scholarly journals Synergies of Human Umbilical Vein Endothelial Cell-Laden Calcium Silicate-Activated Gelatin Methacrylate for Accelerating 3D Human Dental Pulp Stem Cell Differentiation for Endodontic Regeneration

Polymers ◽  
2021 ◽  
Vol 13 (19) ◽  
pp. 3301
Author(s):  
Wei-Yun Lai ◽  
Tzu-Hsin Lee ◽  
Jian-Xun Chen ◽  
Hooi-Yee Ng ◽  
Tsui-Hsien Huang ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Vascular Endothelial Cells ◽  
Umbilical Vein ◽  
Stem Cell Differentiation ◽  
Cell Block ◽  
Fish Gelatin ◽  
Alizarin Red ◽  
Covalent Bonds ◽  
Human Dental Pulp ◽  
Dentin Regeneration

According to the Centers for Disease Control and Prevention, tooth caries is a common problem affecting 9 out of every 10 adults worldwide. Dentin regeneration has since become one of the pressing issues in dentistry with tissue engineering emerging as a potential solution for enhancing dentin regeneration. In this study, we fabricated cell blocks with human dental pulp stem cells (hDPSCs)-laden alginate/fish gelatin hydrogels (Alg/FGel) at the center of the cell block and human umbilical vascular endothelial cells (HUVEC)-laden Si ion-infused fish gelatin methacrylate (FGelMa) at the periphery of the cell block. 1H NMR and FTIR results showed the successful fabrication of Alg/FGel and FGelMa. In addition, Si ions in the FGelMa were noted to be bonded via covalent bonds and the increased number of covalent bonds led to an increase in mechanical properties and improved degradation of FGelMa. The Si-containing FGelMa was able to release Si ions, which subsequently significantly not only enhanced the expressions of angiogenic-related protein, but also secreted some cytokines to regulate odontogenesis. Further immunofluorescence results indicated that the cell blocks allowed interactions between the HUVEC and hDPSCs, and taken together, were able to enhance odontogenic-related markers’ expression, such as alkaline phosphatase (ALP), dentin matrix phosphoprotein-1 (DMP-1), and osteocalcin (OC). Subsequent Alizarin Red S stain confirmed the benefits of our cell block and demonstrated that such a novel combination and modification of biomaterials can serve as a platform for future clinical applications and use in dentin regeneration.

scholarly journals ANALYSIS OF LYSATE PLATELET-RICH FIBRIN EFFECTS ON HUMAN DENTAL PULP STEM CELL DIFFERENTIATION THROUGH DENTIN SIALOPHOSPHOPROTEIN EXPRESSION

2020 ◽  
pp. 34-37
Author(s):  
SILVIANA SWASTININGTYAS ◽  
ANGGRAINI MARGONO ◽  
DINI ASRIANTI ◽  
RUNY OKTAYANI ◽  
INDAH YULIANTO
Keyword(s):  
Dental Pulp ◽  
Culture Media ◽  
Stem Cell Differentiation ◽  
Alizarin Red ◽  
Platelet Rich Fibrin ◽  
Dentin Sialophosphoprotein ◽  
Human Dental Pulp ◽  
Stable Growth ◽  
And Control

Objective: In vitro, the culture media in which human dental pulp stem cells (hDPSCs) are grown are supplemented with specific growth factors thatinduce cell cycle entry and differentiation. Lysate platelet-rich fibrin (L-PRF) is a unique and stable growth factor supplement produced from plateletslysed by freezing-thawing. In this study, we aimed to analyze the potential effects of L-PRF on hDPSC differentiation.Methods: We divided hDPSCs isolated from human third molars at the second passage into five culture media groups treated with 1%, 5%, 10%,and 25% L-PRF or 10% fetal bovine serum (control). After 7 days, we evaluated hDPSC differentiation using an enzyme-linked immunosorbent assayspecific for dentin sialophosphoprotein and Alizarin-Red staining.Results: None of our analyses revealed any significant differences between the L-PRF- and control-treated cells.Conclusion: L-PRF could potentially induce the differentiation of hDPSCs in vitro.

scholarly journals Evaluation of Resin-Based Material Containing Copaiba Oleoresin (Copaifera Reticulata Ducke): Biological Effects on the Human Dental Pulp Stem Cells

