ABSTRACT
Epigenetic modulation, including histone modification, alters gene expression and controls cell fate. Histone deacetylases (HDACs) are identified as important regulators of dental pulp cell (DPC) mineralisation processes. Currently, there is a paucity of information regarding the nature of histone modification and HDAC expression in the dentine-pulp complex during dentinogenesis. The aim of this study was to investigate post-translational histone modulation and HDAC expression during DPC mineralisation and the expression of Class I/II HDACs during tooth development and in adult teeth. HDAC expression (isoforms -1 to -6) was analysed in mineralising primary rat DPCs using qRT-PCR and Western blot with mass spectrometry being used to analyse post-translational histone modifications. Maxillary molar teeth from postnatal and adult rats were analysed using immunohistochemical (IHC) staining for HDACs (1-6). HDAC-1, -2, and -4 protein expression increased until days 7 and 11, but decreased at days 14 and 21, while other HDAC expression increased continuously for 21 days. The Class II mineralisation-associated HDAC-4 was strongly expressed in postnatal sample odontoblasts and DPCs, but weakly in adult teeth, while other Class II HDACs (-5, -6) were relatively strongly expressed in postnatal DPCs and adult odontoblasts. Among Class I HDACs, HDAC-1 showed high expression in postnatal teeth, notably in ameloblasts and odontoblasts. HDAC-2 and -3 had extremely low expression in the rat dentine-pulp complex. Significant increases in acetylation were noted during DPC mineralisation processes, while trimethylation H3K9 and H3K27 marks decreased, and the HDAC-inhibitor suberoylanilide hydroxamic acid (SAHA) enhanced H3K27me3. These results highlight a dynamic alteration in histone acetylation during mineralisation and indicate the relevance of Class II HDAC expression in tooth development and regenerative processes.
Subject(s)
Dental Pulp , Dentin , Dentinogenesis , Histone Deacetylases , Animals , Acetylation , Rats , Histone Deacetylases/metabolism , Histone Deacetylases/genetics , Dentin/metabolism , Dental Pulp/metabolism , Dental Pulp/cytology , Dental Pulp/growth & development , Protein Processing, Post-Translational , Histones/metabolism , Molar/metabolism , Molar/growth & development , Odontoblasts/metabolism , MaleABSTRACT
Pulpotomy is an effective treatment for retaining vital pulp after pulp exposure caused by caries removal and/or trauma. The expression of alpha smooth muscle actin (α-SMA) is increased during the wound-healing process, and α-SMA-positive fibroblasts accelerate tissue repair. However, it remains largely unknown whether α-SMA-positive fibroblasts influence pulpal repair. In this study, we established an experimental rat pulpotomy model and found that the expression of α-SMA was increased in dental pulp after pulpotomy relative to that in normal dental pulp. In vitro results showed that the expression of α-SMA was increased during the induction of odontogenic differentiation in dental pulp stem cells (DPSCs) compared with untreated DPSCs. Moreover, α-SMA overexpression promoted the odontogenic differentiation of DPSCs via increasing mitochondrial function. Mechanistically, α-SMA overexpression activated the mammalian target of rapamycin (mTOR) signaling pathway. Inhibition of the mTOR signaling pathway by rapamycin decreased the mitochondrial function in α-SMA-overexpressing DPSCs and suppressed the odontogenic differentiation of DPSCs. Furthermore, we found that α-SMA overexpression increased the secretion of transforming growth factor beta-1 (TGF-ß1). In sum, our present study demonstrates a novel mechanism by which α-SMA promotes odontogenic differentiation of DPSCs by increasing mitochondrial respiratory activity via the mTOR signaling pathway.
