1,25(OH)2D3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential*
Periodontal dental ligament mesenchymal stem cells (PDLMSCs) play a major role in periodontal tissue regeneration by the neoformation of root cementum and alveolar bone. These cells are highly heterogeneous, and many present low potential to renovate the hard tissue damaged by periodontal disease. A...
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University of São Paulo
2024-11-01
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| Series: | Journal of Applied Oral Science |
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| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-77572024000100461&lng=en&tlng=en |
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| _version_ | 1846119102540677120 |
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| author | Bruno Cazotti PEREIRA Catharina Marques SACRAMENTO Enilson Antonio SALLUM Mabelle de Freitas MONTEIRO Renato Corrêa Viana CASARIN Marcio Zaffalon CASATI Karina Gonzales SILVÉRIO |
| author_facet | Bruno Cazotti PEREIRA Catharina Marques SACRAMENTO Enilson Antonio SALLUM Mabelle de Freitas MONTEIRO Renato Corrêa Viana CASARIN Marcio Zaffalon CASATI Karina Gonzales SILVÉRIO |
| author_sort | Bruno Cazotti PEREIRA |
| collection | DOAJ |
| description | Periodontal dental ligament mesenchymal stem cells (PDLMSCs) play a major role in periodontal tissue regeneration by the neoformation of root cementum and alveolar bone. These cells are highly heterogeneous, and many present low potential to renovate the hard tissue damaged by periodontal disease. A previous study found that the low osteoblast/cementoblast (O/C) differentiation potential of PDLMSCs is related to high asporin (ASPN) expression, which was identified as a negative regulator of PDL cells differentiation and mineralization, suppressing BMP-2-induced O/C differentiation. Objective This study aimed to investigate whether 1,25(OH)2D3 treatment could stimulate the O/C differentiation of periodontal ligament mesenchymal progenitor cells characterized as low osteoblast potential (LOP), by asporin and bone morphogenetic protein-2 alteration. Methodology Three LOP cell populations were cultured in standard medium (CONTROL), osteogenic medium (OM), and osteogenic medium associated with 1 nM of 1,25(OH)2D3 (OM + VD). The following assays were performed: 1) MTT to evaluate metabolic activity; 2) gene expression for asporin (ASPN), bone morphogenetic protein-2 (BMP-2), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), and vitamin D receptor (VDR) using qRT-PCR; 3) BMP-2 extracellular expression; and 4) quantification of mineralized nodule deposition by Alizarin Red Staining. Data were subjected to two-way ANOVA and Tukey’s test (P<0.05). Results The results showed that the 1,25(OH)2D3 treatment did not affect the cell viability, as demonstrated by metabolic activity increase over the 10 days in culture. After 14 days of 1,25(OH)2D3 treatment, the mRNA levels for ASPN and VDR decreased (P<0.05), while BMP-2 transcripts and extracellular expression increased (P<0.05). In parallel, RUNX2, ALP, and OCN gene expression was upregulated by 1,25(OH)2D3 treatment, resulting in an increase of mineral nodule deposition in vitro (P<0.05). Conclusions These data show that 1,25(OH)2D3 improves osteoblast/cementoblast differentiation of low osteoblast potential accompanied by alterations in ASPN and BMP-2 expression. |
| format | Article |
| id | doaj-art-4b6823bd8a754b46a86d7cd058b5a652 |
| institution | Kabale University |
| issn | 1678-7765 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | University of São Paulo |
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| series | Journal of Applied Oral Science |
