Dental Pulp Vascular Permeability Changes Induced by Dental Bleaching

Aiming to compare the effect of different light sources for dental bleaching on vascular permeability of dental pulps, forty-eight incisors were used. The bleaching agent (35% hydrogen peroxide) was activated by halogen light; LED (Light Emitting Diode) or LED, followed by laser phototherapy (LPT) (lambda = 780 nm; 3 J/cm(2)). After the bleaching procedures, the animals received an intra-arterial dye injection and one hour later were sacrificed. The teeth were diaphanized and photographed. The amount of blue stain content of each dental pulp was quantified using a computer imaging program. The data was statistically compared (p <= 0.05). The results showed a significant higher (p <= 0.01) dye content in the groups bleached with halogen light, compared with the control, LED and LED plus LPT groups. Thus, tooth bleaching activated by LED or LED plus LPT induces lesser resulted in increased vascular permeability than halogen light.

Dental pulp stem cells express proteins involved in the local invasiveness of odontogenic myxoma

Little is known about the histogenesis of the odontogenic myxoma (OM). Dental pulp stem cells could be candidate precursors of OM because both OM and the dental pulp share the same embryological origin: the dental papilla. For the purpose of comparing OM and stem cells, this study analyzed the expression of two proteins related to OM invasiveness (MMP-2 and hyaluronic acid) in human immature dental pulp stern cells (hIDPSCs). Three lineages of hIDPSCs from deciduous and permanent teeth were used in this study. Immunofluorescence revealed positive reactions for MMP-2 and hyaluronic acid (HA) in all hIDPSCs. MMP-2 appeared as dots throughout the cytoplasm, whereas HA appeared either as diffuse and irregular dots or as short fibrils throughout the cytoplasm and outside the cell bodies. The gene expression profile of each cell lineage was evaluated using RT-PCR analysis, and HA was expressed more intensively than MMP-2. HA expression was similar among the three hIDPSCs lineages, whereas MMP-2 expression was higher in DL-1 than in the other cell lines. The expression of proteins related to OM invasiveness in hIDPSCs could indicate that OM originates from dental pulp stem cells.

Effects of 3-Dimensional Conformal or Intensity-modulated Radiotherapy on Dental Pulp Sensitivity during and after the Treatment of Oral or Oropharyngeal Malignancies

Introduction: Radiation therapy (RT) of malignant tumors in the head and neck area may have damaging effects on surrounding tissues. The aim of this investigation was to evaluate the effects of RI delivered by 3-dimensional conformal radiotherapy (3D-RT) or intensity-modulated radiotherapy (IMRT) on dental pulp sensitivity. Methods: Twenty patients with oral or oropharyngeal cancer receiving RT with 3D-RT or IMRT underwent cold thermal pulp sensitivity testing (PST) of 2 teeth each at 4 time points: before RT (TP1), the beginning of RT with doses between 30 and 35 Gy (TP2), the end of RT with doses between 60 and 70 Gy (TP3), and 4 to 5 months after the start of RT (TP4). Results: All 40 teeth showed positive responses to PST at TP1 (100%) and 9 at TP2 (22.5%; 3/16 [18.8%] for 3D-RT and 6/24 [25.0%] for IMRT). No tooth responded to PST at TP3 and TP4 (0%). A statistically significant difference existed in the number of positive pulp responses between different time points (TP1 through TP4) for all patients receiving RT (P <= .05), IMRT (P <= .05), and 3D-RT (P <= .05). No statistically significant differences in positive sensitivity responses were found between 3D-RT and IMRT at any time point (TP1, TP3, TP4, P = 1.0; TP2, P = .74). A statistically significant correlation existed between the location of the tumor and PST at TP2 for IMRT (P <= .05) but not for 3D-RT (P = .14). Conclusions: RT decreased the number of teeth responding to PST after doses greater than 30 to 35 Gy. The type of RT (3D-RT or IMRT) had no influence on the pulp responses to PST after the conclusion of RT. (J Endod 2012;38:148-152)

Early transplantation of human immature dental pulp stem cells from baby teeth to golden retriever muscular dystrophy (GRMD) dogs: Local or systemic?

