Influence of low-level laser associated with osteogenic proteins recombinant human BMP-2 and Hevea brasiliensis on bone repair in Wistar rats

This study analyzed the newly formed bone tissue after application of recombinant human BMP-2 (rhBMP-2) and P-1 (extracted from Hevea brasiliensis) proteins, 2 weeks after the creation of a critical bone defect in male Wistar rats treated or not with a low-intensity laser (GaAlAs 780 nm, 60 mW of power, and energy density dose of 30 J/cm(2)). The animals were divided into two major groups: (1) bone defect plus low-intensity laser treatment and (2) bone defect without laser irradiation. The following subgroups were also analyzed: (a) 5 μg of pure rhBMP-2; (b) 5 μg of pure P-1 fraction; (c) 5 μg of rhBMP-2/monoolein gel; (d) 5 μg of P-1 fraction/monoolein gel; (e) pure monoolein gel. Comparisons of the groups receiving laser treatment with those that did not receive laser irradiation show differences in the areas of new bone tissue. The group treated with 5 μg of rhBMP-2 and laser irradiation was not significantly different (P >0.05) than the nonirradiated group that received the same treatment. The irradiated, rhBMP-2/monoolein gel treatment group showed a lower area of bone formation than the nonirradiated, rhBMP-2/gel monoolein treatment group (P < 0.001). The area of new bone tissue in the other nonirradiated and irradiated groups was not significantly different (P > 0.05). Furthermore, the group that received the 5 μg of rhBMP-2 application showed the greatest bone formation. We conclude that the laser treatment did not interfere with the area of new bone tissue growth and that the greatest stimulus for bone formation involved application of the rhBMP-2 protein.     

Feasibility of detecting trabecular bone around percutaneous titanium implants in rabbits by in vivo microfocus computed tomography

Objectives: The goal of this study was to examine the feasibility of in vivo imaging of trabecular bone around titanium implants by means of microfocus computed tomography (micro‐CT) and the use of rabbits for this purpose. Materials and Methods: Ten male rabbits type Hollander, received a titanium implant (1.7 mm diameter and 10 mm length) in the trabecular bone of the left tibia. Seven weeks later a micro‐CT scan was taken. Four rabbits were used to monitor potential harmful effects from X‐ray absorption until 4 weeks after scanning. A second group of six rabbits was used for testing the hypothesis that a good correlation exists between in vivo micro‐CT images and histological images of trabecular bone around titanium implants. The six rabbits were scanned and sacrificed immediately. The tibias were extracted and submitted to standard histological procedures. This resulted in a total of 12 histological sections and their corresponding 12 micro‐CT images. Bone area measurements were performed at the left and right side of the implant in three regions: 0–500, 500–1000 and 1000–1500 μm distance from the implant interface. Intra‐class correlations (ICC) were calculated between both techniques. Results: The four rabbits did not show any sign of radiodermatitis 4 weeks after scanning. In the micro‐CT images of the group of six rabbits, trabeculae are visible, but not well defined, due to the presence of noise in the image. The ICC for the right implant side were 0.44 for zone 0–500 μm, 0.48 for zone 500–1000 μm and 0.40 for zone 1000–1500 μm. The ICC for the left implant side could not be calculated. Conclusion: A low agreement was found between the bone measurements from histology and in vivo micro‐CT images. The use of the in vivo micro‐CT for trabecular bone imaging around metallic implants should be restricted to track tendencies in follow‐up studies.     

Systemically alendronate was incorporated into dental tissues but did not cause morphological or mechanical changes in rats teeth

This study evaluated the effect of the systemic use of sodium alendronate in rats in vivo. Forty‐five Wistar rats aged 36 to 42 days and weighing 200 to 230 g were randomly assigned to a control group (n = 20), which received distilled water, and an experimental group (n = 25), which received 2 weekly doses of 1 mg/kg of chemically pure sodium alendronate. The animals were killed after 60 days of treatment. The tibias were removed for analysis of bone mineral density by dual‐energy X‐ray absorptiometry (DXA). Then, the maxillary incisors were extracted for analysis of the mineralized dental tissues using fluorescence spectroscopy (FS), scanning electron microscopy (SEM), bright field microscopy (BFM), and cross‐sectional microhardness (CSMH) testing. DXA and CSMH data were subjected to statistical analysis by Kruskal‐Wallis test (5% significance level). The experimental group presented higher bone mineral density than the control group by DXA. FS analysis revealed presence of alendronate in the mineralized dental tissues of the specimens of the experimental group. Significant morphological differences were not found by SEM and BFM. Enamel and dentin (100 and 300 μm from the dentinoenamel junction) CSMH data did not show significant difference between the control and experimental groups. Based on the obtained results, we conclude that while alendronate increased the bone mineral density and was incorporated into the mineralized dental tissues it did not cause significant alterations in the morphology and microhardness of rat incisor enamel and dentin. Microsc. Res. Tech. 75:1265–1271, 2012. © 2012 Wiley Periodicals, Inc.     

