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.     

Histological and histochemical evaluations on the effects of high incubation temperature on the embryonic development of tibial growth plate in broiler chickens

The effects of experimentally induced high incubation temperature on the embryonic development of growth plate of the chicken were investigated by means of histological and enzyme histochemical methods. In the experiments, 250 fertile eggs of Ross‐308 broiler strain were divided into two groups, the control eggs were maintained under optimal conditions (37.8°C and 65% ± 2% relative humidity, rh) during the whole incubation period. Heat‐stress imposed eggs were maintained under normal conditions (37.8°C and 65% ± 2% rh) until the 10th day of incubation, and then, continuously (24 h per day) exposed to high temperature (38.8°C and 65% ± 2% rh). Tissue samples were taken from 10 animals of each group at the 11th, 13th, 15th, 18th, 21st days of incubation. Tissue samples were processed by enzyme histochemical methods in addition to routine histological techniques. The relative tibia weights and tibia length were lower in the heat‐stress group compared to the control group. The results of the measurements of the growth plate showed that the proliferative zone was narrowed whereas, the transitional and hypertrophic zone were thickened in the heat stress group. Alkaline phosphatase (ALP) density was significantly decreased in the heat‐stress group compared to the control group. In conclusion, bone formation and growth plate formation are crucial for embryo development and 1°C higher from optimum may increase the incidence of skeletal disorders and leg problems in broiler chickens which is one of the major animal welfare concerns for the poultry industry. Microsc. Res. Tech. 79:106–110, 2016. © 2016 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.     

A comparison of ultrasonic and X-ray methods for imaging the growth plate

The purpose of this study was to assess a systematic and scientific method for measuring children's growth development, in which the accuracy of the existing diagnostic method has not yet been concretely examined. The most popular method for diagnosis of children's growth is to analyse the opening degree of the growth plate in each joint by an X-ray image. However, the X-ray method has some disadvantages; it is impossible to measure the diagnosis of growth periodically and repeatedly due to the radiation problem. Hence, this study introduced a profile analysis and the algorithm of analysing the image of the growth plate using the broadband ultrasound attenuation (BUA) of calcaneus, to verify the possibility of using an alternative ultrasonic method harmless to the human body. The images of the growth plate in the proximal tibiae, phalanges, and calcanei of 269 children (7-16 years old) were obtained using X-ray. The image of the growth plate in the calcanei was also obtained from these children using ultrasound. The results showed that the time of the opening degree of the growth plate in each joint was almost consistent between X-ray and ultrasonic images. Also, the images of the growth plate measured using X-ray and ultrasound showed a high correlation. Therefore, it is expected that the algorithm of ultrasonic profile analysis introduced in this study can replace the existing X-ray method to measure the growth plate correctly.     

Influence of anti-inflammatory administration in collagen maturation process during orthodontic tooth movement

Bone formation is essential to orthodontic tooth movement and bone is formed by collagen. To analyze the collagen maturation process on bone matrix neoformed under nonsteroidal and steroidal treatment during orthodontic tooth movement by polarized microscopy, male Wistar rats (n = 90) were randomly divided into three groups (n = 30): C (control), NSAID (potassium diclofenac) and SAID (disodic phosphate dexamethasone). The animals of the C group received 0.9% saline solution; NSAID group received 5 mg/kg potassium diclofenac (CATAFLAM®); and SAID group received 2 mg/kg phosphate dissodic dexamethasone (DEXANIL®). Animals were sacrificed 3, 7 or 14 days after the placement of orthodontic appliances and the upper first molars were processed histologically and stained with picrosirius. Bone formation was evaluated under polarized light microscopy and 4.5 Image Pro‐Plus® software calculated the percentage of immature/mature collagen present in the groups. On the third days after force application, SAID and NSAID groups showed greater proportion of immature collagen than C group. On the seventh and fourteenth days, there was a lower proportion of mature collagen only in the SAID group (P < 0.001). These data demonstrate that dexamethasone delays the collagen maturation process in established bone matrix. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.     

