PMID- 35798352 OWN - NLM STAT- MEDLINE DCOM- 20220823 LR - 20220927 IS - 1544-0591 (Electronic) IS - 0022-0345 (Linking) VI - 101 IP - 10 DP - 2022 Sep TI - HIF-1alpha Stabilization Boosts Pulp Regeneration by Modulating Cell Metabolism. PG - 1214-1226 LID - 10.1177/00220345221091528 [doi] AB - Stem cell-based therapeutics is a promising strategy in dental pulp regeneration. However, low cell viability after transplantation in vivo due to the ischemic microenvironment is still a critical challenge for future clinical application. With the aim of improving postimplantation cell survival and pulp tissue regeneration, stem cells from human exfoliated deciduous teeth (SHED) were preconditioned to a hypoxic condition by hypoxia-inducible factor 1alpha (HIF-1alpha) stabilization via knockdown of prolyl hydroxylase domain-containing protein 2 (PHD2) using lentiviral short hairpin RNA. HIF-1alpha-stabilized SHED were encapsulated in PuraMatrix hydrogel, injected into root canals of human tooth fragments, and implanted in the subcutaneous space of immunodeficient mice. After 28 d, enhanced dental pulp-like tissue formation was observed with a significantly higher level of vascularization, which could be attributed to both endothelial differentiation of SHED and recruitment of host blood vessels. Furthermore, dentin-like tissue formation in vivo and accelerated odontogenic/osteogenic differentiation both in vivo and in vitro were observed. At 7 d postimplantation, significantly less DNA damage and higher Ki67 expression were detected in the HIF-1alpha-stabilized SHED group compared with the control SHED. Accordingly, cell viability assay and staining for Ki67 and apoptotic cells in vitro showed that HIF-1alpha stabilization could decrease cell apoptosis and enhance cell survival significantly. We demonstrated that PI3K/AKT pathway activation had resulted in low caspase 3 expression in HIF-1alpha-stabilized SHED in hypoxic conditions. Furthermore, we found that HIF-1alpha-induced cell survival could also be attributed to the upregulated expression of PDK1, HK2, and Glut1, which contributes to the maintenance of reactive oxygen species homeostasis and metabolic adaptation in hypoxia. In addition, we identified Smad7 as 1 of the top 3 upregulated genes through RNA sequencing in HIF-1alpha-stabilized SHED and demonstrated its essential role in HK2 and Glut1 upregulation. Taken together, HIF-1alpha stabilization enhances cell survival of SHED through modulating various target genes and potential signaling pathways, as well as odontogenic tissue formation during dental pulp regeneration, which could benefit stem cell-based therapy in general. FAU - Han, Y AU - Han Y AUID- ORCID: 0000-0001-6006-9825 AD - Applied Oral Sciences & Community Dental Care, Faculty of Dentistry, The University of Hong Kong, Pokfulam, Hong Kong. FAU - Koohi-Moghadam, M AU - Koohi-Moghadam M AD - Applied Oral Sciences & Community Dental Care, Faculty of Dentistry, The University of Hong Kong, Pokfulam, Hong Kong. FAU - Chen, Q AU - Chen Q AD - Applied Oral Sciences & Community Dental Care, Faculty of Dentistry, The University of Hong Kong, Pokfulam, Hong Kong. FAU - Zhang, L AU - Zhang L AD - Applied Oral Sciences & Community Dental Care, Faculty of Dentistry, The University of Hong Kong, Pokfulam, Hong Kong. FAU - Chopra, H AU - Chopra H AD - Periodontics and Oral Medicine, School of Dentistry, University of Michigan, Ann Arbor, MI, USA. FAU - Zhang, J AU - Zhang J AD - Department of Orthodontics, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China. FAU - Dissanayaka, W L AU - Dissanayaka WL AUID- ORCID: 0000-0002-3621-4866 AD - Applied Oral Sciences & Community Dental Care, Faculty of Dentistry, The University of Hong Kong, Pokfulam, Hong Kong. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220707 PL - United States TA - J Dent Res JT - Journal of dental research JID - 0354343 RN - 0 (Glucose Transporter Type 1) RN - 0 (Hypoxia-Inducible Factor 1, alpha Subunit) RN - 0 (Ki-67 Antigen) RN - EC 1.14.11.29 (Hypoxia-Inducible Factor-Proline Dioxygenases) SB - IM MH - Animals MH - Cell Hypoxia MH - *Dental Pulp MH - Glucose Transporter Type 1 MH - Humans MH - Hypoxia MH - Hypoxia-Inducible Factor 1, alpha Subunit MH - Hypoxia-Inducible Factor-Proline Dioxygenases/genetics MH - Ki-67 Antigen MH - Mice MH - Osteogenesis MH - *Phosphatidylinositol 3-Kinases MH - Regeneration OTO - NOTNLM OT - cell survival OT - cellular metabolism OT - dental pulp regeneration OT - dental stem cell OT - hypoxia-inducible factor 1alpha OT - hypoxic preconditioning EDAT- 2022/07/08 06:00 MHDA- 2022/08/24 06:00 CRDT- 2022/07/07 20:23 PHST- 2022/07/08 06:00 [pubmed] PHST- 2022/08/24 06:00 [medline] PHST- 2022/07/07 20:23 [entrez] AID - 10.1177/00220345221091528 [doi] PST - ppublish SO - J Dent Res. 2022 Sep;101(10):1214-1226. doi: 10.1177/00220345221091528. Epub 2022 Jul 7.