Current therapeutics are not effective for orofacial neuropathic pain, and better options are needed. The present study used inferior orbital nerve (ION)-injured mice to investigate the effect of inhibiting monoacylglycerol lipase (MAGL), an enzyme that degrades the major endocannabinoid 2-arachydonoylgycerol (2-AG) in orofacial neuropathic pain. The head-withdrawal threshold to mechanical stimulation of the whisker pad was reduced on days 3, 5, and 7 after ION injury. Injection of JZL184, a selective inhibitor of MAGL, on day 7 after ION injury attenuated the reduction in head-withdrawal threshold at 2 h after administration. Moreover, the numbers of MAGL-immunoreactive neurons in the trigeminal subnucleus caudalis (Vc) and upper cervical spinal cord (C1-C2) were significantly greater in ION-injured mice than in sham-operated mice but were reduced after administration of JZL184. The increase in MAGL immunoreactivity suggests that increased 2-AG production is followed by rapid enzymatic degradation of 2-AG. JZL184 inhibited this degradation and thus increased 2-AG concentration in the brain, particularly in the Vc and C1-C2 regions, thus attenuating pain. Our findings suggest that inhibition of 2-AG degradation by MAGL inhibitors is a promising therapeutic option for treatment of orofacial neuropathic pain.
Previous in vitro studies have suggested that simvastatin can be used as a direct pulp capping material due to its ability to induce odontoblastic differentiation and angiogenesis. The aim of this animal study was to evaluate the pulpal response to mineral trioxide aggregate (MTA) and four concentrations of simvastatin/MTA in combination. The study was conducted in two stages using four different simvastatin concentrations and MTA as a capping material for rat maxillary molars. The grades of inflammation and continuity of dentin formation were evaluated in hematoxylin and eosin (HE)-stained samples. Dentin thickness was determined by histomorphometric analysis, and the data were subjected to statistical analysis. On day 3, mild inflammation was observed in all groups. On day 7, the simvastatin groups showed a slightly higher rate of chronic inflammation. Inflammation was not present on day 30. Discontinuous dentin was present in all methylcellulose (control) samples. Continuous dentin was formed in all of the samples treated with 1.5% simvastatin. The greatest dentin thickness was observed after treatment with 1.5% simvastatin and MTA, followed by 0.5% simvastatin. Statistical analysis demonstrated no significant differences in dentin thickness and continuity between MTA and simvastatin at 0.5% and 1.5% (P > 0.05).
The greater palatine foramen (GPF) is an important anatomical landmark and has substantial clinical relevance in dental surgery. Knowledge of its precise location and dimensions is required for proper planning of surgical procedures involving the posterior maxilla. We used microfocus computed tomography to determine the location and dimensions of the GPF, and any sex and race variations in those measurements, in 77 human skulls scanned at the South African Nuclear Energy Corporation. Specialized software was used for three-dimensional rendering, segmentation, and visualization of the reconstructed volume data. GPF location ranged from adjacent to the first molar to distal of the third molar. The most common GPF location was near the third molar (66.7% of skulls), and the GPF was as close as 6.31 mm (mean distance 12.75 ± 3 mm). The mean GPF dimensions were 5.22 mm on the anterior-posterior axis and 2.81 mm on the lateral-medial axis. We noted no significant differences in relation to race, sex, or age in the sample. The GPF was adjacent or posterior to the third maxillary molar in most skulls.
Dedifferentiated fat (DFAT) cells were isolated from mature adipocytes using the ceiling culture method. Recently, we successfully isolated DFAT cells from adipocytes with a relatively small size (<40 μm). DFAT cells have a higher osteogenic potential than that of medium adipocytes. Therefore, the objective of this study was to determine the optimal concentration of collagenase solution for isolating small adipocytes from human buccal fat pads (BFPs). Four concentrations of collagenase solution (0.01%, 0.02%, 0.1%, and 0.5%) were used, and their effectiveness was assessed by the number of small adipocytes and DFAT cells isolated. The total number of floating adipocytes that dissociated with 0.02% collagenase was 2.5 times of that dissociated with 0.1% collagenase. The number of floating adipocytes with a diameter of ≤29 μm that dissociated with 0.02% collagenase was thrice of those dissociated with 0.1% and 0.5% collagenase. The number of DFAT cells that dissociated with 0.02% collagenase was 1.5 times of that dissociated with 0.1% collagenase. In addition, DFAT cells that dissociated with 0.02% collagenase had a higher osteogenic differentiation potential than those that dissociated with 0.1% collagenase. These results suggest that 0.02% is the optimal collagenase concentration for isolating small adipocytes from BFPs.
