NLRP3 signalling pathway, and nucleus pulposus pyroptosis. Furthermore, the mechanism of action of small interfering DDIT4 (siDDIT4) on NPCs in vitro was validated. A triplex hydrogel named siDDIT4@G5-P-HA was created by adsorbing siDDIT4 onto fifth-generation polyamidoamine (PAMAM) dendrimer using van der Waals interactions, and then coating it with hyaluronic acid (HA). In addition, we established a rat puncture IVDD model to decipher the hydrogel's mechanism in IVDD.

Results: A correlation between DDIT4 expression levels and disc degeneration was shown with human nucleus pulposus and needle-punctured rat disc specimens. We confirmed that DDIT4 was responsible for activating the ROS-TXNIP-NLRP3 axis during oxidative stress-induced pyroptosis in rat nucleus pulposus in vitro. Mitochondria were damaged during oxidative stress, and DDIT4 contributed to mitochondrial damage and ROS production. In addition, siDDIT4@G5-P-HA hydrogels showed good delivery activity of siDDIT4 to NPCs. In vitro studies illustrated the potential of the siDDIT4@G5-P-HA hydrogel for alleviating IVDD in rats.

Conclusion: DDIT4 is a key player in mediating pyroptosis and IVDD in NPCs through the ROS-TXNIP-NLRP3 axis. Additionally, siDDIT4@G5-P-HA hydrogel has been found to relieve IVDD in rats. Our research offers an innovative treatment option for IVDD.

加入缓释注射水凝胶的 siRNA 可持续沉默 DDIT4,并通过 ROS/TXNIP/NLRP3 轴调节髓核细胞的热解,从而缓解椎间盘退变。
目的:在这项研究中,我们对髓核细胞(NPCs)施加了氧化应激,认识到DNA损伤诱导转录本4(DDIT4)是椎间盘变性(IVDD)的一个组成部分,并设计了一种能够向IVDD传递小干扰RNA(siRNA)的水凝胶:方法:建立了氧化应激诱导的 NPCs 损伤体外模型,以阐明 DDIT4 表达上调、活性氧(ROS)-硫氧还蛋白相互作用蛋白(TXNIP)-NLRP3 信号通路激活以及髓核热解的机制。此外,还在体外验证了小干扰 DDIT4(siDDIT4)对鼻咽癌的作用机制。我们利用范德华相互作用将 siDDIT4 吸附在第五代聚酰胺胺(PAMAM)树枝状聚合物上,然后涂上透明质酸(HA),制成了一种名为 siDDIT4@G5-P-HA 的三重水凝胶。此外,我们还建立了大鼠穿刺 IVDD 模型,以解读水凝胶在 IVDD 中的作用机制:结果:在人类髓核和针刺大鼠椎间盘标本中显示出 DDIT4 表达水平与椎间盘退变之间的相关性。我们证实,在氧化应激诱导的体外大鼠髓核热解过程中,DDIT4 负责激活 ROS-TXNIP-NLRP3 轴。线粒体在氧化应激过程中受损,而 DDIT4 对线粒体的损伤和 ROS 的产生做出了贡献。此外,siDDIT4@G5-P-HA 水凝胶显示了 siDDIT4 对神经核团的良好输送活性。体外研究表明,siDDIT4@G5-P-HA 水凝胶具有缓解大鼠 IVDD 的潜力:结论:DDIT4是通过ROS-TXNIP-NLRP3轴介导NPCs热休克和IVDD的关键角色。此外,siDDIT4@G5-P-HA 水凝胶还能缓解大鼠的 IVDD。我们的研究为 IVDD 提供了一种创新的治疗方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Bone & Joint Research
Bone & Joint Research CELL & TISSUE ENGINEERING-ORTHOPEDICS
CiteScore
7.40
自引率
23.90%
发文量
156
审稿时长
12 weeks
期刊介绍: The gold open access journal for the musculoskeletal sciences. Included in PubMed and available in PubMed Central.
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