Subject(s)
Actins , Dental Pulp , Odontogenesis , Animals , Rats , Actins/metabolism , Actins/pharmacology , Cell Differentiation , Cell Proliferation , Cells, Cultured , Dental Pulp/cytology , Dental Pulp/growth & development , Stem Cells , TOR Serine-Threonine Kinases/metabolism , TOR Serine-Threonine Kinases/pharmacology , PulpotomyABSTRACT
Tooth regeneration is an important issue. The purpose of this study was to explore the feasibility of using adult dental pulp stem cells on polylactic acid scaffolds for tooth regeneration. Three teeth were extracted from each side of the lower jaws of two adult dogs. In the experimental group, dental pulp stem cells were isolated and seeded in the 3D-printed hydroxyapatite/polylactic acid (HA/PLA) scaffolds for transplantation into left lower jaw of each dog. The right-side jaw of each dog was transplanted with cell-free scaffolds as the control group. Polychrome sequentially labeling was performed for observation of mineralization. Dental cone beam computed tomography (CBCT) irradiation was used for assessment. Nine months after surgery, dogs were euthanized, and the lower jaws of dogs were sectioned and fixed for histological observation with hematoxylin and eosin staining. The results showed that the degree of mineralization in the experimental group with cells seeded in the scaffolds was significantly higher than that of the control group transplanted with cell-free scaffolds. However, the HA/PLA scaffolds were not completely absorbed in both groups. It is concluded that dental pulp stem cells are important for the mineralization of tooth regeneration. A more rapid absorbable material was required for scaffold design for tooth regeneration.
Subject(s)
Dental Pulp/growth & development , Durapatite/chemistry , Regeneration/drug effects , Tooth/growth & development , Animals , Dogs , Durapatite/pharmacology , Polyesters/chemistry , Printing, Three-Dimensional , Regeneration/genetics , Stem Cells/cytology , Tissue ScaffoldsABSTRACT
Elucidating the mechanisms underlying human pain sensation requires the establishment of an in vitro model of pain reception comprising human cells expressing pain-sensing receptors and function properly as neurons. Human dental pulp stem cells (hDPSCs) are mesenchymal stem cells and a promising candidate for producing human neuronal cells, however, the functional properties of differentiated hDPSCs have not yet been fully characterized. In this study, we demonstrated neuronal differentiation of hDPSCs via both their expression of neuronal marker proteins and their neuronal function examined using Ca2+ imaging. Moreover, to confirm the ability of nociception, Ca2+ responses in differentiated hDPSCs were compared to those of rat dorsal root ganglion (DRG) neurons. Those cells showed similar responses to glutamate, ATP and agonists of transient receptor potential (TRP) channels. Since TRP channels are implicated in nociception, differentiated hDPSCs provide a useful in vitro model of human peripheral neuron response to stimuli interpreted as pain.
Subject(s)
Dental Pulp/cytology , Mesenchymal Stem Cells/physiology , Neurons/cytology , Animals , Calcium/metabolism , Cell Differentiation , Cells, Cultured , Dental Pulp/growth & development , Dental Pulp/physiology , Fluorescent Antibody Technique , Hippocampus/cytology , Humans , Microscopy, Confocal , Neurons/physiology , Nociception/physiology , Rats , Rats, WistarABSTRACT
Age estimation of living individuals has become a crucial part of the forensic practice, especially due to the global increase in cross-border migration. The low rate of birth registration in many countries, hence of identification documents of migrants, especially in Africa and Asia, highlights the importance of reliable methods for age estimation of living individuals. Despite the fact that a number of skeletal and dental methods for age estimation have been developed, their main limitation is that they are based on specific reference samples and there is still no consensus among researchers on whether these methods can be applied to all populations. Though this issue remains still unsolved, population information at a glance could be useful for forensic practitioners dealing with such issues. This study aims at presenting a scoping review and mapping of the current situation concerning population data for skeletal (hand-wrist and clavicle) and dental methods (teeth eruption and third molar formation) for age estimation in the living. Two hundred studies on the rate of skeletal maturation and four hundred thirty-nine on the rate of dental maturation were found, covering the period from 1952 and 2020 for a total of ninety-eight countries. For most of the western and central African countries there are currently no data on the rate of skeletal and dental maturation. The same applies to the countries of the Middle East, as well as the eastern European countries, especially as regard the skeletal development.
