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1.
Braz Oral Res ; 33: e117, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31939498

RESUMO

The aim of this study was to evaluate the effect of mineral trioxide aggregate (MTA) and Brazilian propolis on the cell viability, mineralization, anti-inflammatory ability, and migration of human dental pulp cells (hDPCs). The cell viability was evaluated with CCK-8 kit after 1, 5, 7, and 9 days. The deposition of calcified matrix and the expression of osteogenesis-related genes were evaluated by Alizarin Red staining and real-time PCR after incubation in osteogenic medium for 21 days. The expression of inflammation-related genes in cells was determined after exposure to 1 µg/mL LPS for 3 h. Finally, the numbers of cells that migrated through the permeable membranes were compared during 15 h. Propolis and MTA significantly increased the viability of hDPCscompared to the control group on days 7 and 9. In the propolis group, significant enhancement of osteogenic potential and suppressed expression of IL-1ß and IL-6 was observed after LPS exposure compared to the MTA and control groups. The number of migration cells in the propolis group was similar to that of the control group, while MTA significantly promoted cell migration. Propolis showed comparable cell viability to that of MTA and exhibited significantly higher anti-inflammatory and mineralization promotion effects on hDPCs.


Assuntos
Compostos de Alumínio/farmacologia , Anti-Inflamatórios/farmacologia , Compostos de Cálcio/farmacologia , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Óxidos/farmacologia , Própole/farmacologia , Silicatos/farmacologia , Antraquinonas , Brasil , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Combinação de Medicamentos , Humanos , Interleucina-1beta/análise , Interleucina-6/análise , Odontoblastos/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Estatísticas não Paramétricas , Fator de Necrose Tumoral alfa/análise
2.
J Appl Oral Sci ; 28: e20190215, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31939521

RESUMO

OBJECTIVE: This study evaluated the angiogenesis-enhancing potential of a tricalcium silicate-based mineral trioxide aggregate (ProRoot MTA), Biodentine, and a novel bioceramic root canal sealer (Well-Root ST) in human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPLSCs), and human tooth germ stem cells (hTGSCs). METHODOLOGY: Dulbecco's modified Eagle's medium was conditioned for 24 h by exposure to ProRoot MTA, Biodentine, or Well-Root ST specimens (prepared according to the manufacturers' instructions). The cells were cultured in these conditioned media and their viability was assessed with 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium (MTS) on days 1, 3, 7, 10, and 14. Angiogenic growth factors [platelet-derived growth factor (PDGF), basic fibroblast growth factor (FGF-2), and vascular endothelial growth factor (VEGF)] were assayed by sandwich enzyme-linked immunosorbent assay (ELISA) on days 1, 7, and 14. Human umbilical vein endothelial cell (HUVEC) migration assays were used to evaluate the vascular effects of the tested materials at 6-8 h. Statistical analyses included Kruskal-Wallis, Mann-Whitney U, and Friedman and Wilcoxon signed rank tests. RESULTS: None of tricalcium silicate-based materials were cytotoxic and all induced a similar release of angiogenic growth factors (PDGF, FGF-2, and VEGF) (p>0.05). The best cell viability was observed for hDPSCs (p<0.05) with all tricalcium silicate-based materials at day 14. Tube formation by HUVECs showed a significant increase with all tested materials (p<0.05). CONCLUSION: The tricalcium silicate-based materials showed potential for angiogenic stimulation of all stem cell types and significantly enhanced tube formation by HUVECs.


Assuntos
Indutores da Angiogênese/farmacologia , Compostos de Cálcio/farmacologia , Cerâmica/farmacologia , Materiais Restauradores do Canal Radicular/farmacologia , Silicatos/farmacologia , Células-Tronco/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Fator 2 de Crescimento de Fibroblastos/análise , Fator 2 de Crescimento de Fibroblastos/efeitos dos fármacos , Citometria de Fluxo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Teste de Materiais , Neovascularização Fisiológica/efeitos dos fármacos , Ligamento Periodontal/citologia , Ligamento Periodontal/efeitos dos fármacos , Fator de Crescimento Derivado de Plaquetas/análise , Fator de Crescimento Derivado de Plaquetas/efeitos dos fármacos , Reprodutibilidade dos Testes , Estatísticas não Paramétricas , Germe de Dente/citologia , Germe de Dente/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos
3.
J Appl Oral Sci ; 28: e20190105, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31800873

RESUMO

Calcium aluminate cement (CAC) has been highlighted as a promising alternative for endodontic use aiming at periapical tissue repair. However, its effects on dental pulp cells have been poorly explored. OBJECTIVE: This study assessed the impact of calcium chloride (CaCl2) and bismuth oxide (Bi2O3) or zinc oxide (ZnO) additives on odontoblast cell response to CAC. METHODOLOGY: MDPC-23 cells were exposed for up to 14 d: 1) CAC with 2.8% CaCl2 and 25% ZnO (CACz); 2) CAC with 2.8% CaCl2 and 25% Bi2O3 (CACb); 3) CAC with 10% CaCl2 and 25% Bi2O3 (CACb+); or 4) mineral trioxide aggregate (MTA), placed on inserts. Non-exposed cultures served as control. Cell morphology, cell viability, gene expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), and dentin matrix protein 1 (DMP-1), ALP activity, and extracellular matrix mineralization were evaluated. Data were compared using ANOVA (α=5%). RESULTS: Lower cell density was detected only for MTA and CACb+ compared with Control, with areas showing reduced cell spreading. Cell viability was similar among groups at days one and three (p>0.05). CACb+ and MTA showed the lowest cell viability values at day seven (p>0.05). CACb and CACb+ promoted higher ALP and BSP expression compared with CACz (p<0.05); despite that, all cements supported ALP activity. Matrix mineralization were enhanced in CACb+ and MTA. CONCLUSION: In conclusion, CAC with Bi2O3, but not with ZnO, supported the expression of odontoblastic phenotype, but only the composition with 10% CaCl2 promoted mineralized matrix formation, rendering it suitable for dentin-pulp complex repair.


