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Biomedical subjects

Imad About

Publications and source records attributed to Imad About.

13 recordsLinked to original sources

Lack of correlation between ex vivo apical dye penetration and presence of apical radiolucencies.

OBJECTIVE: The aim of this study was to determine if there is a significant correlation between the in vivo presence of periapical radiolucency and ex vivo apical dye penetration on the same human teeth. STUDY DESIGN: Eighty-four endodontically filled teeth that were scheduled for extraction were classified into 2 groups according to the presence or absence of a periapical radiolucency and further divided into 2 subgroups according to the quality of the root canal filling. After extraction, the apical filling was evaluated by a dye penetration method. RESULTS: The dye extraction evaluation showed no correlation between apical dye penetration and the presence of a periapical radiolucency (not significant), but a statistically significant correlation with the quality of the root canal filling (P = .03). CONCLUSION: The results of the dye penetration study were correlated to the quality of the root canal filling but had no predictive value for the development of periapical radiolucency.

Coloring Agents↗

Activation of human dental pulp progenitor/stem cells in response to odontoblast injury.

In restorative dentistry, whilst moderate carious lesion treatment does not significantly compromise odontoblast cell survival, deep cavity preparation may lead to a partial death of these cells. However, newly formed odontoblast-like cells can replace the necrotic odontoblasts and secrete a reparative dentine matrix. Although several lines of evidence strongly suggest the presence of resting progenitor or stem cells in the dental pulp, little is known about the activation and migration of these cells in response to injury. Human immature third molars extracted for orthodontic reasons were used in this work to study the activation of progenitor/stem cells and their migration after deep cavity preparation involving in pulpal exposure using 5-bromo-2'-deoxyuridine labelling (BrdU). After incubation for 1 day, the BrdU was localised to the nuclei of cells in the perivascular area. The BrdU-immunolabelling exhibited a gradient. It was strong in the blood vessels surrounding the pulpal cavity and decreased in those away from the cavity. After incubation for 2 weeks, labelled cells were seen in the vicinity of the cavity. At 4 weeks, the immunolabelling was localised to the cavity area only. Control teeth without cavities or with shallow dentine cavities did not show any perivascular labelling after culture. These results clearly demonstrate that perivascular progenitor/stem cells can proliferate in response to odontoblast injury. They also show that these proliferating cells can migrate to the pulpal injury site in their tissue of origin simulating the situation in vivo.

Biomarkers↗

Polymerized bonding agents and the differentiation in vitro of human pulp cells into odontoblast-like cells.

OBJECTIVES: Odontoblasts are highly differentiated post-mitotic cells, which under pathological conditions such as carious lesions and dental injuries may degenerate and be replaced by other pulp cells. We have recently shown that this physiological event can be reproduced in an in vitro assay system, but is highly modified by the presence of unpolymerized resinous monomers. Our hypothesis was that the photopolymerization of the monomers in the bonding agents might abolish these negative effects. The purpose of this study was to evaluate the effects of polymerized dentin bonding agents, through dentin slices, on odontoblast differentiation in vitro. METHODS: Pulp cells were obtained from human third molars. They were used to study the effects of four dentin bonding agents through 0.7 mm dentin slices which served as a barrier between the bonding agents and the culture medium. The media containing the bonding agents' extracts were added at non-toxic concentrations onto the cultured cells. Immunohistochemistry was performed to study the differentiation of pulp fibroblasts into odontoblasts under these conditions by evaluating the expression of several odontoblast specific genes. RESULTS: Pulp fibroblasts cultivated under these conditions synthesized type I collagen, osteonectin, dentin sialoprotein and nestin at the same level as in control cultures. Moreover, pulp cells synthesized a mineralized nodular extracellular matrix. Expression of these proteins was higher in the cells contributing to the nodule formation. In addition, except nestin, all these proteins were expressed in the mineral nodules. SIGNIFICANCE: This work shows the lack of effects of photopolymerized bonding agents, through dentin slices, on cytodifferentiation of secondary odontoblasts.

Adolescent↗

Role of injured endothelial cells in the recruitment of human pulp cells.

