Analysis of The Cells Isolated From Epithelial Cell Rests of Malassez Through Single-cell Limiting Dilution

2021 ◽  
Author(s):  
Syed Taufiqul Islam ◽  
Yoshihito Kurashige ◽  
Dembereldorj Bolortsetseg ◽  
Erika Minowa ◽  
Yunosuke Okada ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Single Cell ◽  
Alveolar Bone ◽  
Pcr Analysis ◽  
Cytokeratin 19 ◽  
Dilution Method ◽  
Cell Marker ◽  
Epithelial Cell Marker ◽  
Human Periodontal Ligament Fibroblast ◽  
Limiting Dilution

Abstract The epithelial cell rests of Malassez (ERM) play a pivotal role in preventing ankylosis between the alveolar bone and the tooth. Although several functions of ERM has been reported, the mechanism behind preventing dentoalveolar ankylosis remains unclear. In this study, 18 clones were isolated from ERM (CRUDE) using the single-cell limiting dilution method. Among them, ERM-2 and -3, which exhibited the highest and lowest proliferation rates, respectively, were selected. ERM-2, ERM-3, and CRUDE ERM were stained with epithelial markers, including cytokeratin-wide and cytokeratin-19, via immunofluorescence. The qRT-PCR analysis revealed increased expression levels of p75 (ameloblast marker), amelogenin, and sfrp5 (inner enamel epithelial cell marker) in the ERM-2 cells. Alternatively, ameloblastin and ck-14 (outer enamel epithelial cell marker) were highly expressed in ERM-3 cells. The mineralization of human periodontal ligament fibroblast (HPDLF) was inhibited when co-cultured with ERM-2, ERM-3, and CRUDE ERM cells. The addition of an anti-amelogenin antibody restored the mineralization of HPDLF cells. Transplanted rat molar cultured in ERM-2 (high amelogenin secretive clone) cell-derived supernatant resulted in significantly smaller bone formation than those cultured in the CRUDE ERM and ERM-3 cell-derived supernatants. These findings indicate that amelogenin produced by ERM cells might be involved in preventing dentoalveolar ankylosis.

scholarly journals Analysis of the cells isolated from epithelial cell rests of Malassez through single-cell limiting dilution

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Syed Taufiqul Islam ◽  
Yoshihito Kurashige ◽  
Erika Minowa ◽  
Koki Yoshida ◽  
Durga Paudel ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Single Cell ◽  
Periodontal Ligament ◽  
Alveolar Bone ◽  
Clonal Diversity ◽  
Periodontal Ligament Fibroblasts ◽  
Antibody Staining ◽  
Ngs Data ◽  
Limiting Dilution

AbstractThe epithelial cell rests of Malassez (ERM) are essential in preventing ankylosis between the alveolar bone and the tooth (dentoalveolar ankylosis). Despite extensive research, the mechanism by which ERM cells suppress ankylosis remains uncertain; perhaps its varied population is to reason. Therefore, in this study, eighteen unique clones of ERM (CRUDE) were isolated using the single-cell limiting dilution and designated as ERM 1–18. qRT-PCR, ELISA, and western blot analyses revealed that ERM-2 and -3 had the highest and lowest amelogenin expression, respectively. Mineralization of human periodontal ligament fibroblasts (HPDLF) was reduced in vitro co-culture with CRUDE ERM, ERM-2, and -3 cells, but recovered when an anti-amelogenin antibody was introduced. Transplanted rat molars grown in ERM-2 cell supernatants produced substantially less bone than those cultured in other cell supernatants; inhibition was rescued when an anti-amelogenin antibody was added to the supernatants. Anti-Osterix antibody staining was used to confirm the development of new bones. In addition, next-generation sequencing (NGS) data were analysed to discover genes related to the distinct roles of CRUDE ERM, ERM-2, and ERM-3. According to this study, amelogenin produced by ERM cells helps to prevent dentoalveolar ankylosis and maintain periodontal ligament (PDL) space, depending on their clonal diversity.

scholarly journals The plasma membrane-actin linking protein, ezrin, is a glomerular epithelial cell marker in glomerulogenesis, in the adult kidney and in glomerular injury