Assuntos
Compostos de Alumínio/química , Compostos de Alumínio/farmacologia , Compostos de Cálcio/química , Compostos de Cálcio/farmacologia , Cimentos Dentários/química , Cimentos Dentários/farmacologia , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Fosfatase Alcalina/análise , Fosfatase Alcalina/efeitos dos fármacos , Animais , Bismuto/química , Bismuto/farmacologia , Cloreto de Cálcio/química , Cloreto de Cálcio/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Combinação de Medicamentos , Expressão Gênica/efeitos dos fármacos , Teste de Materiais , Camundongos , Odontoblastos/efeitos dos fármacos , Óxidos/química , Óxidos/farmacologia , Reprodutibilidade dos Testes , Silicatos/química , Silicatos/farmacologia , Fatores de Tempo , Óxido de Zinco/química , Óxido de Zinco/farmacologia
4.
Arch Oral Biol ; 109: 104572, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31600663

RESUMO

OBJECTIVE: To compare three different xeno-free protocols for neural differentiation of human dental pulp stem cells (DPSC). METHODS: DPSC were treated with three different media to induce neural differentiation namely N1 (DMEM for 5 days), N2 (PSC neural induction media for 7 days) and N3 (neural media with B27 supplement, 40 ng/ml bFGF and 20 ng/ml EGF for 21 days). Cell proliferation (MTS assay), morphology, gene (qPCR for NESTIN, VIMENTIN, TUB-3, ENO2, NF-M and NF-H) and protein expression (flow cytometry) of neurogenic markers were assessed at different time points and compared to untreated cells (DMEM supplemented with 10% FBS). Statistical analysis was performed with global significance level of 5%. RESULTS: N1 and N2 formulations increased the genetic expression of two out of six genes TUB-3, NF-M and TUB-3, NF-H, respectively, whereas N3 elevated the expression of all genes by the late stage. N3 also stimulated protein expression for NESTIN, TUB-3 and NF-H. Cells treated with both N2 and N3 presented neuron-like morphology, decreased proliferation and expression of stemness genes at protocol end point. CONCLUSION: N3 was the most effective formulation in promoting a neurogenic shift in gene and protein expression. Cells provided with the N3 formulation exhibited neuron-like morphology, elaborating axonal-like projections concomitant with cell cycle withdrawal and reduced expression of stemness genes indicating greater commitment to a neurogenic lineage.


Assuntos
Técnicas de Cultura de Células , Diferenciação Celular , Polpa Dentária/citologia , Neurogênese , Células-Tronco/citologia , Células Cultivadas , Meios de Cultura , Humanos , Neurônios/citologia
5.
Arch Oral Biol ; 109: 104582, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31605918

RESUMO

OBJECTIVE: The aim of this study was to evaluate the proliferation and odontogenic differentiation of human dental pulp cells (hDPCs) and human umbilical cord mesenchymal stem cells (hUCMSCs) in three-dimensional co-culture system which was established with the help of bone morphogenetic protein-2 (BMP-2) and hydrogel. METHODS: hDPCs and hUCMSCs were cultured in different concentrations of hydrogel to explore the more suitable concentrations for subsequent experiments. hUCMSCs and hDPCs induced by BMP-2 were co-cultured in the hydrogel. MTT assay was used to measure the cell viability. The differentiation into odontoblast-like cells were measured by the mRNA expression of dentin salivary phosphoprotein (DSPP), dentin matrix protein-1 (DMP-1), alkaline phosphatase and osteocalcin. Alizarin red staining was performed for the formation of mineralized nodules. RESULTS: hUCMSCs and hDPCs could grow and proliferate in hydrogel scaffold. The growth rate of cells in lower concentrations hydrogels were higher than that of high concentrations hydrogels (P < 0.05). The study showed that 0.25% hydrogel scaffold was more suitable for subsequent experiments than other groups. Compared with hUCMSCs-monoculture and hDPCs-monoculture, the co-culture groups exhibited more proliferative potential, alkaline phosphatase activity and mineralization nodule formation (P < 0.05). The mRNA expression in co-culture groups were higher than that of hUCMSCs-monoculture, closed to or even higher than that of hDPCs-monoculture. CONCLUSION: 0.25% hydrogel was the suitable concentration in co-culture system for subsequent experiments. The co-culture groups had stronger abilities of odontoblastic differentiation and mineralization than cells-monoculture groups, indicated that the co-culture conditions could regulate cell proliferation and differentiation within a certain range.


