scholarly journals Leukotriene B 4 Loaded in Microspheres Regulate the Expression of Genes Related to Odontoblastic Differentiation and Biomineralization by Dental Pulp Stem Cells

2021 ◽  
Author(s):  
Francine Lorencetti Silva ◽  
Giuliana Campos Chaves Lamarque ◽  
Fernanda Maria Machado Pereira Cabral de Oliveira ◽  
Paulo Nelson-Filho ◽  
Léa Assed Bezerra Silva ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Differentiation ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Lipid Mediator ◽  
Cell Phenotype ◽  
Nodule Formation ◽  
Pulp Cell ◽  
Pulp Inflammation ◽  
Dental Pulp Cell

Abstract Background: Leukotriene B4 (LTB4) is a potent lipid mediator that stimulate the immune response. Because dental pulp inflammation and dentin repair are intrinsically related responses, the aim of this research was to investigate the potential of LTB4 in inducing differentiation of dental pulp stem cells. Methods: Microspheres (MS) loaded with LTB4 were prepared using an oil emulsion solvent extraction evaporation process and sterility, characterization, efficiency of LTB4 encapsulation and in vitro LTB4 release assay were investigated. Mouse dental pulp stem cells (OD-21) were stimulated with soluble LTB4 or MS loaded with LTB4 (0.01 and 0.1 μM). Cytotoxicity and cell viability was determined by lactate dehydrogenase (LDH) and MTT (methylthiazol tetrazolium) assays . Gene expression were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) after 3, 6, 24, 48 and 72 h. Results: Mineralized nodule formation was assessed after 28 days of OD-21 cell stimulation with LTB4. Groups were compared using the one-way ANOVA test followed by Dunnett's post-test (α = 0.05). Treatment with LTB4 or MS loaded with LTB4 (0.01 and 0.1 micrometer- μM) were not cytotoxic to OD-21 cells. Treatment with LTB4 modulated the expression of the Ibsp (integrin binding sialoprotein) and Runx2 (runt-related transcription factor 2) genes differently depending on the experimental period analyzed. Interestingly LTB4 loaded in microspheres (0.1 μM) allowed long term dental pulp cell differentiation and biomineralization. LTB4 loaded in MS was not cytotoxic and induced an odontoblastic cell phenotype differentiation. Conclusion: These findings shed light on a novel pharmacological strategy to induce dental pulp cell differentiation.

Differential expression of long noncoding RNAs in SDF-1α-induced dental pulp stem cells

2021 ◽  
Author(s):  
min xiao ◽  
Bo Yao ◽  
Xiaohan Mei ◽  
yu bai ◽  
Jueyu Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Rho Gtpase ◽  
Noncoding Rnas ◽  
Dental Pulp Stem Cells ◽  
Long Noncoding Rnas ◽  
Nodule Formation ◽  
Related Factor ◽  
Altered Expression ◽  
Odontogenic Differentiation

Abstract Background SDF-1α cotreatment was shown to have synergistic effects on BMP-2-induced odontogenic differentiation of human apical dental papillary stem cells (SCAP) both in vitro and in vivo. Long noncoding RNAs (lncRNAs) have an important role in the odontogenic differentiation of dental pulp stem cells (DPSCs). Methods We examined the altered expression of lncRNAs in SDF-1α-induced odontogenic differentiation of DPSCs by lncRNA microarray and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analyses. Alterations in lncRNA expression during odontogenic differentiation of DPSCs were identified. Moreover, bioinformatic analysis [Gene Ontology (GO) analysis and coding-noncoding gene coexpression (CNC) analysis] was conducted to predict the interactions of lncRNAs and identify core regulatory factors in SDF-1α-induced odontogenic differentiation of DPSCs. Results The microarray analysis identified 206 differentially expressed lncRNAs (134 lncRNAs with upregulated expression and 72 with downregulated expression) at 7 days post‑treatment. The data demonstrated that one lncRNA, AC080037.1, regulates SDF-1α-induced odontogenic differentiation of DPSCs. Our data showed that lncRNA AC080037.1 siRNA suppresses DPSCs migration and the expression of Rho GTPase induced by SDF-1α. Moreover, AC080037.1 knockdown significantly affected SDF-1α- and BMP-2-induced mineralized nodule formation and strongly suppressed Runt-related factor-2 (RUNX-2), DMP-1 and DSPP expression in DPSCs. Conclusions Our

