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2.
Int Endod J ; 57(4): 464-476, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38279773

RESUMO

AIM: To investigate novel diagnostic markers for pulpitis and validate by clinical samples from normal and inflamed pulp. To explore the relationship between diagnostic markers and immune cells or their phenotypes during pulp inflammation. METHODOLOGY: Two microarray datasets, GSE77459 and GSE92681, and identified differential expression genes were integrated. To understand immune features, gene functions, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Disease Ontology (DO) and ImmuneSigDB Gene Set Enrichment Analysis (GSEA) were analysed. For predictive purposes, machine learning techniques were applied to detect diagnostic markers. Immune infiltration in inflamed pulp was studied using CIBERSORT. The relationship between diagnostic markers and immune cells was investigated and validated their gene expression in clinical samples from the normal or inflamed pulp by qRT-PCR. Finally, the correlation between one marker, secreted phosphoprotein 1 (SPP1), encoding osteopontin (OPN), and dendritic cells (DCs)/macrophages was identified via HE staining and multiplex immunohistochemistry. An in vitro inflammatory dental pulp microenvironment model of THP-1 macrophages cocultured with dental pulp cells derived conditioned media (DPCs-CM) to investigate OPN production and macrophage phenotypes was established. RESULTS: Analysis revealed unique immunologic features in inflamed pulp. Three diagnostic markers for pulpitis: endothelin-1 (EDN1), SPP1, and purine nucleoside phosphorylase (PNP), and validated them using qRT-PCR were predicted. Multiplex immunohistochemistry demonstrated OPN co-localized with activated DCs and M2 macrophages during pulp inflammation. In vitro experiments showed that THP-1 macrophages produced the highest levels of OPN when stimulated with DPCs-CM derived from the 20 µg/mL LPS pre-conditioned group, suggesting an M2b-like phenotype by increasing surface marker CD86 and expression of IL6, TNFα, IL10, and CCL1 but not CCL17 and MerTK. Levels of CCL1 and IL10 elevated significantly in the macrophages' supernatant from the 20 µg/mL LPS pre-conditioned CM group. OPN was proven co-localizing with CD86 in the inflamed pulp by immunofluorescence. CONCLUSIONS: The current findings suggest that OPN can serve as a promising biomarker for pulpitis, correlated with DCs and macrophages. OPN+ macrophages in the inflamed pulp are associated with M2b-like phenotypes. These insights offer the potential for improved diagnosis and targeted therapy.


Assuntos
Pulpite , Humanos , Pulpite/metabolismo , Osteopontina , Interleucina-10/metabolismo , Lipopolissacarídeos/metabolismo , Inflamação/metabolismo , Macrófagos , Biomarcadores/metabolismo , Perfilação da Expressão Gênica , Células Dendríticas/metabolismo , Polpa Dentária/metabolismo
3.
PeerJ ; 11: e15605, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37397017

RESUMO

Background: Human microbiome dysbiosis is related to various human diseases, and identifying robust and consistent biomarkers that apply in different populations is a key challenge. This challenge arises when identifying key microbial markers of childhood caries. Methods: We analyzed unstimulated saliva and supragingival plaque samples from children of different ages and sexes, performed 16S rRNA gene sequencing, and sought to identify whether consistent markers exist among subpopulations by using a multivariate linear regression model. Results: We found that Acinetobacter and Clostridiales bacterial taxa were associated with caries in plaque and saliva, respectively, while Firmicutes and Clostridia were found in plaque isolated from children of different ages in preschool and school. These identified bacterial markers largely differ between different populations, leaving only Saccharibacteria as a significant caries-associated phylum in children. Saccharibacteria is a newly identified phylum, and our taxonomic assignment database could not be used to identify its specific genus. Conclusion: Our data indicated that, in a South China population, oral microbial signatures for dental caries show age and sex differences, but Saccharibacteria might be a consistent signal and worth further investigation, considering the lack of research on this microbe.