1998 ◽  
Vol 54 (6) ◽  
pp. 1934-1944 ◽  
Author(s):  
Christian Hugo ◽  
Masaomi Nangaku ◽  
Stuart J. Shankland ◽  
Raimund Pichler ◽  
Katherine Gordon ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Epithelial Cell ◽  
Cell Marker ◽  
Glomerular Injury ◽  
Glomerular Epithelial Cell ◽  
Epithelial Cell Marker ◽  
Adult Kidney

scholarly journals Tracheal occlusion stimulates cell cycle progression and type I cell differentiation in lungs of fetal rats

2003 ◽  
Vol 285 (2) ◽  
pp. L344-L353 ◽  
Author(s):  
Jyoji Yoshizawa ◽  
Cheryl J. Chapin ◽  
Lourenço Sbragia ◽  
Robert Ertsey ◽  
Jorge A. Gutierrez ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Epithelial Cell ◽  
Cell Cycle Progression ◽  
Type I ◽  
Type Ii ◽  
Cell Marker ◽  
Lung Weight ◽  
Cycle Progression ◽  
Epithelial Cell Marker ◽  
Fetal Rats

Fetal tracheal occlusion (TO) has been reported to stimulate lung growth but decreases number and maturation of type II cells, effects that vary with gestational age and duration of TO. We examined effects of a novel method of TO (unipolar microcautery to seal the trachea) produced at 19.5–20 days (d) of gestation in fetal rats; fetuses were delivered at term, 22 d. Controls were sham operated and unoperated littermates. TO increased wet lung weight but not dry lung weight or lung DNA and protein. To evaluate further the effects of TO, we examined the cell cycle regulators, cyclins D1 and A, in fetal lungs. Cyclin D1 increased with TO ( P < 0.005). TO also increased expression of the type I epithelial cell marker RTI40 (mRNA and protein). TO decreased mRNA for surfactant proteins (SP)-A and -C but did not affect protein levels of SP-A and -B and of RTII70, a type II epithelial cell marker. We conclude that TO by microcautery, even of short duration, has diverse pulmonary effects including stimulating increased levels of cyclin D1 with probable cell cycle progression, type I cell differentiation, and possibly inhibiting type II cell function.

scholarly journals Mapping the immune microenvironment for mandibular alveolar bone homeostasis at single-cell resolution

Bone Research ◽  
2021 ◽  
Vol 9 (1) ◽  
Author(s):  
Weimin Lin ◽  
Qiwen Li ◽  
Danting Zhang ◽  
Xiaohan Zhang ◽  
Xingying Qi ◽  
...  
Keyword(s):  
Single Cell ◽  
Alveolar Bone ◽  
Immune Cell ◽  
Adipogenic Differentiation ◽  
Oncostatin M ◽  
Long Bone ◽  
Bone Homeostasis ◽  
Oral Diseases ◽  
Immune Microenvironment ◽  
Occlusal Force

AbstractAlveolar bone is the thickened ridge of jaw bone that supports teeth. It is subject to constant occlusal force and pathogens invasion, and is therefore under active bone remodeling and immunomodulation. Alveolar bone holds a distinct niche from long bone considering their different developmental origin and postnatal remodeling pattern. However, a systematic explanation of alveolar bone at single-cell level is still lacking. Here, we construct a single-cell atlas of mouse mandibular alveolar bone through single-cell RNA sequencing (scRNA-seq). A more active immune microenvironment is identified in alveolar bone, with a higher proportion of mature immune cells than in long bone. Among all immune cell populations, the monocyte/macrophage subpopulation most actively interacts with mesenchymal stem cells (MSCs) subpopulation. Alveolar bone monocytes/macrophages express a higher level of Oncostatin M (Osm) compared to long bone, which promotes osteogenic differentiation and inhibits adipogenic differentiation of MSCs. In summary, our study reveals a unique immune microenvironment of alveolar bone, which may provide a more precise immune-modulatory target for therapeutic treatment of oral diseases.

