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European Journal of Histochemistry : EJH Sep 2019Ferritin, an iron-binding protein, is composed of two subunits, a heavy chain and a light chain. It regulates many biological functions, such as proliferation,...
Ferritin, an iron-binding protein, is composed of two subunits, a heavy chain and a light chain. It regulates many biological functions, such as proliferation, angiogenesis, and immunosuppression. The objective of this study was to determine the expression and distribution of ferritin in the periodontal tissuesof primates.First, we assessed the expression of ferritin in primary cultured cells isolated from human periodontal tissues using the polymerase chain reaction and immunofluorescent staining. Second, we investigated the expression and distribution of ferritin in the periodontal tissues of Macaca fascicularis, human gingival tissues, and human gingival carcinoma tissues using immunohistochemistry.Both protein and mRNA of ferritin were constitutively present in human primary cultured cells, including those from the dental apical papilla, periodontal ligament, dental pulp, and gingival epithelium, as well as gingival fibroblasts. In M. fascicularistissues, the immunohistochemical staining was particularly strong in blood vessel and mineralizing areas of the dental pulp and periodontal ligament. Ferritin heavy chain exhibited specific immunopositivity in in the stratum basale of the epithelium in human gingival tissue and strong immunostaining was found in peripheral regions of gingival carcinoma sites. Ferritin is constitutivelypresent andwidelydistributed in the periodontal tissues of primates. Ferritin may play roles in epithelial proliferation, vascular angiogenesis, and mineralization in these tissues.
Topics: Animals; Cells, Cultured; Ferritins; Gingiva; Gingival Neoplasms; Humans; Macaca fascicularis; Male; Periodontium
PubMed: 31505926
DOI: 10.4081/ejh.2019.3046 -
Journal of Oral Biology and... 2021Although dental pulp and apical papilla are originated from neural crest cells, these tissues exhibit distinct characteristics. Notch signaling is one of the known...
Although dental pulp and apical papilla are originated from neural crest cells, these tissues exhibit distinct characteristics. Notch signaling is one of the known signaling pathways regulating stemness and behaviors of stem cells. The aim of this study was to examine Notch signaling related gene expression profile comparing between coronal pulp tissues and apical pulp complex. Results demonstrated that coronal pulp tissue had higher expression levels of various genes in Notch pathway. However, , , , and mRNA levels were significantly lower in coronal pulp tissue than those of apical pulp complex. Furthermore, dental pulp stem cells (DPSCs) and stem cells isolated from apical papilla (SCAPs) were isolated and characterized. These two cell types exhibited similar mesenchymal stem cell surface markers. DPSCs expressed higher mRNA levels of and . In addition, SCAPs demonstrated higher colony formation and cell proliferation than DPSCs. In summary, cells and tissues from dental pulp and apical papilla exhibited the distinct gene expression profile of Notch related genes. This could be of one the signaling participated in control of DPSCs and SCAPs cells behaviors.
PubMed: 33996433
DOI: 10.1016/j.jobcr.2021.04.004 -
C-Jun N-terminal kinase (JNK) pathway activation is essential for dental papilla cells polarization.PloS One 2021During tooth development, dental papilla cells differentiate into odontoblasts with polarized morphology and cell function. Our previous study indicated that the C-Jun...
During tooth development, dental papilla cells differentiate into odontoblasts with polarized morphology and cell function. Our previous study indicated that the C-Jun N-terminal kinase (JNK) pathway regulates human dental papilla cell adhesion, migration, and formation of focal adhesion complexes. The aim of this study was to further examine the role of the JNK pathway in dental papilla cell polarity formation. Histological staining, qPCR, and Western Blot suggested the activation of JNK signaling in polarized mouse dental papilla tissue. After performing an in vitro tooth germ organ culture and cell culture, we found that JNK inhibitor SP600125 postponed tooth germ development and reduced the polarization, migration and differentiation of mouse dental papilla cells (mDPCs). Next, we screened up-regulated polarity-related genes during dental papilla development and mDPCs or A11 differentiation. We found that Prickle3, Golga2, Golga5, and RhoA were all up-regulated, which is consistent with JNK signaling activation. Further, constitutively active RhoA mutant (RhoA Q63L) partly rescued the inhibition of SP600125 on cell differentiation and polarity formation of mDPCs. To sum up, this study suggests that JNK signaling has a positive role in the formation of dental papilla cell polarization.
