• Title/Summary/Keyword: Conservative gene

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Epigenetics: general characteristics and implications for oral health

  • Seo, Ji-Yun;Park, Yoon-Jung;Yi, Young-Ah;Hwang, Ji-Yun;Lee, In-Bog;Cho, Byeong-Hoon;Son, Ho-Hyun;Seo, Deog-Gyu
    • Restorative Dentistry and Endodontics
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    • v.40 no.1
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    • pp.14-22
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    • 2015
  • Genetic information such as DNA sequences has been limited to fully explain mechanisms of gene regulation and disease process. Epigenetic mechanisms, which include DNA methylation, histone modification and non-coding RNAs, can regulate gene expression and affect progression of disease. Although studies focused on epigenetics are being actively investigated in the field of medicine and biology, epigenetics in dental research is at the early stages. However, studies on epigenetics in dentistry deserve attention because epigenetic mechanisms play important roles in gene expression during tooth development and may affect oral diseases. In addition, understanding of epigenetic alteration is important for developing new therapeutic methods. This review article aims to outline the general features of epigenetic mechanisms and describe its future implications in the field of dentistry.

DETECTION OF BLACK-PIGMENTED BACTERIA IN INFECTED ROOT CANALS (감염근관에서 Black-Pigmented Bacteria의 동정에 대한 연구)

  • Kwon, Eun-Kyoung;Kim, Eun-Sook;Kwak, Ju-Seog;Lee, Hwang;Lee, Su-Jong;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.27 no.1
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    • pp.54-65
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    • 2002
  • Black-pigmented bacteria have been implicated in the endodontic infections. This group of microorganisms includes Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. The organisms display a wide variety of virulence factors that may be pertinent to acute endodontic infections. The aim of this study was to identify P. endodontalis, P. gingivalis, P. intermedia, and P. nigrescens by using special potency disk test, filter paper spot test, 165 rRNA gene-directed PCR, and API 32A. Microbial samples were collected from root canals of 33 intact teeth with necrotic pulp and/or apical periodontitis. Conventional laboratory methods were used for identification of the strains of black pigmented bacteria. Eighteen of 33 samples were positive for the growth of black-pigmented bacteria Five colonies were cultured from each pure cultured colonies from Brucella agar plate. Seventy seven colonies were positive for the growth of black-pigmented bacteria. Thirty three of 77(42.6%) were identifed as P. nigrescens, 10 of 77(12.9%)were P. gingivalis, 6 of 77(7.8%) were P. endodontalis, 10 of 77(12.9%) were P. intermedia. On the contrary the reference strains of P. nigrescens, experimental strains of P nigrescens was sensitive to kanamycin in special potency disk test. 165 rRNA gene PCR and API test after rapid presumptative identification methods, such as special potency disk test and filter paper spot test, would be accurate detection methods for black-pigemented bacteria.

Study on Biocompatibility and Mineralization Potential of Capseal

  • Bae, Kwang Shik;Chang, Seok Woo;Kum, Kee Yeon;Lee, Woo Cheol
    • Journal of Korean Dental Science
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    • v.7 no.1
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    • pp.1-5
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    • 2014
  • Purpose: Capseal I and Capseal II are calcium silicate and calcium phosphate based experimental root canal sealers. This study sought to evaluate the biocompatibility and mineralization potential of Capseal I and Capseal II. Materials and Methods: The biocompatibility and mineralization related gene expression (alkaline phosphatase [ALP], bone sialoprotein [BSP], and osteocalcin) of Capseal I and Capseal II were compared using methylthiazol tetrazolium assay and reverse transcription-polymerization chain reaction analysis, respectively. The results were analyzed by Kruskal-Wallis test. A P-value of <0.05 was considered significant. Result: Both Capseal I and Capseal II were favorable in terms of biocompatibility, influencing the messenger RNA expression of ALP and BSP. Conclusion: Within the limitation of this study, Capseal is biocompatible, with mineralization promoting potential; thus, it could be a promising root canal sealer.