Histological analysis of the alterations on cortical bone channels network after radiotherapy: A rabbit study

The aim of this study was to evaluate the effects of radiotherapy in cortical bone channels network. Fourteen rabbits were divided in two groups and test group received single dose of 15 Gy cobalt‐60 radiation in tibia, bilaterally. The animals were sacrificed and a segment of tibia was removed and histologically processed. Histological images were taken and had their bone channels segmented and called regions of interest (ROI). Images were analyzed through developed algorithms using the SCILAB mathematical environment, getting percentage of bone matrix, ROI areas, ROI perimeters, their standard deviations and Lacunarity. The osteocytes and empty lacunae were also counted. Data were evaluated using Kolmogorov‐Smirnov, Mann Whitney, and Student's t test (P < 0.05). Significant differences in bone matrix percentage, area and perimeters of the channels, their respective standard deviations and lacunarity were found between groups. In conclusion, the radiotherapy causes reduction of bone matrix and modifies the morphology of bone channels network. Microsc. Res. Tech. 73:1015–1018, 2010. © 2010 Wiley‐Liss, Inc.     

Investigation of the mechanical properties of bone using ultrasound

This paper examines the serial use of ultrasonic velocity measurement to monitor fracture healing. New Zealand White rabbit tibiae were fractured using a constant-energy technique and the ultrasonic velocity along the bone measured in animals sacrificed at 16 day intervals up to 96 days from fracture. In parallel with these measurements the mechanical performance of the healed tibiae were determined using a three-point bending test. Regression analysis failed to show a sufficiently good correlation between ultrasonic velocity measurements and the bending properties of healing fractures for the method to be of use clinically.     

Histological analysis of biocompatibility of different surgical adhesives in subcutaneous tissue

The focus of this study was to test the hypothesis that there would be no difference between the biocompatibility of cyanoacrylate‐based adhesives in rat subcutaneous tissues. In total, 60 male Wistar rats were used, and divided into four groups (n = 15): Group C (control, PVA‐polyvinyl alcohol sponge), Group NO (N‐butyl‐2‐octylcyanoacrylate), Group NH (n‐hexyl‐cyanoacrylate), and Group EC (Ethyl‐cyanoacrylate). The animals were sacrificed after time intervals of 7, 15, and 30 days and tissues were analyzed under optical microscope as regards the events of inflammatory infiltrate, edema, necrosis, granulation tissue, giant cells, young fibroblasts, and collagen formation. The results were statistically analyzed by the Kruskal–Wallis and Dunn tests (p < .05). Significant inflammatory infiltrate was shown for all the adhesives in the time intervals of 7 (p = .004) and 15 days (p = .003). In the time interval of 30 days, moderate inflammatory infiltrate was observed in Groups NH and EC, with significant difference from Control (p = .001). The quantity of collagen fibers in all the experimental groups showed significant difference compared with Control in the time intervals of 7 (p = .002) and 15 days (p = .001), at 30 days only Group EC showed a smaller quantity of collagen fibers in comparison with Control (p = .001). The hypothesis was rejected. The adhesive N‐butyl‐2‐octylcyanoacrylate had less influence on the inflammatory intensity of multinucleated giant cells. Ethyl‐cyanoacrylate demonstrated the lowest level of biocompatibility among the adhesives, but its use in clinical practice may be promising for coaptation of smaller edges of superficial tissue. Surgical adhesives were shown to be feasible for clinical use in substitution of conventional suturing. Ethyl‐cyanoacrylate should be used with caution due to its greater influence on tissues.     

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.     