Assuntos
Diferenciação Celular , Polpa Dentária/citologia , Hidrogéis , Células-Tronco Mesenquimais/citologia , Fosfatase Alcalina/genética , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura , Proteínas da Matriz Extracelular/genética , Humanos , Odontoblastos/citologia , Osteocalcina/genética , Fosfoproteínas/genética , Sialoglicoproteínas/genética , Cordão Umbilical/citologia
6.
J Photochem Photobiol B ; 203: 111727, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31862637

RESUMO

Blindness and vision impairment are caused by irremediable retinal degeneration in affected individuals worldwide. Cell therapy for a retinal replacement can potentially rescue their vision, specifically for those who lost the light sensing photoreceptors in the eye. As such, well-characterized retinal cells are required for the replacement purposes. Stem cell-based therapy in photoreceptor and retinal pigment epithelium transplantation is well received, however, the drawbacks of retinal transplantation is the limited clinical protocols development, insufficient number of transplanted cells for recovery, the selection of potential stem cell sources that can be differentiated into the target cells, and the ability of cells to migrate to the host tissue. Dental pulp stem cells (DPSC) belong to a subset of mesenchymal stem cells, and are recently being studied due to its high capability of differentiating into cells of the neuronal lineage. In this review, we look into the potential uses of DPSC in treating retinal degeneration, and also the current data supporting its application.


Assuntos
Polpa Dentária/citologia , Degeneração Retiniana/terapia , Transplante de Células-Tronco , Humanos , Células Fotorreceptoras/fisiologia , Retina/fisiologia , Células-Tronco/citologia
7.
Cient. dent. (Ed. impr.) ; 16(3): 201-207, sept.-dic. 2019. tab, ilus
Artigo em Espanhol | IBECS | ID: ibc-ET2-3541

RESUMO

Introducción: El empleo de dientes autógenos, como material de injerto, es una opción terapéutica actual en casos de regeneración ósea. Su obtención se ha facilitado con la introducción de dispositivos capaces de procesar los dientes. El objetivo de este trabajo es realizar, a propósito de un caso clínico, una revisión de la literatura sobre el uso de dientes autólogos como material de injerto óseo y los dispositivos para su procesamiento. Caso clínico: Paciente varón de 18 años que acude a consulta presentando un cordal inferior retenido. El diagnóstico determinó la necesidad de extraer el diente y se informó al paciente de la posibilidad de utilizarlo como material de regeneración ósea. Tras la exodoncia, el diente procesado con el dispositivo Tooth Transformer(R) (Imbiodent), fue utilizado como material de injerto autólogo. El postoperatorio no presentó ninguna complicación y la evaluación radiográfica, tras 8 días y tras 10 semanas, mostró una evolución favorable del tratamiento. Discusión: La dentina desmineralizada es un material orgánico cuyo potencial reside en los factores de crecimiento que contiene para estimular la formación y reparación ósea. No obstante, no existe consenso sobre el grado de desmineralización o tamaño de partícula ideal. La reciente introducción de dispositivos, capaces de procesar dientes, facilita la obtención de un material de injerto dental para su uso en terapias de regeneración ósea. Conclusión: El uso de dientes autólogos constituye una alternativa prometedora en el campo de los injertos óseos. La técnica de transformación del diente es sencilla con el empleo de los dispositivos actuales


Introduction: The use of autogenous teeth, as graft material, is a current therapeutic option in cases of bone regeneration. Its obtention has been facilitated by the introduction of devices capable of processing teeth. The aim of this article is to perform, based on a clinical case, a review of the literature about the use of autologous teeth as bone graft material and the devices for its processing. Clinical case: Male patient, 18 years of age, who comes to the dental office presenting a lower wisdom retained. Extraction of the tooth was determined by diagnosis and the patient was informed about the possibility of using it as bone regeneration material. After the extraction, the tooth was processed by the Tooth Transformer(R) (Imbiodent) device and was used as autologous graft material. No postoperative complications were presented and the radiographic evaluation, at 8 days and 10 weeks, showed a favorable evolution of the treatment. Discusion: Demineralized dentin is a organic material whose potential relies in the growth factors it contains to stimulate bone formation and repair. However, there is no consensus on the degree of demineralization or the ideal particle size. The recent introduction of devices, capable of processing teeth, enables the obtention of a dental graft material for bone regeneration therapies. Conclusion: The use of autologous teeth is a promising alternative in the bonev grafts field. The technique of tooth transformation is simple with the use of the current devices


Assuntos
Humanos , Masculino , Adolescente , Regeneração Óssea , Polpa Dentária/citologia , Dentina , Carga Imediata em Implante Dentário/métodos , Transplante Ósseo/métodos , Procedimentos Cirúrgicos Reconstrutivos , Desmineralização do Dente , Esmalte Dentário/química , Procedimentos Cirúrgicos Bucais/métodos
8.
Braz Oral Res ; 33: e059, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31664357

RESUMO

We recently demonstrated that a co-culture system of human umbilical vein endothelial cells (HUVECs) and human dental pulp stem cells (hDPSCs) could enhance angiogenesis ability in vitro. However, whether tumor necrosis factor α (TNF-α) could promote blood vessel formation during pulp regeneration remained unknown. The aim of this study was to investigate the effects of TNF-α on the formation of endothelial tubules and vascular networks in a co-culture system of hDPSCs and HUVECs. hDPSCs were co-cultured with HUVECs at a ratio of 1:5. The Matrigel assay was performed to detect the total tubule branching lengths and numbers of branches, and the Cell-Counting Kit 8 assay was performed to examine the effect of TNF-α on cell proliferation. Real-time polymerase chain reactions and western blot were used to detect vascular endothelial growth factor (VEGF) mRNA and protein expression. The Matrigel assay showed significantly greater total branching lengths and numbers of branches formed in the experimental groups treated with different concentrations of TNF-α compared with the control group. The decomposition times of the tubule structures were also significantly prolonged (P < 0.05). Treatment with 50 ng/ml TNF-α did not significantly change the proliferation of co-cultured cells, but it significantly increased the VEGF mRNA and protein expression levels (p < 0.05). In addition, the migration abilities of HUVECs and hDPSCs increased after co-culture with TNF-α (p < 0.05). TNF-α enhanced angiogenic ability in vitro in the co-culture system of hDPSCs and HUVECs.