VARIATION OF UREASE GENES(ureC) FROM Streptococcus salivarius (Streptococcus salivarisu의 요소분해효소 유전자 변이에 관한 연구)

  • Choi, Hye-Jin;Lee, Jin-Yong;Choi, Ho-Young
    • Restorative Dentistry and Endodontics
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    • v.24 no.4
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    • pp.535-545
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    • 1999
  • Crease of Streptococcus salivarius is believed to play a critical role in bacterial ecology and pH homeostasis in the mouth, and consequently affect the pathogenesis of dental caries and periodontal diseases. Expression of the urease gene is greatly enhanced by low p. f. excess of Carbohydrate, and faster growth. It was observed that urease activity of the strains of S. salivarius that exhibited no of low urease activity was not increased even in low pH condition. In this study, it was hypothesized that the urease gene of the strains is absent, defected, or greatly changed by genetic combination. In order to prove this hypothesis, chromosomes were obtained from 28 S. salivarius strains which had been isolated from normal teeth and carious lesions, subjected to polymerase chain reaction (PCR) using primers encoding highly conserved sequence from ureC, and then the obtained PCR products were compared. The results were as follows: 1. After PCR the strains generated either one of 0.54- and 1.3-kbp PCR products, or none. 2. All 16 strains having a higher urease activity(<50${\mu}mol/min/mg$) produced 0.54-kbp PCR products. 3. Twelve strains without urease activity and with a lower urease activity(<50${\mu}mol/min/mg$) yield either one of 0.54 and 1.3-kbp PCR products, or none. 4. The DNA sequence of the 0.54-kbp PCR product (pCAP-0.54) exhibited 95% identity to the ureC of S. salivarus 57.I; 30bp were found to be different, which led to difference of only 2 amino acids in the sequence. 5. The DNA sequence of the 1.3-kbp PCR product(pCAP-1.3) was found to be highly homologous to the aminopeptidase C gene of Streptococcus thermophilus. Overall results indicate that there are considerable variations of the urease genes from S. salivarus strains and the variations may affect the uncolytic activity of the bacteria directly of indirectly.

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Analysis of gene expression during odontogenic differentiation of cultured human dental pulp cells

  • Seo, Min-Seock;Hwang, Kyung-Gyun;Kim, Hyong-Bum;Baek, Seung-Ho
    • Restorative Dentistry and Endodontics
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    • v.37 no.3
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    • pp.142-148
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    • 2012
  • Objectives: We analyzed gene-expression profiles after 14 day odontogenic induction of human dental pulp cells (DPCs) using a DNA microarray and sought candidate genes possibly associated with mineralization. Materials and Methods: Induced human dental pulp cells were obtained by culturing DPCs in odontogenic induction medium (OM) for 14 day. Cells exposed to normal culture medium were used as controls. Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR). We also performed a gene set enrichment analysis (GSEA) of the microarray data. Results: Six hundred and five genes among the 47,320 probes on the BeadChip differed by a factor of more than two-fold in the induced cells. Of these, 217 genes were upregulated, and 388 were down-regulated. GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated. Conclusions: Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.

Effect of dentin treatment on proliferation and differentiation of human dental pulp stem cells