RT-PCR analysis of ABC, SLC and SLCO drug transporters in human lung epithelial cell models

2009 ◽  
Vol 61 (5) ◽  
pp. 583-591 ◽  
Author(s):  
Sibylle Endter ◽  
Danielle Francombe ◽  
Carsten Ehrhardt ◽  
Mark Gumbleton
Keyword(s):  
Epithelial Cell ◽  
Human Lung ◽  
Drug Transporters ◽  
Lung Epithelial Cell ◽  
Pcr Analysis ◽  
Cell Models ◽  
Rt Pcr ◽  
Human Lung Epithelial Cell ◽  
Lung Epithelial

scholarly journals Early megakaryocyte lineage-committed progenitors in adult mouse bone marrow

2021 ◽  
Author(s):  
Zixian Liu ◽  
Jinhong Wang ◽  
Miner Xie ◽  
Peng Wu ◽  
Yao Ma ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Single Cell ◽  
Pcr Analysis ◽  
Mouse Bone Marrow ◽  
Hematopoietic Stem ◽  
Colony Assay ◽  
Megakaryocyte Progenitors ◽  
Cell Colony

Hematopoietic stem cells (HSCs) have been considered to progressively lose their self-renewal and differentiation potentials prior to the commitment to each blood lineage. However, recent studies have suggested that megakaryocyte progenitors are generated at the level of HSCs. In this study, we newly identified early megakaryocyte lineage-committed progenitors (MgPs) in CD201-CD48- cells and CD48+ cells separated from the CD150+CD34-Kit+Sca-1+Lin- HSC population of the bone marrow in C57BL/6 mice. Single-cell transplantation and single-cell colony assay showed that MgPs, unlike platelet-biased HSCs, had little repopulating potential in vivo, but formed larger megakaryocyte colonies in vitro (on average eight megakaryocytes per colony) than did previously reported megakaryocyte progenitors (MkPs). Single-cell RNA-sequencing supported that these MgPs lie between HSCs and MkPs along the megakaryocyte differentiation pathway. Single-cell colony assay and single-cell RT-PCR analysis suggested the coexpression of CD41 and Pf4 is associated with megakaryocyte colony-forming activity. Single-cell colony assay of a small number of cells generated from single HSCs in culture suggested that MgPs are not direct progeny of HSCs. In this study, we propose a differentiation model in which HSCs give rise to MkPs through MgPs.

Abstract 180: Increased Atherosclerosis Associated with Periodontal Disease Is Mediated by CD36

2012 ◽  
Vol 32 (suppl_1) ◽  
Author(s):  
Maria Febbraio ◽  
Paul M Brown
Keyword(s):  
Periodontal Disease ◽  
Alveolar Bone ◽  
Inflammatory Responses ◽  
Disease Risk ◽  
Plasma Cholesterol ◽  
Atherosclerotic Lesion ◽  
Pcr Analysis ◽  
Modified Ldl ◽  
Oxidatively Modified

We previously showed that inflammation, and not hyperlipidemia alone, was necessary for CD36 dependent atherogenesis. Chronic periodontal disease is characterized by a persistent inflammatory state and is epidemiologically associated with cardiovascular disease. We hypothesize that CD36 is an essential link between periodontal disease and atherosclerosis. Low density lipoprotein receptor knock out (LDLR KO) mice and CD36/LDLR double KO mice were infected with the periodontal disease associated bacteria, Porphyromonas gingivalis (Pg), by oral lavage and fed a Western diet for 12 weeks (n = 7-14/group). We assessed periodontal disease, risk factors associated with atherosclerosis, and lesion burden. We conducted studies in isolated macrophages to understand mechanistic differences between the groups. Wild type and CD36 KO macrophages equally phagocytosed bacteria. We measured the cemento-enamel junction of each molar to assess periodontal disease and found that it was significantly increased in infected mice compared with uninfected controls. Histological analysis showed neutrophil, osteoclast and macrophage infiltrates in the alveolar bone of infected mice. Differences in plasma cholesterol, triacylglycerol, insulin resistance and weight gain did not necessarily track with atherosclerosis burden, however blood neutrophils and cytokines were increased in infected LDLR KO mice compared with all other groups. Infected LDLR KO mice had significantly increased atherosclerotic lesion burden compared with uninfected LDLR KO mice, and all of the increased lesion was CD36 dependent. PCR analysis found no evidence for direct infection of atherosclerotic lesions by Pg. In vitro macrophage studies showed that heat killed Pg, lipopolysaccharide (LPS) derived from Pg, oxidatively modified LDL or plasma from infected mice, could not activate the NALP3 inflammasome. Combining heat killed Pg or Pg LPS with oxidatively modified LDL or plasma from infected mice, however, led to significant IL-1 beta secretion that was CD36 and NFkB dependent. Our data suggest that atherosclerosis associated with periodontal disease is mediated by cellular inflammatory responses involving both CD36 and Toll-like receptor.

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