High dose glucocorticoid hampers bone formation and resorption after bone marrow ablation in rat

We analyzed the effect of glucocorticoid on bone regeneration after bone marrow ablation in tibiae of 8-week-old rats. Methylprednisolone sodium succinate (MPSS) was injected intramuscularly at a dose of 100 mg/kg/day for 3 days. Tibiae on days 1, 3, 5, 7, 10, 12, and 14 after ablation were subjected to tartrate-resistant acid phosphatase staining, immunohistochemistry, in situ hybridization, and transmission electron microscopy (TEM), and measurement of the volume of newly-formed bone and the osteoclast number. MPSS significantly decreased the newly-formed bone volume on day 7, and immature bone still remained on day 10 in the MPSS-treated group. The volume of this bone was significantly higher than that in the control group. However, there were no differences between the groups in the osteoclast number, the expression of mRNAs for osteoblast differentiation markers, and alkaline phosphatase and cathepsin K judged by immunohistochemistry. TEM findings showed no difference in the form of osteoblasts, whereas osteoclasts in the MPSS-treated group had less developed ruffled borders, compared to those in the control group. These results suggest that MPSS treatment affects neither the differentiation nor the shape of osteoblasts, and does not change the osteoclast number or the cathepsin K level. However, high dose MPSS inhibits both bone formation and resorption during bone regeneration after rat tibial bone marrow ablation, and inhibits ruffled border formation in osteoclasts. These data will be useful to develop bone regenerative therapies for bone diseases due to high dose steroid administration.     

Effect of treatment with simvastatin on bone microarchitecture of the femoral head in an osteoporosis animal model

The objective of this study was to evaluate the microarchitecture and trabecular bone strength at the distal region of the femur, and its biomechanical properties with simvastatin administration with two different doses in ovariectomized (OVX) rats. Ninety rats were divided into six groups to evaluate treatment with the simvastatin drug (n = 15): SH (Sham surgery), SH‐5 (5 mg simvastatin), SH‐20 (20 mg simvastatin), OVX, OVX‐5, and OVX‐20. Euthanasia was performed at three different times, five animals per period: 7, 14, and 28 days. The effectiveness of the treatments was evaluated by mechanical testing and histomorphometric analysis of the femurs. The results of analysis by the linear model of mixed effects showed 20 mg of simvastatin results in increased trabecular bone after 14 days (P = 0.039) of ingestion in ovariectomized animals. However, ingestion of 5 mg of simvastatin is able to sensitize the trabecular bone only at 28 days (P = 0.005) of ingestion. In the mechanical tests stiffness improves within 28 days (P = 0.003). Regarding maximum strength, no statistical differences were observed. According to these results, it can be concluded that for a decrease in oral intake, longer treatment times are required. Microsc. Res. Tech. 79:684–690, 2016. © 2016 Wiley Periodicals, 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.     

Alcohol extract of Bauhinia forficata link reduces lipid peroxidation in the testis and epididymis of adult Wistar rats

Bauhinia forficata is a medicinal plant that has flavonoid components with hypoglycemic, antioxidant, hepatoprotective, antibacterial, antiviral and anti‐inflammatory action. Aim of this study is to evaluate the action of B. forficata alcoholic extract in the male genital system of adult male Wistar rats. For that, 20 adult male Wistar rats were distributed into two experimental groups: the B. forficata group, receiving B. forficata alcoholic extract (0.1 ml/10 g body weight/day) on alternate days, and the control group, receiving just the vehicle for 30 days straight both via gavage. On the 31st day, the animals were euthanized, and the testis and epididymis were collected for histopathological, biochemical, morphometric, and sperm count analysis. Mass spectrometry identified new compounds in the extract: trans‐caffeic acid, liquiritigenin, gallocatechin, and 2,4,6‐trihydroxyphenanthren‐2‐glycoside. Biochemical analysis showed higher total cholesterol levels in the testis and lower malondialdehyde levels in the testis and epididymis, in the B. forficata group. The mast cell count showed a reduction in degranulated mast cells in the caput region of the epididymis, in the B. forficata group. The luminal compartment of the caput and the epithelial of the epididymis cauda were reduced, whereas the stromal region of the epididymis caput was increased in the B. forficata group, compared with the control group. The testicular tissue was less impaired, considering that all the histological analyses were similar to the control. We believe that B. forficata alcoholic extract in the male genital system showed antioxidant action, especially in the epididymal tissue.     

