Células madre en odontología: nuevas perspectivas / Stem cells in dentistry: new perspectives

En el campo de la odontología, prevalecen actualmente alternativas terapéuticas con una filosofía conservadora. Sin embargo, con el advenimiento de los tratamientos con células madre (CM), se amplían las posibilidades terapéuticas, que buscan la combinación y el equilibrio entre la intervención tradicional y las posibilidades de reposición de estructuras anatómicas dañadas, a través de la regeneración de tejidos utilizando células madre o sus derivados (AU)

In the dentistry field, therapeutic alternatives with a conservative philosophy currently prevail. However, with the advent of stem cell (SC) treatments, therapeutic possibilities are expanding, seeking a combination and balance between traditional intervention and the pos- sibility of replacing damaged anatomical structures through tissue regeneration, using stem cells or their derivatives (AU)

Delivery of affordable and scalable encapsulated allogenic/autologous mesenchymal stem cells in coagulated platelet poor plasma for dental pulp regeneration.

The main goal of regenerative endodontics procedures (REPs) is to revitalize teeth by the regeneration of healthy dental pulp. In this study, we evaluated the potential of combining a natural and accessible biomaterial based on Platelet Poor Plasma (PPP) as a support for dental pulp stem cells (DPSC) and umbilical cord mesenchymal stem cells (UC-MSC). A comparison study between the two cell sources revealed compatibility with the PPP based scaffold with differences noted in the proliferation and angiogenic properties in vitro. Additionally, the release of growth factors including VEGF, HGF and DMP-1, was detected in the media of cultured PPP and was enhanced by the presence of the encapsulated MSCs. Dentin-Discs from human molars were filled with PPP alone or with MSCs and implanted subcutaneously for 4 weeks in mice. Histological analysis of the MSC-PPP implants revealed a newly formed dentin-like structure evidenced by the expression of Dentin sialophosphoprotein (DSPP). Finally, DPSC induced more vessel formation around the dental discs. This study provides evidence of a cost-effective, xenofree scaffold that is compatible with either autologous or allogenic strategy for dental pulp regeneration. This attempt if successfully implemented, could make REPs treatment widely accessible, contributing in improving global health conditions.

Urgência em Endodontia: diagnóstico e tratamento em casos de pulpite irreversível sintomática / Urgency in Endodontics: Diagnosis and Treatment in Cases of Symptomatic Irreversible Pulpitis

Objetivo: discutir sobre o diagnóstico e a conduta terapêutica em casos de urgência endodôntica em dentes que apresentam pulpite irreversível sintomática. Material e Métodos: realizou-se uma revisão bibliográfica de estudos publicados nos últimos 5 anos (2015- 2020) por meio de busca nas bases de dados: PubMED, BVS (Biblioteca Virtual em Saúde) e Scielo (Scientific Eletronic Library). Para a pesquisa, foram utilizados os seguintes descritores: Pulpite Irreversível (Irreversible Pulpitis), Tratamento (Treatment), Dor (Pain) e Endodontia (Endodontics). Resultados: O diagnóstico é um passo fundamental no tratamento das urgências e emergências de origem endodôntica, pois é a partir do correto diagnóstico que será instituído o tratamento correto, reestabelecendo o conforto do paciente. Quando o profissional dispõe de tempo suficiente para realizar a remoção do tecido pulpar e o preparo do canal radicular, esse é o tratamento de escolha para os casos de pulpite irreversível sintomática, o qual pode ser realizado em sessão única ou em múltiplas sessões. Quando o profissional não dispõe de tempo suficiente para realizar o tratamento endodôntico convencional, a opção de tratamento é realizar apenas o atendimento de urgência para retirar o paciente do quadro de dor aguda presente, e em um momento futuro realizar o tratamento endodôntico completo. Conclusão: As urgências endodônticas sempre estão presentes nos consultórios odontológicos, os profissionais devem estar sempre preparados para realizar um correto diagnóstico e tratamento para cada caso, trazendo assim conforto ao paciente.

