Scielo RSS <![CDATA[Journal of Applied Oral Science]]> vol. 23 num. 1 lang. pt <![CDATA[SciELO Logo]]> <![CDATA[Bone biology & pathology moves on: from bone resorption to formation, the rise of new therapeutic opportunities and experimental tools]]> <![CDATA[Validating of the pre-clinical mouse model for metastatic breast cancer to the mandible]]> Metastatic breast carcinoma has a great tendency to spread to the mandible. It is concomitantly associated with bone destruction, food intake disorder, and a poorer prognosis. Appropriate animal models need to be developed for a better understanding of the mechanisms underlying the metastatic process of breast cancer cells to mandible and to test the effects of potential lead compounds. Here, we assessed the metastasis model of intracardiac injection using luciferase-transfected metastatic breast cancer cells (MDA-MB-231Luc+) by determining the incidences of metastasis, mCT images, and histopathological results. A high bioluminescence signal mainly detected mandibular lesions with less frequent distal femora and proximal tibiae lesions. Extensive mandibular bone destruction occurred in nude mice grafted with metastatic breast cancer cells. This type of animal model might be a useful tool in assessing therapeutic implications and the efficacy of anti-cancer drugs for osteolytic cancers. <![CDATA[Bone tissue response to plasma-nitrided titanium implant surfaces]]> A current goal of dental implant research is the development of titanium (Ti) surfaces to improve osseointegration. Plasma nitriding treatments generate surfaces that favor osteoblast differentiation, a key event to the process of osteogenesis. Based on this, it is possible to hypothesize that plasma-nitrided Ti implants may positively impact osseointegration. Objective The aim of this study was to evaluate the in vivo bone response to Ti surfaces modified by plasma-nitriding treatments. Material and Methods Surface treatments consisted of 20% N2 and 80% H2, 450°C and 1.5 mbar during 1 h for planar and 3 h for hollow cathode. Untreated surface was used as control. Ten implants of each surface were placed into rabbit tibiae and 6 weeks post-implantation they were harvested for histological and histomorphometric analyses. Results Bone formation was observed in contact with all implants without statistically significant differences among the evaluated surfaces in terms of bone-to-implant contact, bone area between threads, and bone area within the mirror area. Conclusion Our results indicate that plasma nitriding treatments generate Ti implants that induce similar bone response to the untreated ones. Thus, as these treatments improve the physico-chemical properties of Ti without affecting its biocompatibility, they could be combined with modifications that favor bone formation in order to develop new implant surfaces. <![CDATA[TiF<sub>4</sub> and NaF varnishes as anti-erosive agents on enamel and dentin erosion progression <em>in vitro</em>]]> Objective This study assessed the effect of fluoride varnishes on the progression of tooth erosion in vitro. Material and Methods: Forty-eight enamel and 60 root dentin samples were previously demineralized (0.1% citric acid, pH 2.5, 30 min), leading to a baseline and erosive wear of 12.9 and 11.4 µm, respectively. The samples were randomly treated (6 h) with a 4% TiF4 varnish (2.45%F-, pH 1.0), a 5.42% NaF varnish (2.45%F-, pH 5.0), a placebo varnish and no varnish (control). The samples were then subjected to erosive pH cycles (4x90 s/day in 0.1% citric acid, intercalated with artificial saliva) for 5 days. The increment of the erosive tooth wear was calculated. In the case of dentin, this final measurement was done with and without the demineralized organic matrix (DOM). Enamel and dentin data were analyzed using ANOVA/Tukey’s and Kruskal-Wallis/Dunn tests, respectively (p&lt;0.05). Results The TiF4 (mean±s.d: 1.5±1.1 µm) and NaF (2.1±1.7 µm) varnishes significantly reduced enamel wear progression compared to the placebo varnish (3.9±1.1 µm) and control (4.5±0.9 µm). The same differences were found for dentin in the presence and absence of the DOM, respectively: TiF4 (average: 0.97/1.87 µm), NaF (1.03/2.13 µm), placebo varnish (3.53/4.47 µm) and control (3.53/4.36 µm). Conclusion The TiF4 and NaF varnishes were equally effective in reducing the progression of tooth erosion in vitro. <![CDATA[New methodology for evaluating osteoclastic activity induced by orthodontic load]]> Orthodontic tooth movement (OTM) is a dynamic process of bone modeling involving osteoclast-driven resorption on the compression side. Consequently, to estimate the influence of various situations on tooth movement, experimental studies need to analyze this cell. Objectives The aim of