Energy dispersive X‐ray spectrometry study of the protective effects of fluoride varnish and gel on enamel erosion

Background: Dental erosion is a risk factor for dental health, introduced by today's lifestyle. Topical fluoride applications in the form of varnishes and gel may lead to deposition of fluoride on enamel. Purpose: This in vitro study aimed to evaluate the effect of two fluoride varnishes and one fluoride gel on the dissolution of bovine enamel by acids. Methods: Enamel samples (72) were divided (n = 8): artificial saliva (control‐G1), Pepsi Twist® (G2), orange juice (G3), Duraphat® + Pepsi Twist® (G4), Duraphat® + orange juice (G5), Duofluorid® + Pepsi Twist® (G6), Duofluorid® + orange juice (G7), fluoride gel + Pepsi Twist® (G8), and fluoride gel + orange juice (G9). Fluoride gel was applied for 4 min and the varnishes were applied and removed after 6 h. The samples were submitted to six cycles (demineralization: Pepsi Twist® or orange juice, 10 min; remineralization: saliva, 1 h). Samples were analyzed by energy‐dispersive X‐ray fluorescence (144 line‐scanning). Results: The amount of Ca and P decreased significantly in the samples of G2 and G3, and the Ca/P ratio decreased in G3. Mineral gain (Ca) was greater in G9 samples than in G4 > G3 > G5 > G1, and (P) greater in G7 samples than in G9 > G4‐6 > G2‐3. Conclusions: The protective effect of Duofluorid® was significantly lower than fluoride gel against orange juice. The fluoride varnishes can interfere positively with the dissolution of dental enamel in the presence of acidic beverages. Fluoride gel showed the best protection level to extrinsic erosion with low costs. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.     

The role of TiO2 nanotube surface on osseointegration of titanium implants: Biomechanical and histological study in rats

The nanoscale surface of titanium has been studied to improve the cellular recognition of the biological microenvironment and to increase bone–implant interaction. The aim of this study was to analyze the effect of a titanium oxide (TiO₂) nanotube surface with a machined surface on osseointegration tibia implants without primary stability. This study used an experimental design, divided into two groups (n = 16): commercially pure titanium machined implants (Cp‐Ti Ma) and commercially pure titanium anodized implants (Cp‐Ti An). Titanium nanotubes were produced by anodic oxidation, and the topography of surface was analyzed using field emission scanning microscope (FE‐SEM). The implants (2.1 × 2.8 mm Ø) were surgically placed in the right tibia (defects with milling drill 2.5 × 3.2 mm Ø) of 32 Wistar male rats (250–300 g). The animals were euthanized at 7 weeks postoperatively. The maximum value of removal torque was measured (N/cm) in the right tibia half of each group (8 animals/8 tibiae); the other half of each group underwent a nondecalcified protocol, stained with Stevenel blue/Alizarin red, and the formation of bone tissue in close contact to the implant was measured. The obtained data were analyzed statistically (t test). Differences were considered statistically significant for α < 0.05. Cp‐Ti An implants were significantly higher in removal torque and peri‐implant bone healing compared with Cp‐Ti Ma implants (p < .01). Within the limitations of this study, it was observed that the surface modification of titanium by anodization (TiO₂ nanotubes) can improve osseointegration, and this may be very useful to reduce the time required for peri‐implant bone formation.     

