Spheroid Formation Enhances the Regenerative Capacity of Nucleus Pulposus Cells via Regulating N-CDH and ITG beta 1 Interaction

作者全名："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"

作者地址："[Wang, Yiyang; Zhuo, Yunyun; Hu, Yanzhu; Li, Xiaoxiao; Zou, Luetao; Liu, Liehua; Luo, Lei; Zhao, Chen; Li, Pei; Zhou, Qiang] Chongqing Med Univ, Dept Orthoped, Affiliated Hosp 3, Chongqing 401120, Peoples R China; [Wang, Yiyang; Wang, Haoming; Zhuo, Yunyun; Hu, Yanzhu; Li, Xiaoxiao; Zou, Luetao; Li, Pei; Zhou, Qiang] Chongqing Med Univ, Tissue Repairing & Biotechnol Res Ctr, Affiliated Hosp 3, Chongqing 401120, Peoples R China; [Wang, Haoming] Chongqing Univ, Dept Orthoped, Three Gorges Cent Hosp, Chongqing 404000, Peoples R China; [Xu, Yanqin; Sun, Biemin; Liu, Min] Chongqing Univ, Coll Chem & Chem Engn, Chongqing 400044, Peoples R China"

通信作者："Li, P; Zhou, Q (通讯作者)，Chongqing Med Univ, Dept Orthoped, Affiliated Hosp 3, Chongqing 401120, Peoples R China.; Li, P; Zhou, Q (通讯作者)，Chongqing Med Univ, Tissue Repairing & Biotechnol Res Ctr, Affiliated Hosp 3, Chongqing 401120, Peoples R China."

来源：INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:000804481100001

JCR分区：Q1

影响因子：9.2

年份：2022

卷号：18

期号：9

开始页：3676

结束页：3696

文献类型：Article

关键词：Nucleus pulposus cell (NPC); extracellular matrix (ECM); tissue regeneration; microtissue; hydrogel

摘要："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 Integrin beta 1 (ITG beta 1), 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 ITG beta 1 interaction-mediated ECM homeostesis. Moreover, the strategy of hydrogel-assisted NPC spheroids transplantation may potentially be used in the future treatment of IVD degeneration."

基金机构："Natural Science Foundation of China [81974346, 81772378]; National Key Research and Develop-ment Program of China [2018YFC1105800]; Postdoctoral Science Foundation of China [2021M6 93925]; Natural Science Foundation of Chongqing, China [cstc2020jcyj-msxmX0148]; Project of Innovative Science Research for Postgraduate of Chongqing Municipal Education Committee, Chongqing, China [CYB20168, CYS20226]"

基金资助正文："This work was supported by grants from the Natural Science Foundation of China (No.81974346, No.81772378) ; National Key Research and Develop-ment Program of China (No.2018YFC1105800) ; Postdoctoral Science Foundation of China (2021M6 93925) ; Natural Science Foundation of Chongqing, China (No. cstc2020jcyj-msxmX0148) ; Project of Innovative Science Research for Postgraduate of Chongqing Municipal Education Committee, Chongqing, China (CYB20168, CYS20226) ."