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Journal of Dentistry (Shiraz, Iran) Sep 2023The occurrence of papillary defects adjacent to teeth or dental implants causes both the dental staff and the patients to be concerned about the esthetic issues....
STATEMENT OF THE PROBLEM
The occurrence of papillary defects adjacent to teeth or dental implants causes both the dental staff and the patients to be concerned about the esthetic issues. Interdental papilla reconstruction surgery is one of the most difficult and unpredictable mucogingival surgeries.
PURPOSE
The present study aimed to investigate the efficacy of hyaluronic acid injection in the reconstruction of the interdental papilla.
MATERIALS AND METHOD
This clinical trial study was conducted on four patients with 20 deficient interdental papillae who met the inclusion criteria. At first, local anesthesia was applied. Afterward, 0.2 mL of 1.6% hyaluronic acid (HA) gel was injected (at the tip of the papilla and 2-3 mm below the tip of the papilla) three times every two weeks. At baseline, three, and six months later, clinical photography was taken under standard conditions. The papilla height (the distance between the interdental papilla tip and the basis), black triangle area, and the distance between the interdental papilla tip and contact point of adjacent teeth were all measured using Image J software.
RESULTS
The effectiveness of using HA gel in reducing the black triangle area was 85.06%. Furthermore, the papilla length increased by 70.256% while contact to papilla distance decreased by 83.026%. At different times, the values of the studied variables in the three levels were significantly different (< 0.05).
CONCLUSION
Injection of HA with 1.6% concentration at two points of the interdental papilla was effective in interdental papilla reconstruction at the aesthetic zone, especially in long-term, follow-ups (especially 6 months).
PubMed: 37727351
DOI: 10.30476/dentjods.2022.94766.1808 -
Journal of Endodontics Jun 2021The goal of regenerative endodontic procedures is to preserve and stimulate stem cells from the apical papilla (SCAPs) to develop the pulp-dentin complex using various...
Vascular Endothelial Growth Factor and/or Nerve Growth Factor Treatment Induces Expression of Dentinogenic, Neuronal, and Healing Markers in Stem Cells of the Apical Papilla.
INTRODUCTION
The goal of regenerative endodontic procedures is to preserve and stimulate stem cells from the apical papilla (SCAPs) to develop the pulp-dentin complex using various growth factors and scaffolds. We hypothesized that the treatment of SCAPs with vascular endothelial growth factor (VEGF) or nerve growth factor (NGF) may impact the expression of osteogenic and dentinogenic markers.
METHODS
The optimum concentration of VEGF and NGF on SCAP viability was assessed and introduced to SCAPs for 6-24 hours. SCAPs were also challenged with Escherichia coli lipopolysaccharide (LPS). Messenger RNA (mRNA) expression of DSPP, DMP1, TGFB1, OCN, SP7, and TWIST1 was examined via quantitative reverse transcription polymerase chain reaction. Immunohistochemistry was used to verify protein expression. In addition, total RNA from NGF-treated SCAPs in the presence or absence of LPS was extracted for RNA sequencing.
RESULTS
Compared with untreated cells, NGF-treated SCAPs showed markedly higher levels of DSPP, DMP1, and TGFB1 mRNAs (>9-fold change, P < .05), and SCAPs treated with both VEGF and NGF showed a significant increase of DSPP and TGFB1 mRNAs (P < .05). In addition, in LPS-challenged SCAPs, treatment with these growth factors also exhibited increased expression of DSPP, DMP1, and TGFB1 mRNAs, with the most significant change induced by VEGF (P < .05). Immunohistochemistry confirmed increased dentin sialophosphoprotein, dentin matrix acidic phosphoprotein 1, and transforming growth factor beta 1 protein expression in treated SCAPs. RNA sequencing revealed multiple pathways regulated by NGF, including TGF-β and neurogenic pathways.
CONCLUSIONS
VEGF- and NGF-induced dentinogenic/neuronal/healing marker expression in SCAPs indicates the potential value of applying these growth factors in regenerative endodontic procedures.
Topics: Cell Differentiation; Cell Proliferation; Cells, Cultured; Dental Papilla; Nerve Growth Factor; Osteogenesis; Stem Cells; Vascular Endothelial Growth Factor A
PubMed: 33652017
DOI: 10.1016/j.joen.2021.02.011 -
International Journal of Bioprinting 2022Three-dimensional (3D) bioprinting is an emerging method for tissue regeneration. However, promoting the epithelial-mesenchymal interaction (EMI), while maintaining the...