Objective: to discuss the diagnosis and therapeutic management in cases of endodontic urgency in teeth with symptomatic irreversible pulpitis. Material and Methods: a bibliographic review of studies published in the last 5 years (2015-2020) was carried out by searching the databases: PubMED, BVS (Virtual Health Library) and Scielo (Scientific Electronic Library). For the research, the following descriptors were used: Irreversible Pulpitis, Treatment, Pain and Endodontics. Results: The diagnosis is a fundamental step in the treatment of urgencies and emergencies of endodontic origin, as it is from the correct diagnosis that the correct treatment will be instituted, reestablishing the patient's comfort. When the professional has enough time to remove the pulp tissue and prepare the root canal, this is the treatment of choice for cases of symptomatic irreversible pulpitis, which can be performed in a single session or in multiple sessions. When the professional does not have enough time to carry out the conventional endodontic treatment, the treatment option is to perform only emergency care to remove the patient from the present acute pain condition, and at a future time to carry out the complete endodontic treatment. Conclusion: Endodontic emergencies are always present in dental offices, professionals must always be prepared to carry out a correct diagnosis and treatment for each case, thus bringing comfort to the patient.

TNF-alpha increases angiogenic potential in a co-culture system of dental pulp cells and endothelial cells.

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.

TNF-alpha increases angiogenic potential in a co-culture system of dental pulp cells and endothelial cells

Abstract 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.

Apical Papilla Cells Are Capable of Forming a Pulplike Tissue with Odontoblastlike Cells without the Use of Exogenous Growth Factors.

INTRODUCTION: Dental pulp is a complex tissue with highly differentiated cells, which makes its reconstruction a challenging task. The apical papilla is an undifferentiated tissue considered as the remnant of the dental papilla that forms the dentin-pulp complex. Aiming to analyze morphologic features of the tissue formed in an in vivo pulp model, we used human apical papilla as a cell source without the use of exogenous growth factors. METHODS: A construct was built using newborn mice molar crowns treated with TrypLE (Fisher Scientific, Loughborough, UK) and EDTA. The crowns were filled with PuraMatrix (Corning Inc, Corning, NY) and a pool population of human apical papilla cells. As a control, we used crowns filled only with PuraMatrix and empty crowns. The constructs were transplanted under severe combined immunodeficient mice kidney capsules. Immunohistochemistry for lamin A, dentin sialophosphoprotein, and dentin matrix protein 1 was performed. RESULTS: Morphologic analysis of all transplanted crowns showed the formation of a loose connective tissue of variable cellularity with the presence of well-formed functional vessels. In the study group, lamin A-positive cells represented the majority of cells within the pulp chamber and a few cells in the vessel lining. We also found positivity for dentin sialophosphoprotein and dentin matrix protein 1, an indicator of odontoblast differentiation. CONCLUSIONS: In our study model, human transplanted apical papilla cells mixed with the host cells and formed a vascularized viable tissue, and these cells were able to differentiate into odontoblastlike cells without the use of exogenous growth factors.

There is no pulp necrosis or calcific metamorphosis of pulp induced by orthodontic treatment: biological basis.

To biologically explain why the orthodontic treatment does not induce pulp necrosis and calcific metamorphosis of the pulp, this paper presents explanations based on pulp physiology, microscopy and pathology, and especially the cell and tissue phenomena that characterize the induced tooth movement. The final reflections are as follows: 1) the orthodontic movement does not induce pulp necrosis or calcific metamorphosis of the pulp; 2) there is no literature or experimental and clinical models to demonstrate or minimally evidence pulp alterations induced by orthodontic movement; 3) when pulp necrosis or calcific metamorphosis of the pulp is diagnosed during orthodontic treatment or soon after removal of orthodontic appliances, its etiology should be assigned to concussion dental trauma, rather than to orthodontic treatment; 4) the two pulp disorders that cause tooth discoloration in apparently healthy teeth are the aseptic pulp necrosis and calcific metamorphosis of the pulp, both only induced by dental trauma; 5) the concussion dental trauma still requires many clinical and laboratory studies with pertinent experimental models, to increasingly explain its effects on the periodontal and pulp tissues.

Intracardiac Injection of Dental Pulp Stem Cells After Neonatal Hypoxia-Ischemia Prevents Cognitive Deficits in Rats.

