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 Distinguish fatty acids impact survival, differentiation and cellular function of periodontal ligament fibroblasts

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Judit Symmank ◽  
Martin Chorus ◽  
Sophie Appel ◽  
Jana Marciniak ◽  
Isabel Knaup ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Periodontal Ligament ◽  
Alveolar Bone ◽  
Tooth Movement ◽  
Orthodontic Tooth Movement ◽  
Serum Levels ◽  
Osteoblastic Differentiation ◽  
Mechanical Stimuli ◽  
Periodontal Ligament Fibroblasts

Abstract Alveolar bone (AB) remodeling is necessary for the adaption to mechanical stimuli occurring during mastication and orthodontic tooth movement (OTM). Thereby, bone degradation and assembly are strongly regulated processes that can be altered in obese patients. Further, increased fatty acids (FA) serum levels affect bone remodeling cells and we, therefore, investigated whether they also influence the function of periodontal ligament fibroblast (PdLF). PdLF are a major cell type regulating the differentiation and function of osteoblasts and osteoclasts localized in the AB. We stimulated human PdLF (HPdLF) in vitro with palmitic (PA) or oleic acid (OA) and analyzed their metabolic activity, growth, survival and expression of osteogenic markers and calcium deposits. Our results emphasize that PA increased cell death of HPdLF, whereas OA induced their osteoblastic differentiation. Moreover, quantitative expression analysis of OPG and RANKL revealed altered levels in mechanically stimulated PA-treated HPdLF. Furthermore, osteoclasts stimulated with culture medium of mechanical stressed FA-treated HPdLF revealed significant changes in cell differentiation upon FA-treatment. For the first time, our results highlight a potential role of specific FA in the function of HPdLF-modulated AB remodeling and help to elucidate the complex interplay of bone metabolism, mechanical stimulation and obesity-induced alterations.

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 PPARγ-Induced Global H3K27 Acetylation Maintains Osteo/Cementogenic Abilities of Periodontal Ligament Fibroblasts

2021 ◽  
Vol 22 (16) ◽  
pp. 8646
Author(s):  
Hang Yuan ◽  
Shigeki Suzuki ◽  
Shizu Hirata-Tsuchiya ◽  
Akiko Sato ◽  
Eiji Nemoto ◽  
...  
Keyword(s):  
Periodontal Ligament ◽  
Alveolar Bone ◽  
Key Factors ◽  
Active Chromatin ◽  
Periodontal Ligament Fibroblasts ◽  
Chromatin Region ◽  
Transcriptional Alterations ◽  
Pparγ Agonists

The periodontal ligament is a soft connective tissue embedded between the alveolar bone and cementum, the surface hard tissue of teeth. Periodontal ligament fibroblasts (PDLF) actively express osteo/cementogenic genes, which contribute to periodontal tissue homeostasis. However, the key factors maintaining the osteo/cementogenic abilities of PDLF remain unclear. We herein demonstrated that PPARγ was expressed by in vivo periodontal ligament tissue and its distribution pattern correlated with alkaline phosphate enzyme activity. The knockdown of PPARγ markedly reduced the osteo/cementogenic abilities of PDLF in vitro, whereas PPARγ agonists exerted the opposite effects. PPARγ was required to maintain the acetylation status of H3K9 and H3K27, active chromatin markers, and the supplementation of acetyl-CoA, a donor of histone acetylation, restored PPARγ knockdown-induced decreases in the osteo/cementogenic abilities of PDLF. An RNA-seq/ChIP-seq combined analysis identified four osteogenic transcripts, RUNX2, SULF2, RCAN2, and RGMA, in the PPARγ-dependent active chromatin region marked by H3K27ac. Furthermore, RUNX2-binding sites were selectively enriched in the PPARγ-dependent active chromatin region. Collectively, these results identified PPARγ as the key transcriptional factor maintaining the osteo/cementogenic abilities of PDLF and revealed that global H3K27ac modifications play a role in the comprehensive osteo/cementogenic transcriptional alterations mediated by PPARγ.

scholarly journals In Vitro Compression Model for Orthodontic Tooth Movement Modulates Human Periodontal Ligament Fibroblast Proliferation, Apoptosis and Cell Cycle

Biomolecules ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 932
Author(s):  
Julia Brockhaus ◽  
Rogerio B. Craveiro ◽  
Irma Azraq ◽  
Christian Niederau ◽  
Sarah K. Schröder ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Periodontal Ligament ◽  
Environmental Changes ◽  
Tooth Movement ◽  
Orthodontic Tooth Movement ◽  
Compressive Force ◽  
Periodontal Ligament Fibroblasts ◽  
Human Periodontal Ligament Fibroblasts

Human Periodontal Ligament Fibroblasts (hPDLF), as part of the periodontal apparatus, modulate inflammation, regeneration and bone remodeling. Interferences are clinically manifested as attachment loss, tooth loosening and root resorption. During orthodontic tooth movement (OTM), remodeling and adaptation of the periodontium is required in order to enable tooth movement. hPDLF involvement in the early phase-OTM compression side was investigated for a 72-h period through a well-studied in vitro model. Changes in the morphology, cell proliferation and cell death were analyzed. Specific markers of the cell cycle were investigated by RT-qPCR and Western blot. The study showed that the morphology of hPDLF changes towards more unstructured, unsorted filaments under mechanical compression. The total cell numbers were significantly reduced with a higher cell death rate over the whole observation period. hPDLF started to recover to pretreatment conditions after 48 h. Furthermore, key molecules involved in the cell cycle were significantly reduced under compressive force at the gene expression and protein levels. These findings revealed important information for a better understanding of the preservation and remodeling processes within the periodontium through Periodontal Ligament Fibroblasts during orthodontic tooth movement. OTM initially decelerates the hPDLF cell cycle and proliferation. After adapting to environmental changes, human Periodontal Ligament Fibroblasts can regain homeostasis of the periodontium, affecting its reorganization.

Cultivation of Periodontal Ligament Fibroblasts on Dental Implant Materials and Enzymatically Debrided Teeth

1974 ◽  
Vol 53 (6) ◽  
pp. 1368-1376 ◽  
Author(s):  
T.K. Huard ◽  
L.F. Arnold ◽  
P. Baram
Keyword(s):  
Rhesus Monkey ◽  
Methyl Methacrylate ◽  
Dental Implant ◽  
Electron Micrographs ◽  
Periodontal Ligament ◽  
Scanning Electron Micrographs ◽  
Periodontal Ligament Fibroblasts ◽  
Implant Materials ◽  
Scanning Electron

Rhesus monkey periodontal ligament-derived fibroblasts were cultured on glass, Vitallium, poly(methyl methacrylate) and enzymatically debrided teeth. Scanning electron micrographs of these preparations and of the periodontal ligament surrounding normal and replanted teeth were compared. The fibroblasts cultured in vitro could organize on implant material and enzymatically debrided teeth to produce a network with fibers resembling those that are seen in the normal periodontal ligament.

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