this study was to test and validate a new method for evaluating osteoclastic activity stimulated by mechanical loading based on the fractal analysis of the periodontal ligament (PDL)-bone interface. Material and Methods The mandibular right first molars of 14 rabbits were tipped mesially by a coil spring exerting a constant force of 85 cN. To evaluate the actual influence of osteoclasts on fractal dimension of bone surface, alendronate (3 mg/Kg) was injected weekly in seven of those rabbits. After 21 days, the animals were killed and their jaws were processed for histological evaluation. Osteoclast counts and fractal analysis (by the box counting method) of the PDL-bone interface were performed in histological sections of the right and left sides of the mandible. Results An increase in the number of osteoclasts and in fractal dimension after OTM only happened when alendronate was not administered. Strong correlation was found between the number of osteoclasts and fractal dimension. Conclusions Our results suggest that osteoclastic activity leads to an increase in bone surface irregularity, which can be quantified by its fractal dimension. This makes fractal analysis by the box counting method a potential tool for the assessment of osteoclastic activity on bone surfaces in microscopic examination. <![CDATA[Osteopontin expression in reactive lesions of gingiva]]> Reactive proliferations of the gingiva comprise lesions such as pyogenic granuloma (PG), inflammatory fibroepithelial hyperplasia (IFH), peripheral ossifying fibroma (POF), and peripheral giant cell lesion. Osteopontin (OPN) has a dual role, it promotes mineralization when it is bound to solid substrate, and on the other hand, it inhibits mineralization when it is seen in association with solution. Objectives The study aimed to evaluate the expression of osteopontin in normal gingival tissue and different types of focal reactive proliferations of gingival tissue, and its role in the development of calcification within it. Material and Methods The presence and distribution of osteopontin was assessed using immunohistochemistry in five cases of normal gingival tissue and 30 cases of focal reactive proliferations of gingiva. Results There was no expression of osteopontin in normal subjects. Few cases of pyogenic granuloma, inflammatory fibroepithelial hyperplasia, and all the cases of peripheral ossifying fibroma showed positivity for osteopontin in the inflammatory cells, stromal cells, extracellular matrix, and in the calcifications. Conclusion The expression of osteopontin in all the cases of peripheral ossifying fibroma speculates that the majority of the cases of peripheral ossifying fibroma originate from the periodontal ligament cells. The treatment modalities for peripheral ossifying fibroma should differ from other focal reactive proliferations of gingiva. <![CDATA[Effect of sumac extract on serum oxidative status, RANKL/OPG system and alveolar bone loss in experimental periodontitis in rats]]> Objectives Sumac (Rhus coriaria L.) is widely used spice which has several properties such as antioxidant, anti-inflammatory and antimicrobial. The purpose of this animal study was to evaluate the effects of sumac extract on levels of receptor activator of nuclear factor-kappa B ligand (RANKL), osteoprotegerin (OPG) expression, serum oxidative status, and alveolar bone loss in experimental periodontitis. Material and Methods Twenty-four Wistar rats were separated into three groups: non-ligated (NL, n=8), ligature only (LO, n=8), and ligature and treated with sumac extract (S, n=8) (20 mg/kg per day for 11 days). A 4/0 silk suture was placed around the mandibular right first molars subgingivally; after 11 days, the rats were sacrificed, and alveolar bone loss was histometrically measured. The detection of RANKL and OPG were immunohistochemically performed. Levels of serum total antioxidant status (TAS)/total oxidant status (TOS), and oxidative stress index (OSI) were also analyzed. Results Alveolar bone loss was significantly greater in the LO group compared to the S and NL groups (p&lt;0.05). The number of inflammatory cell infiltrate (ICI) and osteoclasts in the LO group was significantly higher than that of the NL and S groups (p&lt;0.05). The number of osteoblasts in the LO and S groups was significantly higher than that of the NL group (p&lt;0.05). There were significantly more RANKL-positive cells in the LO group than in the S and NL groups (p&lt;0.05). OPG-positive cells were higher in S group than in LO and NL groups (p&lt;0.05). TOS and OSI levels were significantly reduced in S group compared to LO group (P&lt;0.05) and TAS levels were similar in S and NL group (p&gt;0.05). Conclusions The present study showed that systemic administration