In restorative dentistry, deep cavity preparation may lead to partial destruction of the odontoblastic layer. However, newly formed odontoblast-like cells can replace the necrotic odontoblasts and secrete a reparative dentine matrix. While growth factors such as transforming growth factor beta1 (TGFbeta1) and bone morphogenetic proteins (BMP-2 and BMP-4) seem to be involved in the proliferation and differentiation of pulp cells, little is known about the migration of the newly proliferating stem cells to the injury site. Our hypothesis was that endothelial cell injury may be involved in directing these cells towards the injury site. For this study, human pulp fibroblasts and L929 cells were fluorescence-labeled by transduction with the Enhanced Green Fluorescent Protein (EGFP). Similarly, human umbilical vein endothelial cells (HUVEC) were labeled with the Discosoma Red Fluorescent Protein-2 (DsRed2). Cell migration was then studied in an insert cell culture system. The HUVEC cells were cultured in the lower compartment while the human pulp fibroblasts or L929 were in the upper compartment. After artificial injury to the HUVEC cells, only human pulp fibroblasts migrated to the lower compartment. At early time periods (4 days), migrating cells were randomly localized on the HUVEC layer. However, after 14 and 20 days, they were perfectly aligned along the injury site. In the absence of injury, no migration was observed. These results suggest that, the endothelial injury is involved in the recruitment of odontoblast-like cells at the injury site.

Cell Death↗

Apical leakage of four endodontic sealers.

The purpose of this study was to evaluate the sealing properties of four root canal sealers. Forty-eight maxillary central incisors were instrumented with Profile rotary instruments. They were randomly divided into four groups (n = 12) and filled using lateral condensation with one of the four sealers: Sealapex, Pulp Canal Sealer, AH 26, and Ketac-Endo. The apical leakage was measured with a fluid filtration method and expressed as L s(-1) KPa(-1). The teeth filled with Sealapex displayed a higher apical leakage (8.42 +/- 4.2 10(-11) L s(-1) KPa(-1)) than those filled with AH 26 (2.10 +/- 1.39 10(-11) L s(-1) KPa(-1)), Pulp Canal Sealer (0.17 +/- 0.09 10(-11) L s(-1) KPa(-1)) or Ketac-Endo (0.32 +/- 0.24 10(-1) L s(-1) KPa(-1)) (p < 0.01). No statistically significant difference was found among AH 26, Pulp Canal Sealer, and Ketac-Endo. No correlation was found between the sealing efficiency of the four sealers and their adhesive properties recorded in a previous study.

Adhesiveness↗

Cytotoxicity testing of endodontic sealers: a new method.

The purpose of this study was to compare ISO standards versus a new technique for in vitro evaluation of cytotoxicity of root canal sealers. The cytotoxicity of AH Plus, Cortisomol, and Sealapex was first recorded according to ISO standards on L 929 fibroblasts by the MTT assay. In parallel, 30 single-rooted teeth were cut at the cementum enamel junction (CEJ), and the roots were prepared and sterilized before filling with the lateral condensation using one of three sealers (n = 10). The apexes of the roots were dipped into 1 ml of minimum essential medium for 1, 2, and 30 days renewing the medium every other day. After 24-h contact between the medium and the filled roots, the medium was used to measure the cytotoxicity on L 929 with the MTT assay. ISO standards always gave a statistically higher cytotoxicity than the root-dipping technique (p < 0.0001), whatever the sealer and the exposure time. The ISO standards showed statistically significant differences among the sealers (p < 0.0001). AH Plus was noncytotoxic, Cortisomol showed a high cytotoxicity decreasing over time (p < 0.001), and Sealapex displayed a high cytotoxicity that did not decrease over time (NS). The new technique showed statistically significant differences among the sealers (p = 0.001), but the differences were so small that they were likely not clinically relevant. The high cytotoxicity of Sealapex decreased over time but the cytotoxicity of AH Plus and Cortisomol did not. The results show that the ISO standards may strongly over-evaluate the cytotoxicity of the endodontic sealers, emphasize the difference among the sealers, and may clinically correspond to a large overfilling. The new technique reduces the discrimination of the test and may clinically correspond to a classical filling. Therefore, both methods might be considered as clinically relevant, corresponding to classical and overfilling conditions.

Analysis of Variance↗

Dentin permeability and eugenol diffusion after full crown preparation.

