OBJECTIVE: To explore the effects of two methods of smear layer removal on the surface properties of dentin. METHODS: Sixty extracted sound third molars were collected in this study, and were prepared as uniform dentin specimens with smear layer. All specimens were randomly divided into three groups: Control group, ultrasonic treatment (UT) group and etched treatment (ET) group. Scanning electron microscope (SEM) were used to observe the surface micromorphology of all three groups. Then, the surface elements, mineral phases and functional groups were analyzed by energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD) and flourier transformed infrared spectrometer (FTIR) respectively. The mechanical properties, hydrophilicity and biocompatibility were also further evaluated. RESULTS: It was revealed that dentin tubules of UT and ET groups were exposed, but lots of dentin debris piled up on the surface of the control one which covered up dentin tubules on the surface. The EDX results should that the weaker peak value of calcium and phosphorus in ET group than control and UT groups. Characteristic peaks of hydroxyapatite could be seen by XRD in all of the three groups, but lower distinctive peaks of amide Ⅰ, Ⅱ and Ⅲ bands of collagen of the dentin surface in control group than in ET and UT groups. The microhardness results showed that ET group was lower than control and UT groups, the difference was significant (P < 0.05). Better hydrophilicity of ET group was investigated (P < 0.05) than control group and UT group. Cells could be observed to adhere normally to dentin surface of each group which meant that all of the three groups had good biocompatibility. CONCLUSION Both UT and ET could effectively remove the smear layer on the surface of dentin and had no adverse effect of the dentin micromorphology and biocompatibility. The ultrasonic removal of the smear layer did not influence the mineral structure, hydrophilicity and mechanical properties of dentin surface. Although ET can effectively improve the hydrophilicity of dentin but decreased mechanical properties and the content of calcium and phosphorus.
Dentin/ultrastructure* ; Humans ; Surface Properties ; Smear Layer ; Molar, Third ; Microscopy, Electron, Scanning ; Dental Etching/methods*

OBJECTIVE: To explore the histological structure of the deciduous teeth and the tooth germs of Tibetan miniature pigs for studies of dental tissue diseases and tooth regeneration. METHODS: The structure of the deciduous teeth of Tibetan miniature pigs was observed by X-ray. The ultrastructure of the enamel and dentin of deciduous teeth was characterized by scanning electron microscopy. The jaws and teeth were three-dimensionally reconstructed using Mimics software based on Micro-CT scanning of the deciduous teeth. Image J software was used to calculate the gray value and the mineralization density of the deciduous teeth. Hisotological structure of the tooth germ and the pulp tissue of Tibetan miniature pigs was observed using HE staining. RESULTS: The deciduous teeth of Tibetan miniature pigs were composed of enamel, dentin and medullary pulp tissue. The permanent tooth germ were formed during the deciduous dentition. The enamel and dentin ultrastructure of deciduous teeth were consistent with that of human deciduous teeth. The enamel and dentin mineralization densities were 2.47±0.09 g/cm and 1.72±0.07 g/cm, respectively. The pathological structures of tooth germ and pulp tissue were similar to those of human teeth, and the pulp tissue of the deciduous teeth was in an undifferentiated state. CONCLUSIONS The deciduous teeth of Tibetan miniature pig have similar anatomy, ultrastructure and histopathological structure to human teeth and can serve as a good animal model for studying human dental tissue diseases and the mechanisms of tooth regeneration.
Animals ; Dental Enamel ; ultrastructure ; Dental Pulp ; Dentin ; ultrastructure ; Swine ; Swine, Miniature ; Tibet ; Tooth Germ ; Tooth, Deciduous ; anatomy & histology