scholarly journals Reactionary Versus Reparative Dentine in Deep Caries

2019 ◽  
Vol 25 (1) ◽  
pp. 15-21
Author(s):  
Iliescu Alexandru-Andrei ◽  
Gheorghiu Irina-Maria ◽  
Tănase Mihaela ◽  
Iliescu Andrei ◽  
Mitran Loredana ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Complex Response ◽  
Pulp Inflammation ◽  
Proliferation And Differentiation ◽  
Molecular Signals

Abstract The dentine-pulp complex response in deep caries is histological characterized by tertiary formation and mild chronic pulp inflammation. The quiescent primary odontoblasts are reactivated, laying down reactionary tertiary dentine. In more severe carious damage the primary odontoblasts die and reparative tertiary dentine is secreted by odontoblast-like cells, which are differentiated in adult teeth mainly from dental pulp stem cells DPSC. Though associated with reversible pulpitis DPSC still preserve in deep caries the capability of migration, proliferation and differentiation. Some common mechanisms of molecular signals involved in tertiary dentine formation might also explain the balance between inflammation and regeneration of dentine-pulp complex.

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.

scholarly journals Prostaglandin E2 Induces Expression of Mineralization Genes by Undifferentiated Dental Pulp Cells

2019 ◽  
Vol 30 (3) ◽  
pp. 201-207 ◽  
Author(s):  
Francine Lorencetti-Silva ◽  
Priscilla Aparecida Tartari Pereira ◽  
Alyne Fávero Galvão Meirelles ◽  
Lúcia Helena Faccioli ◽  
Francisco Wanderley Garcia Paula-Silva
Keyword(s):  
Stem Cells ◽  
Prostaglandin E2 ◽  
Dental Pulp ◽  
Colorimetric Assay ◽  
Dental Pulp Stem Cells ◽  
Lipid Mediator ◽  
Water Emulsion ◽  
Mtt Colorimetric Assay ◽  
Oil In Water Emulsion

Abstract Prostaglandin E2 (PGE2) is a lipid mediator usually released during inflammation. This study aimed to investigate the potential of soluble or microsphere-loaded PGE2 on inducing differentiation of dental pulp stem cells. PGE2-loaded microspheres (MS) were prepared using an oil-in-water emulsion solvent extraction-evaporation process and were characterized. Mouse dental pulp stem cells (OD-21) were stimulated with soluble or PGE2-loaded MS (0.01 and 0.1 µM). Cell viability was determined by MTT colorimetric assay. Ibsp, Bmp2 and Runx2 expression was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) after 3, 6, and 24 h. The results showed that the soluble PGE2 reduced dental pulp stem cells viability after 24 h of stimulation whereas PGE2-loaded MS did not. Soluble PGE2 up-regulated Ibsp and Bmp2 at 3 h, differently from PGE2-loaded MS. On the other hand, PGE2-MS induced Bmp2 and Runx2 at 6 h and Ibsp at 24 h. In conclusion, our in vitro results show that PGE2, soluble or loaded in MS are not cytotoxic and modulateIbsp,Bmp2, andRunx2gene expression in cultured OD-21 cells.

scholarly journals Differential expression of long noncoding RNAs in SDF-1α-induced odontogenic differentiation of human dental pulp stem cells

2021 ◽  
Author(s):  
Bo Yao ◽  
Xiaogang Cheng ◽  
Xiaohan Mei ◽  
Jun Chou ◽  
Beidi Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Rho Gtpase ◽  
Noncoding Rnas ◽  
Dental Pulp Stem Cells ◽  
Long Noncoding Rnas ◽  
Nodule Formation ◽  
Related Factor ◽  
Altered Expression ◽  
Odontogenic Differentiation

