Decellularized Disc Hydrogels for hBMSCs tissue-specific differentiation and tissue regeneration

Yizhong Peng; Xiangcheng Qing; Hui Lin; Donghua Huang; Jinye Li; Shuo Tian; Sheng Liu; Xiao Lv; Kaige Ma; Rui Li; Zilong Rao; Ying Bai; Songfeng Chen; Ming Lei; Daping Quan; Zengwu Shao

doi:10.1016/j.bioactmat.2021.03.014

Decellularized Disc Hydrogels for hBMSCs tissue-specific differentiation and tissue regeneration

Bioact Mater. 2021 Mar 22;6(10):3541-3556. doi: 10.1016/j.bioactmat.2021.03.014. eCollection 2021 Oct.

Authors

Yizhong Peng¹, Xiangcheng Qing¹, Hui Lin¹, Donghua Huang², Jinye Li¹, Shuo Tian¹, Sheng Liu¹, Xiao Lv¹, Kaige Ma¹, Rui Li³, Zilong Rao³, Ying Bai⁴, Songfeng Chen⁵, Ming Lei¹, Daping Quan^{3

4}, Zengwu Shao¹

Affiliations

¹ Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
² Musculoskeletal Tumor Center, Department of Orthopedics, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
³ School of Chemistry, Sun Yat-sen University, Guangzhou, 510127, China.
⁴ School of Materials Science and Engineering, Sun Yat-sen University, Guangzhou, 510127, China.
⁵ Department of Orthopaedic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou City, 450052, China.

Abstract

Tissue specificity, a key factor in the decellularized tissue matrix (DTM), has shown bioactive functionalities in tuning cell fate-e.g., the differentiation of mesenchymal stem cells. Notably, cell fate is also determined by the living microenvironment, including material composition and spatial characteristics. Herein, two neighboring tissues within intervertebral discs, the nucleus pulposus (NP) and annulus fibrosus (AF), were carefully processed into DTM hydrogels (abbreviated DNP-G and DAF-G, respectively) to determine the tissue-specific effects on stem cell fate, such as specific components and different culturing methods, as well as in vivo regeneration. Distinct differences in their protein compositions were identified by proteomic analysis. Interestingly, the fate of human bone marrow mesenchymal stem cells (hBMSCs) also responds to both culturing methods and composition. Generally, hBMSCs cultured with DNP-G (3D) differentiated into NP-like cells, while hBMSCs cultured with DAF-G (2D) underwent AF-like differentiation, indicating a close correlation with the native microenvironments of NP and AF cells, respectively. Furthermore, we found that the integrin-mediated RhoA/LATS/YAP1 signaling pathway was activated in DAF-G (2D)-induced AF-specific differentiation. Additionally, the activation of YAP1 determined the tendency of NP- or AF-specific differentiation and played opposite regulatory effects. Finally, DNP-G and DAF-G specifically promoted tissue regeneration in NP degeneration and AF defect rat models, respectively. In conclusion, DNP-G and DAF-G can specifically determine the fate of stem cells through the integrin-mediated RhoA/LATS/YAP1 signaling pathway, and this tissue specificity is both compositional and spatial, supporting the utilization of tissue-specific DTM in advanced treatments of intervertebral disc degeneration.

Keywords: 2D, two-dimensional; 3D, three-dimensional; AF, annulus fibrosus; Col I–S, collagen type I solution; DAF, decellularized annulus fibrosus; DAF-G, decellularized annulus fibrosus hydrogel; DAF-S, decellularized annulus fibrosus solution; DNP, decellularized nucleus pulposus; DNP-G, decellularized nucleus pulposus hydrogel; DNP-S, decellularized nucleus pulposus solution; DTM, decellularized tissue matrix; Decellularized tissue matrix; Differentiation; ECM, extracellular matrix; FAF, fresh annulus fibrosus; FNP, fresh nucleus pulposus; IDD, intervertebral disc degeneration; Intervertebral disc; MSC, mesenchymal stem cell; NP, nucleus pulposus; Tissue specificity; YAP1; YAP1, yes-associated protein 1.