Reduced RANKL expression impedes osteoclast activation and tooth eruption in alendronate-treated rats.

The creation of the eruption pathway requires the resorption of the occlusal alveolar bone by osteoclasts and signaling events between bone and dental follicle are necessary. The aim of the present study has been to evaluate the effect of alendronate on osteoclastogenesis and the expression of the regulator proteins of osteoclast activation, namely RANK, RANKL and OPG, in the bone that covers the first molar germ. Newborn Wistar rats were treated daily with 2.5 mg/kg alendronate for 4, 8, 14, 21 and 28 days, whereas controls received sterile saline solution. At the time points cited, maxillae were fixed, decalcified and processed for light and electron microscopic analysis. TRAP histochemistry was performed on semi-serial sections and the osteoclasts in the occlusal half of the bony crypt surface were counted. TUNEL analysis was carried out on paraffin sections. The occlusal bone that covers the upper first molar was removed in additional 4- and 8-day-old alendronate-treated and control rats in which the expression of RANK, RANKL and OPG was analyzed by SDS-polyacrylamide gel electrophoresis and Western blotting. TRAP-positive osteoclasts were more numerous in the alendronate group at all time points, despite their unactivated phenotype and the presence of apoptotic cells. RANKL expression in the alendronate specimens was inhibited at all time points, unlike in controls. Our findings indicate that the expression of RANKL in the occlusal portion of the bony crypt is unrelated to osteoclast recruitment and differentiation but is crucial to their activation during the creation of the eruption pathway.

Tubule density and diameter in coronal dentin from primary and permanent human teeth.

This study compared dentinal tubule density and diameter of human primary and permanent teeth at different depths of the coronal dentin. Crowns of eight primary second molars and eight permanent third molars were serially sectioned into three disks of ~0.5 mm thickness (superficial, middle, and deep layers), perpendicular to the long axis. Tubule density and diameter were evaluated in 2,000× and 3,000× magnifications by scanning electron microscopy. Data obtained were subjected to two-way repeated measures ANOVA and Tukey's post hoc test (α = 0.05). Tubule density was greater in primary teeth compared with permanent ones, regardless of depth (primary: 124,329 ± 43,594 mm2; permanent: 45,972 ± 21,098 mm2). In general, the tubule density increased as the dentin depth increased, except to the superficial and middle layers from permanent teeth. Tubule diameter was larger in the dentin layer close to the pulp chamber (superficial: 2.4 ± 0.07 µm; middle: 3.70 ± 0.06 µm; deep: 4.28 ± 0.04 µm). No difference was observed between primary (3.48 ± 0.81 µm) and permanent teeth (3.47 ± 0.73 µm). The tubule diameter increases as the dentin depth increases for primary and permanent teeth; however, the tubule density is higher in primary teeth.

Influence of heat treatment and oxygen doping on the mechanical properties and biocompatibility of titanium-niobium binary alloys.

The most commonly used titanium (Ti)-based alloy for biological applications is Ti-6Al-4V, but some studies associate the vanadium (V) with the cytotoxic effects and adverse reactions in tissues, while aluminum (Al) has been associated with neurological disorders. Ti-Nb alloys belong to a new class of Ti-based alloys with no presence of Al and V and with elasticity modulus values that are very attractive for use as a biomaterial. It is well known that the presence of interstitial elements (such as oxygen, for example) changes the mechanical properties of alloys significantly, particularly the elastic properties, the same way that heat treatments can change the microstructure of these alloys. This article presents the effect of heat treatment and oxygen doping in some mechanical properties and the biocompatibility of three alloys of the Ti-Nb system, characterized by density measurements, X-ray diffraction, optical microscopy, Vickers microhardness, in vitro cytotoxicity, and mechanical spectroscopy.

Scanning electron microscopy in vitro study on the effect of Carisolv application on periodontally diseased root surfaces.