Background:The golden retriever muscular dystrophy (GRMD) dogs represent the best available animal model for therapeutic trials aiming at the future treatment of human Duchenne muscular dystrophy (DMD). We have obtained a rare litter of six GRMD dogs (3 males and 3 females) born from an affected male and a carrier female which were submitted to a therapeutic trial with adult human stem cells to investigate their capacity to engraft into dogs muscles by local as compared to systemic injection without any immunosuppression. Methods Human Immature Dental Pulp Stem Cells (hIDPSC) were transplanted into 4 littermate dogs aged 28 to 40 days by either arterial or muscular injections. Two non-injected dogs were kept as controls. Clinical translation effects were analyzed since immune reactions by blood exams and physical scores capacity of each dog. Samples from biopsies were checked by immunohistochemistry (dystrophin markers) and FISH for human probes. Results and Discussion We analyzed the cells' ability in respect to migrate, engraftment, and myogenic potential, and the expression of human dystrophin in affected muscles. Additionally, the efficiency of single and consecutive early transplantation was compared. Chimeric muscle fibers were detected by immunofluorescence and fluorescent in situ hybridisation (FISH) using human antibodies and X and Y DNA probes. No signs of immune rejection were observed and these results suggested that hIDPSC cell transplantation may be done without immunosuppression. We showed that hIDPSC presented significant engraftment in GRMD dog muscles, although human dystrophin expression was modest and limited to several muscle fibers. Better clinical condition was also observed in the dog, which received monthly arterial injections and is still clinically stable at 25 months of age. Conclusion Our data suggested that systemic multiple deliveries seemed more effective than local injections. These findings open important avenues for further researches.

Clinical, radiographic and histologic analysis of the effects of pulp capping materials used in pulpotomies of human primary teeth

Aim: To compare the clinical, radiographic and histological responses of the pulp to mineral trioxide aggregate (MTA), calcium hydroxide (CH) and Portland cement (PC) when used as a pulpotomy agent in human primary teeth. Study design: Forty-five mandibular primary molar teeth were randomly assigned to CH, MTA or PC groups and treated by pulpotomy technique. Methods: The teeth were treated by conventional pulpotomy technique, differing only in the capping material for each group. Clinical and radiographic evaluations were recorded at 6-, 12- and 24-month follow-up. Teeth in the regular exfoliation period were further processed for histologic analysis. Statistics: The teeth were treated by conventional pulpotomy technique, differing only in the capping material for each group. Clinical and radiographic evaluations were recorded at 6-, 12- and 24-month follow-up. Teeth in the regular exfoliation period were further processed for histologic analysis. Statistics: Clinically and radiographically, the MTA and PC groups showed 100 % success rates at 6, 12 and 24 months. In CH group, several teeth presented clinical and radiographic failures detected throughout the follow-up period, and internal resorption was a frequent radiographic finding. Histologic analysis revealed the presence of dentine-like mineralised material deposition obliterating the root canal in the PC and MTA groups. CH group presented, in most of the sections, necrotic areas in the root canals. Conclusions: MTA and PC may serve as effective materials for pulpotomies of primary teeth as compared to CH. Although our results are very encouraging, further studies and longer follow-up assessments are needed in order to determine the safe clinical indication of Portland cement.

CCL3 and CXCL12 production in vitro by dental pulp fibroblasts from permanent and deciduous teeth stimulated by Porphyromonas gingivalis LPS

Objective: The aim of this study was to compare the production of the chemokines CCL3 and CXCL12 by cultured dental pulp fibroblasts from permanent (PDPF) and deciduous (DDPF) teeth under stimulation by Porphyromonas gingivalis LPS (PgLPS). Material and Methods: Primary culture of fibroblasts from permanent (n=3) and deciduous (n=2) teeth were established using an explant technique. After the fourth passage, fibroblasts were stimulated by increasing concentrations of PgLPS (0 – 10 µg/mL) at 1, 6 and 24 h. The cells were tested for viability through MTT assay, and production of the chemokines CCL3 and CXCL12 was determined through ELISA. Comparisons among samples were performed using One-way ANOVA for MTT assay and Two-way ANOVA for ELISA results. Results: Cell viability was not affected by the antigen after 24 h of stimulation. PgLPS induced the production of CCL3 by dental pulp fibroblasts at similar levels for both permanent and deciduous pulp fibroblasts. Production of CXCL12, however, was significantly higher for PDPF than DDPF at 1 and 6 h. PgLPS, in turn, downregulated the production of CXCL12 by PDPF but not by DDPF. Conclusion: These data suggest that dental pulp fibroblasts from permanent and deciduous teeth may present a differential behavior under PgLPS stimulation.

Dental pulp vascular permeability changes induced by dental bleaching

Aiming to compare the effect of different light sources for dental bleaching on vascular permeability of dental pulps, forty-eight incisors were used. The bleaching agent (35 % hydrogen peroxide) was activated by halogen light; LED (Light Emitting Diode) or LED, followed by laser phototherapy (LPT) (λ = 780 nm; 3 J/cm²). After the bleaching procedures, the animals received an intra-arterial dye injection and one hour later were sacrificed. The teeth were diaphanized and photographed. The amount of blue stain content of each dental pulp was quantified using a computer imaging program. The data was statistically compared (p < 0.05). The results showed a significant higher (p < 0.01) dye content in the groups bleached with halogen light, compared with the control, LED and LED plus LPT groups. Thus, tooth bleaching activated by LED or LED plus LPT induces lesser resulted in increased vascular permeability than halogen light.