Tooth development is a complex process that involves precise and time-dependent orchestration of multiple genetic, molecular, and cellular interactions. Ameloblastin (AMBN, also named "amelin" or "sheathlin") is the second most abundant enamel matrix protein known to have a key role in amelogenesis. Amelogenesis imperfecta (AI [MIM: 104500]) refers to a genetically and phenotypically heterogeneous group of conditions characterized by inherited developmental enamel defects. The hereditary dentin disorders comprise a variety of autosomal-dominant genetic symptoms characterized by abnormal dentin structure affecting either the primary or both the primary and secondary teeth. The vital role of Ambn in amelogenesis has been confirmed experimentally using mouse models. Only two cases have been reported of mutations of AMBN associated with non-syndromic human AI. However, no AMBN missense mutations have been reported to be associated with both human AI and dentin disorders. We recruited one kindred with autosomal-dominant amelogenesis imperfecta (ADAI) and dentinogenesis imperfecta/dysplasia characterized by generalized severe enamel and dentin defects. Whole exome sequencing of the proband identified a novel heterozygous C-T point mutation at nucleotide position 1069 of the AMBN gene, causing a Pro to Ser mutation at the conserved amino acid position 357 of the protein. Exfoliated third molar teeth from the affected family members were found to have enamel and dentin of lower mineral density than control teeth, with thinner and easily fractured enamel, short and thick roots, and pulp obliteration. This study demonstrates, for the first time, that an AMBN missense mutation causes non-syndromic human AI and dentin disorders.
Adult ; Amelogenesis Imperfecta ; genetics ; Cells, Cultured ; China ; Codon ; Dentin ; abnormalities ; ultrastructure ; Female ; Humans ; Male ; Microsatellite Repeats ; Microscopy, Electron, Scanning ; Middle Aged ; Mutation, Missense ; Pedigree ; RNA ; analysis ; Transfection ; Whole Exome Sequencing

The time domain entombment of bacteria by intratubular mineralization following orthograde canal obturation with mineral trioxide aggregate (MTA) was studied by scanning electron microscopy (SEM). Single-rooted human premolars (n=60) were instrumented to an apical size #50/0.06 using ProFile and treated as follows: Group 1 (n=10) was filled with phosphate buffered saline (PBS); Group 2 (n=10) was incubated with Enterococcus faecalis for 3 weeks, and then filled with PBS; Group 3 (n=20) was obturated orthograde with a paste of OrthoMTA (BioMTA, Seoul, Korea) and PBS; and Group 4 (n=20) was incubated with E. faecalis for 3 weeks and then obturated with OrthoMTA-PBS paste. Following their treatments, the coronal openings were sealed with PBS-soaked cotton and intermediate restorative material (IRM), and the roots were then stored in PBS for 1, 2, 4, 8 or 16 weeks. After each incubation period, the roots were split and their dentin/MTA interfaces examined in both longitudinal and horizontal directions by SEM. There appeared to be an increase in intratubular mineralization over time in the OrthoMTA-filled roots (Groups 3 and 4). Furthermore, there was a gradual entombment of bacteria within the dentinal tubules in the E. faecalis inoculated MTA-filled roots (Group 4). Therefore, the orthograde obturation of root canals with OrthoMTA mixed with PBS may create a favorable environment for bacterial entombment by intratubular mineralization.
Aluminum Compounds ; therapeutic use ; Calcification, Physiologic ; physiology ; Calcium Compounds ; therapeutic use ; Crystallization ; Dental Pulp Cavity ; microbiology ; Dentin ; microbiology ; Drug Combinations ; Enterococcus faecalis ; ultrastructure ; Humans ; Methylmethacrylates ; therapeutic use ; Microscopy, Electron, Scanning ; Oxides ; therapeutic use ; Root Canal Filling Materials ; therapeutic use ; Root Canal Obturation ; methods ; Root Canal Preparation ; instrumentation ; Silicates ; therapeutic use ; Time Factors ; Zinc Oxide-Eugenol Cement ; therapeutic use