Neonatal hypoxia-ischemia (HI) is associated to cognitive and motor impairments and until the moment there is no proven treatment. The underlying neuroprotective mechanisms of stem cells are partially understood and include decrease in excitotoxicity, apoptosis and inflammation suppression. This study was conducted in order to test the effects of intracardiac transplantation of human dental pulp stem cells (hDPSCs) for treating HI damage. Seven-day-old Wistar rats were divided into four groups: sham-saline, sham-hDPSCs, HI-saline, and HI-hDPSCs. Motor and cognitive tasks were performed from postnatal day 30. HI-induced cognitive deficits in the novel-object recognition test and in spatial reference memory impairment which were prevented by hDPSCs. No motor impairments were observed in HI animals. Immunofluorescence analysis showed human-positive nuclei in hDPSC-treated animals closely associated with anti-GFAP staining in the lesion scar tissue, suggesting that these cells were able to migrate to the injury site and could be providing support to CNS cells. Our study evidence novel evidence that hDPSC can contribute to the recovery following hypoxia-ischemia and highlight the need of further investigation in order to better understand the exact mechanisms underlying its neuroprotective effects.

There is no pulp necrosis or calcific metamorphosis of pulp induced by orthodontic treatment: biological basis

ABSTRACT To biologically explain why the orthodontic treatment does not induce pulp necrosis and calcific metamorphosis of the pulp, this paper presents explanations based on pulp physiology, microscopy and pathology, and especially the cell and tissue phenomena that characterize the induced tooth movement. The final reflections are as follows: 1) the orthodontic movement does not induce pulp necrosis or calcific metamorphosis of the pulp; 2) there is no literature or experimental and clinical models to demonstrate or minimally evidence pulp alterations induced by orthodontic movement; 3) when pulp necrosis or calcific metamorphosis of the pulp is diagnosed during orthodontic treatment or soon after removal of orthodontic appliances, its etiology should be assigned to concussion dental trauma, rather than to orthodontic treatment; 4) the two pulp disorders that cause tooth discoloration in apparently healthy teeth are the aseptic pulp necrosis and calcific metamorphosis of the pulp, both only induced by dental trauma; 5) the concussion dental trauma still requires many clinical and laboratory studies with pertinent experimental models, to increasingly explain its effects on the periodontal and pulp tissues.

RESUMO Para fundamentar biologicamente por que o tratamento ortodôntico não induz necrose pulpar e metamorfose cálcica da polpa, apresentou-se explicações com base na fisiologia, microscopia e patologia pulpar, bem como, e principalmente, nos fenômenos celulares e teciduais que caracterizam a movimentação dentária induzida. As reflexões finais foram: 1) o movimento ortodôntico não induz necrose pulpar ou metamorfose cálcica da polpa; 2) não há literatura e modelos experimentais e clínicos que comprovem ou minimamente evidenciem alterações pulpares induzidas pelo movimento ortodôntico; 3) quando a necrose pulpar ou metamorfose cálcica da polpa for diagnosticada durante o tratamento ortodôntico ou logo após a remoção dos aparelhos ortodônticos, a sua etiologia deve ser atribuída ao traumatismo dentário do tipo concussão, e não ao tratamento ortodôntico; 4) as duas doenças pulpares que levam ao escurecimento coronário em dentes aparentemente hígidos são a necrose pulpar asséptica e a metamorfose cálcica da polpa, ambas induzidas exclusivamente pelo traumatismo dentário; 5) o traumatismo dentário do tipo concussão requer, ainda, muitos estudos clínicos e laboratoriais, com modelos experimentais pertinentes, para fundamentar cada vez mais os seus efeitos sobre os tecidos periodontais e pulpares.

Biological Analysis of Simvastatin-releasing Chitosan Scaffold as a Cell-free System for Pulp-dentin Regeneration.

