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Anatomical Record (Hoboken, N.J. : 2007) May 2024PIEZO1 and PIEZO2 are essential components of mechanogated ion channels, which are required for mechanotransduction and biological processes associated with mechanical...
PIEZO1 and PIEZO2 are essential components of mechanogated ion channels, which are required for mechanotransduction and biological processes associated with mechanical stimuli. There is evidence for the presence of PIEZO1 and PIEZO2 in teeth and periodontal ligaments, especially in cell lines and mice, but human studies are almost nonexistent. Decalcified permanent human teeth and mouse molars were processed for immunohistochemical detection of PIEZO1 and PIEZO2. Confocal laser microscopy was used to examine the co-localization of PIEZO 1 and PIEZO2 with vimentin (a marker of differentiated odontoblasts) in human teeth. In the outer layer of the human dental pulp, abundant PIEZO1- and PIEZO2-positive cells were found that had no odontoblast morphology and were vimentin-negative. Based on their morphology, location, and the absence of vimentin positivity, they were identified as dental pulp stem cells or pre-odontoblasts. However, in mice, PIEZO1 and PIEZO2 were ubiquitously detected and colocalized in odontoblasts. Intense immunoreactivity of PIEZO1 and PIEZO2 has been observed in human and murine periodontal ligaments. Our findings suggest that PIEZO1 and PIEZO2 may be mechanosensors/mechanotransducers in murine odontoblasts, as well as in the transmission of forces by the periodontal ligament in humans and mice.
Topics: Humans; Mice; Animals; Periodontal Ligament; Vimentin; Mechanotransduction, Cellular; Dental Pulp; Ion Channels
PubMed: 37975162
DOI: 10.1002/ar.25351 -
Journal of Dental Research Apr 2021WW domain-containing E3 Ub-protein ligase 2 (WWP2) belongs to the homologous to E6AP C-terminus (HECT) E3 ligase family. It has been explored to regulate osteogenic...
WW domain-containing E3 Ub-protein ligase 2 (WWP2) belongs to the homologous to E6AP C-terminus (HECT) E3 ligase family. It has been explored to regulate osteogenic differentiation, chondrogenesis, and palatogenesis. Odontoblasts are terminally differentiated mesenchymal cells, which contribute to dentin formation in tooth development. However, it remained unknown whether WWP2 participated in odontoblast differentiation. In this study, WWP2 was found to be expressed in mouse dental papilla cells (mDPCs), odontoblasts, and odontoblastic-induced mDPCs by immunohistochemistry and Western blotting. Besides, WWP2 expression was decreased in the cytoplasm but increased in the nuclei of differentiation-induced mDPCs. When was knocked down, the elevated expression of odontoblast marker genes ( and ) in mDPCs induced by differentiation medium was suppressed. Meanwhile, a decrease of alkaline phosphatase (ALP) activity was observed by ALP staining, and reduced formation of mineralized matrix nodules was demonstrated by Alizarin Red S staining. Overexpression of WWP2 presented opposite results to knockdown experiments, suggesting that WWP2 promoted odontoblastic differentiation of mDPCs. Further investigation found that WWP2 was coexpressed and interacted with KLF5 in the nuclei, leading to ubiquitination of KLF5. The PPPSY (PY2) motif of KLF5 was essential for its physical binding with WWP2. Also, cysteine 838 (Cys838) of WWP2 was the active site for ubiquitination of KLF5, which did not lead to proteolysis of KLF5. Then, KLF5 was confirmed to be monoubiquitinated and transactivated by WWP2, which promoted the expression of KLF5 downstream genes and . Deletion of the PY2 motif of KLF5 or mutation of Cys838 of WWP2 reduced the upregulation of and . Besides, lysine (K) residues K31, K52, K83, and K265 of KLF5 were verified to be crucial to WWP2-mediated KLF5 transactivation. Taken together, WWP2 promoted odontoblastic differentiation by monoubiquitinating KLF5.