Osseointegration of hydroxyapatite and remodeling‐resorption of tricalciumphosphate ceramics

Background: Cancellous bone defects surrounded by still intact bone structures never heal. Ceramics offer a solution providing osteoconductive scaffolds. Purpose: The purpose of the study is to evaluate whether structured β‐TCP and HA implants can reconstruct cancellous bone defects, which role micro‐ and macro‐porosity, stiffness and surface area play; finally the indication for both materials based on its resorbability. Material & Methods: 10 German Shepard dogs were operated on both tibial heads implanting shell‐like fully interconnected ceramic cylinders, using a wet grinding hollow drill coated with diamonds. β‐TCP was compared with HA. A polychromatic sequential labelling with 4 different fluorochromes controlled bone formation dynamics. Non‐decalcifying histology after perfusion fixation and vessel casting was performed. μ‐CT was combined with high resolution microradiography and histology on thin ground crossections. The stages after 6 weeks, 2, 3, 4 months and 15 months were evaluated. Results: In spite of osseointegration of HA and β‐TCP, the osseointegration of both materials was completely different. Both shell‐like bone void fillers were osseointegrated in a sandwich‐like manner. HA yielded primarily a reinforcement of the recipient's cancellous‐bone bed and full osseointegration after 4 months, whereas β‐TCP‐implants were fully osseointegrated after 6 weeks. HA did not show signs of resorption. The resorption of the β‐TCP resulted during remodelling. The final stage showed restitution “ad integrum” of the β‐TCP defects with a physiological architecture, whereas HA was integrated in the cancellous bone construction providing 600 μm measuring macropores showing osteoinductive properties. Microsc. Res. Tech. 76:370–380, 2013. © 2013 Wiley Periodicals, Inc.     

A rabbit maxillary sinus model with simultaneous customized‐implant placement: Comparative microscopic analysis for the evaluation of surface‐treated implants

We describe the use of a rabbit maxillary sinus model, characterized by thin osseous tissue and low bone density, for the evaluation of surface‐treated implants by histologically and histomorphometrically comparing the osseointegration patterns depending on the surface treatment methods. Twenty rabbits were randomly assigned to two groups of 10 animals, one receiving 5 × 3 mm customized implants (machined, MA or sandblasted and acid etched, SLA) placed in sinus and the other receiving implants placed in a tibia. Histological observation of the implant placed in sinus shows relatively more active new bone formation, characterized by trabecular bone pattern underneath the cortical bone in sinus as compared with that in tibia. Histomorphometric analysis in the rabbits receiving implants in a tibia, the NBIC (%) associated with the SLA surface implant was greater than that associated with the MA implant at 2 weeks (55.63 ± 8.65% vs. 47.87 ± 10.01%; P > 0.05) and at 4 weeks (61.76 ± 9.49% vs. 42.69 ± 10.97%; P < 0.05). Among rabbits receiving implants in a sinus, the NBIC (%) associated with the SLA surface implant was significantly greater than that associated with the MA surface implant both at 2 weeks (37.25 ± 7.27% vs. 20.98 ± 6.42%; P < 0.05) and at 4 weeks (48.82 ± 6.77% vs. 31.51 ± 9.14%; P < 0.05). As a result, we suggest that the maxillary sinus model is an appropriate animal model for assessing surface‐treated implants and may be utilized for the evaluation of surface‐treated implants in poor bone quality environment. Microsc. Res. Tech. 78:697–706, 2015. © 2015 Wiley Periodicals, Inc.     

Analysis of efficacy of the self‐assembling peptide‐based remineralization agent on artificial enamel lesions