Three-dimensional (3D) bioprinting is an emerging method for tissue regeneration. However, promoting the epithelial-mesenchymal interaction (EMI), while maintaining the characteristics of epithelial cells has always been a challenge in tissue engineering. Since EMI acts as a critical factor in bone regeneration, this study aims to promote EMI by recombining epithelial and mesenchymal cells through 3D bioprinting. Hertwig's epithelial root sheath (HERS) is a transient structure appeared in the process of tooth root formation. Its epithelial characteristics are easy to attenuate under appropriate culture environment. We recombined HERS cells and dental papilla cells (DPCs) through 3D bioprinting to simulate the micro-environment of cell-cell interaction . HERS cells and DPCs were mixed with gelatin methacrylate (GelMA) separately to prepare bio-inks for bioprinting. The cells/GelMA constructs were transplanted into the alveolar socket of Sprague-Dawley rats and then observed for 8 weeks. Hematoxylin and eosin staining, Masson staining, and immunohistochemical analysis showed that dimensional cultural pattern provided ideal environment for HERS cells and DPCs to generate mineralization texture and promote alveolar bone regeneration through their interactions. 3D bioprinting technology provides a new way for the co-culture of HERS cells and DPCs and this study is inspiring for future research on EMI model.
PubMed: 36105141
DOI: 10.18063/ijb.v8i3.512 -
BioMed Research International 2020This study is aimed at evaluating the effects of platelet-rich plasma (PRP) on proliferation, viability, and odontogenic differentiation of neural crest stem-like cells...
OBJECTIVE
This study is aimed at evaluating the effects of platelet-rich plasma (PRP) on proliferation, viability, and odontogenic differentiation of neural crest stem-like cells (NCSCs) derived from human dental apical papilla.
MATERIALS AND METHODS
Cells from apical papillae were obtained and then induced to form neural spheres. The expression of NCSC markers p75NTR and HNK-1 in neural sphere cells was detected by immunofluorescence staining. Human PRP was prepared by a 2-step centrifugation method and activated by CaCl and thrombin. The concentrations of PDGF-BB and TGF-1 in whole blood and PRP were measured by an ELISA kit. PRP in five different concentrations (0%, 2.5%, 5%, 10%, and 25%) was applied to culture NCSCs. On the 1, 3, 5, and 7 days, cell proliferation was evaluated by CCK8. Cell viability was tested by a live/dead staining kit. mRNA and protein expression of DSPP and BMP4 were analyzed by RT-qPCR and western blot, respectively. Statistical analysis was performed by a one-way analysis of variance (ANOVA) test or -test.
RESULTS
Dental apical papilla cells formed neural spheres, from which cells displayed positive expression of p75NTR and HNK-1. The concentrations of PDGF-BB and TGF-1 in PRP were about 3.5-fold higher than those in whole blood. 5% and 10% PRP significantly promoted proliferation of NCSCs, while 25% and 50% PRP inhibited cell proliferation from Day 3 to Day 7. Low-concentration (2.5%, 5%, and 10%) PRP slightly improved viability of NCSCs on Day 7. On the other hand, high-concentration (25% and 50%) PRP significantly inhibited viability of NCSCs from Day 3 to Day 7. RT-qPCR and western blot results indicated that 10% PRP could promote odontogenic differentiation of NCSCs on Day 7. mRNA and protein expression of DSPP and BMP4 were significantly upregulated in the 10% PRP group compared to those in the control group ( < 0.05).
CONCLUSIONS
PRP is a simply acquirable blood derivative which contains high concentration of growth factors like PDGF-BB and TGF-1. PRP in a proper concentration could promote proliferation, viability, and odontogenic differentiation of NCSCs derived from human dental apical papilla.
Topics: Biological Products; Cell Proliferation; Cell Survival; Cells, Cultured; Dental Papilla; Humans; Neural Crest; Neural Stem Cells; Odontogenesis; Platelet-Rich Plasma
PubMed: 32461990
DOI: 10.1155/2020/4671989 -
Journal of Dental Research Jun 2021A comprehensive study of odontoblastic differentiation is essential to understand the process of tooth development and to achieve the ability of tooth regeneration in...
A comprehensive study of odontoblastic differentiation is essential to understand the process of tooth development and to achieve the ability of tooth regeneration in the future. Zinc finger E-box-binding homeobox 1 () is a transcription factor expressed in various neural crest-derived tissues, including the mesenchyme of the tooth germ. However, its role in odontoblastic differentiation remains unknown. In this study, we found the expression of gradually increased during odontoblast differentiation in vivo, as well as during induced differentiation of cultured primary murine dental papilla cells (mDPCs) in vitro. In addition, the differentiation of mDPCs was repressed in silenced cells. We used RNA sequencing (RNAseq) to identify the transcriptome-wide targets of and used assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) to explore the direct targets of in both the early stage (embryonic day 16.5; E16.5) and the late stage (postnatal day 0; PN0) of tooth development. We identified the motifs of transcription factors enriched in -dependent accessible chromatin regions and observed that only in the early stage of mDPCs could significantly change the accessibility of chromatin regions. In vivo and in vitro experiments confirmed that silencing of at E16.5 inhibited dentinogenesis. Analysis of RNA-seq and ATAC-seq resulted in the identification of , a gene directly regulated by during early odontoblast differentiation. enhances the expression of by binding to its -elements, and ZEB1 interacts with RUNX2. In the late stage of tooth development, we found that ZEB1 could directly bind to and increase the enhancer activity of an element upstream of and promote dentinogenesis. In this study, for the first time, we revealed that ZEB1 promoted odontoblast differentiation in the early stage by altering chromatin accessibility of -elements near genes such as , while in the late stage, it directly enhanced transcription, thereby performing a dual role.