The purpose of this study was to evaluate the characteristics of diseased root surfaces treated by the association of scaling and the application of Carisolv. Twenty-four uniradicular periodontally involved teeth were used in this study. The teeth were divided randomly into three groups: eight teeth were scaled and root planed until there was a complete visible removal of calculus (group 1), Carisolv was applied on the root surfaces of eight teeth twice for 30 seconds before scaling with a sharp curette (group 2), and eight teeth received the same treatment as in group 2 but with a blunt curette (group 3). Specimens were examined using scanning electron microscopy. The superficial aspect of the roots from group 1 presented scratches that mirrored the curette cutting edge, and the smear layer completely covered the surface. Root surfaces from groups 2 and 3 also presented a smear layer that covered the surface completely, but it was somewhat smoother than group 1. The use of Carisolv as an adjunct to scaling and root planing presented no advantage for smear layer removal over scaling alone, suggesting that no benefit is obtained by the use of Carisolv during periodontal mechanical treatment.

Comparison of dentin root canal permeability and morphology after irradiation with Nd:YAG, Er:YAG, and diode lasers.

The aim of this study was to compare the effects of Nd:YAG, Er:YAG, and diode lasers on the morphology and permeability of root canal walls. The three laser wavelengths mentioned interact differently with dentin and therefore it is possible that the permeability changes caused will determine different indications during endodontic treatment. Twenty-eight human single-rooted teeth were instrumented up to ISO 40 and divided into four groups: group C, control (GC), non-laser irradiated; group N (GN), irradiated with Nd:YAG laser; group E (GE), with Er:YAG laser and group D (GD) with diode laser. After that, the roots were filled with a 2% methylene blue dye, divided into two halves and then photographed. The images were analyzed using Image J software and the percentage of dye penetration in the cervical, middle, and apical root thirds were calculated. Additional scanning electron microscopy (SEM) analyses were also performed. The analysis of variance (ANOVA) showed significant permeability differences between all groups in the middle and cervical thirds (p < 0.05). The Tukey test showed that in the cervical third, GN presented means of dye penetration statistically significantly lower than all of the other groups. In the middle third, GE and GD showed statistically higher dye penetration means than GC and GN. SEM analysis showed melted surfaces for GN, clean wall surfaces with open dentinal tubules for GE, and mostly obliterated dentinal tubules for GD. Er:YAG (2,094 nm) laser and diode laser (808 nm) root canal irradiation increase dentinal permeability and Nd:YAG (1,064 nm) laser decreases dentin permeability, within the studied parameters.

Adhesives bonded to erbium:yttrium-aluminum-garnet laser-irradiated dentin: transmission electron microscopy, scanning electron microscopy and tensile bond strength analyses.

The aim of this in vitro study was to investigate the effect of erbium:yttrium-aluminum-garnet (Er:YAG) laser irradiation on dentinal collagen by transmission electron microscopy and to analyze the resin-dentin interface by scanning electron microscopy. A tensile bond strength test was also applied. Specimens from 69 sound human third molars were randomly divided into three groups: control (no laser), and two irradiated groups, laser 250 (250 mJ/2 Hz) and laser 400 (400 mJ/4 Hz). Then, specimens were restored with two adhesive systems, an etch-and-rinse or a self-etch system. Although ultrastructural examination showed a modified surface in the irradiated dentin, there was no statistical difference in bond strength values between the laser groups and controls (P < 0.05). In conclusion, the use of Er:YAG laser for ablating human dentin did not alter the main adhesion parameters when compared with those obtained by conventional methods, thus reinforcing its use in restorative dentistry.

Ultrastructure of the interface between periodontal tissues and titanium mini-implants.

OBJECTIVE: To describe the ultrastructure of the interface between periodontal tissues and titanium mini-implants in rat mandibles. MATERIALS AND METHODS: A titanium mini-implant was placed between the buccal roots of the mandibular first molar of 24 adult rats. After 21, 30, 45, 60, 90, and 120 days of implantation, the mandibular portion was removed and fixed in cacodylate-buffered 2% glutaraldehyde + 2.5% formaldehyde. The material was decalcified and processed for scanning and transmission electron microscopy. RESULTS: Ultrastructural analysis revealed a thin cementum-like layer at longer times after implantation at the areas in which the periodontal ligament was in contact with the implant. CONCLUSIONS: The alveolar bone and the periodontal ligament reorganized their constituents around the implant, and a thin cementum-like layer was formed at longer times after implantation at the areas in which the periodontal ligament was in contact with the implant.

Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) for the Examination of Dental Hard Tissues.

This chapter describes laboratory protocols for TEM and SEM approaches allowing the examination of the dental hard tissues' constituents at the ultrastructural level. TEM has the highest resolution power to examine the cellular and extracellular matrix ultrastructure inside a given sample, detecting the presence, location, and quantification of organelles related to the metabolism of the cell type as well as membrane specializations. SEM allows the observation of the sample surface, for examining dimensional topography and distribution of exposed features.