Assuntos
Indutores da Angiogênese/farmacologia , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Adolescente , Adulto , Western Blotting , Contagem de Células , Ensaios de Migração Celular , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Células Cultivadas , Colágeno , Polpa Dentária/fisiologia , Combinação de Medicamentos , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Laminina , Neovascularização Fisiológica/fisiologia , Proteoglicanas , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Reprodutibilidade dos Testes , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Adulto Jovem
9.
Cell Prolif ; 52(6): e12675, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31553127

RESUMO

OBJECTIVES: To evaluate the regenerative potential of human dental pulp stem cells (hDPSCs) in an animal model of stress urinary incontinence (SUI). SUI, an involuntary leakage of urine, is due to physical stress involving an increase in bladder pressure and a damage of external urethral sphincter affecting muscles and nerves. Conventional therapies can only relieve the symptoms. Human DPSCs are characterized by peculiar stemness and immunomodulatory properties and might provide an alternative tool for SUI therapy. MATERIALS AND METHODS: In vitro phase: hDPSCs were induced towards the myogenic commitment following a 24 hours pre-conditioning with 5-aza-2'-deoxycytidine (5-Aza), then differentiation was evaluated. In vivo phase: pudendal nerve was transected in female rats to induce stress urinary incontinence; then, pre-differentiated hDPSCs were injected in the striated urethral sphincter. Four weeks later, urethral sphincter regeneration was assayed through histological, functional and immunohistochemical analyses. RESULTS: Human DPSCs were able to commit towards myogenic lineage in vitro and, four weeks after cell injection, hDPSCs engrafted in the external urethral sphincter whose thickness was almost recovered, committed towards myogenic lineage in vivo, promoted vascularization and an appreciable recovery of the continence. Moreover, hDPSCs were detected within the nerve, suggesting their participation in repair of transected nerve. CONCLUSIONS: These promising data and further investigations on immunomodulatory abilities of hDPSCs would allow to make them a potential tool for alternative therapies of SUI.


Assuntos
Polpa Dentária/efeitos dos fármacos , Células-Tronco/citologia , Uretra/efeitos dos fármacos , Incontinência Urinária por Estresse/tratamento farmacológico , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Polpa Dentária/citologia , Modelos Animais de Doenças , Feminino , Humanos , Ratos , Uretra/citologia
10.
Artif Cells Nanomed Biotechnol ; 47(1): 3431-3437, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31411067

RESUMO

Presently, tissue engineering has been developed as an effective option in the restoration and repair of tissue defects. One of the tissue engineering strategies is to use both biodegradable scaffolds and stimulating factors for enhancing cell responses. In this study, the effect of zeolite was assessed on cell viability, proliferation, osteo/odontogenic differentiation, and mineralization of human dental pulp stem cells (hDPSCs) cultured on poly (ε-coprolactone) - poly (ethylene glycol)-poly (ε-caprolactone) (PCL-PEG-PCL) nanofibers. For this purpose, PCL-PEG-PCL nanofibrous scaffolds incorporated with zeolite were prepared via electrospinning. Both PCL-PEG-PCL and PCL-PEG-PCL/Zeolite nanofibrous scaffolds revealed bead-less constructions with average diameters of 430 nm and 437 nm, respectively. HDPSCs were transferred to PCL-PEG-PCL nanofibrous scaffolds containing zeolite nanoparticles. Cell adhesion and proliferation of hDPSCs and their osteo/odontogenic differentiation on these scaffolds were evaluated using MTT assay, Alizarin red S staining, and qRT-PCR assay. The results revealed that PCL-PEG-PCL/Zeolite nanofibrous scaffolds could support better cell adhesion, proliferation and osteogenic differentiation of hDPSCs and as such is expected to be a promising scaffold for bone tissue engineering applications.


Assuntos
Polpa Dentária/citologia , Nanofibras/química , Osteogênese/efeitos dos fármacos , Poliésteres/farmacologia , Polietilenoglicóis/farmacologia , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Zeolitas/química , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Poliésteres/química , Polietilenoglicóis/química , Células-Tronco/metabolismo , Engenharia Tecidual/métodos , Tecidos Suporte/química
11.
Int J Mol Sci ; 20(15)2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31370244