of sumac extract may reduce alveolar bone loss by affecting RANKL/OPG balance, TOS and OSI levels in periodontal disease in rats. <![CDATA[Rat subcutaneous tissue response to calcium silicate containing different arsenic concentrations]]> Objective: To evaluate the response of rat subcutaneous tissue in implanted polyethylene tubes that were filled with GMTA Angelus and Portland cements containing different arsenic concentrations. Material and Methods: Atomic absorption spectrophotometry was utilized to obtain the values of the arsenic concentration in the materials. Thirty-six rats were divided into 3 groups of 12 animals for each experimental period. Each animal received two implants of polyethylene tubes filled with different test cements and the lateral of the tubes was used as a control group. After 15, 30 and 60 days of implantation, the animals were killed and the specimens were prepared for descriptive and morphometric analysis considering: inflammatory cells, collagen fibers, fibroblasts, blood vessels and other components. The results were analyzed utilizing the Kuskal-Wallis test and the Dunn's Multiple test for comparison (p&lt;0.05). Results: The materials showed, according to atomic absorption spectrophotometry, the following doses of arsenic: GMTA Angelus: 5.01 mg/kg, WPC Irajazinho: 0.69 mg/kg, GPC Minetti: 18.46 mg/kg and GPC Votoran: 10.76 mg/kg. In a 60-day periods, all specimens displayed a neoformation of connective tissue with a structure of fibrocellular aspect (capsule). Control groups and MTA Angelus produced the lower amount of inflammatory reaction and GPC Minetti, the highest reaction. Conclusions: There was no direct relationship between the concentration of arsenic present in the composition of the materials and the intensity of the inflammatory reactions. Higher values, as 18.46 mg/kg of arsenic in the cement, produce characteristics of severe inflammation reaction at the 60-day period. The best results were found in MTA angelus. <![CDATA[Novel biological activity of ameloblastin in enamel matrix derivative]]> Objective Enamel matrix derivative (EMD) is used clinically to promote periodontal tissue regeneration. However, the effects of EMD on gingival epithelial cells during regeneration of periodontal tissues are unclear. In this in vitro study, we purified ameloblastin from EMD and investigated its biological effects on epithelial cells. Material and Methods Bioactive fractions were purified from EMD by reversed-phase high-performance liquid chromatography using hydrophobic support with a C18 column. The mouse gingival epithelial cell line GE-1 and human oral squamous cell carcinoma line SCC-25 were treated with purified EMD fraction, and cell survival was assessed with a WST-1 assay. To identify the proteins in bioactive fractions of EMD, we used proteome analysis with two-dimensional gel electrophoresis followed by identification with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Results Purified fractions from EMD suppressed proliferation of GE-1 and SCC-25. LC-MS/MS revealed that ameloblastin in EMD is the component responsible for inhibiting epithelial cell proliferation. The inhibitory effect of ameloblastin on the proliferation of GE-1 and SCC-25 was confirmed using recombinant protein. Conclusion The inhibitory effects of EMD on epithelial cell proliferation are caused by the biological activities of ameloblastin, which suggests that ameloblastin is involved in regulating epithelial downgrowth in periodontal tissues. <![CDATA[Comparison between 3D volumetric rendering and multiplanar slices on the reliability of linear measurements on CBCT images: an <em>in vitro</em> study]]> OBJECTIVE: The purpose of this study was to determine the accuracy and reliability of two methods of measurements of linear distances (multiplanar 2D and tridimensional reconstruction 3D) obtained from cone-beam computed tomography (CBCT) with different voxel sizes. MATERIAL AND METHODS: Ten dry human mandibles were scanned at voxel sizes of 0.2 and 0.4 mm. Craniometric anatomical landmarks were identified twice by two independent operators on the multiplanar reconstructed and on volume rendering images that were generated by the software Dolphin®. Subsequently, physical measurements were performed using a digital caliper. Analysis of variance (ANOVA), intraclass correlation coefficient (ICC) and Bland-Altman were used for evaluating accuracy and reliability (p&lt;0.05). RESULTS: Excellent intraobserver reliability and good to high precision interobserver reliability values were found for linear measurements from CBCT 3D and multiplanar images. Measurements performed on multiplanar reconstructed images were more accurate than measurements in volume rendering compared with the