Phototherapy is unable to exert beneficial effects on orthodontic tooth movement in rat molars.

OBJECTIVES: To investigate the effects of irradiation lasers and light-emitting diode (LED) light on root resorption in rat molars during orthodontic tooth movement (OTM). MATERIALS AND METHODS: Twenty-one 12-week-old Wistar male rats were divided into three groups: OTM only (control [CG]), OTM and LED irradiation (DG), and OTM and low laser irradiation therapy (LG). The distance between the first and second molars was used to evaluate the amount of tooth movement. The mesial surfaces of the distopalatal roots were analyzed by scanning electron microscopy, and the area of the resorption was calculated. RESULTS: Statistical analysis showed a decreased amount of tooth movement in the exposed DG or LG compared with CG, which was statistically significant (P = .031 and P = .004, respectively). However, when the DG and LG groups were compared, no statistically significant differences were found (P = .504). The root resorption areas were similar between CG and DG. However, statistically significant differences were found between LG and CG (P = .014) and LG and DG (P = .038). CONCLUSIONS: Phototherapy did not enhance tooth movement, while infrared laser irradiation did increase root resorption.

Roughness of human enamel surface submitted to different prophylaxis methods.

The purpose of this in vitro study was to evaluate alterations in the surface roughness and micromorphology of human enamel submitted to three prophylaxis methods. Sixty-nine caries-free molars with exposed labial surfaces were divided into three groups. Group I was treated with a rotary instrument set at a low speed, rubber cup and a mixture of water and pumice; group II with a rotary instrument set at a low speed, rubber cup and prophylaxis paste Herjos-F (Vigodent S/A Indústria e Comércio, Rio de Janeiro, Brazil); and group III with sodium bicarbonate spray Profi II Ceramic (Dabi Atlante Indústrias Médico Odontológicas Ltda, Ribeirão Preto, Brazil). All procedures were performed by the same operator for 10 s, and samples were rinsed and stored in distilled water Pre and post-treatment surface evaluation was completed using a surface profilometer (Perthometer S8P, Marh, Perthen, Germany) in 54 samples. In addition, the other samples were coated with gold and examined in a scanning electron microscope (SEM). The results of this study were statistically analyzed with the paired t-test (Student), the Kruskal-Wallis test and the Dunn (5%) test. The sodium bicarbonate spray led to significantly rougher surfaces than the pumice paste. The use of prophylaxis paste showed no statistically significant difference when compared with the other methods. Based on SEM analysis, the sodium bicarbonate spray presented an irregular surface with granular material and erosions. Based on this study, it can be concluded that there was an increased enamel surface roughness when teeth were treated with sodium bicarbonate spray when compared with teeth treated with pumice paste.

A comprehensive analysis of human dental pulp cell spheroids in a three-dimensional pellet culture system.

OBJECTIVE: Three-dimensional (3D) cell culture methods are of high importance to studies of biological processes. This is particularly the case with spheroid cultures, which create 3D cell aggregates without the use of exogenous materials. Compared to conventional monolayer cultures, cellular spheroid cultures have been demonstrated to improve multilineage potential and extracellular matrix production. To address this issue in depth, we present a more comprehensive analysis of 3D human dental pulp cell (hDPC) spheroids. DESIGN: hDPC spheroids were fabricated by the pellet culture method and were cultured without adding any reagent to induce differentiation. The gene-expression profiles of the 3D and two-dimensional (2D) cultured hDPCs were compared by complementary DNA microarray analysis. Odontoblastic and osteoblastic differentiation marker gene expression was evaluated by quantitative real-time PCR (RT-qPCR). Hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM) were applied to examine the morphology of hDPC spheroids and extracellular matrix components. RESULTS: Compared with 2D monolayer culture, microarray analysis identified 405 genes and 279 genes with twofold or greater differential expression after 3 days and 28 days of 3D culture, respectively. In 3D hDPC spheroids, gene ontology analysis revealed upregulation of extracellular matrix-related genes and downregulation of cell growth-related genes. RT-qPCR analysis showed higher expression levels of osteocalcin, dentin sialophosphoprotein, and alkaline phosphatase. TEM revealed the morphological characteristics of the fibrillar collagen-rich matrix and cell-cell interactions. CONCLUSIONS: The present findings provide clues to understanding the mechanisms of pellet-cultured hDPCs and contribute to future research in the comparative studies of different 3D culture methods.