Subject(s)
Age Determination by Skeleton/methods , Age Determination by Teeth/methods , Clavicle/growth & development , Dental Pulp/growth & development , Hand Bones/growth & development , Humans , Molar, Third/growth & development , Osteogenesis , Tooth Calcification , Tooth EruptionABSTRACT
Hyaluronic acid (HA) and dental pulp stem cells (DPSCs) are attractive research topics, and their combined use in the field of tissue engineering seems to be very promising. HA is a natural extracellular biopolymer found in various tissues, including dental pulp, and due to its biocompatibility and biodegradability, it is also a suitable scaffold material. However, low molecular weight (LMW) fragments, produced by enzymatic cleavage of HA, have different bioactive properties to high molecular weight (HMW) HA. Thus, the impact of HA must be assessed separately for each molecular weight fraction. In this study, we present the effect of three LMW-HA fragments (800, 1600, and 15,000 Da) on DPSCs in vitro. Discrete biological parameters such as DPSC viability, morphology, and cell surface marker expression were determined. Following treatment with LMW-HA, DPSCs initially presented with an acute reduction in proliferation (p < 0.0016) and soon recovered in subsequent passages. They displayed significant size reduction (p = 0.0078, p = 0.0019, p = 0.0098) while maintaining high expression of DPSC markers (CD29, CD44, CD73, CD90). However, in contrast to controls, a significant phenotypic shift (p < 0.05; CD29, CD34, CD90, CD106, CD117, CD146, CD166) of surface markers was observed. These findings provide a basis for further detailed investigations and present a strong argument for the importance of HA scaffold degradation kinetics analysis.
Subject(s)
Cell Proliferation/drug effects , Dental Pulp/growth & development , Hyaluronic Acid/pharmacology , Stem Cells/cytology , Cell Differentiation/drug effects , Cells, Cultured , Dental Pulp/drug effects , Humans , Molecular Weight , Stem Cells/drug effects , Tissue EngineeringABSTRACT
Dental pulp is exposed to resin monomers leaching from capping materials. Toxic doses of the monomer, triethyleneglycol dimethacrylate (TEGDMA), impact cell growth, enhance inflammatory and oxidative stress responses, and lead to tissue necrosis. A therapeutic agent is required to rescue growth-arrested tissues by continuing their development and modulating the exacerbated responses. The functionality of N-Acetyl Cysteine (NAC) as a treatment was assessed by employing a 3D dental pulp microtissue platform. Immortalized and primary microtissues developed and matured in the extracellular matrix (ECM). TEGDMA was introduced at various concentrations. NAC was administered simultaneously with TEGDMA, before or after monomer addition during the development and after the maturation stages of the microtissue. Spatial growth was validated by confocal microscopy and image processing. Levels of inflammatory (COX2, NLRP3, IL-8) and oxidative stress (GSH, Nrf2) markers were quantified by immunoassays. NAC treatments, in parallel with TEGDMA challenge or post-challenge, resumed the growth of the underdeveloped microtissues and protected mature microtissues from deterioration. Growth recovery correlated with the alleviation of both responses by decreasing significantly the intracellular and extracellular levels of the markers. Our 3D/ECM-based dental pulp platform is an efficient tool for drug rescue screening. NAC supports compromised microtissues development, and immunomodulates and maintains the oxidative balance.
Subject(s)
Acetylcysteine/pharmacology , Dental Pulp/drug effects , Extracellular Matrix/drug effects , Inflammation/drug therapy , Cell Proliferation/drug effects , Composite Resins/pharmacology , Dental Pulp/growth & development , Dental Pulp/metabolism , Drug Evaluation, Preclinical , Extracellular Matrix/genetics , Gene Expression Regulation, Developmental/drug effects , Humans , Inflammation/chemically induced , Inflammation/genetics , Inflammation/pathology , Interleukin-8/genetics , NF-E2-Related Factor 2/genetics , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , Oxidative Stress/drug effects , Polyethylene Glycols/pharmacology , Polymethacrylic Acids/pharmacologyABSTRACT
Odontoblasts and pulp stroma cells are embedded within supramolecular networks of extracellular matrix (ECM). Fibrillin microfibrils and associated proteins are crucial constituents of these networks, serving as contextual scaffolds to regulate tissue development and homeostasis by providing both structural and mechanical properties and sequestering growth factors of the TGF-ß superfamily. EMILIN-1, -2, and -3 are microfibril-associated glycoproteins known to modulate cell behaviour, growth factor activity, and ECM assembly. So far their expression in the various cells of the dentin-pulp complex during development, in the adult stage, and during inflammation has not been investigated. Confocal immunofluorescence microscopy and western blot analysis of developing and adult mouse molars and incisors revealed an abundant presence of EMILINs in the entire dental papilla, at early developmental stages. Later in development the signal intensity for EMILIN-3 decreases, while EMILIN-1 and -2 staining appears to increase in the pre-dentin and in the ECM surrounding odontoblasts. Our data also demonstrate new specific interactions of EMILINs with fibulins in the dentin enamel junction. Interestingly, in dentin caries lesions the signal for EMILIN-3 was significantly increased in inflamed odontoblasts. Overall our findings point for the first time to a role of EMILINs in dentinogenesis, pulp biology, and inflammation.