PURPOSE: To compare teeth prepared to receive a metallic versus a metal-ceramic crown with respect to (1) in vitro dentin permeability before and after using a desensitizing agent and (2) pulpal eugenol concentration after sealing a temporary crown with a zinc oxide-eugenol based cement. MATERIALS AND METHODS: The roots of 20 human mandibular molars were separated and the crowns were prepared to receive a metallic crown. The hydraulic conductance was recorded before and after using Protect dentin desensitizer. The crowns were then further reduced to receive a metal-ceramic crown and the hydraulic conductance was recorded under the same conditions, on deeper dentin, before and after using the desensitizing agent. Twenty additional teeth were prepared: 10 to receive a metallic crown and 10 teeth to receive a metal-ceramic crown. A tube filled with 1 mL phosphate buffered saline (PBS) was sealed to the cementoenamel junction. A temporary crown was sealed onto the preparation with Temp Bond. The amount of eugenol that diffused across dentin into PBS was spectrofluorimetrically measured at Day 1 and Day 7. The crown preparations were vertically sectioned and the dentin remaining thickness was recorded. RESULTS: The hydraulic conductance of teeth prepared for a metal-ceramic crown was twice as high as teeth prepared for a metallic crown (P < 0.001). The desensitizing agent reduced the hydraulic conductance in both groups (P< 0.001). The two groups showed the same hydraulic conductance after using Protect (ns). No significant difference was found in the amount of eugenol diffusion between the two groups of teeth although eugenol diffusion decreased with time (P < 0.01). No correlation was found between eugenol diffusion and remaining dentin thickness.

Analysis of Variance↗

Influence of resinous monomers on the differentiation in vitro of human pulp cells into odontoblasts.

Odontoblasts are highly differentiated postmitotic cells, which under pathological conditions such as carious lesions and dental injuries may degenerate and be replaced by other pulp cells. A recent work showed that this physiological event can be reproduced in an in vitro assay system. The purpose of the present study was to evaluate the effects of resinous monomers on odontoblast differentiation in vitro. Pulp cores from extracted human third molars were cultured with beta-glycerophosphate (2 mM) and used to evaluate the effects of TEGDMA, HEMA, UDMA, and Bis-GMA on the differentiation of pulp fibroblasts into odontoblasts. The effect of the monomers was studied by evaluating the expression of several odontoblast specific genes. In the absence of monomers, mineral nodule formation was observed. Pulp cells contributing to the nodule formation synthesized type I collagen, osteonectin, and dentin sialoprotein (DSP). In addition, Fourier transform infrared microspectroscopy showed that the mineral and organic composition of the nodules were characteristic of dentin. When the monomers were added at nontoxic concentrations, the effects of HEMA and Bis-GMA were more evident than that of TEGDMA and UDMA on collagen 1, osteonectin, and DSP expression. However, all monomers significantly decreased DSP expression and completely inhibited the mineral nodule formation.

Acrylates↗

E- and N-cadherin distribution in developing and functional human teeth under normal and pathological conditions.

Cadherins are calcium-dependent cell adhesion molecules involved in the regulation of various biological processes such as cell recognition, intercellular communication, cell fate, cell polarity, boundary formation, and morphogenesis. Although previous studies have shown E-cadherin expression during rodent or human odontogenesis, there is no equivalent study available on N-cadherin expression in dental tissues. Here we examined and compared the expression patterns of E- and N-cadherins in both embryonic and adult (healthy, injured, carious) human teeth. Both proteins were expressed in the developing teeth during the cap and bell stages. E-cadherin expression in dental epithelium followed an apical-coronal gradient that was opposite to that observed for N-cadherin. E-cadherin was distributed in proliferating cells of the inner and outer enamel epithelia but not in differentiated cells such as ameloblasts, whereas N-cadherin expression was up-regulated in differentiated epithelial cells. By contrast to E-cadherin, N-cadherin was also expressed in mesenchymal cells that differentiate into odontoblasts and produce the hard tissue matrix of dentin. Although N-cadherin was not detected in permanent intact teeth, it was re-expressed during dentin repair processes in odontoblasts surrounding carious or traumatic sites. Similarly, N-cadherin re-expression was seen in vitro, in cultured primary pulp cells that differentiate into odontoblast-like cells. Taken together these results suggest that E- and N-cadherins may play a role during human tooth development and, moreover, indicate that N-cadherin is important for odontoblast function in normal development and under pathological conditions.

Adolescent↗

The effect of etching on bacterial microleakage of an adhesive composite restoration.