RESUMO

Aneurysmal subarachnoid hemorrhage (aSAH), characterized by the extravasation of blood into the subarachnoid space caused by an intracranial aneurysm rupture, may lead to neurocognitive impairments and permanent disability and usually carries poor outcome. Dental or gingiva-derived stem cells have been shown to contribute to immune modulation and neuroregeneration, but the underlying mechanisms are unclear. In the present study, we sought to investigate whether dental pulp stem cells (DPSCs) secrete certain factor(s) that can ameliorate the neural damage and other manifestations in a rat aSAH model. Twenty-four hours after the induction of aSAH, microthrombosis, cortical vasoconstriction, and the decrease in microcirculation and tissue oxygen pressure were detected. Intrathecal administration of DPSC-derived conditioned media (DPSC-CM) ameliorated aSAH-induced vasoconstriction, neuroinflammation, and improved the oxygenation in the injured brain. Rotarod test revealed that the aSAH-induced cognitive and motor impairments were significantly improved by this DPSC-CM administration. Cytokine array indicated the major constituent of DPSC-CM was predominantly insulin growth factor-1 (IGF-1). Immunohistochemistry staining of injured brain tissue revealed the robust increase in Iba1-positive cells that were also ameliorated by DPSC-CM administration. Antibody-mediated neutralization of IGF-1 moderately deteriorated the rescuing effect of DPSC-CM on microcirculation, Iba1-positive cells in the injured brain area, and the cognitive/motor impairments. Taken together, the DPSC-derived secretory factors showed prominent therapeutic potential for aSAH. This therapeutic efficacy may include improvement of microcirculation, alleviation of neuroinflammation, and microglial activation; partially through IGF-1-dependent mechanisms.


Assuntos
Isquemia Encefálica/tratamento farmacológico , Meios de Cultivo Condicionados/farmacologia , Transtornos Neurocognitivos/tratamento farmacológico , Fármacos Neuroprotetores/farmacologia , Transtornos Psicomotores/tratamento farmacológico , Hemorragia Subaracnóidea/tratamento farmacológico , Trombose/tratamento farmacológico , Animais , Isquemia Encefálica/genética , Isquemia Encefálica/metabolismo , Isquemia Encefálica/fisiopatologia , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Meios de Cultivo Condicionados/química , Polpa Dentária/citologia , Polpa Dentária/metabolismo , Modelos Animais de Doenças , Expressão Gênica , Injeções Espinhais , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Masculino , Microcirculação/efeitos dos fármacos , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Transtornos Neurocognitivos/genética , Transtornos Neurocognitivos/metabolismo , Transtornos Neurocognitivos/fisiopatologia , Fármacos Neuroprotetores/química , Consumo de Oxigênio/efeitos dos fármacos , Transtornos Psicomotores/genética , Transtornos Psicomotores/metabolismo , Transtornos Psicomotores/fisiopatologia , Ratos , Ratos Wistar , Teste de Desempenho do Rota-Rod , Células-Tronco/química , Células-Tronco/citologia , Células-Tronco/metabolismo , Hemorragia Subaracnóidea/genética , Hemorragia Subaracnóidea/metabolismo , Hemorragia Subaracnóidea/fisiopatologia , Trombose/genética , Trombose/metabolismo , Trombose/fisiopatologia , Vasoconstrição/efeitos dos fármacos
12.
Braz Oral Res ; 33: e084, 2019 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-31460610

RESUMO

This study aimed to evaluate the role of photobiomodulation (PBM) in apexification and apexogenesis of necrotic rat molars with an open apex. Rat molars were exposed to the oral environment for 3 weeks. Canals were rinsed with 2.5% NaOCl and 17% EDTA, filled with antibiotic paste and sealed. After 7 days, canals were rinsed and divided into six groups (n=6): mineral trioxide aggregate (MTA); blood clot (BC); human dental pulp stem cells (hDPSC); MTA+PBM; BC+PBM; and hDPSC+PBM. In hDPSC groups, a 1% agarose gel scaffold was used. Two groups were not exposed: healthy tooth+PBM (n = 6), healthy tooth (n = 3); and one was exposed throughout the experiment: necrotic tooth (n = 3). In PBM groups, irradiation was performed with aluminum gallium indium phosphide (InGaAlP) diode laser for 30 days within 24-h intervals. After that, the specimens were processed for histological and immunohistochemical analyses. Necrotic tooth showed greater neutrophil infiltrate (p < 0.05). Necrotic tooth, healthy tooth, and healthy tooth+PBM groups showed absence of a thin layer of fibrous condensation in the periapical area. All the other groups stimulated the formation of a thicker layer of fibers (p < 0.05). All groups formed more mineralized tissue than necrotic tooth (p < 0.05). PBM associated with MTA, BC, or hDPSC formed more mineralized tissue (p < 0.05). MTA+PBM induced apexification (p < 0.05). Rabbit polyclonal anti-bone sialoprotein (BSP) antibody confirmed the histological findings of mineralized tissue formation, and hDPSC groups exhibited higher percentage of BSP-positive cells. It can be concluded that PBM improved apexification and favored apexogenesis in necrotic rat molars with an open apex.


Assuntos
Apexificação/métodos , Cavidade Pulpar/efeitos da radiação , Necrose da Polpa Dentária/radioterapia , Lasers Semicondutores/uso terapêutico , Terapia com Luz de Baixa Intensidade/métodos , Ápice Dentário/efeitos da radiação , Odontopatias/radioterapia , Compostos de Alumínio/uso terapêutico , Animais , Compostos de Cálcio/uso terapêutico , Polpa Dentária/citologia , Cavidade Pulpar/patologia , Necrose da Polpa Dentária/patologia , Combinação de Medicamentos , Imuno-Histoquímica , Sialoproteína de Ligação à Integrina/análise , Óxidos/uso terapêutico , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos Testes , Silicatos/uso terapêutico , Células-Tronco , Ápice Dentário/patologia , Odontopatias/patologia , Resultado do Tratamento
13.
Cell Prolif ; 52(6): e12676, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31424140