  • Park, Minjeong;Pang, Nan-Sim;Jung, Il-Young
    • Restorative Dentistry and Endodontics
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    • v.40 no.4
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    • pp.290-298
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    • 2015
  • Objectives: Sodium hypochlorite (NaOCl) is an excellent bactericidal agent, but it is detrimental to stem cell survival, whereas intracanal medicaments such as calcium hydroxide ($Ca[OH]_2$) promote the survival and proliferation of stem cells. This study evaluated the effect of sequential NaOCl and $Ca(OH)_2$ application on the attachment and differentiation of dental pulp stem cells (DPSCs). Materials and Methods: DPSCs were obtained from human third molars. All dentin specimens were treated with 5.25% NaOCl for 30 min. DPSCs were seeded on the dentin specimens and processed with additional 1 mg/mL $Ca(OH)_2$, 17% ethylenediaminetetraacetic acid (EDTA) treatment, file instrumentation, or a combination of these methods. After 7 day of culture, we examined DPSC morphology using scanning electron microscopy and determined the cell survival rate with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. We measured cell adhesion gene expression levels after 4 day of culture and odontogenic differentiation gene expression levels after 4 wk using quantitative real-time polymerase chain reaction. Results: DPSCs did not attach to the dentin in the NaOCl-treated group. The gene expression levels of fibronectin-1 and secreted phosphoprotein-1 gene in both the $Ca(OH)_2$- and the EDTA-treated groups were significantly higher than those in the other groups. All $Ca(OH)_2$-treated groups showed higher expression levels of dentin matrix protein-1 than that of the control. The dentin sialophosphoprotein level was significantly higher in the groups treated with both $Ca(OH)_2$ and EDTA. Conclusions: The application of $Ca(OH)_2$ and additional treatment such as EDTA or instrumentation promoted the attachment and differentiation of DPSCs after NaOCl treatment.

Chemical Constitution, Morphological Characteristics, and Biological Properties of ProRoot Mineral Trioxide Aggregate and Ortho Mineral Trioxide Aggregate

  • Kum, Kee Yeon;Yoo, Yeon Jee;Chang, Seok Woo
    • Journal of Korean Dental Science
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    • v.6 no.2
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    • pp.41-49
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    • 2013
  • Purpose: This study sought to compare the elemental constitution, morphological characteristics, particle size distribution, biocompatibility, and mineralization potential of Ortho MTA (OMTA) and ProRoot MTA (PMTA). Materials and Methods: OMTA and PMTA were compared using energy-dispersive spectrometry, particle size analysis, and scanning electron microscopy. The biocompatibility and mineralization-related gene expression (osteonectin and osteopontin) of both MTAs were also compared using methylthiazol tetrazolium assay and reverse transcription-polymerization chain reaction analysis, respectively. The results were analyzed by Kruskal-Wallis test with Bonferroni correction. P-value of <0.05 was considered significant. Result: The morphology of OMTA powders was similar to that of PMTA. The constituent elements of both MTAs were calcium, silicon, and aluminum. The mean particle sizes of OMTA and PMTA were 4.60 and 3.34 mm, respectively. Both MTAs had equally favorable in vitro biocompatibility and affected the messenger RNA expression of osteonectin and osteopontin. Conclusion: Within the limitations of this study, OMTA could be a promising biomaterial in clinical endodontics.

Analysis of Conservative Genes in Thermophilic and Hyperthermophilic Bacteria (고온성과 초고온성 세균의 보존적 유전자 분석)

  • Lee Dong-Geun;Lee Jae-Hwa;Ha Bae Jin;Ha Jong-Myung;Lee Jung-Hyun;Kim Sang-Jin;Lee Sang Hyeon
    • KSBB Journal
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    • v.20 no.5 s.94
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    • pp.387-391
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    • 2005
  • Totally 16,299 conservative genes, commonly found in 13 thermophilic and hyperthermophilic bacteria, were analyzed. All genes were belong to W 67 COGs (clusters of orthologous groups of proteins). COGs related to protein metabolism were 80 among 167 COGs. Conservative genes were not limited only thermophiles and hyperthermophiles, meaning thermal stability is independent of specific protein. However reverse gyrase was only found in all hyperthermophilic archaebacteria and eubacteria, meaning DNA stability is important in hyperthermophiles. Hyperthermophilic eubacteria and thermophilic archaebacteria had different position between phylogenetic tree of gene content and 165 rRNA gene. Thermophilic archaebacteria hyperthermophilic eubacteria and archaebacteria had similar values by the statistical analysis of distance values with 167 COGs in each organism.