Assuntos
Cárie Dentária , Criança , Humanos , Pré-Escolar , Masculino , Feminino , RNA Ribossômico 16S/genética , Cárie Dentária/epidemiologia , Suscetibilidade à Cárie Dentária , Bactérias/genética , Firmicutes/genética , China/epidemiologia
4.
Front Med (Lausanne) ; 10: 832878, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36910498

RESUMO

Introduction: Little attention has been given to the factors associated with basilar artery (BA) dolichosis. This study aims to elucidate the prevalence and associated factors of BA dolichosis in patients with acute cerebral infarction (ACI). Methods: We collected the clinical and laboratory data of 719 patients with ACI admitted to our department. Magnetic resonance angiography was used to evaluate the geometric parameters of the BA and intracranial vertebral arteries (VAs). A BA curve length > 29.5 mm or bending length (BL) > 10 mm was identified as BA dolichosis. Univariate and multivariate logistic regression were performed to determine the factors associated with BA dolichosis. Results: Among 719 patients with ACI, 238 (33.1%) demonstrated BA dolichosis, including 226 (31.4%) with simple BA dolichosis and 12 (1.7%) with basilar artery dolichoectasia (BADE). Pearson correlation analyses showed that BA curve length was positively correlated with BL (r = 0.605). Multivariate logistic regression analysis demonstrated that current smoking (OR = 1.50, 95% CI: 1.02-2.21, p = 0.039), diabetes mellitus (OR = 1.66, 95% CI: 1.14-2.41, p = 0.008), BA diameter (OR = 3.04, 95% CI: 2.23-4.13, p < 0.001), BA bending (OR = 4.24, 95% CI: 2.91-6.17, p < 0.001) and BL (OR = 1.45, 95% CI: 1.36-1.55, p < 0.001) were significantly associated with BA dolichosis. Conclusion: This study suggests that BA dolichosis was common in patients with ACI, and the morphological parameters of the vertebrobasilar artery and acquired risk factors (including smoking and diabetes) were risk factors for BA dolichosis.

5.
Lasers Med Sci ; 38(1): 86, 2023 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-36932298

RESUMO

Macrophages are the main mediators of the inflammatory response and play a major role in the onset and maintenance of periodontitis. Studies revealed that photobiomodulation (PBM) can change the polarization state of macrophages and inflammation reduction, although the cellular mechanisms are not fully elucidated. Here, the present study explored the effect of PBM (980 nm) on undifferentiated and M1-type macrophages and the underlying mechanism. RAW264.7 cells were exposed to laser irradiation under different laser parameters (0.5, 5.0, and 10.0 J/cm2) with or without LY294002 (an inhibitor of PI3K pathway). Then, confocal laser microscopy was used to observe cell differentiation; qPCR was performed to examine the gene expression and western blotting was used to detect the protein in the PI3K/AKT/mTOR pathway and activated macrophage markers. The obtained results revealed that 980 nm PBM increased the mRNA expression of iNOS, Il-10, Arg1, and Il-12 along with the inflammatory cytokines Tnfα, IL-1ß, and Il-6 in M0-type macrophages in dose-dependent manner. More interestingly, PBM at 5 J/cm2 decreased the mRNA expression of iNOS, Il-12, Tnfα, IL-1ß, and Il-6 and increased the expression of Arg1 and Il-10 by M1-type macrophages, along with the elevated expression of phosphorylation of AKT and mTOR. Moreover, PBM-induced M1-type macrophage polarization was significantly attenuated via LY294002 treatment. These suggest that 980 nm PBM could activate M0-type macrophages and increase M2/M1 ratio via the PI3K/AKT/mTOR pathway.


Assuntos
Interleucina-10 , Proteínas Proto-Oncogênicas c-akt , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Interleucina-12/farmacologia , Interleucina-6/metabolismo , Macrófagos/metabolismo , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Animais , Camundongos , Células RAW 264.7
8.
Inflammation ; 46(3): 876-891, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36598593

