PAX9 Is Involved in Periodontal Ligament Stem Cell-like Differentiation of Human-Induced Pluripotent Stem Cells by Regulating Extracellular Matrix

Risa Sugiura; Sayuri Hamano; Atsushi Tomokiyo; Daigaku Hasegawa; Shinichiro Yoshida; Hideki Sugii; Shoko Fujino; Orie Adachi; Masataka Kadowaki; Daiki Yamashita; Hidefumi Maeda

doi:10.3390/biomedicines10102366

PAX9 Is Involved in Periodontal Ligament Stem Cell-like Differentiation of Human-Induced Pluripotent Stem Cells by Regulating Extracellular Matrix

Biomedicines. 2022 Sep 22;10(10):2366. doi: 10.3390/biomedicines10102366.

Authors

Risa Sugiura¹, Sayuri Hamano^{1

2}, Atsushi Tomokiyo^{3

4}, Daigaku Hasegawa³, Shinichiro Yoshida³, Hideki Sugii¹, Shoko Fujino¹, Orie Adachi¹, Masataka Kadowaki¹, Daiki Yamashita¹, Hidefumi Maeda^{1

3}

Affiliations

¹ Department of Endodontology and Operative Dentistry, Faculty of Dental Science, Kyushu University, Fukuoka 812-8582, Japan.
² OBT Research Center, Faculty of Dental Science, Kyushu University, Fukuoka 812-8582, Japan.
³ Department of Endodontology, Kyushu University Hospital, Fukuoka 812-8582, Japan.
⁴ DDR Research Center, Faculty of Dental Science, Kyushu University, Fukuoka 812-8582, Japan.

Abstract

Periodontal ligament stem cells (PDLSCs) play central roles in periodontal ligament (PDL) tissue homeostasis, repair, and regeneration. Previously, we established a protocol to differentiate human-induced pluripotent stem cell-derived neural crest-like cells (iNCs) into PDLSC-like cells (iPDLSCs) using human PDL cell-derived extracellular matrix (ECM). However, it remained unclear what factors principally regulate the differentiation of iNCs into iPDLSCs. In this study, we aimed to identify the transcription factor regulating production of human PDL cell-derived ECM, which is responsible for the generation of iPDLSCs. We cultured iNCs on ECMs of two human PDL cell lines (HPDLC-3S and HPDLC-3U) and of human dermal fibroblasts (HDF). iNCs cultured on HPDLC-3U demonstrated higher iPDLSC-associated gene expression and mesenchymal differentiation capacity than cells cultured on HDF or HPDLC-3S. The transcription factor PAX9 was highly expressed in HPDLC-3U compared with HDF and HPDLC-3S. iNCs cultured on siPAX9-transfected HPDLC-3U displayed downregulation of iPDLSC-associated marker expression and adipocytic differentiation capacity relative to controls. Our findings suggest that PAX9 is one of the transcription factors regulating ECM production in human PDL cells, which is responsible for the differentiation of iNCs into iPDLSCs.

Keywords: PAX9; extracellular matrix; induced pluripotent stem cells; periodontal ligament stem cells; transcription factor.

Grants and funding

This work was financially supported by Grants-in-Aid for Scientific Research (Project Nos. JP17H01598, JP18K19651, JP21K09876, JP21K19608, and JP22H03271) from the Japan Society for the Promotion of Science.