Ion-releasing dental restorative composites containing functionalized brushite nanoparticles for improved mechanical strength.

OBJECTIVES: This study describes the synthesis of brushite nanoparticles (CaHPO4·2H2O) functionalized with triethylene glycol dimethacrylate (TEGDMA) and their application in dental restorative composites with remineralizing capabilities. METHODS: Nanoparticles were synthesized, with TEGDMA being added to one of the precursor solutions at three different molar ratios (0:1, 0.5:1 and 1:1, in relation to the ammonium phosphate precursor). Then, they were added (10 vol%) to a photocurable dimethacrylate matrix containing 50 vol% of reinforcing glass particles. The resulting composites were tested for degree of conversion, biaxial flexural strength and elastic modulus (after 24h and 28days in water), and ion release (over a 28-day period). Commercial composites (one microhybrid and one microfilled) were tested as controls. RESULTS: The final TEGDMA content in the functionalizing layer was modulated by the molar ratio added to the precursor solution. Functionalization reduced nanoparticle size, but did not reduce agglomeration. Improved mechanical properties were found for the composite containing nanoparticles with higher TEGDMA level in comparison to the composite containing non-functionalized nanoparticles or those with a low TEGDMA level. All brushite composites presented statistically significant reductions in strength after 28 days in water, but only the material with high-TEGDMA nanoparticles retained strength similar to the microhybrid commercial control. Overall, ion release was not affected by functionalization and presented steady levels for 28 days. SIGNIFICANCE: Though agglomeration was not reduced by functionalization, the improvement in the matrix-nanoparticle interface allowed for a stronger material, without compromising its remineralizing potential.

TGF-beta1 enhances the expression of alpha-smooth muscle actin in cultured human pulpal fibroblasts: immunochemical and ultrastructural analyses.

Transforming growth factor-beta 1 (TGF-beta1) has been related to induce the expression of alpha-smooth muscle actin (alpha-SMA) in fibroblasts during repair. Because pulpal fibroblasts seem to be somewhat different from other fibroblasts, the present study investigated in vitro whether TGF-beta1 enhances the expression of alpha-SMA in human pulpal fibroblasts. TGF-beta1 was added in doses between 5-10 ng/mL to cultures of both dental pulp and gingival human fibroblasts. The expression of alpha-SMA was analyzed by immunofluorescence and Western blotting, whereas the ultrastructure was evaluated by electron microscopy. In addition, the expression of tenascin, osteonectin, and vimentin was also investigated. Both cell types were immunoreactive for alpha-SMA even without TGF-beta1. When TGF-beta1 was added to cell cultures, the expression of alpha-SMA increased dramatically in pulpal fibroblasts, independent of the concentration used. It was confirmed by the Western blotting analysis. Ultrastructure revealed myofilaments and indented nuclei in both fibroblasts treated with TGF-beta1. Tenascin and osteonectin were only immunolabeled in pulpal fibroblasts treated or not with TGF-beta1. Both fibroblast types were positive for vimentin. The present findings showed that TGF-beta1 up-regulated the expression of alpha-SMA, thus inducing pulpal fibroblasts to acquire the myofibroblast phenotype.

Density and diameter of dentinal tubules in etched and non-etched bovine dentine examined by scanning electron microscopy.

Bovine teeth have been widely used in studies focusing adhesion to dentine over the last years. However, little is known about main structural aspects of bovine dentine, especially regarding density and diameter of its tubules. Thirty bovine incisors were randomly divided into two groups. In group I, teeth were cross-sectioned at three depths: outer, middle and inner. The dentinal surfaces were etched with 35% phosphoric acid for 90s. In group II, the teeth were fractured at the same three depths. All the specimens were processed and examined in a Jeol 6100 SEM. Fifteen micrographs from each depth were obtained for the two groups at a magnification of 2500x. To determine the diameter of the dentinal tubules, the LEICA Q500 MC software was used. All the results were statistically processed in the EXCEL software. The dentinal tubules were wider at the outer regions (GI: 5.21+/-0.64microm; GII: 2.30+/-0.09microm) than that at the inner regions (GI: 2.71+/-0.72microm; GII: 1.77+/-0.06microm). Tubular density was higher at the inner regions (50310+/-11178tubules/mm(2)) than that at the outer regions (18772+/-2951tubules/mm(2)). In addition, the peritubular dentine was wider at the outer depth than at the inner one. Our results showed that bovine dentine possesses some structural aspects different from those previously reported for human dentine, mainly related to the diameter of dentinal tubules and to the thickness of peritubular dentine at the several depths.