RESUMO

Macrophages actively participate in immunomodulatory processes throughout periodontal inflammation. Regulation of M1/M2 polarization affects macrophage chemokine and cytokine secretion, resulting in a distinct immunological status that influences prognosis. Semaphorin 3A (Sema3A), a neurite growth factor, exerts anti-inflammatory effects. In this study, we investigated the immunomodulation of Sema3A on macrophage-related immune responses in vivo and in vitro. Topical medications of Sema3A in mice with periodontitis alleviated inflammatory cell infiltration into gingival tissue and reduced areas with positive IL-6 and TNFα expression. We observed that the positive area with the M2 macrophage marker CD206 increased and that of the M1 macrophage marker iNOS decreased in Sema3A-treated mice. It has been postulated that Sema3A alleviates periodontitis by regulating alternative macrophage activation. To understand the mechanism underlying Sema3A modulation of macrophage polarization, an in vitro macrophage research model was established with RAW264.7 cells, and we demonstrated that Sema3A promotes LPS/IFNγ-induced M1 macrophages to polarize into M2 macrophages and activates the PI3K/AKT/mTOR signaling pathways. Inhibition of the PI3K signaling pathway activation might reduce anti-inflammatory activity and boost the expression of the inflammatory cytokines, iNOS, IL-12, TNFα, and IL-6. This study indicated that Sema3A might be a feasible drug to regulate alternative macrophage activation in the inflammatory response and thus alleviate periodontitis.


Assuntos
Periodontite , Semaforina-3A , Camundongos , Animais , Semaforina-3A/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Ativação de Macrófagos , Interleucina-6/farmacologia , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo , Anti-Inflamatórios/farmacologia , Periodontite/tratamento farmacológico
9.
Photochem Photobiol ; 99(4): 1181-1192, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36437584

RESUMO

Increasing evidence suggests stem cells from human exfoliated deciduous teeth (SHEDs) serve as desirable sources of dentin regeneration. Photobiomodulation (PBM) has shown great potential in enhancing the proliferation and osteogenesis of human bone marrow mesenchymal stem cells (hBMMSCs). However, the specific role of PBM in odontogenic differentiation of SHEDs is little know, and we further investigated potential mechanism of PBM osteo/odontogenisis. A 980 nm diode laser with different energy densities of (0.5, 5, 10 J cm-2 ) in a 100-mW continuous wave was used for irradiation every 24 h. Osteo/odontogenic differentiation of SHEDs was achieved by performing alkaline phosphatase (ALP) and alizarin red staining (ARS) and osteo/odontogenic markers were also evaluated by qRT-PCR and western blotting. Additionally, western blot and immunohistochemical staining were performed to evaluate the levels of BMP/Smad and Wnt/ß-catenin signaling-related proteins. We found that PBM at 5 J cm-1 increased mineral deposition and upregulated the expression of related osteo/odontogenic markers along with the elevated expression of ß-catenin and phosphorylation level of Smad1/5/9. Furthermore, Wnt signaling inhibition using DKK1 and BMP signaling inhibition using noggin inhibited PBM-induced osteo/odontogenic marker expression when used individually or jointly. In conclusion, PBM induces the osteo/odontogenic differentiation of SHEDs through cross talk between BMP/Smad and Wnt/ß-catenin signaling pathways.


Assuntos
Via de Sinalização Wnt , beta Catenina , Humanos , Diferenciação Celular/fisiologia , Células-Tronco , Dente Decíduo , Células Cultivadas , Proliferação de Células
10.
Neurol Sci ; 44(4): 1273-1280, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36564659