Our previous studies showed that biomodification of demineralized dentin collagen with proanthocyanidin (PA) for a clinically practical duration improves the mechanical properties of the dentin matrix and the immediate resin-dentin bond strength. The present study sought to evaluate the ability of PA biomodification to reduce collagenase-induced biodegradation of demineralized dentin matrix and dentin/adhesive interfaces in a clinically relevant manner. The effects of collagenolytic and gelatinolytic activity on PA-biomodified demineralized dentin matrix were analysed by hydroxyproline assay and gelatin zymography. Then, resin-/dentin-bonded specimens were prepared and challenged with bacterial collagenases. Dentin treated with 2% chlorhexidine and untreated dentin were used as a positive and negative control, respectively. Collagen biodegradation, the microtensile bond strengths of bonded specimens and the micromorphologies of the fractured interfaces were assessed. The results revealed that both collagenolytic and gelatinolytic activity on demineralized dentin were notably inhibited in the PA-biomodified groups, irrespective of PA concentration and biomodification duration. When challenged with exogenous collagenases, PA-biomodified bonded specimens exhibited significantly less biodegradation and maintained higher bond strengths than the untreated control. These results suggest that PA biomodification was effective at inhibiting proteolytic activity on demineralized dentin matrix and at stabilizing the adhesive/dentin interface against enzymatic degradation, is a new concept that has the potential to improve bonding durability.
Chlorhexidine ; chemistry ; pharmacology ; Collagenases ; pharmacology ; Dental Bonding ; Dental Cements ; chemistry ; Dental Stress Analysis ; instrumentation ; Dentin ; drug effects ; ultrastructure ; Dentin-Bonding Agents ; chemistry ; Gelatinases ; pharmacology ; Humans ; Hydroxyproline ; analysis ; Matrix Metalloproteinase 8 ; pharmacology ; Matrix Metalloproteinase Inhibitors ; chemistry ; pharmacology ; Proanthocyanidins ; chemistry ; pharmacology ; Stress, Mechanical ; Surface Properties ; Tensile Strength ; Tooth Demineralization ; pathology ; physiopathology

This study examined the adhesive strength of two self-adhesive methacrylate resin-based sealers (MetaSEAL and RealSeal SE) to root dentin and compared them with RealSeal and AH Plus in properties. A total of 48 extracted human single-rooted teeth were used to prepare the 0.9-mm thick longitudinal tooth slice (each per tooth). Standardized simulated canal spaces of uniform dimensions were prepared in the middle of radicular dentin. After treated with 5.25% sodium hypochlorite (NaOCl) and 17% EDTA, tooth slices were allocated randomly to four groups (n=12) in terms of different sealers used: MetaSEAL, RealSeal SE, RealSeal, and AH plus groups. The simulated canal spaces were obturated with different sealers in each group. There were 10 slabs with 20 simulated canal spaces (n=20) used in each group for push-out testing. The failure modes and the ultrastructures of fractured sealer-dentin interfaces were examined. The remaining 2 slabs in each group underwent partial demineralization for observation of the ultrastructure of resin tags. The results showed that the push-out bond strength was 12.01±4.66 MPa in MetaSEAL group, significantly higher than that in the other three groups (P<0.05). Moreover, no statistically significant differences were noted in the push-out bond strength between RealSeal SE (5.43±3.68 MPa) and AH Plus (7.34±2.83 MPa) groups and between RealSeal SE and RealSeal (2.93±1.76 MPa) groups (P>0.05). Mixed failures were predominant in the fractured sealer-dentin interfaces in MetaSEAL and AH Plus groups, while adhesive failures were frequently seen in RealSeal SE and RealSeal groups. In conclusion, after complete removal of the smear layer, MetaSEAL showed superior bond ability to root dentin. The RealSeal SE is applicable in clinical practice, with its adhesive strength similar to that of AH Plus. The self-adhesive methacrylate resin-based sealer holds promise for use in endodontic treatment.
Adhesives ; standards ; Composite Resins ; standards ; Compressive Strength ; Dental Bonding ; Dental Pulp Cavity ; ultrastructure ; Dental Stress Analysis ; methods ; Dentin ; Dentin-Bonding Agents ; standards ; Epoxy Resins ; standards ; Humans ; Materials Testing ; methods ; Methacrylates ; standards ; Microscopy, Electron, Scanning ; Root Canal Filling Materials ; standards ; Root Canal Preparation ; Tooth Root