Topics: Animals; Cell Differentiation; Extracellular Matrix Proteins; Mice; Odontoblasts; Odontogenesis; Phosphoproteins
PubMed: 33419386
DOI: 10.1177/0022034520982249 -
The Journal of Prosthetic Dentistry Nov 2019Polyvinylphosphonic acid (PVPA) could be used as a biomimetic remineralization analog and a matrix metalloproteinases (MMPs) inhibitor. However, studies are lacking...
STATEMENT OF PROBLEM
Polyvinylphosphonic acid (PVPA) could be used as a biomimetic remineralization analog and a matrix metalloproteinases (MMPs) inhibitor. However, studies are lacking regarding the performance of PVPA in dental bonding systems for maintaining the durability of the resin-dentin bond.
PURPOSE
The purpose of this in vitro study was to investigate the effect of PVPA on the durability of resin-dentin bonds and the viability of mouse dental papilla cell-23 (MDPC-23). The mechanical properties of resin-dentin interfaces during long-term storage were analyzed, and the potential application of PVPA as a biomimetic remineralization analog in adhesive dentistry was evaluated.
MATERIAL AND METHODS
Seventy-five extracted noncarious human third molars were collected and randomly divided into 5 groups, and then the microtensile bond strength (μTBS) data and scanning electron microscope (SEM) images were used to evaluate the preservation condition of resin-dentin bonds after 1 day, 6 months, and 1 year of storage. The cytotoxicity of PVPA was detected by cell proliferation assay and cell apoptosis assay.
RESULTS
Compared with the control and chlorhexidine (CHX) groups, the combined group (treated with both 200-μg/mL PVPA and biomimetic remineralization) had excellent bond durability. The exposed collagen fibril from the PVPA-treated groups (included 200-μg/mL and 500-μg/mL PVPA groups and a combined group) still showed integrity after 1 year of storage when compared with the control group. PVPA up to 500 μg/mL showed no cytotoxicity to MDPC-23 and did not inhibit cell growth.
CONCLUSIONS
This study offered evidence that PVPA did not result in cytotoxicity at low concentrations as an MMP inhibitor and a biomimetic remineralization analog. In addition, the application of PVPA improved bond strength and preserved collagen integrity after 1 year of in vitro storage.
Topics: Acid Etching, Dental; Animals; Dental Bonding; Dental Papilla; Dentin; Dentin-Bonding Agents; Humans; Materials Testing; Mice; Resin Cements; Surface Properties; Tensile Strength
PubMed: 31623837
DOI: 10.1016/j.prosdent.2019.08.011 -
International Journal of Molecular... May 2022Stem cells from the apical papilla (SCAP) are a promising resource for use in regenerative endodontic treatment (RET) that may be adversely affected by oral bacteria,...
Stem cells from the apical papilla (SCAP) are a promising resource for use in regenerative endodontic treatment (RET) that may be adversely affected by oral bacteria, which in turn can exert an effect on the success of RET. Our work aims to study the cytokine profile of SCAP upon exposure to oral bacteria and their supernatants- and -as well as to establish their effect on the osteogenic and immunogenic potentials of SCAP. Further, we target the presence of key proteins of the Wnt/β-Catenin, TGF-β, and NF-κB signaling pathways, which play a crucial role in adult osteogenic differentiation of mesenchymal stem cells, using the Western blot (WB) technique. The membrane-based sandwich immunoassay and transcriptomic analysis showed that, under the influence of (both bacteria and supernatant), the production of pro-inflammatory cytokines IL-6, IL-8, and MCP-1 occurred, which was also confirmed at the mRNA level. Conversely, reduced the secretion of the aforementioned cytokines at both mRNA and protein levels. WB analysis showed that SCAP co-cultivation with led to a decrease in the level of the key proteins of the Wnt/β-Catenin and NF-κB signaling pathways: β-Catenin ( = 0.0068 *), LRP-5 ( = 0.0059 **), and LRP-6 ( = 0.0329 *), as well as NF-kB ( = 0.0034 **) and TRAF6 ( = 0.0285 *). These results suggest that oral bacteria can up- and downregulate the immune and inflammatory responses of SCAP, as well as influence the osteogenic potential of SCAP, which may negatively regulate the success of RET.