RESUMO

INTRODUCTION: Limited cross-sectional or case-control studies have identified the relationship between basilar artery (BA) curvature and posterior circulation infarction (PCI). This study aimed to identify the influence of BA curvature severity on the risk of PCI occurrence in patients without vertebrobasilar stenosis through a prospective cohort study. METHODS: In this study, we enrolled 171 patients with BA dolichosis but without vertebrobasilar stenosis. The BA geometric parameters were evaluated on MRA. The primary outcome was the occurrence of PCI, mainly referring to cerebellar and/or brainstem infarction. Cox proportional hazard models were used to detect possible predictors of PCI. RESULTS: Among them, 134 (78.4%) patients were diagnosed with BA curvature, including 124 with moderate curvature and 10 with prominent curvature. The defined PCI occurrence was observed in 32 (18.7%) patients with a median follow-up time of 45.6 months. Cox proportional hazard analysis showed that BA prominent curvature (HR = 6.09; 95% CI: 1.36-27.28; P = 0.018) significantly increased the risk of PCI occurrence, and bending length (BL) was also significantly associated with PCI occurrence, with the adjusted HR per 1-mm increase of BL of 1.09 (95% CI: 1.01-1.18; P = 0.040). In the subgroup analysis stratified by age, BA prominent curvature was highly associated with PCI occurrence in patients aged > 61 years (HR = 11.76; 95% CI: 1.21-113.90; P = 0.033). Additionally, good antiplatelet therapy adherence could significantly reduce the risk of PCI occurrence. CONCLUSION: BA curvature may increase the risk of PCI occurrence, especially in elderly patients with prominent curvature. Improving adherence to antiplatelet therapy can help reduce the risk of PCI occurrence.


Assuntos
Infartos do Tronco Encefálico , Insuficiência Vertebrobasilar , Idoso , Humanos , Pessoa de Meia-Idade , Artéria Basilar/diagnóstico por imagem , Estudos Prospectivos , Constrição Patológica , Estudos Transversais , Inibidores da Agregação Plaquetária/uso terapêutico , Insuficiência Vertebrobasilar/complicações , Insuficiência Vertebrobasilar/diagnóstico por imagem , Insuficiência Vertebrobasilar/epidemiologia , Infartos do Tronco Encefálico/complicações , Infartos do Tronco Encefálico/diagnóstico por imagem , Infartos do Tronco Encefálico/epidemiologia
11.
Front Cell Dev Biol ; 10: 921364, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36035997

RESUMO

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) belongs to the long non-coding RNA (LncRNA) family. LncRNA-MALAT1 is expressed in a variety of tissues and is involved in a variety of diseases and biological processes. Although LncRNA-MALAT1 is upregulated in a high-glucose microenvironment and may participate in odontogenic differentiation, the underlying mechanism is not yet well elucidated. Here, we show that MALAT1 was mainly expressed in the cytoplasm of dental pulp cells (DPCs) in situ hybridization. In addition, high levels of mineralization-related factors, namely, tumor growth factors ß 1 and 2 (TGFß-1 and TGFß-2), bone morphogenetic proteins 2 and 4 (BMP2 and BMP4), bone morphogenetic protein receptor 1 (BMPR1), SMAD family member 2 (SMAD2), runt-related transcription factor 2 (RUNX2), Msh homeobox 2 (MSX2), transcription factor SP7 (SP7), alkaline phosphatase (ALP), dentin matrix acidic phosphoprotein 1 (DMP1), and dentin sialophosphoprotein (DSPP), were expressed, and MALAT1 was significantly overexpressed in DPCs 7 and 14 days after mineralization induction in a high-glucose microenvironment, but only TGFß-1, BMP2, MSX2, SP7, ALP, and DSPP were significantly downregulated in DPCs after MALAT1 inhibition. MALAT1 may participate in the mineralization process of DPCs by regulating multiple factors (TGFß-1, BMP2, MSX2, SP7, ALP, and DSPP).

12.
BMC Ophthalmol ; 22(1): 96, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-35232403

RESUMO

BACKGROUND: The movement of intraventricular silicone oil observed in the supine position is extremely rare. Herein, we describe a patient who presented with dynamically moving silicone oil particles in the ventricle when changing position and provide an updated review of this phenomenon. CASE PRESENTATION: We report a case of a 70-year-old woman who presented with intraventricular hyperdensities that were occasionally found on brain computed tomography (CT). Initial nonenhanced brain CT demonstrated nondependent hyperdensities in the bilateral anterior horns of the lateral ventricles, the third ventricle, and the right suprasellar cistern, mimicking an intraventricular hemorrhage. Further brain magnetic resonance imaging (MRI) in the supine position revealed abnormal signals in the bilateral anterior horns of the lateral ventricles, the posterior horn of the right lateral ventricle, the third ventricle, the right suprasellar cistern, and the bilateral eyeballs, with isosignal intensities surrounded by low-signal chemical shift artifacts on T1-weighted imaging and variable signals (hypo- or hyperintensity) on T2-weighted imaging. The lesion in the anterior horn of the right ventricle largely moved to the posterior horn of the ipsilateral ventricle. The final craniocervical CT angiography showed that the lesion in the posterior horn had moved back to the anterior horn of the right lateral ventricle. These features were consistent with intraventricular silicone oil migration. The final spinal MRI did not demonstrate a migration of silicone oil into the spinal subarachnoid space. DISCUSSION AND CONCLUSIONS: This case report describes a dynamic process of silicone oil displacement in the supine position and provides a comprehensive imaging presentation. The moving pattern and a characteristic chemical shift artifact on MRI are key to the diagnosis and may help prevent unnecessary examinations or intervention.