INTRODUCTION: The improvement of biomaterials capable of driving the regeneration of the pulp-dentin complex mediated by resident cells is the goal of regenerative dentistry. In the present investigation, a chitosan scaffold (CHSC) that released bioactive concentrations of simvastatin (SIM) was tested, aimed at the development of a cell-free tissue engineering system. METHODS: First, we performed a dose-response assay to select the bioactive dose of SIM capable of inducing an odontoblastic phenotype in dental pulp cells (DPCs); after which we evaluated the synergistic effect of this dosage with the CHSC/DPC construct. SIM at 1.0 µmol/L (CHSC-SIM1.0) and 0.5 µmol/L were incorporated into the CHSC, and cell viability, adhesion, and calcium deposition were evaluated. Finally, we assessed the biomaterials in an artificial pulp chamber/3-dimensional culture model to simulate the cell-free approach in vitro. RESULTS: SIM at 0.1 µmol/L was selected as the bioactive dose. This drug was capable of strongly inducing an odontoblastic phenotype on the DPC/CHSC construct. The incorporation of SIM into CHSC had no deleterious effect on cell viability and adhesion to the scaffold structure. CHSC-SIM1.0 led to significantly higher calcium-rich matrix deposition on scaffold/dentin disc assay compared with the control (CHSC). This biomaterial induced the migration of DPCs from a 3-dimensional culture to its surface as well as stimulated significantly higher expressions of alkaline phosphatase, collagen type 1 alpha 1, dentin matrix acidic phosphoprotein 1, and dentin sialophosphoprotein on 3-dimensional-cultured DPCs than on those in contact with CHSC. CONCLUSIONS: CHSC-SIM1.0 scaffold was capable of increasing the chemotaxis and regenerative potential of DPCs.

Disclosing the physiology of pulp tissue for vital pulp therapy.

The discovery that dentine is a reservoir of bioactive molecules that can be recruited on demand has attracted efforts to develop new protocols and materials for vital pulp therapy (VPT). The noncollagenous proteins (NCPs) present in the dentine extracellular matrix (ECM) include growth factors (TGF-ß1, BMP-7, FGF-2, IGF-1 and IGF-2, NGF and GDNF), extracellular matrix molecules (DSP, DPP, BSP, DMP-1 and DSPP) and both anti-inflammatory and pro-inflammatory chemokines and cytokines (TNF-α, IL-1, IL-6 and IL-10). Molecules such as DSP and DPP are mainly expressed by odontoblasts, and they are cleaved products from dentine sialophosphoprotein (DSPP). Some molecules, such as TGF-ß1, specifically interact with decorin/biglycan in dentine. Although TGF-ß1 increases the expression and secretion of NGF in human pulp cells, NGF induces mineralization and increases the expression of DSPP and DMP-1. Furthermore, GDNF may act as a cell survival factor and mitogen during tooth injury and repair. Pulp capping materials, such as MTA and calcium hydroxide, can solubilize bioactive dentine molecules (TGF-ß1, NGF and GDNF) that stimulate tertiary dentinogenesis. The binding of these signalling molecules leads to activation of several signalling transduction pathways involved in dentinogenesis, odontoblast differentiation and inflammatory responses, such as the p38 MAPK, NF-kß and Wnt/ß-catenin signalling pathways. Understanding the cascade of cellular and molecular events underlying the repair and regeneration processes provides a reasonable new approach to VPT through a targeted interaction between tooth tissue and bioactive molecules.

The effect of adjusting the pH of local anaesthetics in dentistry: a systematic review and meta-analysis.

The acidic nature of commercial local anaesthetics (LAs) can cause pain during infiltration and delay the onset of anaesthesia. It is suggested that adjusting the pH of anaesthetic agents could minimize these effects. This systematic review aimed to evaluate the efficacy of buffered LAs in reducing infiltration pain and onset time during dental procedures. MEDLINE, Embase, Scopus and Scielo databases were searched up to April 2017. Randomized controlled trials comparing buffered and unbuffered LAs for intraoral injections were included. Risk of bias was assessed using the Cochrane Collaboration tool. Data upon injection pain and onset time were pooled in a random-effects model. Subgroup analyses compared normal and inflamed tissues, and terminal infiltrations and inferior alveolar nerve (IAN) blocks. Meta-regressions were performed to explain heterogeneity. Fourteen articles were included in this review. Lidocaine with epinephrine was the most used anaesthetic combination. Nonlidocaine studies (n = 2) were not pooled in the meta-analysis. Buffered lidocaine did not result in less pain during intraoral injections: mean difference -6.4 (95% CI -12.81 to 0.01) units in a 0-100 scale. Alkalinized lidocaine did not reduce the onset time in normal tissues when terminal infiltration techniques were used, but resulted in a more rapid onset for IAN blocks (-1.26 min) and in inflamed tissues (-1.37 min); however, this change may not be clinically relevant, considering the time required to prepare the buffered agent. Studies performed using other anaesthetic salts did not show robust and clinically significant results in favour of alkalinization.