Topics: Animals; Cell Differentiation; Dental Pulp; Extracellular Matrix Proteins; Kruppel-Like Transcription Factors; Mesenchymal Stem Cells; Mice; Odontoblasts; Odontogenesis; Osteogenesis; Phosphoproteins; Ubiquitin-Protein Ligases
PubMed: 33164644
DOI: 10.1177/0022034520970866 -
Journal of Endodontics Apr 2023Prostaglandin E (PGE) exerts biological actions through its transport pathway involving intracellular synthesis, extracellular transport, and receptor binding. This...
INTRODUCTION
Prostaglandin E (PGE) exerts biological actions through its transport pathway involving intracellular synthesis, extracellular transport, and receptor binding. This study aimed to determine the localization of the components of the PGE-transporting pathway in human dental pulp and explore the relevance of PGE receptors (EP2/EP4) to angiogenesis and dentinogenesis.
METHODS
Protein localization of microsomal PGE (mPGES)synthase, PGE transporters (multidrug resistance-associated protein-4 [MRP4] and prostaglandin transporter [PGT]), and EP2/EP4 was analyzed using double immunofluorescence staining. Tooth slices from human third molars were cultured with or without butaprost (EP2 agonist) or rivenprost (EP4 agonist) for 1 week. Morphometric analysis of endothelial cell filopodia was performed to evaluate angiogenesis, and real-time polymerase chain reaction was performed to evaluate angiogenesis and odontoblast differentiation markers.
RESULTS
MRP4 and PGT were colocalized with mPGES and EP2/EP4 in odontoblasts and endothelial cells. Furthermore, MRP4 was colocalized with mPGES and EP4 in human leukocyte antigen-DR-expressing dendritic cells. In the tooth slice culture, EP2/EP4 agonists induced significant increases in the number and length of filopodia and mRNA expression of angiogenesis markers (vascular endothelial growth factor and fibroblast growth factor-2) and odontoblast differentiation markers (dentin sialophosphoprotein and collagen type 1).
CONCLUSIONS
PGE-producing enzyme (mPGES), transporters (MRP4 and PGT), and PGE-specific receptors (EP2/EP4) were immunolocalized in various cellular components of the human dental pulp. EP2/EP4 agonists promoted endothelial cell filopodia generation and upregulated angiogenesis- and odontoblast differentiation-related genes, suggesting that PGE binding to EP2/EP4 is associated with angiogenic and dentinogenic responses.
Topics: Humans; Receptors, Prostaglandin E, EP4 Subtype; Receptors, Prostaglandin E, EP2 Subtype; Dental Pulp; Vascular Endothelial Growth Factor A; Endothelial Cells; Dinoprostone; Multidrug Resistance-Associated Proteins; Cells, Cultured
PubMed: 36758673
DOI: 10.1016/j.joen.2023.01.009 -
Current Issues in Molecular Biology May 2021The role of inflammatory mediators in dental pulp is unique. The local environment of pulp responds to any changes in the physiology that are highly fundamental, like... (Review)
Review
The role of inflammatory mediators in dental pulp is unique. The local environment of pulp responds to any changes in the physiology that are highly fundamental, like odontoblast cell differentiation and other secretory activity. The aim of this review is to assess the role of cathelicidins based on their capacity to heal wounds, their immunomodulatory potential, and their ability to stimulate cytokine production and stimulate immune-inflammatory response in pulp and periapex. Accessible electronic databases were searched to find studies reporting the role of cathelicidins in pulpal inflammation and regeneration published between September 2010 and September 2020. The search was performed using the following databases: Medline, Scopus, Web of Science, SciELO and PubMed. The electronic search was performed using the combination of keywords "cathelicidins" and "dental pulp inflammation". On the basis of previous studies, it can be inferred that LL-37 plays an important role in odontoblastic cell differentiation and stimulation of antimicrobial peptides. Furthermore, based on these outcomes, it can be concluded that LL-37 plays an important role in reparative dentin formation and provides signaling for defense by activating the innate immune system.
Topics: Cathelicidins; Cell Differentiation; Dental Pulp; Humans; Immunomodulation; Inflammation; Odontoblasts; Wound Healing
PubMed: 34068275
DOI: 10.3390/cimb43010010 -
Journal of Dental Research Mar 2020Dentin is an important structural component of the tooth. Odontoblast differentiation is an essential biological process that guarantees normal dentin formation, which...