RESUMO

OBJECTIVES: Topographic cues can modulate morphology and differentiation of mesenchymal stem cells. This study aimed to determine how topographic cues of a novel bilayered poly (lactic-co-glycolic acid) (PLGA) scaffold affect osteogenic/odontogenic differentiation of dental pulp stem cells (DPSCs). METHODS: The surface morphology of the scaffolds was visualized by scanning electron microscope, and the surface roughness was measured by profilometry. DPSCs were cultured on each side of the scaffolds. Cell morphology, expression of Yes-associated protein (YAP) and osteogenic/odontogenic differentiation were analysed by immunohistochemistry, real-time polymerase chain reaction, and Alizarin Red S staining. In addition, cytochalasin D (CytoD), an F-actin disruptor, was used to examine the effects of F-actin on intracellular YAP localisation. Verteporfin, a YAP transcriptional inhibitor, was used to explore the effects of YAP signalling on osteogenic/odontogenic differentiation of DPSCs. RESULTS: The closed side of our scaffold showed smaller pores and less roughness than the open side. On the closed side, DPSCs exhibited enhanced F-actin stress fibre alignment, larger spreading area, more elongated appearance, predominant nuclear YAP localization and spontaneous osteogenic differentiation. Inhibition of F-actin alignments was correlated with nuclear YAP exclusion of DPSCs. Verteporfin restricted YAP localisation to the cytoplasm, down-regulated expression of early osteogenic/odontogenic markers and inhibited mineralization of DPSCs cultures. CONCLUSIONS: The surface topographic cues changed F-actin alignment and morphology of DPSCs, which in turn regulated YAP signalling to control osteogenic/odontogenic differentiation.


Assuntos
Actinas/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Diferenciação Celular/fisiologia , Polpa Dentária/citologia , Células-Tronco/citologia , Fatores de Transcrição/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Sinais (Psicologia) , Citoesqueleto/metabolismo , Humanos , Odontogênese/fisiologia , Osteogênese/fisiologia
14.
Cell Prolif ; 52(6): e12680, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31454111

RESUMO

OBJECTIVES: The odontoblastic differentiation of dental pulp stem cells (DPSCs) contributes to tertiary dentin formation. Our previous study indicated that epiregulin (EREG) enhanced odontogenesis potential of dental pulp. Here, we explored the effects of EREG during DPSC odontoblastic differentiation. METHODS: The changes in EREG were detected during tertiary dentin formation. DPSCs were treated with recombinant human EREG (rhEREG), EREG receptor inhibitor gefitinib and short hairpin RNAs. The odontoblastic differentiation was assessed with ALP staining, ALP activity assay, alizarin red S staining and real-time RT-PCR of DSPP, OCN, RUNX2 and OSX. Western blot was conducted to examine the levels of p38 mitogen-activated protein kinase (p38 MAPK), c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase 1/2 (Erk1/2). The expression of EREG and odontoblastic differentiation-related markers was investigated in human dental pulp from teeth with deep caries and healthy teeth. RESULTS: Epiregulin was upregulated during tertiary dentin formation. rhEREG enhanced the odontoblastic differentiation of DPSCs following upregulated p38 MAPK and Erk1/2 phosphorylation, but not JNK, whereas depletion of EREG suppressed DPSC differentiation. Gefitinib decreased odontoblastic differentiation with decreased phosphorylation of p38 MAPK and Erk1/2. And suppression of p38 MAPK and Erk1/2 pathways attenuated DPSC differentiation. In human dental pulp tissue, EREG upregulation in deep caries correlates with odontoblastic differentiation enhancement. CONCLUSION: Epiregulin is released during tertiary dentin formation. And EREG enhanced DPSC odontoblastic differentiation via MAPK pathways.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Polpa Dentária/efeitos dos fármacos , Epirregulina/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Células-Tronco/citologia , Animais , Proliferação de Células/efeitos dos fármacos , Polpa Dentária/citologia , Proteínas da Matriz Extracelular/metabolismo , Masculino , Odontoblastos/citologia , Odontoblastos/efeitos dos fármacos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos
15.
Biomed Res Int ; 2019: 4759060, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31396530

RESUMO

Introduction: Pulp regeneration, as a treatment for pulp necrosis, has significant advantages over root canal therapy for the preservation of living pulp. To date, research on pulp regeneration has mainly focused on the transplantation of pulp stem cells into the root canal, but there is still a lack of research on the migration of pulp cells into the root canal via cell homing. Stem cells from the apical tooth papilla (SCAP) are recognized as multidirectional stem cells, but these cells are difficult to obtain. MicroRNAs are small noncoding RNAs that play crucial roles in regulating normal and pathologic functions. We hypothesized that some types of microRNAs might improve the migration and proliferation function of dental pulp stem cells (DPSCs), which are easily obtained in clinical practice, and as a result, DPSCs might replace SCAP and provide valuable information for regenerative endodontics. Methods: Magnetic activated cell sorting of DPSCs and SCAP was performed. Next-generation sequencing was performed to examine DPSCs and SCAP miRNAs expression and to identify the most significant differentially expressed miRNA. CCK-8 and transwell assays were used to determine the impact of this miRNA on DPSCs proliferation and migration. Results: The most significant differentially expressed miRNA between DPSCs and SCAP was miR-224-5p. Downregulating miR-224-5p promoted DPSCs proliferation and migration; the opposite results were observed when miR-224-5p was upregulated. Conclusion: MiR-224-5p promotes proliferation and migration in DPSCs, a finding that is of great significance for further exploring the role of dental pulp stem cells in regenerative endodontics.