Efficacy of removal of cariogenic bacteria and carious dentin by ablation using different modes of Er:YAG lasers.

The primary objective of this in vitro study was to evaluate the efficiency of removal of cariogenic bacteria and carious dentin by ablation using two lasers: fluorescence-feedback controlled (FFC) Er:YAG laser and different pulses of Er:YAG laser based on variable square pulse technology (VSPt). The secondary objective was to measure the temperature during laser ablation of carious tissue. Seventy-two extracted human molars were used in this study. Sixty teeth with carious dentin were randomly divided into four experimental groups according to the treatment for caries removal: group 1: 400 µs (FFC group); group 2: super short pulse (SSP group, 50 µs pulse); group 3: medium short pulse (MSP group, 100 µs pulse); group 4: short pulse (SP group, 300 µs pulse) and one positive control group with no treatment. Twelve teeth without carious lesion were used as a negative control group. After caries removal, swabs were taken with cotton pellets and real-time PCR analysis was performed. During caries ablation, a thermal infrared camera was used to measure the temperature changes. In all experimental groups, specimens were free of bacterial contamination after the treatment. In the SSP, MSP and SP groups, temperatures measured during caries ablation were significantly higher compared to temperatures in the FFC group (P<0.001). In this in vitro study, laser treatment for removal of carious dentin and cariogenic bacteria was an efficient treatment modality without causing excessive temperatures that might adversely affect pulp vitality.

Efficacy of removal of cariogenic bacteria and carious dentin by ablation using different modes of Er: YAG lasers

The primary objective of this in vitro study was to evaluate the efficiency of removal of cariogenic bacteria and carious dentin by ablation using two lasers: fluorescence-feedback controlled (FFC) Er:YAG laser and different pulses of Er:YAG laser based on variable square pulse technology (VSPt). The secondary objective was to measure the temperature during laser ablation of carious tissue. Seventy-two extracted human molars were used in this study. Sixty teeth with carious dentin were randomly divided into four experimental groups according to the treatment for caries removal: group 1: 400 µs (FFC group); group 2: super short pulse (SSP group, 50 µs pulse); group 3: medium short pulse (MSP group, 100 µs pulse); group 4: short pulse (SP group, 300 µs pulse) and one positive control group with no treatment. Twelve teeth without carious lesion were used as a negative control group. After caries removal, swabs were taken with cotton pellets and real-time PCR analysis was performed. During caries ablation, a thermal infrared camera was used to measure the temperature changes. In all experimental groups, specimens were free of bacterial contamination after the treatment. In the SSP, MSP and SP groups, temperatures measured during caries ablation were significantly higher compared to temperatures in the FFC group (P<0.001). In this in vitro study, laser treatment for removal of carious dentin and cariogenic bacteria was an efficient treatment modality without causing excessive temperatures that might adversely affect pulp vitality.

Determination of the Influence of Chronic Periodontitis on Pulp Sensibility by Means of Electric and Thermal Cold Testing.

INTRODUCTION: Communication between pulp and periodontal tissue has been well established. However, it is unknown when periodontal disease begins to affect the clinical response of pulp tissue. The aim of this study was to assess the influence of periodontal severity on pulp sensibility by means of electric and thermal cold testing. METHODS: The teeth assessed in this study were allocated into 3 groups considering radiographic alveolar bone loss (ABL) as follows: slight periodontitis (SP, ABL ≤7 mm without reaching the apex, n = 25), moderate periodontitis (ABL >7 mm without reaching the apex, n = 23), and severe periodontitis (SvP, ABL >7 reaching the apex, n = 8). Gingival recession (GR), probing depth (PD), and clinical attachment level (CAL) were also measured. RESULTS: The results showed higher levels of PD and CAL in the SvP group compared with the SP group (P < .05), with no significant difference in GR (P > .05). The SvP group showed significant ABL compared with the other groups (P > .05). The SP group showed a significant number of teeth with a positive pulp response, whereas the SvP group showed a significant number of teeth with a negative pulp response (P < .05); no significant differences were observed between the thermal cold and electric tests (P > .05). CONCLUSIONS: Within the limits of this study, it can be concluded that pulp clinical involvement with a negative response to thermal cold and electric testing occurs only in the most advanced stage of chronic periodontitis with apical involvement.

Angiogenic mechanisms of human dental pulp and their relationship with substance P expression in response to occlusal trauma.