Subject(s)
Antigens, Surface/metabolism , Dental Pulp/metabolism , Dentin/metabolism , Membrane Glycoproteins/metabolism , Molar/growth & development , Adolescent , Adult , Animals , Animals, Newborn , Dental Caries/metabolism , Dental Pulp/growth & development , Glycoproteins/metabolism , Humans , Incisor/metabolism , Mice, Inbred C57BL , Molar/embryology , Molar/metabolism , Young AdultABSTRACT
The age threshold of 14 years has become vital in proving legal violations involving children, particularly in cases of suspected child labour, child pornography and the minimum age of criminal responsibility. In recent years, there has been great interest in the evaluation of age in children and sub-adults using analysis of regressive changes in teeth, with a particular focus on age thresholds that are of medico-legal importance. This research aimed to compare the sensitivity and specificity of stages of root pulp visibility by Olze et al. in a sample of South Indian children aged between 12 and 16 years, with an age threshold of 14 years, using receiver operating characteristic curves and the area under the curve (AUC). Among the four stages of root pulp visibility, stage 2 showed the highest AUC in both female and male cohorts. For stage 2, lower sensitivity and higher specificity values were recorded, indicating the possibility of type II errors (i.e. false negatives). In both sexes, stage 2 had a higher AUC (i.e. 0.696 in females and 0.706 in males, respectively). Based on our findings, it can be concluded that this staging method in lower first molars is of limited value in indicating the legal age threshold of 14 years. Future research should validate the proposed approach in a larger sample and consider how to improve predictions in this area.
Subject(s)
Age Determination by Teeth/methods , Dental Pulp/growth & development , Dentin, Secondary/physiology , Molar/anatomy & histology , Molar/diagnostic imaging , Tooth Root/growth & development , Adolescent , Child , Female , Humans , India , Male , Mandible/anatomy & histology , Radiography, Panoramic , Sensitivity and SpecificityABSTRACT
Forensic odontologists often confront with conceptually simple medicolegal question of whether an individual is a juvenile or an adult. The demand for additional research into extending dental age estimation methods in late adolescence is never the less, especially in those who have passed 15 years and are suspected older than 18 years. The present research investigated the regressive dental characteristic, i.e. radiographic visibility of the root pulp in mandibular second molars for the purpose of age assessment, especially for determining the age over 18 years. Nine hundred thirty-six orthopantomograms comprised of 436 males and 500 females aged from 14 to 22 years were examined. Descriptive statistics were performed for each stage in both sexes. The correlation statistics revealed that there is a strong, positive correlation between the chronological age and root pulp visibility stages. The analysis revealed that stage 0 occurred first at the age of 14 years in both sexes. Stage 1 first occurred at the age of 14.26 years in females and 14.74 years in males. Stage 2 was first achieved at the age of 18.6 and 15.2 years in males and females, respectively. The suitability of the studied characteristics in mandibular second molars for age estimation could be confirmed but of limited value. The presence of this stage 2 root pulp visibility in male subjects represents a potential criterion for indicating the age over 18 years. In future studies, the pattern of secondary dentin formation in other tooth types should be investigated.