Immunodetection of bone-related proteins in microcalcifications of human dental pulp

Objective: Root canal obliterations may pose esthetic and clinical problems or may even be a risk factor for tooth survival. Microcalcifications in the pulp can be so extensive that the entire root canal system becomes obliterated. Since bone sialoprotein (BSP) and osteopontin (OPN) are involved in both physiological and pathological mineralization processes, our hypothesis was that these two bone-related noncollagenous proteins are present in microcalcifications of the pulp. The purpose of this study was, therefore, to characterize the nature of microcalcifications in the pulp of aged human teeth. Methods: From a large collection of human teeth, 10 were found to exhibit pulpal microcalcifications. The teeth were extracted for periodontal reasons from 39-60 year old patients. After fixation in aldehydes and decalcification, teeth were processed for embedding in LR White resin for analysis in the light and transmission electron microscope. For the detection of BSP and OPN, post-embedding high resolution immunocytochemistry was applied. Results: The microcalcifications were round or elongated, occasionally coalescing, and intensely stained with toluidine blue. Collagen fibrils were found in most but not all microcalcifications. All microcalcifications were immunoreactive for both antibodies and showed an identical labeling pattern. Gold particle labeling was extensively found throughout the interfibrillar ground substance of the microcalcifications, whereas the dentin matrix lacked immunolabeling. Conclusion: BSP and OPN appear to be major matrix constituents of pulp microcalcifications and may thus, like in other mineralized tissues, be involved in their mineralization process.

Differential inflammatory response of dental pulp explants and fibroblasts to saliva

Abstract AIM: To investigate the inflammatory response of dental pulp fibroblasts and the respective explants to whole saliva. METHODOLOGY: Explants from human and porcine dental pulp tissue and isolated dental pulp fibroblasts were used to investigate the inflammatory response to sterile saliva. Cytokine and chemokine expression was assessed by RT-PCR. Western blot analysis and pharmacologic inhibitors were used to determine the involvement of signalling pathways. RESULTS: Dental pulp explants of human and porcine origin exposed to human saliva exhibited no major changes of IL-6 and IL-8 mRNA expression (P > 0.05). In contrast, isolated porcine and human dental pulp fibroblasts, when stimulated with human saliva, exhibited a vastly increased expression of IL-6 and IL-8 mRNA (P < 0.05). In pulp fibroblasts, saliva also increased the expression of other cytokines and chemokines via activation of NFkappaB, ERK and p38 signalling. Notably, a significantly reduced inflammatory response was elicited when pulp fibroblasts were transiently exposed to saliva. CONCLUSIONS: Saliva has a potential impact on inflammation of dental pulp fibroblasts in vitro but not when cells are embedded in the intrinsic extracellular matrix of the explant tissue.

Effects of Prolyl Hydroxylase Inhibitor L-mimosine on Dental Pulp in the Presence of Advanced Glycation End Products

INTRODUCTION Proangiogenic prolyl hydroxylase (PHD) inhibitors represent a novel approach to stimulate tissue regeneration. Diabetes mellitus involves the accumulation of advanced glycation end products (AGEs). Here we evaluated the impact of AGEs on the response of human pulp tissue to the PHD inhibitor L-mimosine (L-MIM) in monolayer cultures of dental pulp-derived cells (DPCs) and tooth slice organ cultures. METHODS In monolayer cultures, DPCs were incubated with L-MIM and AGEs. Viability was assessed based on formazan formation, live-dead staining, annexin V/propidium iodide, and trypan blue exclusion assay. Vascular endothelial growth factor (VEGF), interleukin (IL)-6, and IL-8 production was evaluated by quantitative polymerase chain reaction and immunoassays. Furthermore, expression levels of odontoblast markers were assessed, and alizarin red staining was performed. Tooth slice organ cultures were performed, and VEGF, IL-6, and IL8 levels in their supernatants were measured by immunoassays. Pulp tissue vitality and morphology were assessed by MTT assay and histology. RESULTS In monolayer cultures of DPCs, L-MIM at nontoxic concentrations increased the production of VEGF and IL-8 in the presence of AGEs. Stimulation with L-MIM decreased alkaline phosphatase levels and matrix mineralization also in the presence of AGEs, whereas no significant changes in dentin matrix protein 1 and dentin sialophosphoprotein expression were observed. In tooth slice organ cultures, L-MIM increased VEGF but not IL-6 and IL-8 production in the presence of AGEs. The pulp tissue was vital, and no signs of apoptosis or necrosis were observed. CONCLUSIONS Overall, in the presence of AGEs, L-MIM increases the proangiogenic capacity, but decreases alkaline phosphatase expression and matrix mineralization.