Angiogenesis is the formation of new blood vessels based on a pre-existing vasculature. It comprises two processes, sprouting of endothelial cells and the division of vessels due to abnormal growth of the microvasculature. It has been demonstrated that substance P (SP) can induce angiogenesis either by modulating endothelial cell growth (direct mechanism) or by attracting cells with angiogenic potential to the injury site (indirect mechanism). Therefore, the purpose of this article is to review the angiogenic mechanisms that regulate mineralized tissue formation in human dental pulp tissue and their relationship with SP expression as a defence response to stimuli such as the masticatory function and occlusal trauma. Articles included in this review were searched in PubMed, Scopus and ISI Web of Science databases, combining the following keywords: human dentine pulp, angiogenesis, angiogenic growth factors, neuropeptides, substance P, neurogenic inflammation, dentine matrix, dentinogenesis, occlusal trauma and dental occlusion. It is concluded that human dental pulp tissue responds to occlusal trauma and masticatory function with a neurogenic inflammatory phenomenon in which SP plays an important role in the direct and indirect mechanisms of angiogenesis by the action evoked via NK1 receptors at different cells, such as fibroblasts, endothelial and inflammatory cells, leading to new blood vessel formation which are needed to stimulate mineralized tissue formation as a defence mechanism.

Influence of dentin thickness on intrapulpal temperature under simulated pulpal pressure during Nd:YAG laser irradiation.

The aim of this study was to evaluate the effects of dentin thickness and pulpal pressure simulation (PPS) on the variation of intrapulpal temperature (∆T) when submitted to an adhesive technique using laser irradiation. Sixty sound human molars were sectioned and randomly divided into two groups (n = 30): group 1-1 mm of dentin thickness; group 2-2 mm of dentin thickness. Each group was divided into two subgroups (n = 15): subgroup A-absence of PPS; subgroup P-presence of PPS (15 cm H2O), sequentially treated with the following: 37 % phosphoric acid, adhesive system (Adper Single Bond), irradiation with Nd:YAG laser (1064 nm, 10 Hz, 60 s) using 60, 80, and 100 mJ/pulse energy parameters and light-curing (10 s). The ∆T was evaluated during the laser irradiation with a digital thermometer. Data were analyzed by three-way ANOVA and Tukey tests (p < 0.05). Three-way ANOVA revealed no significant differences for dentin thickness (p = 0.6512) on ∆T. PPS significantly reduced ∆T (p = 0.0001). The laser energy parameters (p = 0.0027) indicated that 100 mJ presented with significantly greater ∆T when compared to the groups irradiated with 80 and 60 mJ. Dentin thickness did not affect ∆T. The presence of PPS reduced the mean temperature values. The Nd:YAG laser energy parameters had a negative influence on the variation of temperature in the absence of PPS. In the presence of PPS, there was no risk to the pulp, since this study obtained temperature increases below 5.5 °C for all energy parameters, showing the technical viability for in vivo conditions.

Multiwall carbon nanotubes/polycaprolactone scaffolds seeded with human dental pulp stem cells for bone tissue regeneration.

Conventional approaches to bone regeneration rarely use multiwall carbon nanotubes (MWCNTs) but instead use polymeric matrices filled with hydroxyapatite, calcium phosphates and bioactive glasses. In this study, we prepared composites of MWCNTs/polycaprolactone (PCL) for bone regeneration as follows: (a) MWCNTs randomly dispersed on PCL, (b) MWCNTs aligned with an electrical field to determine if the orientation favors the growing of human dental pulp stem cells (HDPSCs), and (c) MWCNTs modified with ß-glycerol phosphate (BGP) to analyze its osteogenic potential. Raman spectroscopy confirmed the presence of MWCNTs and BGP on PCL, whereas the increase in crystallinity by the addition of MWCNTs to PCL was confirmed by X-ray diffraction and differential scanning calorimetry. A higher elastic modulus (608 ± 4.3 MPa), maximum stress (42 ± 6.1 MPa) and electrical conductivity (1.67 × 10(-7) S/m) were observed in non-aligned MWCNTs compared with the pristine PCL. Cell viability at 14 days was similar in all samples according to the live/dead assay, but the 21 day cell proliferation, measured by MTT was higher in MWCNTs aligned with BGP. Von Kossa and Alizarin red showed larger amounts of mineral deposits on MWCNTs aligned with BGP, indicating that at 21 days, this scaffold promotes osteogenic differentiation of HDPSCs.