Subject(s)
Age Determination by Teeth/methods , Dental Pulp/diagnostic imaging , Dental Pulp/growth & development , Molar/diagnostic imaging , Molar/growth & development , Tooth Root/diagnostic imaging , Tooth Root/growth & development , Adolescent , Female , Forensic Dentistry/methods , Humans , Male , Mandible , Radiography, Panoramic , Young AdultABSTRACT
BACKGROUND: The innate immune activation which promotes inflammation responses in the dental pulp tissue leads to the progression of dentin caries. Accordingly, toll-like receptors (TLRs) are key molecules of the innate immune system that identify pathogen-associated molecular patterns (PAMPs) on microorganisms and may have a critical role in a dental injury. Therefore, this study aimed to investigate the expression of TLR2, TLR3, and TLR4 in the human dental pulp of opened and closed apex teeth. METHODS: Human dental pulps were derived from the healthy opened and closed apex premolar, in which extraction was indicated for orthodontic reasons. The extraction of RNA was performed and the gene expression determined by real-time polymerase chain reaction (RT-PCR). The result from real-time PCR was confirmed using western blot analysis. RESULTS: Real-time PCR data analysis showed that the expression TLR2 and TLR4 were significantly increased in closed apex premolar teeth compared to open apex teeth, whereas TLR3 expression was not significantly different in these two groups (p < .05). CONCLUSION: The results of the present study suggested increased expression of TLR2 and TLR4 by the maturation of the apex, which may be due to the presence of microorganisms in the normal or destructed dental pulp tissue. Thus, identifying the expression of TLRs molecules in dental pulp tissue helps to develop a deeper knowledge of the immune responses in the oral cavity.
Subject(s)
Bicuspid/metabolism , Toll-Like Receptors/genetics , Tooth Apex/metabolism , Bicuspid/growth & development , Dental Pulp/growth & development , Dental Pulp/metabolism , Humans , Toll-Like Receptors/metabolism , Tooth Apex/growth & developmentABSTRACT
Sonic hedgehog (Shh) is a secreted protein with important roles in mammalian embryogenesis. During tooth development, Shh is primarily expressed in the dental epithelium, from initiation to the root formation stages. A number of studies have analyzed the function of Shh signaling at different stages of tooth development and have revealed that Shh signaling regulates the formation of various tooth components, including enamel, dentin, cementum, and other soft tissues. In addition, dental mesenchymal cells positive for Gli1, a downstream transcription factor of Shh signaling, have been found to have stem cell properties, including multipotency and the ability to self-renew. Indeed, Gli1-positive cells in mature teeth appear to contribute to the regeneration of dental pulp and periodontal tissues. In this review, we provide an overview of recent advances related to the role of Shh signaling in tooth development, as well as the contribution of this pathway to tooth homeostasis and regeneration.
Subject(s)
Hedgehog Proteins/metabolism , Odontogenesis/physiology , Signal Transduction/physiology , Tooth/growth & development , Animals , Dental Enamel/cytology , Dental Enamel/growth & development , Dental Pulp/growth & development , Epithelium/metabolism , Epithelium/pathology , Homeostasis , Humans , Mesenchymal Stem Cells , Tooth/cytology , Tooth Root/cytology , Tooth Root/growth & development , Zinc Finger Protein GLI1/metabolismABSTRACT
Age estimation has a great importance due to legal requirements. The aim of our study was to determine the applicability of two different methods in age estimation; one of them based on the calculation of the visibility of the root pulp of mandibular third molar teeth and the other based on the calculation of the tooth coronal index (TCI) in the mandibular first and second molar teeth in the 9059 digital orthopantomogram of people aged between 15 and 40 in Bursa. In the first method in which the visibility of the fully mineralized root pulp of the mandibular third molar teeth was evaluated in 4 stages; the stages 0, 1, 2, and 3 were observed regardless of sex at the earliest 17.2; 19.1; 20.1, and 25.1 years, respectively. In the second method in which TCI was regressed on chronological age using measuring crown and coronal pulp cavity heights, the most accurate age estimation model based on simple linear regression for all cases without any sex difference was found to be with right first molar tooth (SEE ±7304 years) and the most accurate age model based on multiple regression model was found to be with bilateral first molar teeth (SEE ±7413 years). In conclusion, we believe that the root pulp visibility of the third molar teeth can be applied safely for stages 1, 2, and 3 at 16, 17, and 21 years of ages, respectively. A correlation was also found between TCI and age, and our findings have shown that both methods are available for forensic purposes.
