Analysis of gene expression during mineralization of cultured human periodontal ligament cells
2
조회
0
다운로드
4
인용
0
참고문헌
키워드
초록
Purpose: Under different culture conditions, periodontal ligament (PDL) stem cells are capable of differentiating into cementoblast-like cells, adipocytes, and collagen-forming cells. Several previous studies reported that because of the stem cells in the PDL, the PDL have a regenerative capacity which, when appropriately triggered, participates in restoring connective tissues and mineralized tissues. Therefore, this study analyzed the genes involved in mineralization during differentiation of human PDL (hPDL) cells, and searched for candidate genes possibly associated with the mineralization of hPDL cells. Methods: To analyze the gene expression pattern of hPDL cells during differentiation, the hPDL cells were cultured in two conditions, with or without osteogenic cocktails (β-glycerophosphate, ascorbic acid and dexamethasone), and a DNA microarray analysis of the cells cultured on days 7 and 14 was performed. Reverse transcription-polymerase chain reaction was performed to validate the DNA microarray data. Results: The up-regulated genes on day 7 by hPDL cells cultured in osteogenic medium were thought to be associated with calcium/iron/metal ion binding or homeostasis (PDE1A, HFE and PCDH9) and cell viability (PCDH9), and the down-regulated genes were thought to be associated with proliferation (PHGDH and PSAT1). Also, the up-regulated genes on day 14 by hPDL cells cultured in osteogenic medium were thought to be associated with apoptosis, angiogenesis (ANGPTL4 and FOXO1A), and adipogenesis (ANGPTL4 and SEC14L2), and the down-regulated genes were thought to be associated with cell migration (SLC16A4). Conclusions: This study suggests that when appropriately triggered, the stem cells in the hPDL differentiate into osteoblasts/cementoblasts, and the genes related to calcium binding (PDE1A and PCDH9), which were strongly expressed at the stage of matrix maturation, may be associated with differentiation of the hPDL cells into osteoblasts/cementoblasts.
Abstract
Purpose: Under different culture conditions, periodontal ligament (PDL) stem cells are capable of differentiating into cementoblast-like cells, adipocytes, and collagen-forming cells. Several previous studies reported that because of the stem cells in the PDL, the PDL have a regenerative capacity which, when appropriately triggered, participates in restoring connective tissues and mineralized tissues. Therefore, this study analyzed the genes involved in mineralization during differentiation of human PDL (hPDL) cells, and searched for candidate genes possibly associated with the mineralization of hPDL cells. Methods: To analyze the gene expression pattern of hPDL cells during differentiation, the hPDL cells were cultured in two conditions, with or without osteogenic cocktails (β-glycerophosphate, ascorbic acid and dexamethasone), and a DNA microarray analysis of the cells cultured on days 7 and 14 was performed. Reverse transcription-polymerase chain reaction was performed to validate the DNA microarray data. Results: The up-regulated genes on day 7 by hPDL cells cultured in osteogenic medium were thought to be associated with calcium/iron/metal ion binding or homeostasis (PDE1A, HFE and PCDH9) and cell viability (PCDH9), and the down-regulated genes were thought to be associated with proliferation (PHGDH and PSAT1). Also, the up-regulated genes on day 14 by hPDL cells cultured in osteogenic medium were thought to be associated with apoptosis, angiogenesis (ANGPTL4 and FOXO1A), and adipogenesis (ANGPTL4 and SEC14L2), and the down-regulated genes were thought to be associated with cell migration (SLC16A4). Conclusions: This study suggests that when appropriately triggered, the stem cells in the hPDL differentiate into osteoblasts/cementoblasts, and the genes related to calcium binding (PDE1A and PCDH9), which were strongly expressed at the stage of matrix maturation, may be associated with differentiation of the hPDL cells into osteoblasts/cementoblasts.