Actualización sobre la evidencia histológica de los tejidos formados mediante terapia de regeneración pulpar guiada / Update on histological evidence of tissue formed by guided pulp tissue regeneration

La finalidad de la endodoncia regenerativa es restablecer la función de la pulpa normal endientes traumatizados, necróticos o infectados, convirtiendo a un diente no vital en uno vital nuevamente. Sin embargo, aún los resultados son imprevisibles. El objetivo de esta revisión fue recopilar y sintetizar la información disponible sobre la evidencia histológicas de los tejidos del complejo pulpo dentinario formados a través de la terapia de regeneración de tejido guiada. Se realizó una investigación basada en la búsqueda en MEDLINE utilizando filtros de tiempo (2011-2016) y palabras clave "Pulp", "Dentin", "Regeneration", "Tissue" and"Histologic". La búsqueda arrojó alrededor de 140 artículos; los de interés fueron seleccionados y descargados a texto completo. Los estudios más alentadores respecto a la regeneración de tejido guiada han sido descritos en reportes de caso de dientes inmaduros con diagnóstico de pulpitis irreversible en el cual al estudio histológico se observaron células tipo odontoblastos. Sin embargo, no existen estudios con seguimiento a largo plazo sobre este tipo de terapia. Algunos protocolos de tratamiento pueden dar lugar a resultados no deseados e impredecibles. Se requieren esfuerzos para mejorar y actualizar las estrategias de endodoncia regenerativa para que sea aplicada de un modo biológicamente eficaz y seguro para salvar los dientes.

The goal of regenerative endodontics is to reinstate normal pulp function in traumatized, necrotic and infected teeth that would result in reestablishment of their functions, but still fail to re-establish real pulp tissue and give unpredictable results. The aim of this review was to compile and synthesize available information on the histological evidence of tissue pulp-dentinal complex formed through guided tissue regeneration. A web-based research on MEDLINE was done using filter terms Review, published in the last 10 years and Dental journals. Keywords used for research were “Pulp", "Dentin", "Regeneratión", "Tissue” and "Histologic". The search yielded about 140 articles; the interest were selected and downloaded in full text. The most encouraging studies regarding guided tissue regeneration have been described in case reports of immature teeth diagnosed with irreversible pulpitis in which the histology odontoblasts type cells were observed. However, there are no studies with long-term follow up on this type of therapy. Some treatment protocols might result in undesired and unpredictable outcomes. Efforts are required to improve and update existing regenerative endodontic strategies to make it an effective, safe, and biological mode to save teeth.

Tooth Tissue Engineering: The Importance of Blood Products as a Supplement in Tissue Culture Medium for Human Pulp Dental Stem Cells.

One of the goals in using cells for tissue engineering (TE) and cell therapy consists of optimizing the medium for cell culture. The present study compares three different blood product supplements for improved cell proliferation and protection against DNA damage in cultured human dental pulp stem cells for tooth TE applications. Human cells from dental pulp were first characterized as adult stem cells (ectomesenchymal mixed origin) by flow cytometry. Next, four different cell culture conditions were tested: I, supplement-free; II, supplemented with fetal bovine serum; III, allogeneic human serum; and IV, autologous human serum. Cultured cells were then characterized for cell proliferation, mineralized nodule formation, and colony-forming units (CFU) capability. After 28 days in culture, the comet assay was performed to assess possible damage in cellular DNA. Our results revealed that Protocol IV achieved higher cell proliferation than Protocol I (p = 0.0112). Protocols II and III resulted in higher cell proliferation than Protocol I, but no statistical differences were found relative to Protocol IV. The comet assay revealed less cell damage in cells cultured using Protocol IV as compared to Protocols II and III. The damage percentage observed on Protocol II was significantly higher than all other protocols. CFUs capability was highest using Protocol IV (p = 0.0018) and III, respectively, and the highest degree of mineralization was observed using Protocol IV as compared to Protocols II and III. Protocol IV resulted in significantly improved cell proliferation, and no cell damage was observed. These results demonstrate that human blood product supplements can be used as feasible supplements for culturing adult human dental stem cells.