gold standard. No statistically significant difference was found between voxel protocols, independently of the measurement method. CONCLUSIONS: Linear measurements on multiplanar images of 0.2 and 0.4 voxel are reliable and accurate when compared with direct caliper measurements. Caution should be taken in the volume rendering measurements, because the measurements were reliable, but not accurate for all variables. An increased voxel resolution did not result in greater accuracy of mandible measurements and would potentially provide increased patient radiation exposure. <![CDATA[Relationship between acetaldehyde concentration in mouth air and tongue coating volume]]> Objective Acetaldehyde is the first metabolite of ethanol and is produced in the epithelium by mucosal ALDH, while higher levels are derived from microbial oxidation of ethanol by oral microflora such as Candida species. However, it is uncertain whether acetaldehyde concentration in human breath is related to oral condition or local production of acetaldehyde by oral microflora. The aim of this pilot study was to investigate the relationship between physiological acetaldehyde concentration and oral condition in healthy volunteers. Material and Methods Sixty-five volunteers (51 males and 14 females, aged from 20 to 87 years old) participated in the present study. Acetaldehyde concentration in mouth air was measured using a portable monitor. Oral examination, detection of oral Candida species and assessment of alcohol sensitivity were performed. Results Acetaldehyde concentration [median (25%, 75%)] in mouth air was 170.7 (73.5, 306.3) ppb. Acetaldehyde concentration in participants with a tongue coating status score of 3 was significantly higher than in those with a score of 1 (p&lt;0.017). After removing tongue coating, acetaldehyde concentration decreased significantly (p&lt;0.05). Acetaldehyde concentration was not correlated with other clinical parameters, presence of Candida species, smoking status or alcohol sensitivity. Conclusion Physiological acetaldehyde concentration in mouth air was associated with tongue coating volume. <![CDATA[Effect of composite surface treatment and aging on the bond strength between a core build-up composite and a luting agent]]> Objective The purpose of this study was to assess the influence of conditioning methods and thermocycling on the bond strength between composite core and resin cement. Material and Methods Eighty blocks (8×8×4 mm) were prepared with core build-up composite. The cementation surface was roughened with 120-grit carbide paper and the blocks were thermocycled (5,000 cycles, between 5°C and 55°C, with a 30 s dwell time in each bath). A layer of temporary luting agent was applied. After 24 h, the layer was removed, and the blocks were divided into five groups, according to surface treatment: (NT) No treatment (control); (SP) Grinding with 120-grit carbide paper; (AC) Etching with 37% phosphoric acid; (SC) Sandblasting with 30 mm SiO2 particles, silane application; (AO) Sandblasting with 50 mm Al2O3 particles, silane application. Two composite blocks were cemented to each other (n=8) and sectioned into sticks. Half of the specimens from each block were immediately tested for microtensile bond strength (µTBS), while the other half was subjected to storage for 6 months, thermocycling (12,000 cycles, between 5°C and 55°C, with a dwell time of 30 s in each bath) and µTBS test in a mechanical testing machine. Bond strength data were analyzed by repeated measures two-way ANOVA and Tukey test (α=0.05). Results The µTBS was significantly affected by surface treatment (p=0.007) and thermocycling (p=0.000). Before aging, the SP group presented higher bond strength when compared to NT and AC groups, whereas all the other groups were statistically similar. After aging, all the groups were statistically similar. SP submitted to thermocycling showed lower bond strength than SP without thermocycling. Conclusion Core composites should be roughened with a diamond bur before the luting process. Thermocycling tends to reduce the bond strength between composite and resin cement. <![CDATA[Differential expression of stem cell-like proteins in normal, hyperplastic and dysplastic oral epithelium]]> Objective The identification of stem cells (SC) remains challenging. In the human oral mucosal epithelium, these cells are believed to be in the basal layer (stem cell niche), but their exact location is unclear. The aim of this study was to examine the dysplastic oral epithelium for these SC-like proteins in order to assess their diagnostic value as biomarkers complementing the histological grading of dysplasia. Material and Methods Thirty oral epithelial dysplasia (OED), 25 oral lichen planus (OLP), 10 oral