Subject(s)
Age Determination by Teeth/methods , Dental Pulp/diagnostic imaging , Molar/diagnostic imaging , Molar/growth & development , Tooth Crown/diagnostic imaging , Tooth Root/diagnostic imaging , Adolescent , Adult , Dental Pulp/growth & development , Female , Forensic Dentistry , Forensic Medicine , Humans , Male , Mandible/diagnostic imaging , Radiography, Dental, Digital , Radiography, Panoramic , Regression Analysis , Sex Characteristics , Tooth Crown/anatomy & histology , Tooth Root/growth & development , Young AdultABSTRACT
INTRODUCTION: The determination of age of majority (the 18-year-old threshold) using the popular Demirjian tooth staging method is unreliable, so other maturity markers are required. This study examines whether the Root Pulp Visibility (RPV) of the mandibular third molar is a useful indicator of age. METHODS: One thousand six dental panoramic radiographs (DPTs) were examined and the left mandibular third molar assessed according to the RPV stages using the method of Olze et al. (Int J Legal Med 124:183-186, 2010) as modified by Lucas et al. (Forensic Sci Int 270:98-102, 2017). Six hundred sixty-two DPTs, 288 males and 374 females, satisfied the inclusion criteria. RESULTS: Individuals who had reached stages RPV-A and RPV-B were represented in a wide range of ages spanning the 18-year-old threshold. Individuals who had reached stages RPV-C and RPV-D were all above the 18-year-old threshold. It was not possible to analyse a large number of DPTs for various reasons. DISCUSSION: Individuals whose mandibular third molar exhibited stages RPV-C and RPV-D may be deemed to have reached the age of majority. These results are consistent with other published studies. Variation in the rate of development of the third molar limits the applicability of this method. CONCLUSIONS: RPV staging is an accurate method of determining the 18-year-old threshold. Substitute methods are required in a large number of cases.
Subject(s)
Age Determination by Teeth/methods , Dental Pulp/diagnostic imaging , Dental Pulp/growth & development , Molar, Third/diagnostic imaging , Molar, Third/growth & development , Tooth Root/diagnostic imaging , Tooth Root/growth & development , Adolescent , Adult , Data Analysis , Data Collection , Female , Forensic Dentistry/methods , Humans , Male , Malta , Mandible/diagnostic imaging , Radiography, Panoramic , Retrospective Studies , Young AdultABSTRACT
Dentin-pulp regeneration requires dental pulp stem cells (DPSCs), but the role of long noncoding RNAs (lncRNAs) during this process remains unclear. Here, we cultured human DPSCs in osteogenic/odontogenic medium for 14 days and analyzed cells via RNA-sequencing. The data were validated by quantitative reverse transcription-polymerase chain reaction and lncRNA-microRNA (miRNA)-messenger RNA (mRNA) networks were constructed to reveal the potential competing endogenous RNA regulatory role of lncRNAs. Kyoto Encyclopedia of Genes and Genomes and Gene Ontology enrichment analysis were performed. One lncRNA, SNHG7, was identified and validated by genetic shRNA silencing. A total of 89 lncRNAs, 1,636 mRNAs, and 113 miRNAs were differentially expressed after differentiation. Bioinformatics identified an array of affected signaling pathways including phosphoinositide-3-kinase-protein kinase B, transforming growth factor-ß, and Wnt. mRNAs were enriched in cell migration, cell differentiation, stem cell development, ossification, and skeletal development. One lncRNA, SNHG7, was indentified to inhibit the odonto/osteogenic differentiation of DPSCs when silenced. In summary, we reveal several lncRNAs that significantly change during DPSC differentiation, including SNHG7. This reveals new targets for dentin-pulp complex regeneration and tissue engineering.