Assuntos
Movimento Celular , Proliferação de Células , Polpa Dentária/metabolismo , Regulação para Baixo , MicroRNAs/biossíntese , Células-Tronco/metabolismo , Adolescente , Adulto , Polpa Dentária/citologia , Feminino , Humanos , Masculino , Células-Tronco/citologia
16.
Arch Oral Biol ; 107: 104485, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31376703

RESUMO

OBJECTIVE: The objectives of this study were (a) to determine the differentially expressed microRNAs that can target heat shock protein B8 (HspB8) during in vitro expansion of dental pulp stem cells (DPSCs); (b) to identify microRNAs involved in posttranscriptional regulation of HspB8 expression; and (c) to determine if HspB8-targeting microRNAs play roles on osteogenic differentiation of DPSCs. DESIGN: DPSCs were established from rat first molars and expanded in vitro until the passage that cells lost osteogenic potential. TargetScan was used to predict the microRNAs that target HspB8 mRNA. Stem-loop quantitative RT-PCR was conducted to identify the HspB8-targeting microRNAs that were upregulated in late passages. The microRNAs mimics were transfected into DPSCs to assess their effects on HspB8 expression and on osteogenic differentiation. RESULTS: let-7b-5p, miR-98-5p, miR-215, miR-219a-1-3p and miR-295-5p were found to consistently increase expression in DPSCs after expansion. HspB8 mRNA and/or protein were significantly decreased in the DPSCs after transfection of miR-215 and miR-219a-1-3p mimics; whereas no significant reduction was seen after transfecting let-7b-5p, miR-98-5p and miR-295-5p mimics. When subjecting the transfected DPSCs to osteogenic induction, reduction of calcium deposition or osteogenic marker expression were observed with miR-215, miR-219a-1-3p and miR-295-5p transfection. CONCLUSIONS: Increased expression of miR-215 and miR-219a-1-3p downregulates HspB8 expression, which contributes to the reduction of osteogenic capability of DPSCs. Increased expression of miR295-5p also causes a reduction of osteogenic differentiation, but not involved in HspB8.


Assuntos
Proteínas de Choque Térmico/genética , MicroRNAs/genética , Osteogênese , Células-Tronco/citologia , Animais , Diferenciação Celular , Células Cultivadas , Polpa Dentária/citologia , Regulação da Expressão Gênica , Ratos
17.
J Photochem Photobiol B ; 198: 111561, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31352000

RESUMO

Blindness and vision loss contribute to irreversible retinal degeneration, and cellular therapy for retinal cell replacement has the potential to treat individuals who have lost light sensitive photoreceptors in the retina. Retinal cells are well characterized in function, and are a subject of interest in cellular replacement therapy of photoreceptors and the retinal pigment epithelium. However, retinal cell transplantation is limited by various factors, including the choice of potential stem cell source that can show variability in plasticity as well as host tissue integration. Dental pulp is one such source that contains an abundance of stem cells. In this study we used dental pulp-derived mesenchymal stem cells (DPSCs) to mitigate sodium iodate (NaIO3) insult in a rat model of retinal degeneration. Sprague-Dawley rats were first given an intravitreal injection of 3 × 105 DPSCs as well as a single systemic administration of NaIO3 (40 mg/kg). Electroretinography (ERG) was performed for the next two months and was followed-up by histological analysis. The ERG recordings showed protection of DPSC-treated retinas within 4 weeks, which was statistically significant (* P ≤ .05) compared to the control. Retinal thickness of the control was also found to be thinner (*** P ≤ .001). The DPSCs were found integrated in the photoreceptor layer through immunohistochemical staining. Our findings showed that DPSCs have the potential to moderate retinal degeneration. In conclusion, DPSCs are a potential source of stem cells in the field of eye stem cell therapy due to its protective effects against retinal degeneration.


Assuntos
Iodatos/toxicidade , Transplante de Células-Tronco Mesenquimais , Degeneração Retiniana/terapia , Epitélio Pigmentado da Retina/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Polpa Dentária/citologia , Modelos Animais de Doenças , Eletrorretinografia , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Células Fotorreceptoras/citologia , Ratos , Ratos Sprague-Dawley , Degeneração Retiniana/etiologia , Epitélio Pigmentado da Retina/patologia
18.
Artif Cells Nanomed Biotechnol ; 47(1): 3058-3066, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31339375

RESUMO

Repairing the lost or damaged mandible is very difficult and time-consuming, so there is a great hope for tissue engineering to accelerate it. At the present study, electrospinning was applied to fabricate polyvinylidene fluoride (PVDF) and PVDF-polyaniline (PANI) composite scaffolds. In addition, extremely low frequency pulsed electromagnetic field (PEMF) was applied for treating the stem cells derived from dental pulp (DPSCs) when cultured on the nanofibrous scaffolds. Osteoinductive property of the fabricated PVDF, PVDF-PANI scaffold at the presence and absence of the PEMF was investigated by evaluating the common osteogenic differentiation markers in seeded-DPSCs on the scaffold. Results demonstrated that cell attachment, protein adsorption and cells viability were increased when PEMF was applied. In addition, ALP activity, calcium content, osteogenic genes and protein evaluations confirmed that PEMF could significantly increase osteoinductivity of the PVDF while composite with PANI. According to the results, the use of polymers with piezoelectricity and conductivity features plus PEMF exposure has a promising potential to improve the current treatment methods in bone and mandibular defects.


