Volume 15, Issue 2 pp. 103-115
RESEARCH ARTICLE

Evidence that TD-198946 enhances the chondrogenic potential of human synovium-derived stem cells through the NOTCH3 signaling pathway

Masato Kobayashi

Masato Kobayashi

Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

Search for more papers by this author
Ryota Chijimatsu

Ryota Chijimatsu

Bone and Cartilage Regenerative Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

Search for more papers by this author
David A. Hart

David A. Hart

McCaig Institute for Bone and Joint Health, University of Calgary, Calgary, Canada

Search for more papers by this author
Shuichi Hamamoto

Shuichi Hamamoto

Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

Search for more papers by this author
George Jacob

George Jacob

Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

Search for more papers by this author
Fumiko Yano

Fumiko Yano

Bone and Cartilage Regenerative Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

Search for more papers by this author
Taku Saito

Taku Saito

Sensory and Motor System Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

Search for more papers by this author
Kazunori Shimomura

Kazunori Shimomura

Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

Search for more papers by this author
Wataru Ando

Wataru Ando

Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

Search for more papers by this author
Ung-il Chung

Ung-il Chung

Center for Disease Biology and Integrative Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

Search for more papers by this author
Sakae Tanaka

Sakae Tanaka

Sensory and Motor System Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

Search for more papers by this author
Hideki Yoshikawa

Hideki Yoshikawa

Orthopaedic Surgery, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

Search for more papers by this author
Norimasa Nakamura

Corresponding Author

Norimasa Nakamura

Institute for Medical Science in Sports, Osaka Health Science University, Osaka, Japan

Correspondence:

Norimasa Nakamura, Osaka Health Science University Graduate School Sports Medical Science Laboratory, 1-9-27, Tennma, Kita-ku, Osaka, Japan.

Email: [email protected]

Search for more papers by this author
First published: 10 November 2020
Citations: 6
Masato Kobayashi and Ryota Chijimatsu contributed equally to this work

Abstract

Human synovium-derived stem cells (hSSCs) are an attractive source of cells for cartilage repair. At present, the quality of tissue and techniques used for cartilage regeneration have scope for improvement. A small compound, TD-198946, was reported to enhance chondrogenic induction from hSSCs; however, other applications of TD-198946, such as priming the cell potential of hSSCs, remain unknown. Our study aimed to examine the effect of TD-198946 pretreatment on hSSCs. HSSCs were cultured with or without TD-198946 for 7 days during expansion culture and then converted into a three-dimensional pellet culture supplemented with bone morphogenetic protein-2 (BMP2) and/or transforming growth factor beta-3 (TGFβ3). Chondrogenesis in cultures was assessed based on the GAG content, histology, and expression levels of chondrogenic marker genes. Cell pellets derived from TD-198946-pretreated hSSCs showed enhanced chondrogenic potential when chondrogenesis was induced by both BMP2 and TGFβ3. Moreover, cartilaginous tissue was efficiently generated from TD-198946-pretreated hSSCs using a combination of BMP2 and TGFβ3. Microarray analysis revealed that NOTCH pathway-related genes and their target genes were significantly upregulated in TD-198946-treated hSSCs, although TD-198946 alone did not upregulate chondrogenesis related markers. The administration of the NOTCH signal inhibitor diminished the effect of TD-198946. Thus, TD-198946 enhances the chondrogenic potential of hSSCs via the NOTCH3 signaling pathway. This study is the first to demonstrate the gradual activation of NOTCH3 signaling during chondrogenesis in hSSCs. The priming of NOTCH3 using TD-198946 provides a novel insight regarding the regulation of the differentiation of hSSCs into chondrocytes.

CONFLICT OF INTEREST

All authors have no conflict of interest to declare.

AUTHOR CONTRIBUTIONS

Masato Kobayashi and Ryota Chijimatsu took responsibility for the integrity of the work as a whole, from inception to the completed manuscript. Conception and design: Masato Kobayashi, Ryota Chijimatsu, Hideki Yoshikawa, and Norimasa Nakamura. Analysis and interpretation of the data, statistical expertise: Masato Kobayashi, Ryota Chijimatsu, David A. Hart, Shuichi Hamamoto, Fumiko Yano, Taku Saito, Kazunori Shimomura, Wataru Ando, Ung-il Chung, Sakae Tanaka, Hideki Yoshikawa, and Norimasa Nakamura. Administrative, technical, or logistic support: Masato Kobayashi and Ryota Chijimatsu, David A. Hart, Shuichi Hamamoto, Fumiko Yano, Taku Saito, Kazunori Shimomura, Wataru Ando, Ung-il Chung, Sakae Tanaka, Hideki Yoshikawa, and Norimasa Nakamura. Collection and assembly of data: Masato Kobayashi and Ryota Chijimatsu, HY, and NN. Final approval of the article: Masato Kobayashi, Ryota Chijimatsu, David A. Hart, Shuichi Hamamoto, Fumiko Yano, Taku Saito, Kazunori Shimomura, Wataru Ando, Ung-il Chung, Sakae Tanaka, Hideki Yoshikawa, and Norimasa Nakamura.

The full text of this article hosted at iucr.org is unavailable due to technical difficulties.