Comparison of gene expression profiles of human dental pulp cells treated with mineral trioxide aggregate and calcium hydroxide (인간치수세포에 Mineral Trioxide Aggregate와 수산화칼슘 제재 적용 시 유전자 발현 양상 비교)

  • Kim, Yong-Beom;Shon, Won-Jun;Lee, Woo-Cheol;Kum, Kee-Yeon;Baek, Seung-Ho;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.36 no.5
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    • pp.397-408
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    • 2011
  • Objectives: This study investigated changes in gene expressions concerning of differentiation, proliferation, mineralization and inflammation using Human-8 expression bead arrays when white Mineral Trioxide Aggregate and calcium hydroxide-containing cement were applied in vitro to human dental pulp cells (HDPCs). Materials and Methods: wMTA (white ProRoot MTA, Dentsply) and Dycal (Dentsply Caulk) in a Teflon tube (inner diameter 10 mm, height 1 mm) were applied to HDPCs. Empty tube-applied HDPCs were used as negative control. Total RNA was extracted at 3, 6, 9 and 24 hr after wMTA and Dycal application. The results of microarray were confirmed by reverse transcriptase polymerase chain reaction. Results: Out of the 24,546 genes, 43 genes (e.g., BMP2, FOSB, THBS1, EDN1, IL11, COL10A1, TUFT1, HMOX1) were up-regulated greater than two-fold and 25 genes (e.g., SMAD6, TIMP2, DCN, SOCS2, CEBPD, KIAA1199) were down-regulated below 50% by wMTA. Two hundred thirty nine genes (e.g., BMP2, BMP6, SMAD6, IL11, FOS, VEGFA, PlGF, HMOX1, SOCS2, CEBPD, KIAA1199) were up-regulated greater than two-fold and 358 genes (e.g., EDN1, FGF) were down-regulated below 50% by Dycal. Conclusions: Both wMTA and Dycal induced changes in gene expressions related with differentiation and proliferation of pulp cells. wMTA induced changes in gene expressions related with mineralization, and Dycal induced those related with angiogenesis. The genes related with inflammation were more expressed by Dycal than by wMTA. It was confirmed that both wMTA and Dycal were able to induce gene expression changes concerned with the pulp repair in different ways.

REGULATION OF PULPAL MICROCIRCULATION BY CALCITONIN GENE-RELATED PEPTIDE (Calcitonin Gene Related Peptide에 의한 치수미세순환 조절)

  • Kim, Sung-Kyo;Kim, Young-Kyung;Jin, Myoung-Uk
    • Restorative Dentistry and Endodontics
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    • v.30 no.6
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    • pp.470-476
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    • 2005
  • The purpose of this Study was to invest)gate the function or calcitonin gene-related peptide (CGRP) in regulatory mechanism of pulpal microcirculation with the aim of elucidating neurogenic inflammation. Experiments were performed on twelve cats under general anesthesia. CGRP was administered through the femoral vein to see the systemic Influence and through the external carotid artery to see the local effect. Sympathetic nerve to the dental pulp was stimulated electrically and pulpal blood flow (PBF) was measured with a laser Doppler flowmeter on the canine teeth to the drug administration. The paired variables of control and experimental data were compared by paired t-test and differences with p < 0.05 were considered statistically significant. Systemic administration of CGRP $(0.3{\mu}g/ka)$ exerted decreases in systemic blood pressure and caused changes in PBF with an initial increase i311owed by decrease and a move marked second increase and decrease. Close intra-arterial (i.a.) injection of CCRP $(0.03{\mu}g/kg)$ resulted in slight PBF increase. The effect of CGRP resulted in no significant increase in PBF in the presence of $CGRP_{8-37}$. The electrical stimulation of the sympathetic nerve alone resulted in PBF decreases. The j.a. administration of CGRP following the electrical stimulation of the sympathetic nerve compensated the decreased PBF. Therefore, CGRP effectively blocked the sympathetic nerve stimulation-induced PBF decrease. Results of the present study have provided evidences that even though the local vasodilatory function of CGRP are weak, CCRP is effectively involved in blocking the vasoconstriction caused by sympathetic nerve stimulation in the feline dental pulp.