Biomolecules ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 972
Author(s):  
Roberta Souza D’Almeida Couto ◽  
Maria Fernanda Setubal Destro Rodrigues ◽  
Leila Soares Ferreira ◽  
Ivana Márcia Alves Diniz ◽  
Fernando de Sá Silva ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Biological Effects ◽  
Dental Pulp Stem Cells ◽  
Nodule Formation ◽  
Alizarin Red ◽  
Pulp Capping ◽  
Hsp 27 ◽  
Dental Pulp Capping ◽  
Human Dental Pulp

The purpose of this study was to analyze in vitro the biological effects on human dental pulp stem cells triggered in response to substances leached or dissolved from two experimental cements for dental pulp capping. The experimental materials, based on extracts from Copaifera reticulata Ducke (COP), were compared to calcium hydroxide [Ca(OH)2] and mineral trioxide aggregate (MTA), materials commonly used for direct dental pulp capping in restorative dentistry. For this, human dental pulp stem cells were exposed to COP associated or not with Ca(OH)2 or MTA. Cell cytocompatibility, migration, and differentiation (mineralized nodule formation (Alizarin red assay) and gene expression (RT-qPCR) of OCN, DSPP, and HSP-27 (genes regulated in biomineralization events)) were evaluated. The results showed that the association of COP reduced the cytotoxicity of Ca(OH)2. Upregulations of the OCN, DSPP, and HSP-27 genes were observed in response to the association of COP to MTA, and the DSPP and HSP-27 genes were upregulated in the Ca(OH)2 + COP group. In up to 24 h, cell migration was significantly enhanced in the MTA + COP and Ca(OH)2 + COP groups. In conclusion, the combination of COP with the currently used materials for dental pulp capping [Ca(OH)2 and MTA] improved the cell activities related to pulp repair (i.e., cytocompatibility, differentiation, mineralization, and migration) including a protective effect against the cytotoxicity of Ca(OH)2.

The Influence of Metalized Graphene Oxide/Reduced Graphene Oxide and Sulfonated Polystyrene on Dental Pulp Stem Cell Differentiation and Protein Adsorption

MRS Advances ◽  
2017 ◽  
Vol 2 (19-20) ◽  
pp. 1059-1070 ◽  
Author(s):  
Rachel Sacks ◽  
Gila Schein ◽  
Rebecca Isseroff ◽  
Vincent Ricotta ◽  
Marcia Simon ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Stem Cell ◽  
Protein Adsorption ◽  
Dental Pulp ◽  
Stem Cell Differentiation ◽  
Sulfonated Polystyrene ◽  
Reduced Graphene ◽  
Human Dental Pulp ◽  
Osteogenic Induction ◽  
Cell Growth And Differentiation

ABSTRACTHuman dental pulp stem cells (DPSCs) can differentiate, showing potential for regenerative medicine. Designing artificial surfaces with properties appropriate for the initiation of extracellular matrix (ECM) adsorption and organization is a critical step in tissue engineering and can greatly impact protein adhesion. Sulfonated polystyrene (SPS), used as a scaffold for tissue development, stimulates protein adsorption due to the increased negative charge of sulfonate.Graphene and graphene oxide (GO) sheets enhance stem cell growth and differentiation because they are soft membranes with “high in-plane” stiffness and have the potential to be transferable and implantable platforms. This project functionalized GO and reduced GO (RGO) with gold or silver nanoparticles, mixing with SPS to investigate their combined impact on DPSC differentiation and protein adsorption, hypothesizing that this combination supplies more charges to better absorb the proteins to the surface and stimulate differentiation.Results indicate that proteins of cells plated on the gold-RGO/SPS surfaces were the most highly adsorbed and most densely packed. Additionally, the cell moduli data indicated that the metal-RGO solutions substantially induced a change in modulus even more than Dexamethasone, a glucocortoid known to enhance this process in DPSCs. This suggests that the metal-RGO solutions may be instrumental in osteogenic induction.