The objective of this in vitro study was to analyze and compare the biomimetic remineralizing efficacy of the self‐assembling peptide (P11‐4) with agents containing casein phoshopeptide‐amorhous calcium phosphate fluoride (CPP‐ACFP) and sodium fluoride (NaF) on artificial caries lesions using DIAGNOdent and micro‐computed tomography (μCT). Artificial enamel lesions were prepared on extracted impacted sound mandibular third molars. The samples were randomly allocated to four groups (n = 8): Group 1, P11‐4 (Curodont Repair, Credentis AG, Switzerland); Group 2, CPP‐ACFP (MI Varnish, GCCo., Japan); Group3, NaF (Duraphat Varnish, Colgate, Colgate‐Palmolive, NY, USA); Group 4, artificial saliva (control). The agents were applied to demineralized surfaces according to manufacturers' instructions; all specimens were stored in artificial saliva for 1 month. Demineralization and remineralization on enamel surfaces were analyzed and quantified by DIAGNOdent (KaVo, Germany) and μCT (SkyScan1174, Belgium) for lesion depth/area/volume/mineral density (MD). The remineralization efficacy of the agents was evaluated by DIAGNOdent on 1st, 7th, 30th days and by μCT on 30th day. Data were statistically analyzed by ANOVA, Kruskal–Wallis, T test, and Wilxocon tests. The highest remineralization efficacy findings in all periods were determined in Group 1, followed by Groups 2, 3, and 4. The remineralization findings for fluorescence, MD, lesion depth in Group 1 were found significantly higher (p < 0.01) than Group 3; and no significant differences (p > 0.05) were found between Groups 1–2 and Groups 2–3. The area and volume change values in Groups 1, 2, and 3 have shown no significancy (p > 0.05). A significant correlation (p < 0.01) was found between μCT and DIAGNOdent methods. The data of this study have demonstrated that P11‐4 has showed the best remineralization efficacy, followed by CPP‐ACFP and NaF. It is concluded that self‐assembling peptide‐based remineralization agent can be used successfully for biomimetic remineralization of enamel subsurface lesions.     

The effect of surface treatments on the microroughness of laser-sintered and vacuum-cast base metal alloys for dental prosthetic frameworks

This study aimed to evaluate the effect of four chemomechanical surface treatments on the surface average microroughness and profile of laser‐sintered and vacuum‐cast dental prosthetic structures. Square‐shaped blocks (10 mm × 10 mm × 1.5 mm) were prepared as follows: (1) laser‐sintered Co? Cr (L) (ST2724G); (2) cast Co? Cr (C) (Gemium‐cn); and (3) cast Ni? Cr? Ti (T) (Tilite). Specimens of each alloy group were randomly divided into five subgroups (n = 10 each), depending on the conditioning method used: (1) no treatment (control); (2) sandblasting (125 μm Al₂O₃‐particles); (3) silica coating (50 μm silica‐modified Al₂O₃‐particles); (4) oxidation; and (5) oxidation plus opacification. Subgroups 2 and 3 represent “inner” pretreatments proposed for ceramometal restorations to improve the metal surface area available for luting cements. Subgroups 4 and 5 are the “outer” pretreatments required for bonding the aesthetic veneering ceramics to the underlying metal frameworks. Average surface roughness (Ra/μm) was determined using a surface profilometer. Data were analyzed by two‐way ANOVA and Student–Newman–Keuls tests (α = 0.05). Metal surface topography was SEM‐analyzed. Despite the inner pretreatment applied, L samples resulted in the highest microroughness (P < 0.001), whereas sandblasting produced a surface‐smoothing effect in cast specimens. After oxidation, a significant increase in surface roughness occurred in all groups compared with controls, L specimens being the roughest (P < 0.001). Opacification caused a flattening effect of all oxidized structures; all opacified groups resulting in similar microroughness. Laser sintering of Co? Cr enhances the roughness of metal structures, which may improve the frameworks' microretention of the cements, and of the opaquer before the copings are veneered with the aesthetic ceramics. Microsc. Res. Tech. 75:1206–1212, 2012. © 2012 Wiley Periodicals, Inc.     

Reflectance confocal‐laser‐scanning microscopy in vivo assessments of cigarette‐induced dynamic alterations of cutaneous microcirculation on histomorphological level

Objective: Until now, high resolution reflectance confocal‐laser‐scanning microscopy (CLSM) was used for observation of cutaneous morphology in vivo and in real time. We hypothesized that CLSM also allows observation of dynamic processes of cutaneous microcirculation. Methods: Reflectance CLSM (Vivascope1500; Lucid, Rochester, NY) was performed in 24 young male habitual smokers (23 years, range: 19–26, body mass index 23.9 ± 4.04) with relatively limited cigarette exposure (mean: 3.1 ± 2.4 pack‐years). Eight matched nonsmokers served as controls. The quantitative blood cell flow and the diameter of capillary loops were determined prior (baseline), during, as well as 5 and 10 min after smoking. Results: Baseline value for blood cell flow was 55.50 ± 2.33 cells/min, and decreased over 45% during smoking (30.43 ± 3.76/min; P = 0.02). They were still 22% lower (43.33 ± 2.45/min; P = 0.01) 5 min after smoking and exceeded baseline values 10 min after smoking by 13% (63.00 ± 3.10/min; P > 0.05). The baseline values for capillary loop diameter (9.03 ± 0.22 μm) decreased by 21% (7.18 ± 0.28 μm; P = 0.03) during smoking, remained about 9% (8.23 ± 0.18 μm; P = 0.01) lower 5 min after smoking and exceeded baseline values insignificantly by 4% (9.38 ± 0.28 μm; P > 0.05) 10 min after smoking. There were no significant differences to the controls. Conclusion: Reflectance CLSM enables qualitative and quantitative observation of dynamic processes of cutaneous microcirculation on histomorphological level. Microsc. Res. Tech., 2009. © 2008 Wiley‐Liss, Inc.     