Assuntos
Migração de Corpo Estranho , Descolamento Retiniano , Idoso , Ventrículos Cerebrais , Feminino , Migração de Corpo Estranho/etiologia , Ventrículos do Coração , Humanos , Imageamento por Ressonância Magnética/métodos , Descolamento Retiniano/patologia , Óleos de Silicone/efeitos adversos
13.
Front Neurol ; 12: 788145, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34966351

RESUMO

Background and Purpose: Patients with basilar artery (BA) dolichosis are at high risk of acute pontine infarction (API), but the association between BA dolichosis and long-term stroke recurrence has received little attention. We aimed to identify the effect of BA dolichosis on the risk of long-term brainstem infarction recurrence in patients with API. Methods: In this prospective cohort study, we enrolled 113 patients with API admitted to our department. BA dolichosis was diagnosed by a BA curve length >29.5 mm or bending length (BL) >10 mm on magnetic resonance angiography. The primary outcome was the occurrence of diffusion-weighted imaging (DWI)-confirmed brainstem infarction. The Cox proportional hazard model was used to detect possible predictors of brainstem infarction recurrence. Results: Among 113 patients with API, 39 (34.5%) patients had BA dolichosis, and DWI-confirmed brainstem infarction recurred in 15 (13.3%) patients with a mean follow-up time of 31.2 months; the estimated 5-year incidence of brainstem infarction recurrence was 23.1% in patients with BA dolichosis, which was significantly higher than the incidence of 8.1% in patients without BA dolichosis. Cox proportional hazard analysis showed that age ≥65 years (hazard ratio (HR) = 3.341, 95% confidence interval (CI): 1.079-10.348, P = 0.036) and BA dolichosis (HR = 3.048, 95% CI: 1.069-8.693, P = 0.037) were significantly associated with a higher risk of brainstem infarction recurrence. In a subgroup analysis stratified by age, the patients aged ≥65 years with BA dolichosis had a higher risk of brainstem infarction recurrence (HR = 7.319, 95% CI: 1.525-35.123, P = 0.013). Conclusions: This study indicates that BA dolichosis may increase the risk of long-term brainstem infarction recurrence in patients with API, especially in elderly patients, and therefore warrants more attention in clinical practice.

14.
Stem Cells Int ; 2021: 6685307, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33936213

RESUMO

BACKGROUND: Exosomes from human dental pulp stem cells (hDPSCs) were indicated to play a positive role in vascular regeneration processes. But the angiogenic capabilities of exosomes from inflammatory hDPSCs and the underlying mechanism remain unknown. In this study, the inflammatory factor lipopolysaccharide (LPS) was used to stimulate hDPSCs, and exosomes were extracted from these hDPSCs. The proangiogenic potential of exosomes was examined, and the underlying mechanism was studied. METHOD: Exosomes were isolated from hDPSCs with or without LPS stimulation (N-EXO and LPS-EXO) and cocultured with human umbilical vein endothelial cells (HUVECs). The proangiogenic potential of exosomes was evaluated by endothelial cell proliferation, migration, and tube formation abilities in vitro. To investigate the proangiogenic mechanism of LPS-EXO, microRNA sequencing was performed to explore the microRNA profile of N-EXO and LPS-EXO. Gene Ontology (GO) analysis was used to study the functions of the predicted target genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was used to estimate the signaling pathways associated with the inflammation-induced angiogenesis process. RESULT: Compared to the uptake of N-EXO, uptake of LPS-EXO activated the angiogenic potential of HUVECs by promoting the proliferation, migration, and tube formation abilities in vitro. The mRNA expression levels of vascular endothelial growth factor (VEGF) and kinase-insert domain-containing receptor (KDR) in the LPS-EXO group were significantly higher than those in the N-EXO group. MicroRNA sequencing showed that 10 microRNAs were significantly changed in LPS-EXO. Pathway analysis showed that the genes targeted by differentially expressed microRNAs were involved in multiple angiogenesis-related pathways. CONCLUSION: This study revealed that exosomes derived from inflammatory hDPSCs possessed better proangiogenic potential in vitro. This is the first time to explore the role of exosomal microRNA from hDPSCs in inflammation-induced angiogenesis. This finding sheds new light on the effect of inflammation-stimulated hDPSCs on tissue regeneration.