Human treated dentin matrices combined with Zn-doped, Mg-based bioceramic scaffolds and human dental pulp stem cells towards targeted dentin regeneration

2016 ◽  
Vol 32 (8) ◽  
pp. e159-e175 ◽  
Author(s):  
Athina Bakopoulou ◽  
Eleni Papachristou ◽  
Maria Bousnaki ◽  
Christina Hadjichristou ◽  
Eleana Kontonasaki ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Human Dental Pulp ◽  
Dentin Regeneration

scholarly journals The Role of Pannexin3-Modified Human Dental Pulp-Derived Mesenchymal Stromal Cells in Repairing Rat Cranial Critical-Sized Bone Defects

2017 ◽  
Vol 44 (6) ◽  
pp. 2174-2188 ◽  
Author(s):  
Fangfang Song ◽  
Hualing Sun ◽  
Liyuan Huang ◽  
Dongjie Fu ◽  
Cui Huang
Keyword(s):  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Dental Pulp ◽  
Underlying Mechanism ◽  
Bone Defects ◽  
Dependent Manner ◽  
Alizarin Red ◽  
Human Dental Pulp

Background/Aims: Human dental pulp-derived mesenchymal stromal cells (hDPSCs) are promising seed cells for tissue engineering due to their easy accessibility and multi-lineage differentiation. Pannexin3 (Panx3) plays crucial roles during bone development and differentiation. The aim of the present study was to investigate the effect of Panx3 on osteogenesis of hDPSCs and the underlying mechanism. Methods: Utilizing qRT-PCR, Western blot, and immunohistochemistry, we explored the change of Panx3 during osteogenic differentiation of hDPSCs. Next, hDPSCs with loss (Panx3 knockdown) and gain (Panx3 overexpression) of Panx3 function were developed to investigate the effects of Panx3 on osteogenic differentiation of hDPSC and the underlying mechanism. Finally, a commercial β-TCP scaffold carrying Panx3-modified hDPSCs was utilized to evaluate bone defect repair. Results: Panx3 was upregulated during osteogenic differentiation in a time-dependent manner. Panx3 overexpression promoted osteogenic differentiation of hDPSCs, whereas depletion of Panx3 resulted in a decline of differentiation, evidenced by upregulated expression of mineralization-related markers, increased alkaline phosphatase (ALP) activity, and enhanced ALP and Alizarin red staining. Panx3 was found to interact with the Wnt/β-catenin signaling pathway, forming a negative feedback loop. However, Wnt/β-catenin did not contribute to enhancement of osteogenic differentiation as observed in Panx3 overexpression. Moreover, Panx3 promoted osteogenic differentiation of hDPSCs via increasing ERK signaling pathway. Micro-CT and histological staining results showed that Panx3-modified hDPSCs significantly improved ossification of critical-sized bone defects. Conclusion: These findings suggest that Panx3 is a crucial modulator of hDPSCs differentiation.

scholarly journals miR-6807-5p Inhibited the Odontogenic Differentiation of Human Dental Pulp Stem Cells Through Directly Targeting METTL7A

2021 ◽  
Vol 9 ◽  
Author(s):  
Ning Wang ◽  
Xiao Han ◽  
Haoqing Yang ◽  
Dengsheng Xia ◽  
Zhipeng Fan
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Binding Protein ◽  
Dental Pulp Stem Cells ◽  
Luciferase Reporter ◽  
Control Group ◽  
Alizarin Red ◽  
Odontogenic Differentiation ◽  
Human Dental Pulp ◽  
Tooth Tissue

Background: Tooth tissue regeneration mediated by mesenchymal stem cells (MSCs) has become the most ideal treatment. Although the known regulatory mechanism and some achievements have been discovered, directional differentiation cannot effectively induce regeneration of tooth tissue. In this study, we intended to explore the function and mechanism of miR-6807-5p and its target gene METTL7A in odontogenic differentiation.Methods: In this study, human dental pulp stem cells (DPSCs) were used. Alkaline phosphatase (ALP), Alizarin red staining (ARS), and calcium ion quantification were used to detect the odontogenic differentiation of miR-6807-5p and METTL7A. Real-time RT-PCR, western blot, dual-luciferase reporter assay, and pull-down assay with biotinylated miRNA were used to confirm that METTL7A was the downstream gene of miR-6807-5p. Protein mass spectrometry and co-immunoprecipitation (Co-IP) were used to detect that SNRNP200 was the co-binding protein of METTL7A.Results: After mineralized induction, the odontogenic differentiation was enhanced in the miR-6807-5p-knockdown group and weakened in the miR-6807-5p-overexpressed group compared with the control group. METTL7A was the downstream target of miR-6807-5p. After mineralized induction, the odontogenic differentiation was weakened in the METTL7A-knockdown group and enhanced in the METTL7A-overexpressed group compared with the control group. SNRNP200 was the co-binding protein of METTL7A. The knockdown of SNRNP200 inhibited the odontogenic differentiation of DPSCs.Conclusion: This study verified that miR-6807-5p inhibited the odontogenic differentiation of DPSCs. The binding site of miR-6807-5p was the 3′UTR region of METTL7A, which was silenced by miR-6807-5p. METTL7A promoted the odontogenic differentiation of DPSCs. SNRNP200, a co-binding protein of METTL7A, promoted the odontogenic differentiation of DPSCs.