Detection of 400-year-old Yersinia pestis DNA in human dental pulp: An approach to the diagnosis of ancient septicemia

Ancient septicemic plague epidemics were reported to have killed millions of people for 2 millenniums. However, confident diagnosis of ancient septicemia solely on the basis of historical clinical observations is not possible. The lack of suitable infected material has prevented direct demonstration of ancient septicemia; thus, the history of most infections such as plague remains hypothetical. The durability of dental pulp, together with its natural sterility, makes it a suitable material on which to base such research. We hypothesized that it would be a lasting refuge for Yersinia pestis, the plague agent. DNA extracts were made from the dental pulp of 12 unerupted teeth extracted from skeletons excavated from 16th and 18th century French graves of persons thought to have died of plague (“plague teeth”) and from 7 ancient negative control teeth. PCRs incorporating ancient DNA extracts and primers specific for the human β-globin gene demonstrated the absence of inhibitors in these preparations. The incorporation of primers specific for Y. pestis rpoB (the RNA polymerase β-subunit-encoding gene) and the recognized virulence-associated pla (the plasminogen activator-encoding gene) repeatedly yielded products that had a nucleotide sequence indistinguishable from that of modern day isolates of the bacterium. The specific pla sequence was obtained from 6 of 12 plague skeleton teeth but 0 of 7 negative controls (P < 0.034, Fisher exact test). A nucleic acid-based confirmation of ancient plague was achieved for historically identified victims, and we have confirmed the presence of the disease at the end of 16th century in France. Dental pulp is an attractive target in the quest to determine the etiology of septicemic illnesses detected in ancient corpses. Molecular techniques could be applied to this material to resolve historical outbreaks.

Temperature changes in dental pulp associated with use of power grinding equipment on equine teeth

Objective To quantify the temperature changes in the dental pulp associated with equine dental procedures using power grinding equipment. Design A matrix experimental design with replication on the same sample was followed to allow the following independent variables to be assessed: horse age (young or old), tooth type (premolar or molar), powered grinding instrument (rotating disc or die grinder), grinding time (15 or 20 seconds) and the presence or absence of water coolant. Procedure Sound premolar and molar teeth from a 6-year-old horse and a 15-year-old horse, which had been removed postmortem, were sectioned parallel to the occlusal plane to allow placement of a miniature thermocouple at the level of the dental pulp. The maximum temperature increase, the time taken to reach this maximum and the cooling time were measured (n=10 in each study). The teeth were placed in a vice and the instrument used on the tooth as per clinical situation. Results Significant differences were recorded for horse age (P < 0.001), instrument type (P < 0.001), grinding time (P < 0.001) and presence or absence of coolant (P < 0.001). There was no significant difference for tooth type. Conclusion Thermal insult to the dental pulp from the use of power instruments poses a significant risk to the tooth. This risk can be reduced or eliminated by appropriate selection of treatment time and by the use of water irrigation as a coolant. The increased dentine thickness in older horses appears to mitigate against thermal injury from frictional heat.

Mineral trioxide aggregate with or without calcium chloride in pulpotomy

The aim of this study was to evaluate the reaction of the pulp tissue against mineral trioxide aggregate (MTA) with or without 10% calcium chloride (CaCl2). Pulpotomies were performed in 4 canines and 8 premolars of two 8-month-old dogs. MTA with or without CaCl2 was applied on the pulp tissue. The animals were killed after 90 days, and the specimens were processed for the microscopic analysis. Pulp tissue response was similar for MTA with and without CaCl2 Pulp vitality was present in all specimens, along with pulp repair with formation of mineralized tissue bridging. The addition of CaCl2 to MTA did not change its biologic properties in formation of mineralized barrier after pulpotomy.

Clinical and radiographic evaluation of pulpotomies employing calcium hydroxide and trioxide mineral aggregate

This study utilized clinical and radiographic examinations to compare the effectiveness of calcium hydroxide paste and mineral trioxide aggregate (MTA) for pulpotomies of primary molars in children. Ninety primary molars that showed clinical and radiographic indications for pulpotomy treatment were selected. The pulpotomies were performed in two sessions, using a corticosteroid/ antibiotic solution as therapeutic dressing. The sample was divided into two groups of 45 teeth, in which the pulpal remains were protected with either calcium hydroxide paste (Group 1) or MTA (Group 2). Radiographs were taken immediately and at 3-, 6-, and 12-month follow-up appointments. Three teeth in Group 1 failed after three months, while two cases failed after six months and one more failed at one year. Two failures were found in Group 2 at the 12-month follow-up. These results indicate that both materials may be utilized for pulpotomies in primary teeth.