Root resorption repair in mandibular first premolars after rotation. A transmission electron microscopy analysis combined with immunolabeling of osteopontin.

INTRODUCTION: A previous study with scanning electron microscopy showed that orthodontic root resorption occurs at the lateral surfaces of premolar roots for 2 to 6 weeks after orthodontic rotation. The purpose of this investigation was to observe how resorbed cementum repairs during rotation movement. METHODS: Twenty-one mandibular first premolars from 12 patients, orthodontically indicated for extraction, were used. They were intra-individually divided into 2 groups: 8 teeth were not moved (control group), and 13 were rotated (experimental group). In the experimental group, a rotational force (25 g both buccally and lingually) with a precise biomechanical model, individually calibrated, was applied for 2, 3, 4, or 6 weeks. After extraction, the teeth were fixed and decalcified, and 8 were conventionally processed for transmission electron microscopy, and 13 teeth were processed for high-resolution immunocytochemistry by using an antibody against osteopontin. The samples were analyzed in a transmission electron microscope. RESULTS: This examination showed areas of repair in previously resorbed lacunae in the experimental group. Both the clastic cells and the root surface showed immunolabeling for osteopontin. In addition to areas of cementum resorption and various degrees of cell and extracellular matrix degeneration, active cementoblasts and fibroblasts in several stages of differentiation and activity appeared adjacent to newly synthesized collagen fibers, thus reestablishing the function of the periodontal ligament. CONCLUSIONS: We concluded that cementum repair occurs after resorption during rotation movement and that noncollagenous matrix protein osteopontin plays a role in both resorbing and repairing.

Polymer-based material containing calcium phosphate particles functionalized with a dimethacrylate monomer for use in restorative dentistry.

Dicalcium phosphate dihydrate particles functionalized with triethyleneglycol dimethacrylate were synthesized and added to a photocurable mixture of bisphenol-A glycidyl dimethacrylate and triethyleneglycol dimethacrylate with the purpose of developing a resin composite capable of releasing calcium and phosphate ions to foster dental remineralization. Particle functionalization would minimize the deleterious effect of adding low cohesive strength nano-structured particles with no chemical interaction with the organic matrix on the material's mechanical properties. The results showed that calcium release over 28 days was not impaired by particle functionalization. A statistically significant 32% increase in strength was recorded with the use of functionalized dicalcium phosphate dihydrate in comparison to the material containing non-functionalized particles. However, the strength of the unfilled resin was not matched by the composite with functionalized particles. Elastic modulus increased with particle incorporation, regardless of functionalization. Degree of conversion and optical properties (total transmittance and color change/ΔE) of the resin-based materials were not affected by the addition of dicalcium phosphate dihydrate particles (functionalized or not).

Histologic and immunohistochemical evaluation of biocompatibility of castor oil polyurethane polymer with calcium carbonate in equine bone tissue.

OBJECTIVE To evaluate the efficacy of castor oil polyurethane polymer with calcium carbonate for use in a unicortical ostectomy on the dorsal surface of the third metacarpal bone of horses. ANIMALS 6 adult horses. PROCEDURES A unicortical ostectomy was created on the dorsal surface of both third metacarpal bones of each horse. Castor bean (Ricinus communis) oil polyurethane polymer with calcium carbonate was implanted into the ostectomy on 1 limb, and the ostectomy of the contralateral limb was left unfilled and served as a control sample. Ostectomy sites were evaluated histologically 120 days later. Biopsy specimens were obtained from the interface of bone and polymer or the interface of bone and newly formed tissue; specimens were processed for histomorphometric evaluation by use of light microscopy, immunohistochemical analysis, histochemical analysis, and transmission electron microscopy. RESULTS Osteoconductive activity of the biomaterial was confirmed by the presence of osteoblasts in the biopsy specimens. Absence of a chronic inflammatory response or foreign body reaction indicated biocompatibility. Expression of osteoblast markers was detected in the newly formed tissue. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that castor oil polyurethane polymer with calcium carbonate could be an acceptable compound for use as a bone substitute in horses with fractures in which bone filling is necessary.