15.
J Dent Sci ; 16(1): 318-326, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33384815

RESUMO

BACKGROUND/PURPOSE: Dental pulp stem cells can be isolated from human teeth with deep caries (cDPSCs), but their biological characteristics are still unclear. The aim of this study was to investigate the angiogenic potential of cDPSCs and compare them to dental pulp stem cells from human normal teeth (nDPSCs). MATERIALS AND METHODS: Cells were isolated from human pulp tissue of normal and infected teeth with deep caries. Basic mesenchymal stem cell (MSC) characterization was conducted. Colony forming units and proliferation ability were evaluated in nDPSCs and cDPSCs. Expression of VEGF in both tissues and cells was examined by immunohistochemical staining. After stimulating nDPSCs and cDPSCs with an angiogenic medium, angiogenic markers were evaluated by qRT-PCR and western blotting. Finally, tube formation assays were used to evaluate the in vitro angiogenesis potential of both cell populations. RESULTS: Both nDPSCs and cDPSCs possessed typical MSC characteristics. cDPSCs had enhanced colony formation and proliferation capacities than nDPSCs did. The expression of VEGF was higher in pulp tissue from teeth with deep caries and cDPSCs than in normal tissue and nDPSCs. When both cell types were grown in vitro under angiogenic conditions, cDPSCs expressed a higher level of angiogenic markers and showed a stronger angiogenesis potential than nDPSCs did. CONCLUSION: cDPSCs maintained MSC traits and presented a higher angiogenesis potential than nDPSCs.

16.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-876366

RESUMO

@#Vital pulp therapy aims to maintain healthy pulp tissue as much as possible to improve the long-term survival of teeth. It has limited indications and uncertain curative effects. The pathological changes in inflamed pulp are the histological basis for the determination of treatment strategies and the treatment outcome; however, pulp sensitivity testing cannot reflect the actual histological status of the pulp. With the development of basic and clinical research on vital pulp therapy, the innovation of modern diagnostic and therapeutic technology and capping material, vital pulp therapy can be used as a treatment of teeth on which it was previously thought pulpectomy was necessary. Based on the evidence-based literature, this paper analyzes and summarizes the pathological changes of pulpitis and clinical research on the treatment of pulpitis. Vital pulp therapy can be a treatment for mature teeth with carious exposure and symptoms of irreversible pulpitis if comprehensive applications, including laser Doppler flowmetry, tissue oxygen monitoring, magnetic resonance imaging and microscopy, are used to determine the degree of pulp retention and if infection control and the use of biocompatible capping material are emphasized. In the future, it will be necessary to improve the success rate of vital pulp therapy for the treatment of pulpitis through research on the mechanism of pulp repair and regeneration, the precise diagnosis of pulpitis, and the development of pulp capping materials.