OBJECTIVE: To explore the effects of three dentine desensitizers on the surface morphology of freshly exposed dentin and to evaluate their occlusion effects on dentinal tubules using scanning electron microscope (SEM). METHODS: A total of 16 isolated human premolar samples, which were prepared to expose dentine, were randomly divided into a control group (n=4), a Hybrid Coat group (n=4), a Prime & Bond NT group (n=4), and a anti-sensitivity toothpaste group (n=4). After treatment with dentine desensitizers, one half of the samples in each group were vertically cleaved. Finally, the surfaces and cross sections of the samples were observed by SEM. RESULTS: The exposed tubule was almost occluded in the Hybrid Coat group and the Prime & Bond NT group, while only the majority of tubules could be sealed in the anti-sensitivity toothpaste group. The cross-section images showed that sediments were visible in all groups except the control group. CONCLUSION Hybrid Coat and Prime & Bond NT are able to effectively seal tubules, while the effect of anti-sensitivity toothpaste is slightly poor.
Dentin ; ultrastructure ; Dentin Desensitizing Agents ; pharmacology ; Dentin Permeability ; drug effects ; Humans ; Microscopy, Electron, Scanning ; Polymethacrylic Acids ; pharmacology ; Toothpastes ; pharmacology

The present investigation assessed the effect of acid etching on marginal adaptation of white- and gray-colored mineral trioxide aggregate (MTA) to apical dentin using microcomputed tomography (micro-CT) and scanning electron microscopy (SEM). Sixty-four extracted single-rooted human maxillary teeth were used. Following root-end resection and apical preparation, the teeth were equally divided into four groups according to the following root end filling materials: (i) white-colored MTA (WMTA), (ii) etched WMTA (EWMTA), (iii) gray-colored MTA (GMTA) and (iv) etched GMTA (EGMTA). After 48 h, the interface between root-end filling materials and the dentinal walls was assessed using micro-CT and SEM. Data were statistically analyzed using the Kruskal-Wallis and Dunn tests. Micro-CT analysis revealed gap volumes between the apical cavity dentin walls and EGMTA, GMTA, EWMTA and WMTA of (0.007 1±0.004) mm(3), (0.053±0.002) mm(3), (0.003 6±0.001) mm(3) and (0.005 9±0.002) mm(3) respectively. SEM analysis revealed gap sizes for EGMTA, WMTA, EWMTA and GMTA to be (492.3±13.8) µm, (594.5±17.12) µm, (543.1±15.33) µm and (910.7±26.2) µm respectively. A significant difference in gap size between root end preparations filled with GMTA and EGMTA was found (P<0.05). No significance difference in gap size between WMTA and EWMTA were found in either SEM or micro-CT analysis. In conclusion, pre-etching of apical dentin can provide a better seal for GMTA but not for WMTA.
Acid Etching, Dental ; methods ; Aluminum Compounds ; chemistry ; Apicoectomy ; methods ; Calcium Compounds ; chemistry ; Dental Bonding ; Dental Marginal Adaptation ; Dental Pulp Cavity ; ultrastructure ; Dentin ; ultrastructure ; Drug Combinations ; Humans ; Materials Testing ; Microscopy, Electron, Scanning ; Oxides ; chemistry ; Retrograde Obturation ; methods ; Root Canal Filling Materials ; chemistry ; Root Canal Preparation ; instrumentation ; methods ; Silicates ; chemistry ; Surface Properties ; Time Factors ; Tooth Apex ; ultrastructure ; X-Ray Microtomography

OBJECTIVETo observe the influence of different bonding process and three different root canal sealing materials on microstructure of root canal dentin bonding interface after fiber post and resin bonding, so as to improve clinical operation steps and to optimize fiber post resin bonding effect.

