Identification of stemness and differentially expressed genes in human cementum-derived cells
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose: Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored. Methods: Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed. Results: The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs. Conclusions: Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs.
Abstract
Purpose: Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored. Methods: Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed. Results: The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs. Conclusions: Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs.
참고문헌 (39)
- Gottlow J, "New attachment formation in the human periodontium by guided tissue regeneration. Case reports", J Clin Periodontol, 13, pp.604~616 (1986)
- Nyman S, "New attachment following surgical treatment of human periodontal disease", J Clin Periodontol, 9, pp.290~296 (1982)
- Tonetti MS, "Generalizability of the added benefits of guided tissue regeneration in the treatment of deep intrabony defects. Evaluation in a multicenter randomized controlled clinical trial", J Periodontol, 69, pp.1183~1192 (1998)
- Tonetti MS, "Factors affecting the healing response of intrabony defects following guided tissue regeneration and access flap surgery", J Clin Periodontol, 23, pp.548~556 (1996)
- Park JY, "Efficacy of periodontal stem cell transplantation in the treatment of advanced periodontitis", Cell Transplant, 20, pp.271~285 (2011)
- Liu Y, "Periodontal ligament stem cell-mediated treatment for periodontitis in miniature swine", Stem Cells, 26, pp.1065~1073 (2008)
- Park JY, "rhBMP-2 pre-treated human periodontal ligament stem cell sheets regenerate a mineralized layer mimicking dental cementum", Int J Mol Sci, 21, pp.3767~ (2020)
- Cho H, "Periodontal ligament stem", progenitor cells with proteinreleasing scaffolds for cementum formation and integration on dentin surface, 2016, pp.57, 488~495
- Grzesik WJ, "Normal human cementumderived cells: isolation, clonal expansion, and in vitro and in vivo characterization", J Bone Miner Res, 13, pp.1547~1554 (1998)
- Liu HW, "A collagenous cementum-derived attachment protein is a marker for progenitors of the mineralized tissue-forming cell lineage of the periodontal ligament", J Bone Miner Res, 12, pp.1691~1699 (1997)
- Alvarez-Pérez MA, "Molecular cloning, expression and immunolocalization of a novel human cementum-derived protein (CP-23)", Bone, 38, pp.409~419 (2006)
- Komaki M, "Cementum protein 1(CEMP1)induces a cementoblastic phenotype and reduces osteoblastic differentiation in periodontal ligament cells", J Cell Physiol, 227, pp.649~657 (2012)
- Bosshardt DD, "Are cementoblasts a subpopulation of osteoblasts or a unique phenotype?", J Dent Res, 84, pp.390~406 (2005)
- Foster BL, "Methods for studying tooth root cementum by light microscopy", Int J Oral Sci, 4, pp.119~128 (2012)
- Matthews BG, "Gene-expression analysis of cementoblasts and osteoblasts", J Periodontal Res, 51, pp.304~312 (2016)
- Salmon CR, "Proteomic analysis of human dental cementum and alveolar bone", J Proteomics, 91, pp.544~555 (2013)
- Kitagawa M, "Characterization of established cementoblast-like cell lines from human cementum-lining cells in vitro and in vivo", Bone, 39, pp.1035~1042 (2006)
- Wu D, "Characterization of a collagenous cementum-derived attachment protein", J Bone Miner Res, 11, pp.686~692 (1996)
- Kim SH, "Alveolar bone regeneration by transplantation of periodontal ligament stem cells and bone marrow stem cells in a canine peri-implant defect model : a pilot study", J Periodontol, 80, pp.1815~1823 (2009)
- Gronthos S, "Postnatal human dental pulp stem cells(DPSCs)in vitro and in vivo", Proc Natl Acad Sci U S A, 97, pp.13625~13630 (2000)
- Seo BM, "Investigation of multipotent postnatal stem cells from human periodontal ligament", Lancet, 364, pp.149~155 (2004)
- Estrela C, "Mesenchymal stem cells in the dental tissues : perspectives for tissue regeneration", Braz Dent J, 22, pp.91~98 (2011)
- Sonoyama W, "Characterization of the apical papilla and its residing stem cells from human immature permanent teeth : a pilot study", J Endod, 34, pp.166~171 (2008)
- Shi S, "Comparison of human dental pulp and bone marrow stromal stem cells by cDNA microarray analysis", Bone, 29, pp.532~539 (2001)
- 김수환, "Gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow", Journal of Periodontal & Implant Science, 41(4), pp.192~200 (2011), 대한치주과학회
- Kawashima N, "Characterisation of dental pulp stem cells: a new horizon for tissue regeneration?", Arch Oral Biol, 57, pp.1439~1458 (2012)
- Taha MF, "Isolation, identification and multipotential differentiation of mouse adipose tissuederived stem cells", Tissue Cell, 42, pp.211~216 (2010)
- Tropel P, "Isolation and characterisation of mesenchymal stem cells from adult mouse bone marrow", Exp Cell Res, 295, pp.395~406 (2004)
- Pittenger MF, "Multilineage potential of adult human mesenchymal stem cells", Science, 284, pp.143~147 (1999)
- Carnes DL, "Cells with osteoblastic phenotypes can be explanted from human gingiva and periodontal ligament", J Periodontol, 68, pp.701~707 (1997)
- Torii D, "Cementogenic potential of multipotential mesenchymal stem cells purified from the human periodontal ligament", Odontology, 103, pp.27~35 (2015)
- Hoz L, "Cementum protein 1(CEMP1)induces differentiation by human periodontal ligament cells under three-dimensional culture conditions", Cell Biol Int, 36, pp.129~136 (2012)
- Nuñez J, "Human cementoblasts express enamelassociated molecules in vitro and in vivo", J Periodontal Res, 45, pp.809~814 (2010)
- Paula-Silva FW, "Calcium hydroxide promotes cementogenesis and induces cementoblastic differentiation of mesenchymal periodontal ligament cells in a CEMP1-and ERK-dependent manner", Calcif Tissue Int, 87, pp.144~157 (2010)
- Gronthos S, "Differential cell surface expression of the STRO-1 and alkaline phosphatase antigens on discrete developmental stages in primary cultures of human bone cells", J Bone Miner Res, 14, pp.47~56 (1999)
- Bronckers AL, "Immunolocalization of osteopontin, osteocalcin, and dentin sialoprotein during dental root formation and early cementogenesis in the rat", J Bone Miner Res, 9, pp.833~841 (1994)
- Pitaru S, "Specific cementum attachment protein enhances selectively the attachment and migration of periodontal cells to root surfaces", J Periodontal Res, 30, pp.360~368 (1995)
- Arzate H, "Cementum proteins: role in cementogenesis, biomineralization, periodontium formation and regeneration", Periodontol 2000, 67, pp.211~233 (2015)
- Arzate H, "Immunolocalization of a human cementoblastoma-conditioned medium-derived protein", J Dent Res, 81, pp.541~546 (2002)