Dentin is an important structural component of the tooth. Odontoblast differentiation is an essential biological process that guarantees normal dentin formation, which is precisely regulated by various proteins. Murine double minute 2 (Mdm2) is an E3 ubiquitin ligase, and it plays a pivotal role in the differentiation of different cell types, such as osteoblasts and myoblasts. However, whether Mdm2 plays a role in odontoblast differentiation remains unknown. Here, we investigated the spatiotemporal expression of Mdm2 by immunostaining and found that Mdm2 was highly expressed in the odontoblasts and slightly in the dental papilla cells of mouse incisors and molars. Gene knockdown and overexpression experiments verified that Mdm2 promoted the odontoblast-like differentiation of mouse dental papilla cells (mDPCs). Intranuclear colocalization and physical interaction between Mdm2 and distal-less 3 (Dlx3), a transcription factor important for odontoblast differentiation, was found during the odontoblast-like differentiation of mDPCs by double immunofluorescence and immunoprecipitation. Mdm2 was proved to monoubiquitinate Dlx3, which enhanced the expression of Dlx3 target gene . In addition, p53, the canonical substrate of Mdm2, was validated to be also ubiquitinated but degraded by Mdm2 during the odontoblast-like differentiation of mDPCs. Gene knockdown experiments confirmed that p53 inhibited the odontoblast-like differentiation of mDPCs. and double knockdown partially rescued the reduced odontoblast-like differentiation by knockdown of alone. Taken together, our study revealed that Mdm2 promoted the odontoblast-like differentiation of mDPCs by ubiquitinating both Dlx3 and p53. On one hand, the monoubiquitination of Dlx3 by Mdm2 led to upregulation of , which is a marker of the odontoblast differentiation. On the other hand, ubiquitination of p53 by Mdm2 resulted in its degradation, which eliminated the inhibitory effect of p53 on the odontoblast-like differentiation of mDPCs.
Topics: Animals; Cell Differentiation; Homeodomain Proteins; Mice; Odontoblasts; Proto-Oncogene Proteins c-mdm2; Transcription Factors; Tumor Suppressor Protein p53
PubMed: 31847675
DOI: 10.1177/0022034519893672 -
Data in Brief Apr 2021Data in this article are associated with our research article "Effect of Myricetin on Odontoblast-like Cells and its Potential to Preserve Resin-Dentin Bonds." Both a...
Data in this article are associated with our research article "Effect of Myricetin on Odontoblast-like Cells and its Potential to Preserve Resin-Dentin Bonds." Both a poor infiltration of resin monomers into the demineralized dentin matrix and hydrolytic degradation of the adhesive could lead to the instability of the resin-dentin interface. The degradation of collagen is caused by matrix metalloproteinases (MMP) and cysteine cathepsins. These collagenolytic enzymes are contained in their latent form as pro-MMPs in the dentinal structure, and undergo activation during the adhesive process. Given that the integrity of the collagen matrix is essential for the preservation of the dentin bond strength in both the medium and long term, the inhibition of these proteases is necessary to improve the durability of adhesive restorations. Among the different strategies suggested to improve both the behavior of the substrate against enzymatic degradation and the biomechanical behavior of the adhesive interface, the use of protease inhibitors and collagen crosslinking agents has been recommended, such as polyphenols. Research has focused on flavonoids such as proanthocyanidins (PAC), a class of phenolic compounds found in a variety of plants such as blueberry and grape whose chemical structure favors their action as cross-linking agents. However, the focus has recently shifted towards myricetin (MYR) due to its chemical structure: a greater amount of hydroxyl groups at the substitution positions, which form bonds with the carbonyl groups of the side chains of collagen amino acids and generate interfiber bonds. Our previous study has shown the efficacy of MYR both as a cross-linking agent and as a MMP inhibitor without any immediate effects on microtensile bond strength (µTBS) and preserving it for six months after storage, and maintaining the odontoblastic phenotype without affecting cell viability. The objective of this article is to present a dataset on the effect of flavonoids PAC and MYR on the resin-dentin interface. Given that durability of the resindentin bond holds great importance for the clinical longevity of adhesive restorations, our data aims to show the effects of these flavonoids on resindentin µTBS after 18-month storage. Test groups for the µTBS assay were set as follows: G1 (negative control), conventional adhesion technique; G2 (vehicle control), 100% ethanol (EtOH) for 120 s; G3, 0.2% chlorhexidine (CHX) for 60 s; G4, 1% glutaraldehyde (GA) for 60 s; and G5, 600 µM myricetin (MYR) for 120 s. Datasets were exported to SPSS software, version 21.0 (SPSS, Chicago, IL, USA) for analysis using the Shapiro-Wilk, a two-way analysis of variance including factor interactions (treatment and storage time). Data are presented as mean ± standard deviation (SD). Differences with p-values < 0.05 were considered significant. Our data can be used as a basis for comparison among other natural and synthetic substances that could work as MMP inhibitors and crosslinking agents. These findings could be useful for designing an effective strategy towards the stabilization of the hybrid layer in a relevant clinical protocol.
