Application and molecular basis of spheroid culture to the deterioration of periodontal ligament cells due to frequent passage

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K10133
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57030:Conservative dentistry-related
• Research Institution: Kyushu Dental College
• Principal Investigator: USUI MICHIHIKO 九州歯科大学, 歯学部, 准教授 (10453630)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: スフェロイド / 歯根膜幹細胞 / 骨分化 / 幹細胞性

Outline of Final Research Achievements

In present study, we performed RNA-seq analysis to find genes specifically expressed in hPDLMSC (human periodontal ligament mesenchymal stem cell) spheroids and to analyze their functions. The hPDLMSC spheroid culture group and monolayer culture group were compared by transcriptome analysis. We found that the transcription factor NR4A2 (Nuclear receptor 4A2) was strongly expressed in the spheroid culture group, and knockdown of NR4A2 significantly increased the expression of osteogenesis-related genes, ALP activity, and calcified nodule formation. These results suggest that NR4A2 negatively regulates osteogenesis in hPDLMSC spheroids.

Free Research Field

歯周病学

Academic Significance and Societal Importance of the Research Achievements

ヒト歯根膜間葉系幹細胞を用いた細胞治療は、次世代の歯周組織再生治療として期待されている。しかし、再生に適した歯根膜幹細胞の培養条件については不明な点が多い。スフェロイド培養は幹細胞の生理的機能を上昇させることが知られているが、今回我々が見出したNR4A2がヒト歯根膜間葉系幹細胞のスフェロイド培養において、幹細胞性の維持・骨分化抑制に関与している可能性が明らかとなったことは学術的に意義深いと考える。