Enhanced osteogenic differentiation of mesenchymal stem cells in ankylosing spondylitis: a study based on a three-dimensional biomimetic environment

Abstract

The mechanism of pathological osteogenesis in Ankylosing spondylitis (AS) is largely unknown. Our previous studies demonstrated that the imbalance between BMP-2 and Noggin secretion induces abnormal osteogenic differentiation of marrow-derived mesenchymal stem cells (MSCs) from AS patients in a two-dimensional culture environment. In this study, HA/β-TCP scaffolds were further used as a three-dimensional (3D) biomimetic culture system to mimic the bone microenvironment in vivo to determine the abnormal osteogenic differentiation of AS-MSCs. We demonstrated that when cultured in HA/β-TCP scaffolds, AS-MSCs had a stronger osteogenic differentiation capacity than that of MSCs from healthy donors (HD-MSCs) in vitro and in vivo. This dysfunction resulted from BMP2 overexpression in AS-MSCs, which excessively activated the Smad1/5/8 and ERK signalling pathways and finally led to enhanced osteogenic differentiation. Both the signalling pathway inhibitors and siRNAs inhibiting BMP2 expression could rectify the enhanced osteogenic differentiation of AS-MSCs. Furthermore, BMP2 expression in ossifying entheses was significantly higher in AS patients. In summary, our study demonstrated that AS-MSCs possess enhanced osteogenic differentiation in HA/β-TCP scaffolds as a 3D biomimetic microenvironment because of BMP2 overexpression, but not Noggin. These results provide insights into the mechanism of pathological osteogenesis, which can aid in the development of niche-targeting medications for AS.

Introduction

Ankylosing spondylitis (AS) is a common autoimmune disease that affects the axial skeleton, and one of its critical pathogenic features is new bone formation at local sites of entheses¹, which is confined to the periosteal bone compartment, outside the cortical bone lining. In recent years, many researchers have investigated the mechanism of pathological osteogenesis in AS^2,3. However, the concrete details of pathological osteogenesis are still controversial, impeding the development of a specific medication for AS. Furthermore, the lack of a specific therapeutic target for pathological osteogenesis results in a high disability rate in AS patients⁴.

Mesenchymal stem cells (MSCs) are one of the most important kinds of multipotential stem cells and possess strong immunoregulatory and trilineage differentiation abilities⁵. Many studies have determined that MSCs are the major origin of osteoblasts⁶. Nevertheless, dysfunction of the osteogenic differentiation ability of MSCs contributes to bone metabolism disorders in rheumatic diseases⁷. For example, although mesenchymal progenitors increased, decreased osteoblast differentiation is progressive with disease development in Interleukin-1 receptor antagonist knock-out mice, which spontaneously develop RA-like disease⁸. The activated NF-kB pathway in MSCs from Systemic lupus erythematosus (SLE) patients inhibits osteoblastic differentiation through BMP/Smad signaling pathway, which may participate in the pathology of osteoporosis in SLE patients⁹. Several histopathologic studies in AS patients prove that endochondral new bone formation during bone erosion involves a series of events from chondrocyte apoptosis to colonization of preosteoblasts, differentiation of the preosteoblasts to osteolasts and bone matrix secretion¹⁰. MSCs as progenitor cells may play an important role in pathophysiological processes.

Recently, our research, as well as that of other researchers, demonstrated that MSCs from AS patients (AS-MSCs) outperformed MSCs from healthy donors (HD-MSCs) in osteogenic differentiation in two-dimensional (2D) culture, which could be an important mechanism of pathological osteogenesis in AS^11,

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Acknowledgements

This study was financially supported by the Engineering Technology Research Center for Comprehensive Diagnosis and Treatment of Ankylosing Spondylitis of Guangdong Province (2015B090903059), the Science and Technology Project of Guangdong Province (2015B020228001, 2017A030310554) and the Science and Technology Project of Guangzhou City (201704020045).

Author information

1. These authors contributed equally: Guan Zheng, Zhongyu **e, Peng Wang

Authors and Affiliations

1. Department of Orthopedics, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 107# Yan Jiang Road West, Guangzhou, 510120, P.R. China

   Guan Zheng, Zhongyu **e, Peng Wang, **teng Li, Ming Li, Shuizhong Cen, Su’an Tang, Wenjie Liu, Guiwen Ye, Yuxi Li & Huiyong Shen

2. Department of Orthopedics, The Eighth Affiliated Hospital, Sun Yat-Sen University, 3025# Shen Nan Road, Shenzhen, 518033, P.R. China

   Zhongyu **e, Peng Wang & Huiyong Shen

3. Center for Biotherapy, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 107# Yan Jiang Road West, Guangzhou, 510120, P.R. China

   Shan Wang, **aohua Wu, Hongjun Su & Yanfeng Wu

Contributions

P.W., Y.W. and H.S. designed the experiments. G.Z., Z.X., J.L., M.L. and S.C. conceived and performed the experiments. S.T., W.L. and G.Y. conceived the experiments and analysed the data. Y.L., S.W., X.W. and H.S. carried out the experiments. All authors were involved in writing the manuscript and approved the submitted and published versions.

Corresponding authors

Correspondence to Yanfeng Wu or Huiyong Shen.

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Edited by D. Aberdam

Supplementary information

Characteristics of the study subjects

The siRNA sequences of BMP2

Characteristics of the study subjects for entheseal biopsy

MSC phenotype identification

BMP4, BMP6, BMP7 and BMP9 expression in HD-MSCs and AS-MSCs in HA/β-TCP scaffolds

Noggin expression was identical at local sites of ossifying entheses in AS patients and non-AS patients

Supplementary figure legends

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Zheng, G., **e, Z., Wang, P. et al. Enhanced osteogenic differentiation of mesenchymal stem cells in ankylosing spondylitis: a study based on a three-dimensional biomimetic environment. Cell Death Dis 10, 350 (2019). https://doi.org/10.1038/s41419-019-1586-1

• Received: 20 December 2018

• Revised: 17 March 2019

• Accepted: 10 April 2019

• Published: 25 April 2019

• DOI: https://doi.org/10.1038/s41419-019-1586-1

• Springer Nature Limited

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