Subject(s)
Dental Pulp/growth & development , MicroRNAs/genetics , Osteogenesis/genetics , RNA, Long Noncoding/genetics , Cell Differentiation/genetics , Computational Biology , Dental Pulp/metabolism , Gene Expression Regulation, Developmental/genetics , Humans , RNA, Messenger/genetics , Stem Cells/metabolismABSTRACT
Gestational diabetes mellitus (GDM) is an important factor involved in the pathogenesis of organ development in the offspring. Here, we analyzed the effects of GDM on odontoblastic differentiation of dental papilla cells (DPCs) and dentin formation in offspring and investigated their underlying mechanisms. A GDM rat model was induced by intraperitoneal injection of streptozotocin and offspring were collected. The results showed that GDM significantly affected odontoblast differentiation and dentin formation in offspring tooth. GDM activated the toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-ĸB) signaling pathway and inhibited SMAD1/5/9 signaling to modulate the odontoblastic differentiation of DPCs in offspring. Inhibition of TLR4 signaling by treated with TAK-242 significantly reverses the suppression of odonto-differentiation of DPCs in diabetic offspring. Taken together, these data indicate GDM activated the offspring DPCs TLR4/NF-ĸB signaling, which suppressed the SMAD1/5/9 phosphorylation and then inhibited odontoblasts differentiation and dentin formation.
Subject(s)
Cell Differentiation/genetics , Dental Papilla/growth & development , Diabetes, Gestational/genetics , Toll-Like Receptor 4/genetics , Animals , Calcification, Physiologic/genetics , Cell Proliferation/drug effects , Dental Papilla/metabolism , Dental Pulp/growth & development , Dental Pulp/pathology , Diabetes, Gestational/pathology , Female , Humans , NF-kappa B/genetics , Odontoblasts/metabolism , Phosphorylation/genetics , Pregnancy , Rats , Signal Transduction/genetics , Smad1 Protein , Sulfonamides/pharmacologyABSTRACT
Pentraxin-3 (PTX3) is recognized as a modulator of inflammation and a mediator of tissue repair. In this study, we characterized the role of PTX3 on some biological functions of human dental pulp stem cells (HDPSCs). The expression level of PTX3 significantly increased during osteogenic/odontogenic differentiation of HDPSCs, whereas the knockdown of PTX3 decreased this differentiation. Silencing of PTX3 in HDPSCs inhibited their migration and C-X-C chemokine receptor type 4 (CXCR4) expression. Our present study indicates that PTX3 is involved in osteogenic/odontogenic differentiation and migration of HDPSCs, and may contribute to the therapeutic potential of HDPSCs for regeneration and repair.
Subject(s)
C-Reactive Protein/metabolism , Cell Differentiation , Cell Movement , Odontogenesis/physiology , Osteogenesis/physiology , Serum Amyloid P-Component/metabolism , C-Reactive Protein/genetics , Dental Pulp/growth & development , Dental Pulp/physiology , Gene Knockdown Techniques , Humans , Receptors, CXCR4/metabolism , Serum Amyloid P-Component/genetics , Stem Cells/physiologyABSTRACT
Secondary dentine deposition is responsible for the decrease in the volume of the pulp cavity with age. Therefore, the volume of the pulp cavity can be considered as a predictor for estimating age. The aims of this study were to investigate the relationship strength between canine pulp volumes and chronological age from homogenous (approximately equal numbers of individuals in each age range) age distribution and to assess the effect of sex as predictor in age estimation. This study was performed on 719 subjects of Pakistani origin. Cone beam computed tomography images of 521 left maxillary and 681 left mandibular canines were collected from 368 females and 349 males aged from 15 to 65 years. Planmeca Romexis® software was used to trace the outline of the pulp cavity and to calculate pulp volumes. Regression analysis was performed to assess the correlation between pulp volumes considering with and without sex as a predictor with chronological age. The obtained results showed that mandibular canine pulp volume and sex have the highest predictive power (R2 = 0.33). The relationship between mandibular canine pulp volume and sex with chronological age demonstrates an odd S-shaped non-linear relationship. A statistically significant difference in volumes of pulp was found (p = 0.000) between males and females. The conclusion was that predictions using the pulp volume of the mandibular canine and sex produced the best estimates of chronological age.