Abstract SDF-1α cotreatment was shown to have synergistic effects on BMP-2-induced odontogenic differentiation of human apical dental papillary stem cells (SCAP) both in vitro and in vivo. Long noncoding RNAs (lncRNAs) have an important role in the odontogenic differentiation of dental pulp stem cells (DPSCs). We examined the altered expression of lncRNAs in SDF-1α-induced odontogenic differentiation of DPSCs by lncRNA microarray and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analyses. Alterations in lncRNA expression during odontogenic differentiation of DPSCs were identified. Moreover, bioinformatic analysis [Gene Ontology (GO) analysis and coding-noncoding gene coexpression (CNC) analysis] was conducted to predict the interactions of lncRNAs and identify core regulatory factors in SDF-1α-induced odontogenic differentiation of DPSCs. The microarray analysis identified 206 differentially expressed lncRNAs (134 lncRNAs with upregulated expression and 72 with downregulated expression) at 7 days post‑treatment. The data demonstrated that one lncRNA, AC080037.1, regulates SDF-1α-induced odontogenic differentiation of DPSCs. Our data showed that lncRNA AC080037.1 siRNA suppresses DPSCs migration and the expression of Rho GTPase induced by SDF-1α. Moreover, AC080037.1 knockdown significantly affected SDF-1α- and BMP-2-induced mineralized nodule formation and strongly suppressed Runt-related factor-2 (RUNX-2), DMP-1 and DSPP expression in DPSCs. Our results highlighted the significant involvement of one lncRNA, AC080037.1, in the positive regulation of the osteo/odontogenic differentiation of DPSCs and indicated that lncRNA AC080037.1 could be a potential target in regenerative endodontics.These findings reveal how lncRNAs are involved in regulating the SDF-1α-induced odontogenic differentiation of DPSCs, which may further advance translational studies of pulp tissue engineering.

scholarly journals Human Dental Pulp Stem Cells via the NF-κB Pathway

2015 ◽  
Vol 36 (5) ◽  
pp. 1725-1734 ◽  
Author(s):  
Shensheng Gu ◽  
Shujun Ran ◽  
Feng Qin ◽  
Dong Cao ◽  
Jia Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Marker Genes ◽  
Nodule Formation ◽  
Dependent Manner ◽  
Odontoblastic Differentiation ◽  
Alp Activity ◽  
Odontogenic Differentiation ◽  
Human Dental Pulp

Background/Aims: Odontogenic differentiation of human dental pulp stem cells (HDPSCs) is regulated by multiple factors and signaling molecules. However, their regulatory mechanisms are not completely understood. In this study, we investigated the role of Zinc finger and BTB domain-containing 20 (ZBTB20) in odontoblastic differentiation of HDPSCs. Methods: HDPSCs were obtained from human third molars and ZBTB20 expression was examined by qRT-PCR and western blot. Their osteo/odontogenic differentiation and the involvement of NF-κB pathway were subsequently investigated. Results: The expression of ZBTB20 is upregulated in a time-dependent manner during odontogenic differentiation of hDPSCs. Inhibition of ZBTB20 reduced osteogenic medium (OM)-induced odontogenic differentiation, reflected in decreased alkaline phosphatase (ALP) activity, mineralized nodule formation and mRNA expression of odonto/osteogenic marker genes. In contrast, overexpression of ZBTB20 enhanced ALP activity, mineralization and the expression of differentiation marker genes. Furthermore, the expression of IκBa was increased by ZBTB20 silencing in HDPSCs, whereas ZBTB20 overexpression decreased IκBa and enhanced nuclear NF-κB p65. Inhibition of the NF-κB pathway significantly suppressed the odontogenic differentiation of HDPSCs induced by ZBTB20. Conclusion: This study shows for the first time that ZBTB20 plays an important role during odontoblastic differentiation of HDPSCs and may have clinical implications for regenerative endodontics.