Scanning Electron Microscopy (SEM) and Light Microscopy (LM)‐based Palyno‐morphological views of Solanaceae in Western Himalaya

In this study, plants belonging to family Solanaceae growing in Western Himalaya region have been observed palynologically under Light Microscope and Scanning electron microscope. Present investigation comprises of 10 genera and 23 species, namely, Atropa acuminata, Capsicum decoraticus, Capsicum frutescens, Cestrum aurantiacum, Cestrum diurnum, Cestrum nocturnum, Datura alba, Datura innoxia, Datura stramonium, Hyoscymus niger, Lycopersicon esculentum, Nicotiana rustica, Nicotiana tabacum, Petunia alba, Petunia hybrida, Solanum erianthum, Solanum melongena, Solanum miniatum, Solanum pseudocapsicum, Solanum surratense, Solanum tuberosum, Withania coagulans, Withania somnifera. Solanaceae is a eurypalynous family. Grains are usually Tricolporate and Tetracolporate, radially symmetrical, isopolar, prolate‐spheroidal to oblate‐spheroidal to oblate‐spheroidal to subprolate to per prolate or suboblate to oblate, size range: 8.55–72 μm, amb circular, semi‐angular or subangular, aperture drop‐type, labrum common‐type, exine usually 2 μm thick, nexine 1–1.5 μm thick. Tectum usually psilate, sexine reticulate, granulate or striato‐reticulate, with obscure pattern, sexine 1–2 μm thick, nexine 1–1.5 μm thick, and intine 0.5–1 μm thick. Most striking variation has been found in the shape class, aperture‐type, and tectal surface. Based on these characters, taxonomic keys have been made for correct identification of members in Solanaceae. However, the grains of this family are usually tricolporate and have direct relationship with certain members of the family Scrophulariaceae. Palyno‐morphological characters of family Solanaceae have been studied for the first time in Western Himalayan region of Pakistan. These palyno‐morphological characters are significant for identification of the members of family Solanaceae.     

CO2 laser emission modes to control enamel erosion

Considering the importance and prevalence of dental erosion, the aim of this in vitro study was to evaluate the influence of different modes of pulse emission of CO₂ laser associated or not to acidulated phosphate fluoride (APF) 1.23% gel, in controlling enamel erosion by profilometry. Ninety‐six fragments of bovine enamel were flattened and polished, and the specimens were subjected to initial erosive challenge with hydrochloric acid (pH = 2). Specimens were randomly assigned according to surface treatment: APF 1.23% gel and gel without fluoride (control), and subdivided according to the modes of pulse CO₂ laser irradiation: no irradiation (control), continuous, ultrapulse, and repeated pulse (n = 12). After surface treatment, further erosive challenges were performed for 5 days, 4 × 2 min/day. Enamel structure loss was quantitatively determined by a profilometer, after surface treatment and after 5 days of erosive challenges. Two‐away ANOVA revealed a significant difference between the pulse emission mode of the CO₂ laser and the presence of fluoride (P ≤ 0.05). The Duncan's test showed that CO₂ laser irradiation in continuous mode and the specimens only received fluoride, promoted lower enamel loss than that other treatments. A lower dissolution of the enamel prisms was observed when it was irradiated with CO₂ laser in continuous mode compared other groups. It can be concluded that CO₂ laser irradiation in continuous mode was the most effective to control the enamel structure loss submitted to erosive challenges with hydrochloric acid. Microsc. Res. Tech. 78:654–659, 2015. © 2015 Wiley Periodicals, Inc.     