17.
Arch Oral Biol ; 113: 104691, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32247880

RESUMO

INTRODUCTION: Angiogenesis is important in pulp-dentin formation. Among the regulatory factors, long noncoding RNA (LncRNA) is a class of functional RNA molecules that are not translated into protein and involved in regulating multiple physiological processes. The different expression of LncRNA and its target gene in dental pulp stem cells (DPSCs) were explored and may provide a theoretical basis for future regulation of dental pulp angiogenesis. METHODS: In this study, we cultured DPSCs from healthy dental pulp tissues and divided them into two groups: the normal DPSCs and the DPSCs cultured in vascular induction medium. In total, 40,173 LncRNA probes and 20,730 protein coding mRNAs were detected through microarray, which were then verified by the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) method. RESULTS: The result of differential expressions measured in LncRNA through microarray showed that 376 LncRNAs increased significantly and 426 were downregulated among the two groups of cells. Moreover, the mRNA microarray in normal cultured DPSCs showed that 629 LncRNAs were significantly upregulated, while 529 of them were downregulated compared with the DPSCs that were cultured in vascular induction medium. Gene ontology (GO) analysis inferred the molecular function of mRNAs. Pathway analysis showed that 52 signaling pathways were involved in the differentiation process of DPSCs. qRT-PCR analysis, conducted for validation, showed results consistent with the microarray analysis. CONCLUSIONS: We found that a number of different regulators are involved in inducing vascular differentiation of DPSCs, which provides a foundation for subsequent experiments.


Assuntos
Polpa Dentária/citologia , Neovascularização Fisiológica , RNA Longo não Codificante/metabolismo , Células-Tronco/metabolismo , Diferenciação Celular , Células Cultivadas , Humanos , RNA Mensageiro
20.
Mol Med Rep ; 20(4): 3924-3932, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31485628

RESUMO

The present study aimed to investigate the effects of vascular endothelial growth factor (VEGF) and insulin­like growth factor­1 (IGF­1) on the proliferation, migration and differentiation of human carious dental pulp stem cells (hCDPSCs), and to elucidate the underlying mechanism(s). Cell counting kit­8 assay was used to detect the effect of different concentrations of IGF­1 and VEGF on the proliferation of hCDPSCs. Transwell assay was used to detect the migratory ability of the hCDPSCs. Alizarin red and alkaline phosphatase (ALP) staining were used to detect the osteogenic ability of hCDPSCs, whereas the angiogenic ability of the hCDPSCs was tested by tube formation assay. Reverse transcription­quantitative polymerase chain reaction (RT­qPCR) and western blotting were used to detect the expression levels of associated genes and proteins. IGF­1 (100 ng/ml) or VEGF (25 ng/ml) alone were revealed to be able to promote proliferation and migration of hCDPSCs; however, the combined use of IGF­1 and VEGF enhanced this effect when compared with the use of either agent in isolation. Alizarin red and ALP staining revealed that the use of either VEGF or IGF­1 alone did not result in any significant effects, whereas their use in combination promoted the osteogenic differentiation of hCDPSCs. In addition, the RT­qPCR and western blotting analyses revealed that the expression levels of Runt­related transcription factor 2 (RUNX2), bone sialoprotein (BSP) and ALP were increased upon combined treatment of the cells with VEGF and IGF­1. The expression levels of VEGF and plateletderived growth factor (PDGF) in hCDPSCs were enhanced upon treatment with either VEGF or IGF­1 in isolation, with greater effects observed when VEGF and IGF­1 were added in combination, indicating that VEGF and IGF­1 may exert a synergistic role in these events. Further experiments revealed that the combination of VEGF and IGF­1 led to an activation of the AKT signaling pathway. The proliferation and angiogenesis of hCDPSCs were also shown to be more effective compared with treatment with either VEGF or IGF­1 in isolation. Taken together, the present study has demonstrated that the combined use of VEGF and IGF­1 leads to an increase in the proliferation, migration, osteogenesis and angiogenesis of hCDPSCs and, furthermore, these signaling molecules may mediate their effects via activation of the AKT signaling pathway.


Assuntos
Polpa Dentária/citologia , Fator de Crescimento Insulin-Like I/metabolismo , Células-Tronco/citologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto , Diferenciação Celular , Movimento Celular , Proliferação de Células , Células Cultivadas , Cárie Dentária/metabolismo , Polpa Dentária/metabolismo , Feminino , Humanos , Masculino , Neovascularização Fisiológica , Osteogênese , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Células-Tronco/metabolismo
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