Topics: Animals; Anthracenes; Cell Differentiation; Cell Movement; Cell Polarity; Cells, Cultured; Dental Papilla; JNK Mitogen-Activated Protein Kinases; MAP Kinase Signaling System; Mice; Mice, Inbred ICR; Mutagenesis; Tooth Germ; rhoA GTP-Binding Protein
PubMed: 33770099
DOI: 10.1371/journal.pone.0233944 -
Low-Level Laser Irradiation Promotes Proliferation and Differentiation on Apical Papilla Stem Cells.Journal of Lasers in Medical Sciences 2021Low-level laser therapy (LLLT) has been reported to improve cell proliferation and differentiation. The stem cells derived from dental apical papilla (SCAPs) are a...
Low-level laser therapy (LLLT) has been reported to improve cell proliferation and differentiation. The stem cells derived from dental apical papilla (SCAPs) are a promising therapy because they are easily obtained from immature human teeth. The effect of LLLT over SCAPs is still unknown. This study aimed to evaluate the proliferation and osteogenic potential of the SCAPs stimulated with LLLT. SCAPs were isolated from the third molars of a healthy donor and characterized according to the minimum established criteria. SCAPs were cultured for 24 hours before being exposed to LLLT. Cells were exposed to different doses, energy, and wavelengths for selecting the irradiation parameters. SCAPs proliferation was evaluated with the MTT assay at 24 hours and 7-day post-laser exposure. VEGF and TGFβ2 expression were assessed with a specific enzyme-linked immunosorbent assay (ELISA). The osteogenic differentiation potential was analyzed with alizarin red staining, and the nodule quantification was performed by the relative optical density (ROD) analysis using ImageJ software. The cells isolated from the apical papilla showed phenotype and stem cell properties. SCAPs irradiated with one dose at 6 J/m and 650 nm exhibited significantly higher proliferation (>0.05) than the controls nonirradiated. LLLT stimulated SCAPs' expression of factors VEGF and TGFβ2. Also, SCAPs irradiated showed higher osteogenic activity (<0.05). LLLT promotes proliferation, osteogenic differentiation, and VEGF and TGFβ2 expression on SCAPs. LLLT is a practical approach for the preconditioning of SCAPs for future regenerative therapies. More studies are needed to determine the underlying molecular processes that determine the mechanism of the LLLT.
PubMed: 35155160
DOI: 10.34172/jlms.2021.75 -
BMC Oral Health Jan 2021Preservation of the interdental papilla is an essential part of the functional and esthetic rehabilitation of dental treatment. It has been described that thicker...
BACKGROUND
Preservation of the interdental papilla is an essential part of the functional and esthetic rehabilitation of dental treatment. It has been described that thicker gingival tissues are more resistant to recession. The main objective of this investigation was to analyze whether a thin gingival phenotype represents a potential risk indicator affecting interdental papilla fill, height, or width in an esthetic region between maxillary central incisors. The secondary goals were: (1) to analyze parameters describing the papilla-fill, height, width, and effect of papilla base width on the vertical papillary dimension; (2) to determine correlation between different non-invasive measurements of gingival thickness; (3) to compare both sexes.
METHODS
A total of 54 periodontally healthy students (20-30 years old) were included in the study. Gingival thickness was measured using Pirop Ultrasonic Biometer. Gingival phenotype was also assessed by gingival probe transparency. Papilla height and width were measured, and the degree of papilla recession was classified.
RESULTS
No significant relationship between papilla fill, height, width and gingival probe transparency or gingival thickness was found. Gingival thickness and gingival probe transparency showed a significant relationship (P < 0.001). There was a significant relationship between papilla height and papilla fill (P = 0.028). A papilla which filled the interdental space completely seemed to be shorter. A strong positive correlation between papilla height and papilla width was found (P < 0.0001). The papilla between maxillary central incisors was significantly higher in males (P = 0.01).
CONCLUSION
The appearance of the interdental papilla may be influenced by various factors. Within the limitations of this study, the results showed that the thin gingival phenotype alone is no potential risk indicator affecting interdental papilla fill, height, or width. It seems that there may be some effect of papilla base width on its vertical dimension. Gingival probe transparency is a simple reliable method of assessment of gingival thickness with a threshold value of 1-mm gingival thickness between the thick and thin phenotypes.
Topics: Adult; Esthetics, Dental; Female; Gingiva; Humans; Male; Maxilla; Odontometry; Phenotype; Young Adult
PubMed: 33485351
DOI: 10.1186/s12903-021-01400-x -
Journal of Clinical and Experimental... Jan 2023Using dental implants to replacing missing teeth and satisfy both functional and aesthetic needs is one of the mainstream dental treatments. New approaches including... (Review)
Review
BACKGROUND
Using dental implants to replacing missing teeth and satisfy both functional and aesthetic needs is one of the mainstream dental treatments. New approaches including computer-aided design and computer-assisted manufacture (CAD/CAM) have been introduced to improve these elements. This systematic review aimed to compare CAD/CAM zirconia (Zr) implant abutments with other available abutments in terms of peri-implant health and aesthetics.