Effect of Chenopodium ambrosioides on the healing process of the in vivo bone tissue

The focus of this double‐blind randomized study was on evaluating the effect of an aqueous extract of Mastruz (Chenopodium ambrosioides L.) on the bone repair process in vivo. In total, 36 male Wistar rats were randomly selected for this study, and divided into 3 groups (n = 12): Group HS (Hemostatic Sponge), Group SM (Hemostatic Sponge with Mastruz) and Group BC (Blood Clot). In each animal, bone defects measuring 2 mm in diameter were performed in both tibias for placement of the substances. After 3 and 10 days, the animals were sacrificed, and the tissues were analyzed under an optical microscope relative to the following events: inflammatory infiltrate; necrosis; young fibroblasts; osteoclastic and osteoblastic activity; endosteal and periosteal bone formation; and bone repair. The results were assessed by using Kruskal–Wallis and Mann–Whitney tests (p < .05). Inflammatory infiltrate demonstrated difference between Groups SM and BC in the time interval of 3 days (p = .004); an event related to the presence of the fibrin sponge and liquid of the extract, which induced a foreign body initial reaction. The presence of young fibroblasts (p = .003), osteoclastic (p = .003), and osteoblastic (p = .020) activity was statistically significant between Groups HS and BC in the time interval of 10 days; performance was related to the presence of the sponge within bone. As regards injured bone tissue repair, Group SM demonstrated a higher level of regenerative capacity (p = 0.004), due to a larger quantities of endosteal and periosteal bone formation, demonstrated in Group SM. The aqueous extract of mastruz stimulated bone neoformation, presenting wound closure with bone tissue at the end of 10 days.     

Morphological alterations of radicular dentine pretreated with different irrigating solutions and irradiated with 980-nm diode laser

Background: The topographical features of intraradicular dentine pretreated with sodium hypochlorite (NaOCl) or ethylenediamine tetraacetic acid (EDTA) followed by diode laser irradiation have not yet been determined. Purpose: To evaluate the alterations of dentine irradiated with 980‐nm diode laser at different parameters after the surface treatment with NaOCl and EDTA. Study design: Roots of 60 canines were biomechanically prepared and irrigated with NaOCl or EDTA. Groups were divided according to the laser parameters: 1.5 W/CW; 1.5 W/100 Hz; 3.0 W/CW; 3.0 W/100 Hz and no irradiation (control). The roots were splited longitudinally and analyzed by scanning electron microscopy (SEM) in a quali‐quatitative way. The scores were submitted to two‐way Kruskal–Wallis and Dunn's tests. Results: The statistical analysis demonstrated that the specimens treated only with NaOCl or EDTA (control groups) were statistically different (P < 0.05) from the laser‐irradiated specimens, regardless of the parameter setting. The specimens treated with NaOCl showed a laser‐modified surface with smear layer, fissures, and no visible tubules. Those treated with EDTA and irradiated by laser presented absence of smear layer, tubules partially exposed and melting areas. Conclusions: The tested parameters of 980‐nm diode laser promoted similar alterations on dentine morphology, dependent to the type of surface pretreatment. Microsc. Res. Tech., 2009. © 2008 Wiley‐Liss, Inc.     