PubMed: 33850990
DOI: 10.1016/j.dib.2021.106984 -
Journal of Biomaterials Applications Aug 2023Pulp-Dentin regeneration is a key aspect of maintain tooth vitality and enabling good oral-systemic health. This study aimed to investigate a nanofibrous scaffold loaded...
Pulp-Dentin regeneration is a key aspect of maintain tooth vitality and enabling good oral-systemic health. This study aimed to investigate a nanofibrous scaffold loaded with a small molecule i.e. Tideglusib to promote odontogenic differentiation. Tideglusib (GSK-3β inhibitor) interaction with GSK-3β was determined using molecular docking and stabilization of β-catenin was examined by confocal microscopy. 3D nanofibrous scaffolds were fabricated through electrospinning and their physicochemical characterizations were performed. Scaffolds were seeded with mesenchymal stem cells or pre-odontoblast cells to determine the cells proliferation and odontogenic differentiation. Our results showed that Tideglusib (TG) binds with GSK-3β at Cys199 residue. Stabilization and nuclear translocation of β-catenin was increased in the odontoblast cells treated with TG. SEM analysis revealed that nanofibers exhibited controlled architectural features that effectively mimicked the natural ECM. UV-Vis spectroscopy demonstrated that TG was incorporated successfully and released in a controlled manner. Both kinds of biomimetic nanofibrous matrices (PCLF-TG100, PCLF-TG1000) significantly stimulated cells proliferation. Furthermore, these scaffolds significantly induced dentinogenic markers (ALP, and DSPP) expression and biomineralization. In contrast to current pulp capping material driving dentin repair, the sophisticated, polymeric scaffold systems with soluble and insoluble spatiotemporal cues described here can direct stem cell differentiation and dentin regeneration. Hence, bioactive small molecule-incorporated nanofibrous scaffold suggests an innovative clinical tool for dentin tissue engineering.
Topics: Tissue Scaffolds; Nanofibers; beta Catenin; Glycogen Synthase Kinase 3 beta; Molecular Docking Simulation; Cells, Cultured; Cell Differentiation; Tissue Engineering; Dental Pulp
PubMed: 37485690
DOI: 10.1177/08853282231190470 -
Journal of Endodontics Apr 2023Pulp calcification (PC) often appears in strong association with nerve fiber bundles, which indicates the important role of dental nerves in the formation of PC....
INTRODUCTION
Pulp calcification (PC) often appears in strong association with nerve fiber bundles, which indicates the important role of dental nerves in the formation of PC. Additionally, given that sensory nerves and calcitonin gene-related peptide (CGRP) secreted from sensory nerve fibers are involved in physiological and pathological bone formation, we aimed to determine whether chronic irritation of sensory nerves can promote the occurrence of PC.
METHODS
A sensory nerve irritation rat model was established via ligation of the inferior alveolar nerve (IAN), and face grooming behavior was analyzed as a measure of pain sensation. Two months post-surgery, PC was determined by imaging and histologic analyses.
