Spheroid Formation Enhances the Regenerative Capacity of Nucleus Pulposus Cells via Regulating N-CDH and ITGβ1 Interaction.
Spheroid Formation Enhances the Regenerative Capacity of Nucleus Pulposus Cells via Regulating N-CDH and ITGβ1 Interaction.
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DOI：
10.7150/ijbs.70903
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发表时间：
2022
影响因子：
9.2
通讯作者：
Zhou, Qiang
中科院分区：
文献类型：
--
作者：
Wang, Yiyang;Wang, Haoming;Zhuo, Yunyun;Hu, Yanzhu;Li, Xiaoxiao;Xu, Yanqin;Sun, Biemin;Liu, Min;Zou, Luetao;Liu, Liehua;Luo, Lei;Zhao, Chen;Li, Pei;Zhou, Qiang
背景资料：髓核退变是椎间盘退变性疾病的核心病理改变，但目前尚无有效的治疗方法。近年来，随着生物材料和组织工程学的迅速发展，生物材料辅助细胞移植成为治疗椎间盘退变的一种很有前途的方法。然而，由于目前不适当的细胞接种或移植方法，NP细胞（NPC）的生物学特性较弱，如增殖能力中等，自我更新能力弱，细胞外基质（ECM）合成能力极低，严重限制了其应用。研究方法：在此，我们开发了一种三维（3D）球化培养方法来构建NPC球体，并在体外和体内研究这些球体的修复和再生潜力。比较NPC球状体和单层培养NPC的体外生物学特性（包括细胞活力和增殖）和体内功能（包括抗变性潜力和诱导组织修复能力）。此外，基于RNA-seq的转录组分析和一系列的功能实验进行了阐明他们的差异，参与组织再生过程的潜在机制。结果：与单层NPC相比，NPC球状体具有明显的上级自我更新和ECM合成能力。在体内，NPC球状体产生更多功能性ECM组分，主要是聚集蛋白聚糖（ACAN）和II型胶原（Col 2），并在部分NP切除诱导的椎间盘退变模型中显著促进NP再生。此外，NPC球体的生物学特性和功能在一定程度上受到NPC表面表达的两种关键的机械敏感ECM受体N-cadherin（N-CDH）和Integrinβ1（ITGβ1）的相互作用的调节。结论：鼻咽癌组织球化培养有利于细胞更新和功能性ECM的生成。参与该再生过程的分子机制与N-CDH和ITGβ1相互作用介导的ECM同源骨形成的调节密切相关。此外，水凝胶辅助的NPC球体移植策略可能用于未来IVD变性的治疗。
Background: Nucleus pulposus (NP) degeneration is the core pathological change of intervertebral disc (IVD) degenerative diseases, but currently, no effective therapy is available. With the rapid development of biomaterials and tissue engineering in recent years, biomaterial-assisted cell transplantation becomes a promising therapy for IVD degeneration. However, the application is severely limited by the weak biological characteristics of NP cells (NPCs), such as a moderate proliferation ability, weak self-renewal capacity, and minimal extracellular matrix (ECM) synthesis capacity, caused by the current inappropriate cell seeding or grafting methods. Methods: Here, we developed a three-dimensional (3D) spheroidizing culture method to construct NPC spheroids and investigated repair and regeneration potential of these spheroids in vitro and in vivo. The in vitro biological characteristics (including cell viability and proliferation), and in vivo functions (including anti-degeneration potential and ability to induce tissue repair) of NPC spheroids and monolayer-cultured NPCs were compared. Furthermore, an RNA-seq-based transcriptome analysis and a series of function experiments were performed to elucidate the potential mechanisms of their differences that were involved in the tissue regeneration process. Results: NPC spheroids exhibited obviously superior self-renewal and ECM synthesis capacities compared to monolayers of NPCs in vitro. In vivo, NPC spheroids generated more functional ECM components, primarily aggrecan (ACAN) and collagen type II (Col2), and markedly promoted NP regeneration in the disc degeneration model induced by partial NP excision. Additionally, the biological characteristics and functions of NPC spheroids were to some extent regulated by the interaction of N-cadherin (N-CDH) and Integrinβ1 (ITGβ1), two key mechanosensing ECM-receptors expressed on NPCs. Conclusions: The NPC spheroidizing culture method is beneficial for cell renewal and the generation of functional ECM in NP tissue. The molecular mechanism involved in this regeneration process is closely associated with the regulation of the N-CDH and ITGβ1 interaction-mediated ECM homeostesis. Moreover, the strategy of hydrogel-assisted NPC spheroids transplantation may potentially be used in the future treatment of IVD degeneration.
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影响因子：
12.7
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Bian Q;Ma L;Jain A;Crane JL;Kebaish K;Wan M;Zhang Z;Edward Guo X;Sponseller PD;Séguin CA;Riley LH;Wang Y;Cao X
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Setton LA