MATERIAL AND METHODS
Five electronic databases (PubMed, Web of Science, Scopus, ProQuest, and Embase) were scoured for clinical studies evaluating Zr abutments reporting on the outcomes of interest including interproximal papilla stability (PS), papilla recession (REC), pink and white esthetic score (PES, WES), marginal bone level (MBL), color, and soft tissue contour. A hand searches in English language journals until September 2020 complemented the search. Two tools of Joanna Briggs Institute and Jaded Score calculation were used for the risk of bias assessment. No quantitative synthesis of the data was done due to high heterogeneity.
RESULTS
A total of six studies from the 412 ones obtained from the search were included. The study designs were either prospective cohort (n=3) or randomized clinical trial (n=3). Papilla fill, WES, PES, and the distance from the bone crest of adjacent teeth to the contact point (CPB) and inter-tooth-implant distance (ITD) was not significantly different between Zr CAD/CAM and Zr stock abutments. However, soft tissue stability and REC index were better in Zr CAD/CAM abutments.
CONCLUSIONS
Higher soft tissue stability can be achieved for Zr compared to titanium abutments with either stock or CAD/CAM abutments. Dental implants, Dental abutment, Computer-Assisted Design, Computer-Aided Manufacturing, Zirconia abutment, Soft tissue stability.
PubMed: 36755676
DOI: 10.4317/jced.59878 -
Journal of Pharmacy & Bioallied Sciences Jul 2022To assess papilla level using different techniques in a second stage dental implant surgery.
OBJECTIVES
To assess papilla level using different techniques in a second stage dental implant surgery.
MATERIALS AND METHODS
Thirty patients who received 45 dental implants were equally divided into 3 groups of 10 each. Group I patients were operated with a scalpel with mid-crestal incision. In group II, dental implants were exposed with a gallium-aluminum-arsenide diode laser. In group III, dental implants were exposed with I shaped incision using a scalpel. Assessment of modified gingival index (mGI), modified plaque index (mPI), and Jemt index were performed at baseline, 3 months, and 6 months. The measurement of FAJI, FAJAdj, ST height, and CP Bone crest was performed.
RESULTS
A significant difference in crestal bone level of FAJ- I, FAJ- adj, ST height, and CP Bone crest was recorded at baseline, 3 months, and 6 months among groups I, II, and III ( < 0.05). At 6 months, both groups II and III exhibited >60% of papilla fill as compared to group I.
CONCLUSION
Diode laser offers maximum papillary fill and resulted in less crestal bone loss as compared to mid-crestal and I shaped incision during a second stage surgery.
PubMed: 36110672
DOI: 10.4103/jpbs.jpbs_115_22 -
Journal of Oral Biosciences Jun 2024This study aimed to investigate the regulatory mechanisms governing dental mesenchymal cell commitment during tooth development, focusing on odontoblast differentiation...
Exploring the Role of DNMT1 in Dental Papilla Cell Fate Specification during Mouse Tooth Germ Development through Integrated Single-Cell Transcriptomics and Bulk RNA Sequencing.
OBJECTIVES
This study aimed to investigate the regulatory mechanisms governing dental mesenchymal cell commitment during tooth development, focusing on odontoblast differentiation and the role of epigenetic regulation in this process.
METHODS
We performed single-cell RNA sequencing (scRNA-seq) of dental cells from embryonic day 14.5 (E14.5) mice to understand the heterogeneity of developing tooth germ cells. Computational analyses including gene regulatory network (GRN) assessment were conducted. We validated our findings using immunohistochemistry (IHC) and in vitro loss-of-function analyses using the DNA methyltransferase 1 (DNMT1) inhibitor Gsk-3484862 in primary dental mesenchymal cells (DMCs) isolated from E14.5 mouse tooth germs. Bulk RNA-seq of Gsk-3484862-treated DMCs was performed to identify potential downstream targets of DNMT1.
RESULTS
scRNA-seq analysis revealed diverse cell populations within the tooth germs, including epithelial, mesenchymal, immune, and muscle cells. Using single-cell regulatory network inference and clustering (SCENIC), we identified Dnmt1 as a key regulator of early odontoblast development. IHC analysis showed the ubiquitous expression of DNMT1 in the dental papilla and epithelium. Bulk RNA-seq of cultured DMCs showed that Gsk-3484862 treatment upregulated odontoblast-related genes, whereas genes associated with cell division and the cell cycle were downregulated. Integrated analysis of bulk RNA-seq data with scRNA-seq SCENIC profiles was used to identify the potential Dnmt1 target genes.