The use of an Er:YAG laser to remove demineralized dentin and its influence on dentin permeability

The purpose of this study was to analyze, correlate, and compare the demineralization and permeability of dentin remaining after caries removal with either an Er:YAG laser, a bur, or a curette. Thirty human dentin fragments were immersed in a demineralizing solution for 20 days and were randomly divided into three groups (n = 10) for the removal of the demineralized lesion. The groups were G1—Er:YAG laser (200 mJ/6 Hz; noncontact at 12 mm; spot: 0.63 mm), G2—Bur, and G3—Curette. The specimens were then immersed in a 10% copper sulfate solution, then in a 1% dithiooxamide alcoholic solution for 30 min and kept in ammonia vapor for 7 days. Next, the specimens were examined with optical microscopy. The amount of demineralized dentin and the level of copper ion infiltration in the dentin were quantified in μm using Axion Vision software. Data were analyzed with the Kruskal‐Wallis test (p < 0.05) and Pearson's Correlation test. The analysis revealed no significant differences between the three caries removal methods in terms of their capacity to remove demineralized tissue (G1: 10.6 μm; G2: 8.4 μm; G3: 11 μm), although the laser removal generated more tissue permeability than the others methods (G1: 17.6 μm; G2: 6.6 μm; G3: 5.5 μm). The correlation between the remaining demineralized dentin and the dentin permeability was moderate for the conventional methods and higher for the Er:YAG laser. It can therefore be concluded that the laser produced an increase in permeability that was directly proportional to the amount of demineralized tissue removal. Microsc. Res. Tech. 76:225–230, 2013. © 2012 Wiley Periodicals, Inc.     

Determination of methyl gallate in Bauhinia retusa extract by high-performance liquid and thin-layer chromatography

Two stability-indicating chromatographic methods are reported for the determination of methyl gallate in crude extracts of Bauhinia retusa. Separation by high performance thin layer chromatography was conducted on silica gel aluminum sheets using 9.5:0.5:0.2 (v/v/v) chloroform:methanol:acetic acid at 280 nm. The results from the 2–40 µg/band were used to prepare a linear calibration graph. The limits of detection and quantitation were 0.5 and 1.5 µg/band, respectively. The reverse phase high performance liquid chromatographic isolation of methyl gallate was performed at ambient temperature with an injection volume of 10 μL. The mobile phase consisted of 40:60 (v/v) methanol:0.1% ortho-phosphoric acid. The separation was performed at 1 mL/min using a detection wavelength of 280 nm. The calibration graph for methyl gallate was rectilinear from 0.02–40 µg/mL with limits of detection and quantitation of 0.004 and 0.010 µg/mL, respectively. For both methods, intra-day and inter-day precision were evaluated and the relative standard deviation was less than 2%, indicating good precision. The robustness was evaluated by making small and deliberate changes to appropriate parameters and the calculated relative standard deviation was less than 2%.The chromatographic methods were employed to determine methyl gallate in crude Bauhinia retusa extracts.     

Effects of sodium hypochlorite as dentin deproteinizing agent and aging media on bond strength of two conventional adhesives

This study evaluated the effect of 10% sodium hypochlorite (NaOCl) as deproteinizing agent and storage media on bond strength (BS) of two etch‐and‐rinse adhesive systems to dentin. Twenty‐eight sound extracted human third molars were divided in four groups (n = 7), according to dentin treatment (conventional etching or etching followed by 10% NaOCl application) and adhesive systems (GB—Gluma 2Bond and OS—One‐Step). After dentin treatments and adhesive application, a composite block was built‐up on dentin surface and teeth were serially sectioned to obtain bonded sticks specimens. The sticks were submitted to three aging conditions: (24H) 24 hr in water (immediate), (SH) 3 hr of NaOCl accelerated‐aging or (1Y) 1 year of water storage. Afterward, submitted to microtensile bond strength test (μTBS), failure modes and adhesive interfaces analyzes. Data were analyzed by two‐way analysis of variance (ANOVA) and Tukey's test (α = .05). Dentin deproteinization before bonding significantly reduced μTBS for GB‐treated group (p < .05), regardless the aging conditions. Water storage for 1 year (1Y) and NaOCl accelerated‐aging (SH) decreased μTBS for both adhesives. Yet, the groups stored in NaOCl (SH) exhibited the lowest BS results (p < .05). Bond strength of deproteinized dentin was dependent on the adhesive system composition and NaOCl accelerated‐aging promoted decreased bond strength and further degradation than water storage for 1 year.     