scholarly journals Effect of Vitamins D and E on the Proliferation, Viability, and Differentiation of Human Dental Pulp Stem Cells: An In Vitro Study

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Lina M. Escobar ◽  
Zita Bendahan ◽  
Andrea Bayona ◽  
Jaime E. Castellanos ◽  
María-Clara González
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Dental Pulp ◽  
Morphological Changes ◽  
In Vitro Study ◽  
Osteoblastic Differentiation ◽  
Dental Pulp Stem Cells ◽  
Alizarin Red ◽  
Human Dental Pulp

Introduction. The aim of the present study was to determine the effects of vitamins D and E on the proliferation, morphology, and differentiation of human dental pulp stem cells (hDPSCs). Methods. In this in vitro experimental study, hDPSCs were isolated, characterized, and treated with vitamins D and E, individually and in combination, utilizing different doses and treatment periods. Changes in morphology and cell proliferation were evaluated using light microscopy and the resazurin assay, respectively. Osteoblast differentiation was evaluated with alizarin red S staining and expression of RUNX2, Osterix, and Osteocalcin genes using real-time RT-PCR. Results. Compared with untreated cells, the number of cells significantly reduced following treatment with vitamin D (49%), vitamin E (35%), and vitamins D + E (61%) after 144 h. Compared with cell cultures treated with individual vitamins, cells treated with vitamins D + E demonstrated decreased cell confluence, with more extensive and flatter cytoplasm that initiated the formation of a significantly large number of calcified nodules after 7 days of treatment. After 14 days, treatment with vitamins D, E, and D + E increased the transcription of RUNX2, Osterix, and Osteocalcin genes. Conclusions. Vitamins D and E induced osteoblastic differentiation of hDPSCs, as evidenced by the decrease in cell proliferation, morphological changes, and the formation of calcified nodules, increasing the expression of differentiation genes. Concurrent treatment with vitamins D + E induces a synergistic effect in differentiation toward an osteoblastic lineage.

scholarly journals The Effect of Mesoporous Bioactive Glass Nanoparticles/Graphene Oxide Composites on the Differentiation and Mineralization of Human Dental Pulp Stem Cells

Nanomaterials ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 620 ◽  
Author(s):  
Jae Hwa Ahn ◽  
In-Ryoung Kim ◽  
Yeon Kim ◽  
Dong-Hyun Kim ◽  
Soo-Byung Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
Graphene Oxide ◽  
Bioactive Glass ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Mrna Levels ◽  
Alizarin Red ◽  
Differentiation Potential ◽  
Human Dental Pulp ◽  
Mesoporous Bioactive Glass

The purpose of this study was to investigate the effects of mesoporous bioactive glass nanoparticle (MBN)/graphene oxide (GO) composites on the mineralization ability and differentiation potential of human dental pulp stem cells (hDPSCs). MBN/GO composites were synthesized using the sol-gel method and colloidal processing to enhance the bioactivity and mechanical properties of MBN. Characterization using FESEM, XRD, FTIR, and Raman spectrometry showed that the composites were successfully synthesized. hDPSCs were then cultured directly on the MBN/GO (40:1 and 20:1) composites in vitro. MBN/GO promoted the proliferation and alkaline phosphatase (ALP) activity of hDPSCs. In addition, qRT-PCR showed that MBN/GO regulated the mRNA levels of odontogenic markers (dentin sialophosphoprotein (DSPP), dentine matrix protein 1 (DMP-1), ALP, matrix extracellular phosphoglycoprotein (MEPE), bone morphogenetic protein 2 (BMP-2), and runt-related transcription factor 2 (RUNX-2)). The mRNA levels of DSPP and DMP-1, two odontogenesis-specific markers, were considerably upregulated in hDPSCs in response to growth on the MBN/GO composites. Western blot analysis revealed similar results. Alizarin red S staining was subsequently performed to further investigate MBN/GO-induced mineralization of hDPSCs. It was revealed that MBN/GO composites promote odontogenic differentiation via the Wnt/β-catenin signaling pathway. Collectively, the results of the present study suggest that MBN/GO composites may promote the differentiation of hDPSCs into odontoblast-like cells, and potentially induce dentin formation.