RESULTS
Rats in the IAN-chronic constriction injury (IAN-CCI) group showed spontaneous pain-associated behavior after the operations and pain tolerance on the 60 postoperative day. The imaging and histological analysis showed more calcified particles in the IAN-innervated first and second molars after day 60 of the dental sensory nerve irritation. These calcified masses had a dentin-like structure that contained sparse, irregularly oriented tubules. Compared to the control and sham groups, the odontoblasts located in the periphery of radicular pulp aligned along a thicker layer of predentin; which expressed more nestin with longer and stouter processes in the IAN-CCI group. Additionally, more CGRP-positive nerves were observed in the IAN-CCI group.
CONCLUSIONS
Irritation of sensory nerves promotes PC formation, and the increased density of CGRP-immunolabeled fibers probably contributes to this process. This highlights the significance of dental sensory nerves in the formation of PC.
Topics: Rats; Animals; Calcitonin Gene-Related Peptide; Dental Pulp; Molar; Odontoblasts; Pain
PubMed: 36758674
DOI: 10.1016/j.joen.2023.01.001 -
Metabolites Apr 2023The cellular metabolic processes ensure the physiological integrity of the dentine-pulp complex. Odontoblasts and odontoblast-like cells are responsible for the defence... (Review)
Review
The cellular metabolic processes ensure the physiological integrity of the dentine-pulp complex. Odontoblasts and odontoblast-like cells are responsible for the defence mechanisms in the form of tertiary dentine formation. In turn, the main defence reaction of the pulp is the development of inflammation, during which the metabolic and signalling pathways of the cells are significantly altered. The selected dental procedures, such as orthodontic treatment, resin infiltration, resin restorations or dental bleaching, can impact the cellular metabolism in the dental pulp. Among systemic metabolic diseases, diabetes mellitus causes the most consequences for the cellular metabolism of the dentine-pulp complex. Similarly, ageing processes present a proven effect on the metabolic functioning of the odontoblasts and the pulp cells. In the literature, several potential metabolic mediators demonstrating anti-inflammatory properties on inflamed dental pulp are mentioned. Moreover, the pulp stem cells exhibit the regenerative potential essential for maintaining the function of the dentine-pulp complex.
PubMed: 37110177
DOI: 10.3390/metabo13040520 -
Current Medicinal Chemistry Mar 2022Transient Receptor Potential (TRP) Channels constitute a large family of non-selective permeable ion channels involved in the perception of environmental stimuli with a... (Review)
Review
BACKGROUND
Transient Receptor Potential (TRP) Channels constitute a large family of non-selective permeable ion channels involved in the perception of environmental stimuli with a central and continuously expanding role in oral tissue homeostasis. Recent studies indicate the regulatory role of TRPs in pulp physiology, oral mucosa sensation, dental pain nociception and salivary gland secretion. This review provides an update on the diverse functions of TRP channels in the physiology of the oral cavity, with emphasis on their cellular location, the underlying molecular mechanisms and clinical significance.
METHODS
A structured search of bibliographic databases (PubMed and MEDLINE) was performed for peer-reviewed studies on the function of TRP channels on oral cavity physiology in the last ten years. A qualitative content analysis was performed of screened papers and a critical discussion on the main findings is provided.
RESULTS
TRPs expression has been detected in major cell types of the oral cavity, including odontoblasts, periodontal ligament, oral epithelial, salivary gland cells, and chondrocytes of temporomandibular joints, where they mediate signal perception and transduction of mechanical, thermal, and osmotic stimuli. They contribute to pulp physiology through dentin formation, mineralization, and periodontal ligament formation, along with alveolar bone remodeling in the dental pulp and periodontal ligament cells. TRPs are also involved in oral mucosa sensation, dental pain nociception, saliva secretion, swallowing reflex and temporomandibular joints' development.
CONCLUSION
Various TRP channels regulate oral cavity homeostasis, playing an important role in the transduction of external stimuli to intracellular signals in a cell typespecific manner and presenting promising drug targets for the development of pharmacological strategies to manage oral diseases.
Topics: Humans; Nociception; Odontoblasts; Transient Receptor Potential Channels
PubMed: 34365940
DOI: 10.2174/0929867328666210806113132