Histochemical evidence of osteoclastic degradation of extracellular matrix in osteolytic metastasis originating from human lung small carcinoma (SBC-5) cells

The aim of this study was to assess the dynamics of osteoclast migration and the degradation of unmineralized extracellular matrix in an osteolytic metastasis by examining a well-standardized lung cancer metastasis model of nude mice. SBC-5 human lung small carcinoma cells were injected into the left cardiac ventricle of 6-week-old BALB/c nu/nu mice under anesthesia. At 25-30 days after injection, the animals were sacrificed and their femora and/or tibiae were removed for histochemical analyses. Metastatic lesions were shown to occupy a considerable area extending from the metaphyses to the bone marrow region. Tartrate resistant acid phosphatase (TRAPase)-positive osteoclasts were found in association with an alkaline phosphatase (ALPase)-positive osteoblastic layer lining the bone surface, but could also be localized in the ALPase-negative stromal tissues that border the tumor nodules. These stromal tissues were markedly positive for osteopontin, and contained a significant number of TRAPase-positive osteoclasts expressing immunoreactivity for CD44. We thus speculated that, mediating its affinity for CD44, osteopontin may serve to facilitate osteoclastic migration after their formation associated with ALPase-positive osteoblasts. We next examined the localization of cathepsin K and matrix metallo-proteinase-9 (MMP-9) in osteoclasts. Osteoclasts adjacent to the bone surfaces were positive for both proteins, whereas those in the stromal tissues in the tumor nests showed only MMP-9 immunoreactivity. Immunoelectron microscopy disclosed the presence of MMP-9 in the Golgi apparatus and in vesicular structures at the baso-lateral cytoplasmic region of the osteoclasts found in the stromal tissue. MMP-9-positive vesicular structures also contained fragmented extracellular materials. Thus, osteoclasts appear to either select an optimized function, namely secreting proteolytic enzymes from ruffled borders during bone resorption, or recognize the surrounding extracellular matrix by mediating osteopontin/CD44 interaction, and internalize the extracellular matrices. Microsc.     

Investigation on the properties of gamma irradiated of polytetrafluoroethylene fibers

The physical, thermal, and chemical properties of gamma‐irradiated polytetrafluoroethylene (PTFE) fibers were investigated using X‐ray diffraction (XRD), differential scanning calorimetry (DSC), and Fourier transform infrared (FTIR) spectroscopy. Scanning electron microscopy (SEM) was also used to analyze the surface morphology of irradiated fiber samples. PTFE fiber samples were irradiated by gamma radiation doses ranging from 3 kGy to 40 kGy. The XRD analyses and DSC measurements showed the improvement of crystallinity by gamma irradiation with dose up to 25 kGy reflecting the induced crosslinking with irradiation for PTFE fibers. The crystallinity was found to decrease with higher dose of 40 kGy, reflecting induced amorphization of the polymer sample at the high radiation dose. The calculated crystallite size and XRD parameters showed obvious variations with sample irradiation. The FTIR results showed the liberation of CF₂ groups and the formation of some new chemical bonding with crosslinking‐induced irradiation. The SEM micrographs revealed no variation in the surface morphology of the irradiated fiber samples than the pristine fiber.     

Resin luting materials: Tissue response in dog's teeth

The aim of this study was to evaluate radiographically and histologically the pulpal and periapical response to self‐adhesive (Rely X? Unicem) and self‐etching and self‐curing (Multilink^®) resin‐based luting materials in deep cavities in dogs' teeth. Deep class V cavities (0.5‐mm–thick dentin) were prepared in 60 canine premolars and the following materials were applied on cavity floor: Groups I/V—RelyX? Unicem; Groups II/VI—Multilink^®; Groups III/VII—zinc phosphate cement (control) and; Groups IV/VIII—gutta‐percha (control). Cavities were restored with silver amalgam. Animals were euthanized after 10 days (groups I–IV) and 90 days (groups V–VIII). Tooth/bone blocks were radiographed and processed for histopathological evaluation of pulp and periapical tissue response to the materials. All materials presented similar histopathological features and radiographic findings at both periods. The pulp tissue was intact. The apical and periapical regions and periodontal ligament thickness were normal. No inflammatory cells, resorption of mineralized tissue (dentin, cementum, and alveolar bone) or bacteria were observed. The lamina dura was intact and no areas of periapical bone rarefaction or internal/external root resorption were observed radiographically. It can be concluded that Rely X? Unicem and Multilink^® caused no adverse tissue reactions and may be indicated for cementation of indirect restorations in deep dentin cavities without pulp exposure. Microsc. Res. Tech. 78:1098–1103, 2015. © 2015 Wiley Periodicals, Inc.     