hyperkeratosis and 5 normal oral epithelium (OE) were immunohistochemically examined for four SC markers [integrin β1, neuron-glial-2 (NG2), notch 1 (N1) and keratin 15 (K15)]. Results Three of four SC markers were heterogeneously detected in all samples. K15 overexpression in the lower two-thirds of severe OED suggests an expanded SC niche. Integrin β1 distribution pattern was not measurably different between OEDs and control. NG2 was almost negative to absent in all samples examined. N1 expression was weak and highly variable in normal and dysplastic epithelium, making it an unreliable epithelial stem cell marker. Conclusions Present findings suggest that these markers were unable to identify individual epithelial stem cells. Instead, subpopulations of cells, most probably stem cells and transit amplifying cells with stem cell-like properties were identified in the dysplastic oral epithelium. The characteristic expressions of K15 might be of diagnostic value for oral dysplasia and should be investigated further. <![CDATA[Numerical study of effect of elastomeric stress absorbers on stress reduction in bone-dental implant interface]]> Objective This paper focused on optimal stress distribution in the mandibular bone surrounding a dental implant and is devoted to the development of a modified Osteoplant® implant type in order to minimize stress concentration in the bone-implant interface. Material and Methods This study investigated 0.4 mm thick layers of two elastomeric stress barriers incorporated into the dental implant using 3-D finite element analysis. Results Overall, this proposed implant provoked lower load transfer in bone-implant interface due to the effect of the elastomers as stress absorbers. The stress level in the bone was reduced between 28% and 42% for three load cases: 75 N, 60 N and 27 N in corono-apical, linguo-buccal and disto-mesial direction, respectively. Conclusion The proposed model provided an acceptable solution for load transfer reduction to the mandible. This investigation also permitted to choose how to incorporate two elastomers into the Osteoplant® implant system. <![CDATA[A titration model for evaluating calcium hydroxide removal techniques]]> Objective Calcium hydroxide (Ca(OH)2) has been used in endodontics as an intracanal medicament due to its antimicrobial effects and its ability to inactivate bacterial endotoxin. The inability to totally remove this intracanal medicament from the root canal system, however, may interfere with the setting of eugenol-based sealers or inhibit bonding of resin to dentin, thus presenting clinical challenges with endodontic treatment. This study used a chemical titration method to measure residual Ca(OH)2 left after different endodontic irrigation methods. Material and Methods Eighty-six human canine roots were prepared for obturation. Thirty teeth were filled with known but different amounts of Ca(OH)2 for 7 days, which were dissolved out and titrated to quantitate the residual Ca(OH)2 recovered from each root to produce a standard curve. Forty-eight of the remaining teeth were filled with equal amounts of Ca(OH)2 followed by gross Ca(OH)2 removal using hand files and randomized treatment of either: 1) Syringe irrigation; 2) Syringe irrigation with use of an apical file; 3) Syringe irrigation with added 30 s of passive ultrasonic irrigation (PUI), or 4) Syringe irrigation with apical file and PUI (n=12/group). Residual Ca(OH)2 was dissolved with glycerin and titrated to measure residual Ca(OH)2 left in the root. Results No method completely removed all residual Ca(OH)2. The addition of 30 s PUI with or without apical file use removed Ca(OH)2 significantly better than irrigation alone. Conclusions This technique allowed quantification of residual Ca(OH)2. The use of PUI (with or without apical file) resulted in significantly lower Ca(OH)2 residue compared to irrigation alone. <![CDATA[Management of the Class III malocclusion treated with maxillary expansion, facemask therapy and corrective orthodontic. A 15-year follow-up]]> The facial growth of Class III malocclusion worsens with age, in this case, the early orthopedic treatment, providing facial balance, modifying the maxillofacial growth and development. A 7.6-year old boy presented with Class III malocclusion associated with anterior crossbite; the mandible was shifted to the right and the maxilla had a transversal deficiency. Rapid maxillary expansion followed by facemask therapy was performed, to correct the anteroposterior relationship and improve the facial profile. The patient was followed for a 15-year period, after completion of the treatment, and stability was observed. Growing patients should be monitored following their treatment, so as to prevent malocclusion relapse.