OBJECTIVES: The incidence of bacterial microleakage, pulp inflammation and necrosis associated with dentine etching treatments prior to restoration are not known. Consequently, to resolve some of the controversy surrounding the effects and importance of vital dentine etching, the authors investigated these factors. METHODS: 110 standardised class V cavities were cut into buccal dentine, without exposing the pulp of teeth scheduled for extraction for orthodontic reasons. Cavities were either left unetched, or etched with the non-equivalent treatments of phosphoric acid gel for 60s or Ethylenediaminetetraacetic acid (EDTA) for 30s, prior to placement of composite resin. Teeth were collected and pulp responses were evaluated according to ISO guidelines, using pathohistomorphometric analysis and ANOVA statistics. RESULTS: Etching was found to be correlated to bacterial microleakage (p=0.0001) and tertiary dentine formation (p=0.0023). Bacterial microleakage was correlated to inflammatory activity (p=0.0001). The frequency of bacterial microleakage was: no etching (65%), EDTA (51%) and phosphoric acid (PA) (20%). SIGNIFICANCE: Vital dentine etching treatment is of extreme importance for the placement of RC to minimise bacterial microleakage. PA etching proved to be more effective at preventing bacterial microleakage than non-etching, and etching with EDTA.

Acid Etching, Dental↗

In vivo and in vitro expression of connexin 43 in human teeth.

Gap junctions are composed of transmembrane proteins belonging to the connexin family. These proteins permit the exchange of mall regulatory molecules directly between cells for the control of growth, development and differentiation. Although the presence of gap junctions in teeth has been already evidenced, the involved connexins have not yet been identified in human species. Here, we examined the distribution of connexin 43 (Cx43) in embryonic and permanent intact and carious human teeth. During tooth development, Cx43 localized both in epithelial and mesenchymal dental cells, correlated with cytodifferentiation gradients. In adult intact teeth, Cx43 was distributed in odontoblast processes. While Cx43 expression was downregulated in mature intact teeth, Cx43 appeared to be upregulated in odontoblasts facing carious lesions. In cultured pulp cells, Cx43 expression was related to the formation of mineralized nodules. These results indicate that Cx43 expression is developmentally regulated in human dental tissues, and suggest that Cx43 may participate in the processes of dentin formation and pathology.

Adolescent↗

Cavity remaining dentin thickness and pulpal activity.

PURPOSE: To investigate pulpal injury by measuring odontoblast numbers, and pulp dentin repair activity by measuring reactionary dentin area, in relation to the remaining dentin thickness (RDT) of cavity preparations in 217 human teeth. MATERIALS AND METHODS: Cavities were restored with adhesive bonded composite, resin-modified glass-ionomer cement, zinc oxide-eugenol or calcium hydroxide materials. The teeth were extracted for orthodontic reasons between 20-381 days post-operatively, and odontoblast numbers and reactionary dentin area were analyzed histomorphometrically, and statistically using ANOVA. RESULTS: Reactionary dentin deposition was observed beneath cavities with a RDT above 0.5 mm as well as beneath cavities with a RDT below 0.25 mm; however maximal reactionary dentin appeared to be beneath cavities with an a RDT between 0.5-0.25 mm (P= 0.0001). The area of reactionary repair was also influenced by the choice of restoration material (P= 0.0385) from greatest to least; calcium hydroxide, composite, resin-modified glass-ionomer cement and zinc oxide-eugenol. Odontoblast numbers were maintained beneath cavities with a RDT above 0.25 mm, cavities placed closer to the pulp appeared to injure underlying odontoblasts, reducing their numbers (P= 0.0001). The choice of cavity restoration material also influenced the survival of underlying odontoblasts (P= 0.0061).

Acid Etching, Dental↗

Efficiency and cytotoxicity of resin-based desensitizing agents.

PURPOSE: To compare in vitro the efficacy of five resin-based desensitizing agents at reducing human dentin permeability and to compare their cytotoxicity. The tested hypothesis was that their different curing techniques cause variations in efficiency and cytotoxicity. MATERIALS AND METHODS: Dentin slices (0.5 +/- 0.05 mm thick) were prepared from human third molars (10 per group) and their hydraulic conductance was recorded before and after application of one of the desensitizing agents with a Flodec device. Six desensitizing agents were studied: one light curing agent (Seal and Protect); one self-curing agent (Pain Free); the resin-based agents without any polymerization initiator (Health-Dent, Gluma Desensitizer, Isodan); one oxalate-based agent served as a control (Protect). A MTT assay on L 929 fibroblasts was performed to measure the cytotoxicity of the six desensitizing agents applied onto additional dentin slices (10 per group). RESULTS: All the desensitizing agents resulted in a large decrease in dentin permeability. The best results were obtained with Gluma Desensitizer, Isodan, Pain Free and Protect. A statistically significant difference was found among the materials (P = 0.001). All the materials were non-cytotoxic. Cell viability ranged from 88% for Seal and Protect to 100% for Isodan. No difference was found among their cytotoxicity.

Analysis of Variance↗