scholarly journals Silencing VEGFR-2 Hampers Odontoblastic Differentiation of Dental Pulp Stem Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Kajohnkiart Janebodin ◽  
Rakchanok Chavanachat ◽  
Aislinn Hays ◽  
Morayma Reyes Gil
Keyword(s):  
Stem Cells ◽  
Blood Vessels ◽  
Dental Pulp ◽  
Alizarin Red ◽  
Odontoblast Differentiation ◽  
Dentin Matrix ◽  
Dentin Regeneration

Dental pulp stem cells (DPSCs) are a source of postnatal stem cells essential for maintenance and regeneration of dentin and pulp tissues. Previous in vivo transplantation studies have shown that DPSCs are able to give rise to odontoblast-like cells, form dentin/pulp-like structures, and induce blood vessel formation. Importantly, dentin formation is closely associated to blood vessels. We have previously demonstrated that DPSC-induced angiogenesis is VEGFR-2-dependent. VEGFR-2 may play an important role in odontoblast differentiation of DPSCs, tooth formation and regeneration. Nevertheless, the role of VEGFR-2 signaling in odontoblast differentiation of DPSCs is still not well understood. Thus, in this study we aimed to determine the role of VEGFR-2 in odontoblast differentiation of DPSCs by knocking down the expression of VEGFR-2 in DPSCs and studying their odontoblast differentiation capacity in vitro and in vivo. Isolation and characterization of murine DPSCs was performed as previously described. DPSCs were induced by VEGFR-2 shRNA viral vectors transfection (MOI = 10:1) to silence the expression of VEGFR-2. The GFP+ expression in CopGFP DPSCs was used as a surrogate to measure the efficiency of transfection and verification that the viral vector does not affect the expression of VEGFR-2. The efficiency of viral transfection was shown by significant reduction in the levels of VEGFR-2 based on the Q-RT-PCR and immunofluorescence in VEGFR-2 knockdown DPSCs, compared to normal DPSCs. VEGFR-2 shRNA DPSCs expressed not only very low level of VEGFR-2, but also that of its ligand, VEGF-A, compared to CopGFP DPSCs in both transcriptional and translational levels. In vitro differentiation of DPSCs in osteo-odontogenic media supplemented with BMP-2 (100 ng/ml) for 21 days demonstrated that CopGFP DPSCs, but not VEGFR-2 shRNA DPSCs, were positive for alkaline phosphatase (ALP) staining and formed mineralized nodules demonstrated by positive Alizarin Red S staining. The expression levels of dentin matrix proteins, dentin matrix protein-1 (Dmp1), dentin sialoprotein (Dspp), and bone sialoprotein (Bsp), were also up-regulated in differentiated CopGFP DPSCs, compared to those in VEGFR-2 shRNA DPSCs, suggesting an impairment of odontoblast differentiation in VEGFR-2 shRNA DPSCs. In vivo subcutaneous transplantation of DPSCs with hydroxyapatite (HAp/TCP) for 5 weeks demonstrated that CopGFP DPSCs were able to differentiate into elongated and polarized odontoblast-like cells forming loose connective tissue resembling pulp-like structures with abundant blood vessels, as demonstrated by H&E, Alizarin Red S, and dentin matrix staining. On the other hand, in VEGFR-2 shRNA DPSC transplants, odontoblast-like cells were not observed. Collagen fibers were seen in replacement of dentin/pulp-like structures. These results indicate that VEGFR-2 may play an important role in dentin regeneration and highlight the potential of VEGFR-2 modulation to enhance dentin regeneration and tissue engineering as a promising clinical application.

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