Oral carcinoma develops from squamous epithelial cells by the acquisition of multiple (epi) genetic alterations that target different genes and molecular pathways. Herein, we performed a comprehensive genomic and epigenetic characterization of the HSC-3 cell line through karyotyping, multicolor fluorescence in situ hybridization, array comparative genomic hybridization, and methylation-specific multiplex ligation-dependent probe amplification. HSC-3 turned out to be a near-triploid cell line with a modal number of 61 chromosomes. Banding and molecular cytogenetic analyses revealed that nonrandom gains of chromosomal segments occurred more frequently than losses. Overall, gains of chromosome 1, 3q, 5p, 7p, 8q, 9q, 10, 11p, 11q13, 12, 13, 14, 17, 18p, 20, Yp, and Xq were observed. The largest region affected by copy number loss was observed at chromosome 18q. Several of the observed genomic imbalances and their mapped genes were already associated with oral carcinoma and/or adverse prognosis, invasion, and metastasis in cancer. The most common rearrangements observed were translocations in the centromeric/near-centromeric regions. RARB, ESR1, and CADM1 genes were methylated and showed copy number losses, whereas TP73 and GATA5 presented with methylation and copy number gains. Thus, the current study presents a comprehensive characterization of the HSC-3 cell line; the use of this cell line may contribute to enriching the resources available for oral cancer research, especially for the testing of therapeutic agents.
A study was conducted to evaluate the mechanisms underlying ectopic orofacial pain associated with tooth pulp inflammation in rats. We observed a significant decrease in the head withdrawal threshold (HWT) response to mechanical and heat stimuli applied to the ipsilateral facial skin upon application of complete Freund’s adjuvant (CFA) to the upper first molar (M1TP) in comparison to application of vehicle. A large number of trigeminal ganglion (TG) neurons showed transient receptor potential vanilloid 1 (TRPV1) immunoreactivity (IR), and some of them were retrogradely labeled with fluorogold injected into the facial skin. A large number of cells showing IR for glial fibrillary acidic protein (GFAP) were observed in the 2nd compared to the 1st or 3rd branch regions of the TG, and TG cells innervating the facial skin were also surrounded by GFAP-IR cells. After administration of TRPV1 antagonist into the facial skin of M1TP CFA-treated rats, the decrease of HWTs in response to mechanical and heat stimulation of the facial skin was significantly reversed. The present findings suggest that the excitability of TG neurons is enhanced upon tooth pulp inflammation, leading to overexpression of TRPV1 in TG neurons innervating the facial skin, and that satellite glial cells are also activated, resulting in the development of ectopic orofacial pain.
The present study was conducted to assess the applicability of liquid-based cytology (LBC) using an innovative oral brush, Orcellex. Fifty healthy volunteers were recruited. From each subject, four samples were collected using “Orcellex” from apparently normal oral mucosal sites. A plastic spatula was also used to obtain an additional sample. Data on the tolerability and acceptability of the Orcellex were collected from the subjects, together with assessments of the adequacy of LBC slide preparations for cellularity, preparation quality, and the types of cells observed. The Orcellex brush was well accepted by the volunteers, who reported relatively little pain. Orcellex brush LBC preparations were of good quality in terms of cell morphology and staining, with a clean background. Only two smears (2/200; 1%) were found to be inadequate due to low cellularity. All of the plastic spatula LBC preparations were inadequate. Representative cells from all layers of the different oral epithelia examined were documented. Oral liquid-based cytology using the Orcellex brush may have considerable potential for early detection of oral cancer and precancer.
This study evaluated the simulated localized wear of resin luting cements for universal adhesive systems using different curing modes. Five resin luting cements for universal adhesive systems were evaluated and subsequently subjected to wear challenge in a Leinfelder-Suzuki wear simulation device. Overall, 20 specimens from each resin luting cement were photo-cured for 40 s (dual-cure group), and 20 specimens of each material were not photo-cured (chemical-cure group). Simulated localized wear was generated using a stainless steel ball-bearing antagonist in water slurry of polymethylmethacrylate beads. In addition, scanning electron microscopy (SEM) observations of resin luting cements and wear facets were conducted. Significant differences in simulated wear and SEM observations of wear facets were evident among the materials in the dual- and chemical-cure groups. The simulated wear and SEM observations of wear facets of G-CEM LinkForce and Panavia V5 were not influenced by the curing mode. SEM observations of resin luting cements were material dependent. In most cases, dual curing appears to ensure greater wear resistance of resin luting cements than chemical curing alone. The wear resistance of some resin luting cements appears to be material dependent and is not influenced by the curing mode.