In Situ analysis of CO2 laser irradiation on controlling progression of erosive lesions on dental enamel

The present study aimed to evaluate in situ the effect of CO₂ laser irradiation to control the progression of enamel erosive lesions. Fifty‐six slabs of bovine incisors enamel (5 × 3 × 2.5 mm³) were divided in four distinct areas: (1) sound (reference area), (2) initial erosion, (3) treatment (irradiated or nonirradiated with CO₂ laser), (4) final erosion (after in situ phase). The initial erosive challenge was performed with 1% citric acid (pH = 2.3), for 5 min, 2×/day, for 2 days. The slabs were divided in two groups according to surface treatment: irradiated with CO₂ laser (λ = 10.6 µm; 0.5 W) and nonirradiate. After a 2‐day lead‐in period, 14 volunteers wore an intraoral palatal appliance containing two slabs (irradiated and nonirradiated), in two intraoral phases of 5 days each. Following a cross‐over design during the first intraoral phase, half of the volunteers immersed the appliance in 100 mL of citric acid for 5 min, 3×/day, while other half of the volunteers used deionized water (control). The volunteers were crossed over in the second phase. Enamel wear was determined by an optical 3D profilometer. Three‐way ANOVA for repeated measures revealed that there was no significant interaction between erosive challenge and CO₂ laser irradiation (P = 0.419). Erosive challenge significantly increased enamel wear (P = 0.001), regardless whether or not CO₂ laser irradiation was performed. There was no difference in enamel wear between specimens CO₂‐laser irradiated and non‐irradiated (P = 0.513). Under intraoral conditions, CO₂ laser irradiation did not control the progression of erosive lesions in enamel caused by citric acid. Microsc. Res. Tech. 77:586–593, 2014. © 2014 Wiley Periodicals, Inc.     

Antibacterial efficacy and remineralization capacity of glycyrrhizic acid added casein phosphopeptide‐amorphous calcium phosphate

The aim was to evaluate remineralization capacity and antibacterial efficiency of Tooth Mousse and various amounts of glycyrrhizic acid added Tooth Mousse on primary tooth enamel. Three groups were formed; Group 1 (CPP‐ACP), Group 2 (CPP‐ACP + 5% glycyrrhizic acid), and Group 3 (CPP‐ACP + 10% glycyrrhizic acid) in order to evaluate remineralization capacity. Enamel samples were immersed in demineralization solution and then remineralization agents were applied. Surface microhardness and SEM analyses were performed at the beginning, after demineralization and remineralization. For antibacterial tests, four groups were formed; Group 1, Group 2 and Group 3 and Group 4 (control). Biofilms were then exposed to 10% sucrose eight times per day for 7 days. After biofilm growth period, samples were treated with materials to evaluate antibacterial efficiency except control group. After application of materials, samples were incubated 2 more days at 37°C and at the end of this period, absorbance values of biofilms were determined and data were analyzed. An increase in microhardness values was Group 2 > Group 3 > Group 1, respectively, but there were no significant differences. After remineralization, microhardness values showed significant increases when compared to demineralized groups, but there was no significant difference. All groups showed decreased absorbance value of biofilm when compared with control group but they were insignificant. It was observed that both in Group 2 and Group 3, glycyrrhizic acid did not have a negative effect on remineralization and although they have an increase, it was insignificant. Although glycyrrhizic acid added CPP‐ACP groups showed increased antibacterial activity, they were not statistically significant.