scholarly journals Periodontal and Dental Pulp Cell-Derived Small Extracellular Vesicles: A Review of the Current Status

Nanomaterials ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1858
Author(s):  
Shu Hua ◽  
Peter Mark Bartold ◽  
Karan Gulati ◽  
Corey Stephen Moran ◽  
Sašo Ivanovski ◽  
...  
Keyword(s):  
Stem Cells ◽  
Extracellular Vesicles ◽  
Dental Pulp ◽  
Therapeutic Potential ◽  
Current Status ◽  
Pulp Cell ◽  
Dental Pulp Cell

Extracellular vesicles (EVs) are membrane-bound lipid particles that are secreted by all cell types and function as cell-to-cell communicators through their cargos of protein, nucleic acid, lipids, and metabolites, which are derived from their parent cells. There is limited information on the isolation and the emerging therapeutic role of periodontal and dental pulp cell-derived small EVs (sEVs, <200 nm, or exosome). In this review, we discuss the biogenesis of three EV subtypes (sEVs, microvesicles and apoptotic bodies) and the emerging role of sEVs from periodontal ligament (stem) cells, gingival fibroblasts (or gingival mesenchymal stem cells) and dental pulp cells, and their therapeutic potential in vitro and in vivo. A review of the relevant methodology found that precipitation-based kits and ultracentrifugation are the two most common methods to isolate periodontal (dental pulp) cell sEVs. Periodontal (and pulp) cell sEVs range in size, from 40 nm to 2 μm, due to a lack of standardized isolation protocols. Nevertheless, our review found that these EVs possess anti-inflammatory, osteo/odontogenic, angiogenic and immunomodulatory functions in vitro and in vivo, via reported EV cargos of EV–miRNAs, EV–circRNAs, EV–mRNAs and EV–lncRNAs. This review highlights the considerable therapeutic potential of periodontal and dental pulp cell-derived sEVs in various regenerative applications.

scholarly journals Biocompatibility and Bioactivity of Set Direct Pulp Capping Materials on Human Dental Pulp Stem Cells

Materials ◽  
2020 ◽  
Vol 13 (18) ◽  
pp. 3925
Author(s):  
Yemi Kim ◽  
Donghee Lee ◽  
Dani Song ◽  
Hye-Min Kim ◽  
Sin-Young Kim
Keyword(s):  
Stem Cells ◽  
Cell Migration ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Nodule Formation ◽  
Alizarin Red ◽  
Pulp Capping ◽  
Alp Activity ◽  
Human Dental Pulp ◽  
And Control

In this study, we assessed the biocompatibility and bioactivity of various pulp capping materials—ProRoot MTA (Dentsply Tulsa Dental Specialties), Biodentine (Septodont), TheraCal LC (Bisco), and Dycal (Dentsply Caulk)—on human dental pulp stem cells (hDPSCs). Experimental disks (diameter, 7 mm; height, 4 mm) were stored in a humified incubator at 37 °C for 48 h. Then, the pulp capping materials were tested for cytotoxic effects by methyl-thiazoldiphenyl-tetrazolium and scratch wound healing assays, and for mineralization potential by Alizarin red S (ARS) staining assay and alkaline phosphatase enzyme (ALP) activity. Cell viability and cell migration did not significantly differ between ProRoot MTA, Biodentine, and control (p > 0.05). TheraCal LC exhibited slower cell migration on days 2–4 compared to control (p < 0.05), and Dycal showed no cell migration. ALP activity was highest with Biodentine on days 10 and 14, and was lowered with TheraCal LC and Dycal (p < 0.05). In the ARS assay, hDPSCs grown in ProRoot MTA and TheraCal LC eluates showed significantly increased mineralized nodule formation on day 21 compared to Biodentine, Dycal, and control (p < 0.05). These findings indicate that ProRoot MTA, Biodentine, and TheraCal LC exhibit better biocompatibility and bioactivity than Dycal.

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