Assuntos
Compostos de Anilina/farmacologia , Diferenciação Celular , Campos Eletromagnéticos , Células-Tronco Mesenquimais/citologia , Osteogênese , Polivinil/química , Tecidos Suporte/química , Fosfatase Alcalina/metabolismo , Cálcio/metabolismo , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/efeitos da radiação , Polpa Dentária/citologia , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos da radiação , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/efeitos da radiação , Osteogênese/efeitos dos fármacos , Osteogênese/efeitos da radiação , Resistência à Tração , Engenharia Tecidual
19.
Mater Sci Eng C Mater Biol Appl ; 103: 109736, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31349524

RESUMO

The aim of this study was to prepare a promising biomaterial for dental pulp tissue repair and regeneration. The ultrasmall superparamagnetic iron oxide (USPIO)-labeled hydroxyapatite (HA)/silk fibroin (SF) scaffold loaded dental pulp stem cells (DPSCs) was designed. The odontogenic differentiation of DPSCs was investigated, and USPIO was used as magnetic resonance imaging (MRI) contrast agent. The results indicated that scaffolds freeze-dried from SF solution with 0.025 mg/mL USPIO and 5 mg/mL HA showed stable physical properties, accurate MRI images, and low cytotoxicity in vitro. The composite scaffolds were characterized and implanted to the subcutaneous space under the nude mice with tooth fragment. After 2 W of implantation, the relaxation rate (R2) values increased because of incorporation of USPIO, then decreased with the degradation of scaffolds. The tissue regeneration and scaffolds degradation behaviors were monitored by MRI. The expression of dentin sialophosphoprotein (DSPP) and dentin matrix acidic phosphoprotein 1 (DMP1) indicated that DPSCs differentiated into odontoblast-like cells. Revascularization and mineralization evaluated by HE-stained images and alizarin red staining images showed good formation. The USPIO/HA/SF composite scaffold loaded DPSCs promoted the repair and regeneration of dental pulp tissue and provided the ability of imaging noninvasively.


Assuntos
Polpa Dentária/fisiologia , Durapatita/química , Fibroínas/química , Nanopartículas de Magnetita/química , Regeneração , Células-Tronco/metabolismo , Tecidos Suporte/química , Animais , Polpa Dentária/citologia , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Células-Tronco/citologia
20.
Int J Mol Med ; 44(3): 823-834, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31257457

RESUMO

Clinically, deep decay can lead to inflammation in the dental pulp. Apart from the use of various materials to sooth the inflamed pulp, there is currently no adequate treatment, and the gold standard, calcium hydroxide, that is used to cover the dentin/pulp, has limited effect. Sometimes the pulp will remain infected and cause pulpitis, and ultimately, the pulp will need to be removed. The first principle of oral treatment is to protect the pulp. Therefore, it is necessary to study the immune response and regeneration of pulp cells in conditions of deep decay. Of the terminal complement system proteins, complement 5a (C5a) has the most potent effect compared to complement 3a (C3a) and complement 4a (C4a). C5a is 20- to 2,500-fold stronger than C3a and C4a. The purpose of this study was to elucidate the association between C5a, secreted by complement activation, and the duration of inflammation. Another key goal was to detect the expression of C5a and its receptor, complement 5a receptor (C5aR). To this end, the cells were divided into 4 groups as per stimulation with lipoteichoic acid (LTA) or lipopolysaccharide (LPS) as follows: i) The 1 µg/ml LTA group; ii) the 1 µg/ml LPS group; iii) the 1 µg/ml LTA and 1 µg/ml LPS group; and iv) the PBS-only group, which served as a control. There were 5 time points for all 4 groups: 1, 2, 3, 5 and 7 days. Reverse transcription-quantitative polymerase chain reaction was used to detect the gene expression levels of C5a, C5aR and interleukin (IL)-6 at different time points. Western blot analyses was carried out to detect the expression of C5aR. Transmission electron microscopy was also conducted to assess the ultrastructural features of dental pulp cells. The gene expression trends of C5a and C5aR mRNA were identical. C5a and C5aR mRNA was highly expressed on the second day of LTA or LPS stimulation. However, in the LTA and LPS co-stimulation group, C5a and C5aR mRNA were highly expressed on both the first and second day, with higher levels on the second day. IL-6 expression decreased as time progressed in the LTA only and in the LTA + LPS co-stimulation groups. However, a peak in its expression was observed on the second day in the LPS group. On the whole, this study demonstrates that a 1 µg/ml concentration of LTA and LPS stimulates human dental pulp cells to activate the expression of C5a.


Assuntos
Complemento C5a/genética , Complemento C5a/imunologia , Polpa Dentária/citologia , Polpa Dentária/metabolismo , Regulação da Expressão Gênica , Receptor da Anafilatoxina C5a/genética , Receptor da Anafilatoxina C5a/imunologia , Células-Tronco/metabolismo , Adipócitos/metabolismo , Adolescente , Adulto , Biomarcadores , Diferenciação Celular , Células Cultivadas , Complemento C5a/metabolismo , Citocinas/metabolismo , Feminino , Humanos , Lipopolissacarídeos/imunologia , Masculino , Osteoblastos/citologia , Osteoblastos/metabolismo , Receptor da Anafilatoxina C5a/metabolismo , Células-Tronco/imunologia , Ácidos Teicoicos/imunologia , Adulto Jovem
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