Residues of different gel formulations on dentinal walls: A SEM/EDS analysis

The aim of this study was to evaluate the presence of residues of sodium hypochlorite gel, chlorhexidine gel, and EDTA gel on dentinal walls after canal preparation through chemical SEM‐ elemental chemical microanalysis (EDS) analysis. Forty‐eight single‐rooted teeth were selected. They had their crowns sectioned and were instrumented with a reciprocating system. The canals were irrigated with 5 mL of saline solution during root canal preparation. After instrumentation, the root canals were irrigated with 3 mL 17% EDTA followed by 1 min of ultrasonic passive activation (3× 20 sec) to remove the smear layer, and then irrigated with 3 mL of saline solution. The specimens were randomized into three groups (n = 12) according to the chemical substance that filled the root canal for 30 min: GI: 5.5% sodium hypochlorite gel; GII: 2% chlorhexidine gel; GIII: 24% EDTA gel; Negative control group: no substance was used. Then, the root canals were irrigated with 6 mL of saline solution followed by 1 min of ultrasonic passive activation (3× 20 sec). After ultrasonic activation, the canals were irrigated with 2 mL saline. The roots were sectioned, and the percentage of each chemical element present in the samples was analyzed through chemical SEM‐EDS microanalysis. All experimental groups showed a significantly higher percentage of chemical elements (Na and/or Cl) than the control group (P < 0.03). This in vitro study has shown that, regardless of chemical solutions used even after the final irrigation protocol, chemical residues of different substances remained attached to the root canal walls. Microsc. Res. Tech. 78:495–499, 2015. © 2015 Wiley Periodicals, Inc.     

Effect of final irrigation protocols on microhardness and erosion of root canal dentin

The aim of this study was to evaluate the effect of final irrigation protocols (17% EDTA, BioPure MTAD, SmearClear, and QMiX) on microhardness and erosion of root canal dentin. Fifty roots were sectioned transversely at the cement–enamel junction and each root was sectioned horizontally into 4‐mm‐thick slices. The samples were divided into five groups (n = 10) according to the final irrigation protocol: G1: distilled water (control group); G2: 17% EDTA; G3: BioPure MTAD; G4: SmearClear; and G5: QMiX. The dentin microhardness was then measured with a load of 25 g for 10 s. Initially, the reference microhardness values were obtained for the samples without any etching. The same samples were then submitted to the final irrigation protocols. A new measure was realized and the difference between before and after the procedures was the dentin microhardness reduction. In sequence, the specimens were submitted to SEM analysis to verify the dentinal erosion. The Kruskal Wallis and Dunn tests (α = 5%) were used to compare the results. The dentin microhardness decreased for all final irrigation protocols. There was no significant difference between groups 2, 3, 4, and 5 (P > 0.05), but this groups presented significant dentin microhardness reduction than G1 (P < 0.05). In G2, occurred the highest incidence of dentinal erosion (P < 0.05). 17% EDTA, BioPure MTAD, SmearClear, and QMiX promoted significant dentin microhardness reduction. Dentinal tubules erosion was promoted by 17% EDTA. Microsc. Res. Tech., 76:1079–1083, 2013. © 2013 Wiley Periodicals, Inc.     

Evaluation of the interaction between sodium hypochlorite and several formulations containing chlorhexidine and its effect on the radicular dentin—SEM and push‐out bond strength analysis

The aim of the current study was to evaluate the presence of debris and smear layer after endodontic irrigation with different formulations of 2% chlorhexidine gluconate (CHX) and its effects on the push‐out bond strength of an epoxy‐based sealer on the radicular dentin. One hundred extracted human canines were prepared to F5 instrument and irrigated with 2.5% sodium hypochlorite and 17% ethylenediaminetetraacetic acid. Fifty teeth were divided into five groups (n = 10), according to the final irrigation protocol with different 2% CHX formulations: G1 (control, no final rinse irrigation), G2 (CHX solution), G3 (CHX gel), G4 (Concepsis), and G5 (CHX Plus). In sequence, the specimens were submitted to scanning electron microscopy (SEM) analysis, in the cervical‐medium and medium‐apical segments, to evaluate the presence of debris and smear layer. The other 50 teeth were treated equally to a SEM study, but with the root canals filled with an epoxy‐based endodontic sealer and submitted to a push‐out bond strength test, in the cervical, middle, and apical thirds. G2, G3, G4, and G5 provided higher precipitation of the debris and smear layer than G1 (P < 0.05), but these groups were similar to each other (P > 0.05), in both segments. The values obtained in the push out test did not differ between groups, independent of the radicular third (P > 0.05). The CHXs formulations caused precipitation of the debris and smear layer on the radicular dentin, but these residues did not interfere in the push‐out bond strength of the epoxy‐based sealer. Microsc. Res. Tech. 77:17–22, 2014. © 2013 Wiley Periodicals, Inc.     