Topics: Adult; Bacteria; Cell Differentiation; Cytokines; Dental Papilla; Humans; Mouth; NF-kappa B; Osteogenesis; Protein Array Analysis; RNA, Messenger; Stem Cells; Transcriptome; Wnt Signaling Pathway; beta Catenin
PubMed: 35563488
DOI: 10.3390/ijms23095098 -
Scientific Reports Sep 2020Odontoblasts and pulp stroma cells are embedded within supramolecular networks of extracellular matrix (ECM). Fibrillin microfibrils and associated proteins are crucial...
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-β 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.
Topics: Adolescent; Adult; Animals; Animals, Newborn; Antigens, Surface; Dental Caries; Dental Pulp; Dentin; Glycoproteins; Humans; Incisor; Membrane Glycoproteins; Mice, Inbred C57BL; Molar; Young Adult
PubMed: 32948785
DOI: 10.1038/s41598-020-72123-2 -
Biotechnic & Histochemistry : Official... 2016The histone demethylase, lysine (K)-specific demethylase 2A (Kdm2a), is highly conserved and expressed ubiquitously. Kdm2a can regulate cell proliferation and...
The histone demethylase, lysine (K)-specific demethylase 2A (Kdm2a), is highly conserved and expressed ubiquitously. Kdm2a can regulate cell proliferation and osteo/dentinogenic, adipogenic and chondrogenic differentiation of mesenchymal stem cells (MSCs) derived from dental tissue. We used quantitative real-time RT-PCR analysis and immunohistochemistry to detect Kdm2a expression during development of the murine molar at embryonic days E12, E14, E16 and E17 and postnatal days P3 and P14. Immunohistochemistry results showed no positive staining of Kdm2a at E12. At E14, Kdm2a was expressed weakly in the inner enamel epithelium, stellate reticulum cells and dental sac. At E16, Kdm2a was expressed mainly in the inner and outer enamel epithelium, stratum intermedium and dental sac, but weaker staining was found in cervical loop and dental papilla cells adjacent to the basement membrane. At E17, the strongest Kdm2a staining was detected in the ameloblasts and stronger Kdm2a staining also was detected in the stratum intermedium, outer enamel epithelium and dental papilla cells compared to the expression at E16. Postnatally, we found that Kdm2a was localized in secretory and mature ameloblasts and odontoblasts, and dentin was unstained. Real-time RT-PCR showed that Kdm2a mRNA levels in murine germ cells increased from E12 to E14 and from E14 to E16; no significant change occurred at E16, E17 or P3, then the levels decreased at P14 compared to P3. Kdm2a expression may be closely related to cell proliferation, to ameloblast and odontoblast differentiation and to the secretion of extracellular enamel and dentin during murine tooth development.
Topics: Ameloblasts; Animals; Cell Differentiation; Cell Proliferation; Epithelium; Female; Gene Expression Regulation, Developmental; Jumonji Domain-Containing Histone Demethylases; Male; Mesenchymal Stem Cells; Mice; Odontogenesis; RNA, Messenger; Tooth Germ
PubMed: 26720400
DOI: 10.3109/10520295.2015.1106586 -
Journal of Cellular Physiology Jan 2021The effects of the renin-angiotensin system (RAS) on stem cells isolated from human dental apical papilla (SCAPs) are completely unknown. Therefore, the aim of this...
The effects of the renin-angiotensin system (RAS) on stem cells isolated from human dental apical papilla (SCAPs) are completely unknown. Therefore, the aim of this study was to identify RAS components expressed in SCAPs and the effects of angiotensin (Ang) II and Ang-(1-7) on cell proliferation. SCAPs were collected from third molar teeth of adolescents and maintained in cell culture. Messenger RNA expression and protein levels of angiotensin-converting enzyme (ACE), ACE2, and Mas, Ang II type I (AT1) and type II (AT2) receptors were detected in SCAPs. Treatment with either Ang II or Ang-(1-7) increased the proliferation of SCAPs. These effects were inhibited by PD123319, an AT2 antagonist. While Ang II augmented mTOR phosphorylation, Ang-(1-7) induced ERK1/2 phosphorylation. In conclusion, SCAPs produce the main RAS components and both Ang II and Ang-(1-7) treatments induced cell proliferation mediated by AT2 activation through different intracellular mechanisms.
Topics: Adolescent; Angiotensin I; Angiotensin II; Cell Proliferation; Cells, Cultured; Dental Papilla; Female; Humans; Imidazoles; MAP Kinase Signaling System; Male; Peptide Fragments; Peptidyl-Dipeptidase A; Phosphorylation; Pyridines; RNA, Messenger; Receptor, Angiotensin, Type 1; Renin-Angiotensin System; Stem Cells
PubMed: 32519379
DOI: 10.1002/jcp.29862