Synthesis and characterization of silver phosphate/calcium phosphate mixed particles capable of silver nanoparticle formation by photoreduction.

Silver phosphate is a semi-conductor sensitive to UV-Vis radiation (<530nm). Exposure to radiation removes electrons from the oxygen valence shell, which are scavenged by silver cations (Ag+), forming metallic silver (Ag0) nanoparticles. The possibility of silver nanoparticle formation in situ by a photoreduction process was the basis for the application of mixed calcium phosphate/silver phosphate particles as remineralizing and antibacterial fillers in resin-based dental materials. Mixed phosphate particles were synthesized, characterized and added to a dimethacrylate resin in 20% or 30% mass fractions to investigate their efficacy as ion-releasing fillers for dental remineralization and antibacterial activity. The formation of metallic silver nanoparticles after exposure to visible radiation from a dental curing unit (peak emission: 470nm) was demonstrated by particle X-ray diffraction and scanning electron microscopy analysis of the composite fractured surface. Calcium and phosphate release from materials containing the mixed particles were similar to those containing pure CaP particles, whereas Streptococcus mutans colonies were reduced by three orders of magnitude in relation to the control, which can be attributed to silver release. As expected, the optical properties of the materials containing mixed phosphate particles were compromised by the presence of silver. Nevertheless, materials containing mixed phosphate particles presented higher fracture strength and elastic modulus than those with pure CaP particles.

Immunocytochemical study of amelogenin deposition during the early odontogenesis of molars in alendronate-treated newborn rats.

Newborn rats were treated with sodium alendronate to study how enamel is formed and the effect of alendronate during early odontogenesis. Ultrastructural analysis combined with high-resolution immunocytochemistry for amelogenin was carried out. Twelve rats were subjected to daily SC injections of sodium alendronate (2.5 mg/kg/day) for 3 days on their dorsal region, whereas three rats were daily injected with saline solution as a control. Molar tooth germs from 3-day-old rats were fixed under microwave irradiation in 0.1% glutaraldehyde + 4% formaldehyde buffered at pH 7.2 with 0.1 M sodium cacodylate. The specimens were left undecalcified, postfixed with osmium tetroxide, dehydrated, and embedded in LR White resin. Ultrathin sections were incubated with a chicken anti-24-kDa rat amelogenin antibody, a secondary antibody, and finally with a protein A-gold complex. Large patches of amelogenin were present over the unmineralized mantle dentin and at early secretory ameloblasts. At more advanced stages, they were also detected at the enamel matrix, as well as in the mineralized dentin, at the periodontoblastic space of the dentinal tubules, and at the predentin. It is likely that the main effect of alendronate at early stages of odontogenesis is the increase of synthesis/secretion of amelogenin, promoting its deposition within the forming dentin and enamel.

Swine teeth as potential substitutes for in vitro studies in tooth adhesion: a SEM observation.

Animal teeth are routinely used in studies that test dental materials; however, no studies have yet proposed the use of swine teeth in laboratory tests. The purpose of this study was to compare the effects of acid etching on the dentine and enamel of human and swine teeth, aiming at the use of swine teeth as substitutes for human teeth in adhesion tests. Four human molars and four swine molars free of caries were used. For the study of enamel, two human and two swine teeth were longitudinally sectioned with a dental handpiece and a carborundum disk. For evaluation of dentine, the remaining teeth had the occlusal third of their crowns removed using a low speed diamond saw. All specimens underwent acid etching with 37% phosphoric acid for 30s. The etched surfaces were then washed with air/water spray for 10s. The specimens were then prepared for analysis in a Sterioscan 440 scanning electron microscope. It was observed a similar morphology in human and swine teeth, despite swine enamel was thinner than human enamel. Acid etching of swine enamel promoted a demineralization similar to that observed in human enamel in which the three types of Silverstone's patterns were observed. Acid etching of dentine exposed the opening of dentinal tubules in both swine and human teeth. Peritubular dentine was removed by phosphoric acid, while collagen fibrils appeared discernible in intertubular dentine. Thus, this study showed that enamel and dentine from swine and human teeth share many structural similarities after acid etching.