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Inactivation of PI3K/Akt promotes the odontoblastic differentiation and suppresses the stemness with autophagic flux in dental pulp cells

BACKGROUND/PURPOSE: Autophagy is involved in controlling differentiation of various cell types. The present study aimed to investigate the mechanism related to autophagy in regulating odontogenic differentiation of dental pulp cells. MATERIALS AND METHODS: Human dental pulp cells (HDPCs) were cultur...

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Autores principales: Park, Sam Young, Cho, Heui Seung, Chung, Kyung Hwun, Lee, Bin Na, Kim, Sun Hun, Kim, Won Jae, Jung, Ji Yeon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Association for Dental Sciences of the Republic of China 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8739242/
https://www.ncbi.nlm.nih.gov/pubmed/35028032
http://dx.doi.org/10.1016/j.jds.2021.05.013
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author Park, Sam Young
Cho, Heui Seung
Chung, Kyung Hwun
Lee, Bin Na
Kim, Sun Hun
Kim, Won Jae
Jung, Ji Yeon
author_facet Park, Sam Young
Cho, Heui Seung
Chung, Kyung Hwun
Lee, Bin Na
Kim, Sun Hun
Kim, Won Jae
Jung, Ji Yeon
author_sort Park, Sam Young
collection PubMed
description BACKGROUND/PURPOSE: Autophagy is involved in controlling differentiation of various cell types. The present study aimed to investigate the mechanism related to autophagy in regulating odontogenic differentiation of dental pulp cells. MATERIALS AND METHODS: Human dental pulp cells (HDPCs) were cultured in differentiation inductive medium (DM) and odontoblastic differentiation and mineralization were evaluated by alkaline phosphatase (ALP) staining and Alizarin red S staining, respectively. Tooth cavity preparation was made on the mesial surface of lower first molars in rat. The expression of autophagy-related signal molecules was detected using Western blot analysis and Immunohistochemistry. RESULTS: HDPCs cultured in DM showed increased autophagic flux and declined phosphorylation of phosphoinositide 3-kinases (PI3K), protein kinase B (Akt), and mTOR. Dentin matrix protein-1 (DMP-1) and dentin sialoprotein (DSP), markers of odontoblastic differentiation, were upregulated and autophagic activation showing increased LC3-II and decreased p62 levels was observed during odontogenic differentiation of HDPCs. However, PI3K blocker 3-methyladenine (3MA), lentiviral shLC3 and Akt activator SC79 attenuated the expression of LC3II as well as DMP-1, ALP activity and mineralization enhanced in HDPCs under DM condition. In addition, 3MA, shLC3 and SC79 recovered the expression of pluripotency factor CD146, Oct4 and Nanog downregulated in DM condition. In rat tooth cavity preparation model, the expression of LC3B and DMP-1 was elevated near odontoblast-dentin layer during reparative dentin formation, whereas 3MA significantly reduced the expression of LC3B and DMP-1. CONCLUSION: These findings indicated autophagy promotes the odontogenic differentiation of dental pulp cells modulating stemness via PI3K/Akt inactivation and the repair of pulp.
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spelling pubmed-87392422022-01-12 Inactivation of PI3K/Akt promotes the odontoblastic differentiation and suppresses the stemness with autophagic flux in dental pulp cells Park, Sam Young Cho, Heui Seung Chung, Kyung Hwun Lee, Bin Na Kim, Sun Hun Kim, Won Jae Jung, Ji Yeon J Dent Sci Original Article BACKGROUND/PURPOSE: Autophagy is involved in controlling differentiation of various cell types. The present study aimed to investigate the mechanism related to autophagy in regulating odontogenic differentiation of dental pulp cells. MATERIALS AND METHODS: Human dental pulp cells (HDPCs) were cultured in differentiation inductive medium (DM) and odontoblastic differentiation and mineralization were evaluated by alkaline phosphatase (ALP) staining and Alizarin red S staining, respectively. Tooth cavity preparation was made on the mesial surface of lower first molars in rat. The expression of autophagy-related signal molecules was detected using Western blot analysis and Immunohistochemistry. RESULTS: HDPCs cultured in DM showed increased autophagic flux and declined phosphorylation of phosphoinositide 3-kinases (PI3K), protein kinase B (Akt), and mTOR. Dentin matrix protein-1 (DMP-1) and dentin sialoprotein (DSP), markers of odontoblastic differentiation, were upregulated and autophagic activation showing increased LC3-II and decreased p62 levels was observed during odontogenic differentiation of HDPCs. However, PI3K blocker 3-methyladenine (3MA), lentiviral shLC3 and Akt activator SC79 attenuated the expression of LC3II as well as DMP-1, ALP activity and mineralization enhanced in HDPCs under DM condition. In addition, 3MA, shLC3 and SC79 recovered the expression of pluripotency factor CD146, Oct4 and Nanog downregulated in DM condition. In rat tooth cavity preparation model, the expression of LC3B and DMP-1 was elevated near odontoblast-dentin layer during reparative dentin formation, whereas 3MA significantly reduced the expression of LC3B and DMP-1. CONCLUSION: These findings indicated autophagy promotes the odontogenic differentiation of dental pulp cells modulating stemness via PI3K/Akt inactivation and the repair of pulp. Association for Dental Sciences of the Republic of China 2022-01 2021-06-16 /pmc/articles/PMC8739242/ /pubmed/35028032 http://dx.doi.org/10.1016/j.jds.2021.05.013 Text en © 2021 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Park, Sam Young
Cho, Heui Seung
Chung, Kyung Hwun
Lee, Bin Na
Kim, Sun Hun
Kim, Won Jae
Jung, Ji Yeon
Inactivation of PI3K/Akt promotes the odontoblastic differentiation and suppresses the stemness with autophagic flux in dental pulp cells
title Inactivation of PI3K/Akt promotes the odontoblastic differentiation and suppresses the stemness with autophagic flux in dental pulp cells
title_full Inactivation of PI3K/Akt promotes the odontoblastic differentiation and suppresses the stemness with autophagic flux in dental pulp cells
title_fullStr Inactivation of PI3K/Akt promotes the odontoblastic differentiation and suppresses the stemness with autophagic flux in dental pulp cells
title_full_unstemmed Inactivation of PI3K/Akt promotes the odontoblastic differentiation and suppresses the stemness with autophagic flux in dental pulp cells
title_short Inactivation of PI3K/Akt promotes the odontoblastic differentiation and suppresses the stemness with autophagic flux in dental pulp cells
title_sort inactivation of pi3k/akt promotes the odontoblastic differentiation and suppresses the stemness with autophagic flux in dental pulp cells
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8739242/
https://www.ncbi.nlm.nih.gov/pubmed/35028032
http://dx.doi.org/10.1016/j.jds.2021.05.013
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