CONCLUSIONS
Dnmt1 may negatively affect odontoblast commitment and differentiation during tooth development. These findings contribute to a better understanding of the molecular mechanisms underlying tooth development and future development of hard-tissue regenerative therapies.
PubMed: 38942194
DOI: 10.1016/j.job.2024.06.010 -
Frontiers in Cell and Developmental... 2019Fibroblast activation protein-α (FAPα) is a membrane protein with dipeptidyl-peptidase and type I collagenase activity and is expressed during fetal growth. At the age...
Fibroblast activation protein-α (FAPα) is a membrane protein with dipeptidyl-peptidase and type I collagenase activity and is expressed during fetal growth. At the age of adolescence, FAPα expression is greatly reduced, only emerging in pathologies associated with extracellular matrix remodeling. We determined whether FAPα is expressed in human dental tissue involved in root maturation i.e., dental follicle and apical papilla and in dental pulp tissue. The dental follicle revealed a high concentration of FAPα and vimentin-positive cells within the stromal tissue. A similar observation was made in cell culture and FACS analysis confirmed these as dental follicle stem cells. Within the remnants of the Hertwigs' epithelial root sheath, we observed FAPα staining in the E-cadherin positive and vimentin-negative epithelial islands. FAPα- and vimentin-positive cells were encountered at the periphery of the islands suggesting an epithelial mesenchymal transition process. Analysis of the apical papilla revealed two novel histological regions; the periphery with dense and parallel aligned collagen type I defined as cortex fibrosa and the inner stromal tissue composed of less compacted collagen defined as medulla. FAPα expression was highly present within the medulla suggesting a role in extracellular matrix remodeling. Dental pulp tissue uncovered a heterogeneous FAPα staining but strong staining was noted within odontoblasts. studies confirmed the presence of FAPα expression in stem cells of the apical papilla and dental pulp. This study identified the expression of FAPα expression in dental stem cells which could open new perspectives in understanding dental root maturation and odontoblast function.
PubMed: 32039205
DOI: 10.3389/fcell.2019.00389 -
Stem Cell Research & Therapy Feb 2022Commitment of mouse dental papilla cells (mDPCs) to the odontoblast lineage is critical for dentin formation, and this biological process is regulated by a complex...
BACKGROUND
Commitment of mouse dental papilla cells (mDPCs) to the odontoblast lineage is critical for dentin formation, and this biological process is regulated by a complex transcription factor network. The transcription factor Mycn is a proto-oncogene that plays an important role in tumorigenesis and normal embryonic development. An early study revealed that Mycn is exclusively expressed in dental mesenchymal cells at E15.5, which implies a potential role of Mycn in dentinogenesis. However, the role of Mycn in dentin formation remains elusive. Thus, it is of considerable interest to elucidate the role of Mycn in dentin formation.
METHODS
Mycn; Osr2 (Mycn) and Mycn; K14 (Mycn) transgenic mice were generated, and micro-CT scans were performed to quantitatively analyse the volumetric differences in the molars and incisors of the mutants and their littermates. Mycn was also knocked down in vitro, and alkaline phosphatase (ALP) and alizarin red staining (ARS) were conducted. Cleavage under targets and tagmentation (CUT&Tag) analysis and dual luciferase assays were performed to identify direct downstream targets of Mycn. Immunofluorescence and immunochemistry staining and western blotting (WB) were performed to analyse the expression levels of potential targets. Quantitative PCR, WB, ALP and ARS were performed to test the rescue efficiency.
RESULTS
Mesenchymal ablation of Mycn (Mycn) led to defective dentin formation, while epithelial deletion (Mycn) had no obvious effects on tooth development. ALP and ARS staining revealed that the commitment capacity of mDPCs to the odontoblast lineage was compromised in Mycn mice. CUT&Tag analysis identified Klf4 as a potential direct target of Mycn, and a dual luciferase reporter assay verified that Mycn could bind to the promotor region of Klf4 and directly activate its transcription. Reciprocally, forced expression of Klf4 partially recovered the odontoblastic differentiation capacity of mDPCs with Mycn knockdown.
CONCLUSIONS
Our results elucidated that mesenchymal Mycn modulates the odontoblastic commitment of dental papilla cells by directly regulating Klf4. Our study illustrated the role of Mycn in dentin development and furthers our general comprehension of the transcription factor networks involved in the dentinogenesis process. Thus, these results may provide new insight into dentin hypoplasia and bioengineered dentin regeneration.
Topics: Animals; Cell Differentiation; Kruppel-Like Factor 4; Mice; N-Myc Proto-Oncogene Protein; Odontoblasts; Odontogenesis; Transcription Factors
PubMed: 35193672
DOI: 10.1186/s13287-022-02749-8