Bone Differentiation Ability of CD146-Positive Stem Cells from Human Exfoliated Deciduous Teeth

Regenerative therapy for tissues by mesenchymal stem cell (MSCs) transplantation has received much attention. The cluster of differentiation (CD)146 marker, a surface-antigen of stem cells, is crucial for angiogenic and osseous differentiation abilities. Bone regeneration is accelerated by the trans...

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Main Authors: Ryo Kunimatsu, Kodai Rikitake, Yuki Yoshimi, Nurul Aisyah Rizky Putranti, Yoko Hayashi, Kotaro Tanimoto
Format: Article
Language:English
Published: MDPI AG 2023-02-01
Series:International Journal of Molecular Sciences
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Online Access:https://www.mdpi.com/1422-0067/24/4/4048
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author Ryo Kunimatsu
Kodai Rikitake
Yuki Yoshimi
Nurul Aisyah Rizky Putranti
Yoko Hayashi
Kotaro Tanimoto
author_facet Ryo Kunimatsu
Kodai Rikitake
Yuki Yoshimi
Nurul Aisyah Rizky Putranti
Yoko Hayashi
Kotaro Tanimoto
author_sort Ryo Kunimatsu
collection DOAJ
description Regenerative therapy for tissues by mesenchymal stem cell (MSCs) transplantation has received much attention. The cluster of differentiation (CD)146 marker, a surface-antigen of stem cells, is crucial for angiogenic and osseous differentiation abilities. Bone regeneration is accelerated by the transplantation of CD146-positive deciduous dental pulp-derived mesenchymal stem cells contained in stem cells from human exfoliated deciduous teeth (SHED) into a living donor. However, the role of CD146 in SHED remains unclear. This study aimed to compare the effects of CD146 on cell proliferative and substrate metabolic abilities in a population of SHED. SHED was isolated from deciduous teeth, and flow cytometry was used to analyze the expression of MSCs markers. Cell sorting was performed to recover the CD146-positive cell population (CD146+) and CD146-negative cell population (CD146-). CD146 + SHED without cell sorting and CD146-SHED were examined and compared among three groups. To investigate the effect of CD146 on cell proliferation ability, an analysis of cell proliferation ability was performed using BrdU assay and MTS assay. The bone differentiation ability was evaluated using an alkaline phosphatase (ALP) stain after inducing bone differentiation, and the quality of ALP protein expressed was examined. We also performed Alizarin red staining and evaluated the calcified deposits. The gene expression of ALP, bone morphogenetic protein-2 (BMP-2), and osteocalcin (OCN) was analyzed using a real-time polymerase chain reaction. There was no significant difference in cell proliferation among the three groups. The expression of ALP stain, Alizarin red stain, ALP, BMP-2, and OCN was the highest in the CD146+ group. CD146 + SHED had higher osteogenic differentiation potential compared with SHED and CD146-SHED. CD146 contained in SHED may be a valuable population of cells for bone regeneration therapy.
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spelling doaj.art-0ce06545230545f7b88f69ab12a884ed2023-11-16T21:08:47ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01244404810.3390/ijms24044048Bone Differentiation Ability of CD146-Positive Stem Cells from Human Exfoliated Deciduous TeethRyo Kunimatsu0Kodai Rikitake1Yuki Yoshimi2Nurul Aisyah Rizky Putranti3Yoko Hayashi4Kotaro Tanimoto5Department of Orthodontics and Craniofacial Developmental Biology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, JapanDepartment of Orthodontics and Craniofacial Developmental Biology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, JapanDepartment of Orthodontics and Craniofacial Developmental Biology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, JapanDepartment of Orthodontics and Craniofacial Developmental Biology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, JapanAnalysis Center of Life Science, Natural Science Center for Basic Research and Development, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, JapanDepartment of Orthodontics and Craniofacial Developmental Biology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, JapanRegenerative therapy for tissues by mesenchymal stem cell (MSCs) transplantation has received much attention. The cluster of differentiation (CD)146 marker, a surface-antigen of stem cells, is crucial for angiogenic and osseous differentiation abilities. Bone regeneration is accelerated by the transplantation of CD146-positive deciduous dental pulp-derived mesenchymal stem cells contained in stem cells from human exfoliated deciduous teeth (SHED) into a living donor. However, the role of CD146 in SHED remains unclear. This study aimed to compare the effects of CD146 on cell proliferative and substrate metabolic abilities in a population of SHED. SHED was isolated from deciduous teeth, and flow cytometry was used to analyze the expression of MSCs markers. Cell sorting was performed to recover the CD146-positive cell population (CD146+) and CD146-negative cell population (CD146-). CD146 + SHED without cell sorting and CD146-SHED were examined and compared among three groups. To investigate the effect of CD146 on cell proliferation ability, an analysis of cell proliferation ability was performed using BrdU assay and MTS assay. The bone differentiation ability was evaluated using an alkaline phosphatase (ALP) stain after inducing bone differentiation, and the quality of ALP protein expressed was examined. We also performed Alizarin red staining and evaluated the calcified deposits. The gene expression of ALP, bone morphogenetic protein-2 (BMP-2), and osteocalcin (OCN) was analyzed using a real-time polymerase chain reaction. There was no significant difference in cell proliferation among the three groups. The expression of ALP stain, Alizarin red stain, ALP, BMP-2, and OCN was the highest in the CD146+ group. CD146 + SHED had higher osteogenic differentiation potential compared with SHED and CD146-SHED. CD146 contained in SHED may be a valuable population of cells for bone regeneration therapy.https://www.mdpi.com/1422-0067/24/4/4048stem cells from human exfoliated deciduous teethbone regenerationCD146
spellingShingle Ryo Kunimatsu
Kodai Rikitake
Yuki Yoshimi
Nurul Aisyah Rizky Putranti
Yoko Hayashi
Kotaro Tanimoto
Bone Differentiation Ability of CD146-Positive Stem Cells from Human Exfoliated Deciduous Teeth
International Journal of Molecular Sciences
stem cells from human exfoliated deciduous teeth
bone regeneration
CD146
title Bone Differentiation Ability of CD146-Positive Stem Cells from Human Exfoliated Deciduous Teeth
title_full Bone Differentiation Ability of CD146-Positive Stem Cells from Human Exfoliated Deciduous Teeth
title_fullStr Bone Differentiation Ability of CD146-Positive Stem Cells from Human Exfoliated Deciduous Teeth
title_full_unstemmed Bone Differentiation Ability of CD146-Positive Stem Cells from Human Exfoliated Deciduous Teeth
title_short Bone Differentiation Ability of CD146-Positive Stem Cells from Human Exfoliated Deciduous Teeth
title_sort bone differentiation ability of cd146 positive stem cells from human exfoliated deciduous teeth
topic stem cells from human exfoliated deciduous teeth
bone regeneration
CD146
url https://www.mdpi.com/1422-0067/24/4/4048
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