Subject(s)
Age Determination by Teeth/methods , Cone-Beam Computed Tomography , Cuspid/diagnostic imaging , Dental Pulp/diagnostic imaging , Dental Pulp/growth & development , Adolescent , Adult , Aged , Female , Forensic Dentistry , Humans , Image Processing, Computer-Assisted , Male , Middle Aged , Pakistan , Young AdultABSTRACT
The present study aims to evaluate the relation between chronological age and the ratio of pulp volume (PV) to enamel volume (EV) of impacted mandibular third molars (IMTMs) by using cone-beam computed tomography (CBCT) images and an improved 3D image segmentation technique. A sample of CBCT images of IMTM was collected from 414 northern Chinese subjects (214 male and 200 female clinical patients) ranging in age from 20 to 65 years. The GrowCut effect image segmentation (GCEIS) module algorithm was used to calculate the PV and EV from CBCT images. The total sample was divided into a training group and validation group in a ratio of 7 to 3. The PV/EV ratio (PEr) in the training sample was used to develop a mathematical formula for age estimation as follows: age = - 5.817-21.726 × Ln PEr (p < 0.0001) (Ln, natural logarithm). The mean absolute error (MAE) and root mean square error (RMSE) were used to determine the precision and accuracy of the mathematical formula in the validation group and all samples. The MAEs in the male, female, and pooled gender samples were 9.223, 7.722, and 8.41, respectively, and the RMSEs in the male, female, and pooled gender samples were 10.76, 9.58, and 9.986, respectively. The precise and accurate results indicate that the PEr of IMTM in CBCT images is a potential index for dental age estimation and is possible to be used in forensic medicine.
Subject(s)
Age Determination by Teeth/methods , Dental Enamel/diagnostic imaging , Dental Pulp/diagnostic imaging , Molar, Third/diagnostic imaging , Tooth, Impacted/diagnostic imaging , Adult , Aged , China , Cone-Beam Computed Tomography , Dental Enamel/growth & development , Dental Pulp/growth & development , Female , Forensic Dentistry/methods , Humans , Male , Mandible , Middle Aged , Young AdultABSTRACT
BACKGROUND: Exosomes derived from dental pulp stem cells (DPSCs) can be used as biomimetic tools to induce odontogenic differentiation of stem cells, but the regulatory mechanisms and functions of exosome-encapsulated microRNAs are still unknown. The present study aimed to clarify the role of microRNAs contained in the exosomes derived from human DPSCs and their potential signaling cascade in odontogenic differentiation. METHODS: Exosomes were isolated from human DPSCs cultured undergrowth and odontogenic differentiation conditions, named UN-Exo and OD-Exo, respectively. The microRNA sequencing was performed to explore the microRNA profile contained in UN-Exo and OD-Exo. Pathway analysis was taken to detect enriched pathways associated with the predicted target genes of microRNAs. The regulatory roles of a highly expressed microRNA in OD-Exo were investigated through its inhibition or overexpression (miRNA inhibitors and miRNA mimics). Automated western blot was used to identify the function of exosomal microRNA and the roles of TGFß1/smads pathway in odontogenic differentiation of DPSCs. A luciferase reporter gene assay was used to verify the direct target gene of exosomal miR-27a-5p. RESULTS: Endocytosis of OD-Exo triggered odontogenic differentiation of DPSCs by upregulating DSP, DMP-1, ALP, and RUNX2 proteins. MicroRNA sequencing showed that 28 microRNAs significantly changed in OD-Exo, of which 7 increased and 21 decreased. Pathway analysis showed genes targeted by differentially expressed microRNAs were involved in multiple signal transductions, including TGFß pathway. 16 genes targeted by 15 differentially expressed microRNAs were involved in TGFß signaling. Consistently, automated western blot found that OD-Exo activated TGFß1 pathway by upregulating TGFß1, TGFR1, p-Smad2/3, and Smad4 in DPSCs. Accordingly, once the TGFß1 signaling pathway was inhibited by SB525334, protein levels of p-Smad2/3, DSP, and DMP-1 were significantly decreased in DPSCs treated with OD-Exo. MiR-27a-5p was expressed 11 times higher in OD-Exo, while miR-27a-5p promoted odontogenic differentiation of DPSCs and significantly upregulated TGFß1, TGFR1, p-Smad2/3, and Smad4 by downregulating the inhibitory molecule LTBP1. CONCLUSIONS: The microRNA expression profiles of exosomes derived from DPSCs were identified. OD-Exo isolated under odontogenic conditions were better inducers of DPSC differentiation. Exosomal microRNAs promoted odontogenic differentiation via TGFß1/smads signaling pathway by downregulating LTBP1.