참고문헌 (25)
- Lekic P, "Tenenbaum H, McCulloch CA. Phenotypic comparison of periodontal ligament cells in vivo and in vitro", J Periodontal Res, 36, pp.71~79 (2001)
- Ivanovski S, "Expression of bone matrix protein mRNAs by primary and cloned cultures of the regenerative phenotype of human periodontal fibroblasts", J Dent Res, 80, pp.1665~1771 (2001)
- Murakami Y, "Novel isolation of alkaline phosphatase-positive subpopulation from periodontal ligament fibroblasts", J Periodontol, 74, pp.780~786 (2003)
- Wu L, "Early osteogenic differential protein profile detected by proteomic analysis in human periodontal ligament cells", J Periodontal Res, 44, pp.645~656 (2009)
- Seo BM, "Investigation of multipotent postnatal stem cells from human periodontal ligament", Lancet, 364, pp.149~155 (2004)
- Flores MG, "Periodontal ligament cell sheet promotes periodontal regeneration in athymic rats", J Clin Periodontol, 35, pp.1066~1072 (2008)
- Iwata T, "Periodontal regeneration with multi-layered periodontal ligament-derived cell sheets in a canine model", Biomaterials, 30, pp.2716~2723 (2009)
- Kulterer B, "Gene expression profiling of human mesenchymal stem cells derived from bone marrow during expansion and osteoblast differentiation", BMC Genomics, 8, pp.70~ (2007)
- Yamada S, "Regulation of PLAP-1 expression in periodontal ligament cells", J Dent Res, 85, pp.447~451 (2006)
- Lallier TE, "Use of microarrays to find novel regulators of periodontal ligament fibroblast differentiation", Cell Tissue Res, 327, pp.93~109 (2007)
- 신재희, "cDNA microarray에 의한 치주인대세포의 광물화 결절형성에 관여하는 유전자들의 분석", Journal of Periodontal & Implant Science, 37(5), pp.447~463 (2007), 대한치주과학회
- Dahl LK. A, "simple and sensitive histochemical method for calcium", Proc Soc Exp Biol Med, 통권 : 80, pp.474~479 (1952)
- Bartold PM, "Stem cells and periodontal regeneration", Periodontol 2000, 40, pp.164~172 (2006)
- Nagatomo K, "Stem cell properties of human periodontal ligament cells", J Periodontal Res, 41, pp.303~310 (2006)
- Kobayashi M, "Recombinant human bone morphogenetic protein-2 stimulates osteoblastic differentiation in cells isolated from human periodontal ligament", J Dent Res, 78, pp.1624~1633 (1999)
- 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)
- Michibata H, "Human Ca2+", calmodulin-dependent phosphodiesterase PDE1A: novel splice variants, their specific expression, genomic organization, and chromosomal localization, 2001, pp.1517, 278~287
- Fidock M, "Isolation and differential tissue distribution of two human cDNAs encoding PDE1 splice variants", Cell Signal, 14, pp.53~60 (2002)
- Zayzafoon M, "Calmodulin and calmodulin-dependent kinase IIalpha regulate osteoblast differentiation by controlling c-fos expression", J Biol Chem, 280, pp.7049~7059 (2005)
- Etzrodt J, "Expression of classic cadherins and delta-protocadherins in the developing ferret retina", BMC Neurosci, 10, pp.153~ (2009)
- Gray MJ, "Therapeutic targeting of neuropilin-2 on colorectal carcinoma cells implanted in the murine liver", J Natl Cancer Inst, 100, pp.109~120 (2008)
- Hopwood B, "Gene expression profile of the bone microenvironment in human fragility fracture bone", Bone, 44, pp.87~101 (2009)
- Cho HM, "Nucleotide sequence and differential expression of the human 3-phosphoglycerate dehydrogenase gene", Gene, 245, pp.193~201 (2000)
- Pollari S, "Enhanced serine production by bone metastatic breast cancer cells stimulates osteoclastogenesis", Breast Cancer Res Treat, 125, pp.421~430 (2011)
- Gallagher SM, "Interaction of monocarboxylate transporter 4 with beta1-integrin and its role in cell migration", Am J Physiol Cell Physiol, 296, pp.C414~C421 (2009)