Oral tissue response to ovine grafting biomaterial: Morphological and morphometric study using scanning electron and light microscopy tissue response to ovine grafting biomaterial

OBJECTIVE: To evaluate the oral tissue response to an experimental particle ovine biomaterial by scanning electron microscopy (SEM) and light microscopy (LM). MATERIAL AND METHODS: Forty‐eight rats had surgical periodontal defects treated with either blood clotting (control), bovine biomaterial? (B), or an experimental ovine biomaterial (O). Data from SEM analysis (defect exposure, root surface exposure, diameter of matrix fibers and bundles, and globuli areas; n = 5) were applied to Shapiro–Wilk, Kruskal–Wallis, and Dunn's test, whereas LM analysis (tissue cicatrization characteristics and diameter defect; n = 3) had data applied to two‐way analysis of variance. Animals were monitored for 1 and 3 weeks. RESULTS: By SEM, the O samples showed significant differences from B and C in the area of defect exposure (H_2,15 = 8.66; P < 0.05). In both periods, O and B samples showed similar results for matrix fiber diameters, differently than C samples (H_2,15 = 14.0; P < 0.05). All other SEM variables were considered equivalent among the groups (P > 0.05). Under LM, an acute and chronic granulomatous inflammation was seen in the presence of both biomaterials (B and O, 1 week); both the control and the ovine grafting samples showed mature bone in the repair site (3 weeks); the defect diameter showed similar values among groups, at both monitoring periods (F_2,12 = 1.0401; P > 0.05). CONCLUSION: The ovine particles of this study showed a favorable response to oral tissue repair, demonstrating to be a potential source for the development of bone grafting biomaterials. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Effect of chitosan and Dysphania ambrosioides on the bone regeneration process: A randomized controlled trial in an animal model

The focus of this triple‐blind study was on evaluating the effect of chitosan combined with Dysphania ambrosioides (A) extract on the bone repair process in vivo. In total, 60 male Wistar rats (Rattus norvegicus albinus) weighing between 260 and 270 g were randomly selected for this study and distributed into four groups (n = 15). Group C (chitosan), Group CA5 (chitosan + 5% of D. ambrosioides), Group CA20 (chitosan + 20% of D. ambrosioides), and Group CO (Control‐Blood clot). In each animal, bone defects measuring 2 mm in diameter were performed in both tibias for placement of the substances. After 7, 15, and 30 days, the animals were sedated and sacrificed using the cervical dislocation technique and the tissues were analyzed under optical microscope relative to the following events: inflammatory infiltrate, necrosis, osteoclasts, osteoblasts, fibroblasts, periosteal, and endosteal bone formation. The data were evaluated to verify distribution using the Kolmogorov–Smirnov test, and variance, using the Levene test; as distribution was not normal, data were subjected to the Kruskal–Wallis and Dunn nonparametric tests (p < .05). A significant inflammatory infiltrate was observed in Group CA5 (p = .008) in the time interval of 7 days, and in Group C at 15 (p = .009) and 30 (p = .017) days. Osteoblastic activity was more significant in Group CA20 (p = .027) compared with CA5 in the time interval of 7 days. Group CA20 demonstrated a significantly higher endosteal and periosteal bone formation value in the time interval of 7 (p = .013), 15 (p = .004), and 30 days (p = .008) compared with the other groups. The null hypothesis was refuted, bone regeneration was faster in spheres with an association of chitosan and 20% extract, and complete bone repair occurred clinically at 15 days and histologically at 30 days. The spheres proved to be a promising method for the biostimulation of alveolar bone repair and bone fractures.