Placement of composite resin restorations in deep subgingival cavities can damage surrounding soft tissues. In addition, commonly used resin-based composites (RBCs) might interfere with wound healing and periodontal health. To clarify cellular interactions with RBCs, we used an MTT assay to investigate adhesion of primary human gingival fibroblasts and human osteoblasts (hFOB 1.19) on five RBC materials with and without surface modifications (alumina blasting with 50- or 110-μm Al2O3). In addition, high-performance liquid chromatography (HPLC) was used to determine release of resin monomers from RBCs after 1 h, 1 day, and 7 days. As compared with tissue culture plastics (the control), cellular adhesion was significantly lower (P < 0.001) for human gingival fibroblasts and osteoblasts. Only minor, nonsignificant differences between individual RBCs were identified. HPLC analyses identified the release of three bifunctional methacrylates bisphenol A glycerolate dimethacrylate, triethylene glycol dimethacrylate, and diurethane dimethacrylate from RBCs and showed that monomer release increased between 1 h and 1 day but remained low. The present findings suggest that surface adhesion in the subgingival area is limited for the tested RBCs. Although residual monomer release was low for all tested RBCs, it might be sufficient to adversely affect cell adhesion.
This retrospective study analyzed clinical outcomes of monolithic zirconia restorations (MZRs) and factors related to restoration success. Patient records were searched to identify those provided MZRs (Cercon ht) for premolars or molars between April 2012 and March 2016. All MZRs were placed according to a standardized protocol. Kaplan-Meier analysis was used to assess MZR performance and failure after recall appointments at 1 year or later. In total, 101 patients received 148 MZRs. Mean duration of follow-up was 25.0 ± 9.9 months. Six MZRs required replacement: three because of pulpal complications, one because of root fracture of an abutment tooth, one because of restoration fracture, and one because the tooth was used as an abutment tooth for a fixed partial denture after root fracture of an adjacent tooth. The cumulative MZR survival (success) rate at 3.5 years was 91.5% (95% confidence interval, 82.1% to 100%). The findings of this short-term retrospective study indicate that posterior MZRs are a therapeutic option for certain patients. In addition, several clinical procedures contribute to MZR success, including preparation design and occlusal and adhesive surface treatments.
We used a polymicrobial (PM) biofilm model to examine associations of bacterial adhesiveness with surface characteristics of various dental materials. Four types of dental materials (apatite pellet, zirconia, ceramic, and composite resin) with rough and mirror surfaces were used. Surface roughness, surface free energy, zeta potential, and colony-forming units (CFUs) of the biofilm formations were measured. Biofilms were cultured for 24 h under anaerobic conditions, plated onto blood agar medium, and anaerobically cultured for 4 days. After culturing, CFU per mm2 was calculated, and samples were observed under a scanning electron microscope. Means and standard deviations of the experimental data were estimated, and one-way ANOVA and Tukey multiple comparison assays were performed. Pearson correlation coefficients were obtained for the CFU and surface characteristics. Surface roughness and surface free energy appeared to affect generation of PM biofilms on oral materials, and zeta potential was involved in generation of PM biofilms on mirror-ground oral materials.
Temporomandibular joint disorders (TMJD) involve orofacial pain and functional limitations that may limit important daily activities such as chewing and speaking. This observational case-control study attempted to identify factors associated with TMJD development, particularly inflammation. The study participants were patients treated at Karolinska University Hospital, Stockholm, Sweden. The cases were patients who received a diagnosis of TMJD, chronic closed lock, or painful clicking and were treated surgically during the period from 2007 through 2011. The control group was randomly selected from among patients who had undergone tooth extraction and was matched by age and sex. A total of 146 cases and 151 controls were included in the analyses. The response rate was 55.3% for the case group and 21.8% for the control group. The male:female ratio for patients with TMJD was 1:4.4. TMJD was significantly associated with pneumonia (odds ratio [OR], 2.1), asthma (OR, 2.1), allergies (OR, 1.8), headache (OR, 3.1), general joint hypermobility (OR, 3.8), orofacial trauma (OR, 3.9), rheumatism (OR, 2.5), and orthodontic treatment (OR, 2.4) (P < 0.05 for all outcomes). In conclusion, autoimmune diseases and inflammatory conditions are associated with increased risk of TMJD. Moreover, certain lung disorders may predict subsequent development of TMJD.