METHODSFifteen human single mandibular first premolars were selected. Three were bonded with fiber posts through Relyx Unicem conventional bonding steps after filled with root canal sealing materials of zinc oxide eugenol paste (Group A), and another three were bonded through the same steps after filled with sealing materials of Vitapex (Group B). The other nine were filled with sealing materials of AH Plus, randomly divided into three groups and bonded through different steps as follows: conventional bonding steps only (Group C), etching with 35% phosphoric acid before conventional bonding steps (Group D), and etching and coating with Singlebond 2 adhesive before conventional bonding steps (Group E). After immersed in saline solution for one week, all the roots were cut into three sections of 3 mm in thickness with emery chip and numbered as crown section, middle section and tip section respectively. The samples were observed the resin protrusion in mixed layer of dentin interface and dentinal tubules by scanning electron microscopy.

RESULTSWe observed the resin protrusion in microstructures of the roots bonded through Relyx Unicem after filled with three different root canal sealing materials (Group A, B, C, E): most obvious in the root crown sections, middle in the root middle sections and least in the root tip sections. Differences were observed in roots filled with different sealing materials: little resin protrusion were observed in crown sections only in Group A and B, but large number of resin protrusion were found in crown and middle sections in Group C-E. Compared with Group C, no more resin protrusion were found in Group D. More and elongated resin protrusions were found in Group E.

CONCLUSIONSWe recommend using AH Plus as root canal sealing materials for residual crown and root needed to strengthen by fiber post. It is no need to etch before Relyx Unicem conventional bonding steps. However, coating Singlebond 2 adhesive after acid etching has the potential to increase fiber post cementation.

Acid Etching, Dental ; methods ; Calcium Hydroxide ; Dental Bonding ; methods ; Dental Pulp Cavity ; ultrastructure ; Dentin ; ultrastructure ; Dentin-Bonding Agents ; chemistry ; Epoxy Resins ; Humans ; Mandible ; ultrastructure ; Microscopy, Electron, Scanning ; Molar ; ultrastructure ; Post and Core Technique ; Resin Cements ; Root Canal Filling Materials ; chemistry ; Silicones ; Tooth Root ; ultrastructure ; Zinc Oxide-Eugenol Cement

OBJECTIVETo establish the femtosecond laser experimental platform in vitro for numerical controlled cavity preparation, and to evaluate the roughness quantitatively and observe the microscopic morphology of the cutting surface.

METHODSEnamel and dentin planes were prepared on human third molars. A universal motion controller was used to control the samples to do rectangle wave motion perpendicular to the incident direction of the laser at focus. The surface roughness was observed with confocal laser scanning microscope.

RESULTSPrecise ablation of the dental hard tissues can be achieved with the established femtosecond laser numerical control platform. For enamel, the surface roughness of the cavity inside laser scanning line was 7.173 µm at the bottom and 2.675 µm on the wall of the cavity. The surface roughness of the cavity between laser scanning lines was 13.667 µm at the bottom and 33.927 µm on the wall. For dentin, the surface roughness of the cavity bottom was 51.182 µm and 25.629 µm for the wall. Scanning electron microscope images showed no micro-cracks or carbonization on enamel, while carbonization, cracks and a small amount of crystalline particles were observed on dentin.

CONCLUSIONSPrecise tooth preparation can be achieved with femtosecond laser numerical control flatform. The surface roughness of cavity wall was less than that of the bottom and can meet the clinical needs. Suitable femtosecond laser output power should be set for different cutting objects, otherwise it may result in tissue damages.

Dental Cavity Preparation ; methods ; Dental Enamel ; surgery ; ultrastructure ; Dentin ; surgery ; ultrastructure ; Hardness ; Humans ; Laser Therapy ; methods ; Microscopy, Electron, Scanning ; Molar, Third ; surgery ; ultrastructure ; Surface Properties