| spelling | doaj-art-4b6823bd8a754b46a86d7cd058b5a6522024-12-17T07:46:02ZengUniversity of São PauloJournal of Applied Oral Science1678-77652024-11-013210.1590/1678-7757-2024-01601,25(OH)2D3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential*Bruno Cazotti PEREIRAhttps://orcid.org/0000-0001-5800-6366Catharina Marques SACRAMENTOhttps://orcid.org/0000-0003-2449-8168Enilson Antonio SALLUMhttps://orcid.org/0000-0003-3827-7091Mabelle de Freitas MONTEIROhttps://orcid.org/0000-0001-9333-4349Renato Corrêa Viana CASARINhttps://orcid.org/0000-0003-1743-5855Marcio Zaffalon CASATIhttps://orcid.org/0000-0001-9234-0536Karina Gonzales SILVÉRIOhttps://orcid.org/0000-0001-5879-9095Periodontal dental ligament mesenchymal stem cells (PDLMSCs) play a major role in periodontal tissue regeneration by the neoformation of root cementum and alveolar bone. These cells are highly heterogeneous, and many present low potential to renovate the hard tissue damaged by periodontal disease. A previous study found that the low osteoblast/cementoblast (O/C) differentiation potential of PDLMSCs is related to high asporin (ASPN) expression, which was identified as a negative regulator of PDL cells differentiation and mineralization, suppressing BMP-2-induced O/C differentiation. Objective This study aimed to investigate whether 1,25(OH)2D3 treatment could stimulate the O/C differentiation of periodontal ligament mesenchymal progenitor cells characterized as low osteoblast potential (LOP), by asporin and bone morphogenetic protein-2 alteration. Methodology Three LOP cell populations were cultured in standard medium (CONTROL), osteogenic medium (OM), and osteogenic medium associated with 1 nM of 1,25(OH)2D3 (OM + VD). The following assays were performed: 1) MTT to evaluate metabolic activity; 2) gene expression for asporin (ASPN), bone morphogenetic protein-2 (BMP-2), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), and vitamin D receptor (VDR) using qRT-PCR; 3) BMP-2 extracellular expression; and 4) quantification of mineralized nodule deposition by Alizarin Red Staining. Data were subjected to two-way ANOVA and Tukey’s test (P<0.05). Results The results showed that the 1,25(OH)2D3 treatment did not affect the cell viability, as demonstrated by metabolic activity increase over the 10 days in culture. After 14 days of 1,25(OH)2D3 treatment, the mRNA levels for ASPN and VDR decreased (P<0.05), while BMP-2 transcripts and extracellular expression increased (P<0.05). In parallel, RUNX2, ALP, and OCN gene expression was upregulated by 1,25(OH)2D3 treatment, resulting in an increase of mineral nodule deposition in vitro (P<0.05). Conclusions These data show that 1,25(OH)2D3 improves osteoblast/cementoblast differentiation of low osteoblast potential accompanied by alterations in ASPN and BMP-2 expression.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-77572024000100461&lng=en&tlng=enCalcitriolOsteoblastCementoblastPeriodontal Dental Ligament Mesenchymal Progenitor CellsDifferentiationAsporinBone Morphogenetic Protein 2 |
| spellingShingle | Bruno Cazotti PEREIRA Catharina Marques SACRAMENTO Enilson Antonio SALLUM Mabelle de Freitas MONTEIRO Renato Corrêa Viana CASARIN Marcio Zaffalon CASATI Karina Gonzales SILVÉRIO 1,25(OH)2D3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential* Journal of Applied Oral Science Calcitriol Osteoblast Cementoblast Periodontal Dental Ligament Mesenchymal Progenitor Cells Differentiation Asporin Bone Morphogenetic Protein 2 |
| title | 1,25(OH)2D3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential* |
| title_full | 1,25(OH)2D3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential* |
| title_fullStr | 1,25(OH)2D3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential* |
| title_full_unstemmed | 1,25(OH)2D3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential* |
| title_short | 1,25(OH)2D3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential* |
| title_sort | 1 25 oh 2d3 increase osteogenic potential of human periodontal ligament cells with low osteoblast potential |
| topic | Calcitriol Osteoblast Cementoblast Periodontal Dental Ligament Mesenchymal Progenitor Cells Differentiation Asporin Bone Morphogenetic Protein 2 |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-77572024000100461&lng=en&tlng=en |
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