Development of calcium phosphate/calcium sulfate biphasic biomedical material with hyaluronic acid containing collagenase and simvastatin for vital pulp therapy.

OBJECTIVE: In vital pulp therapy (VPT), a barrier is created with appropriate capping to protect the remaining pulp and thus maintain pulp vitality. Here, we evaluated the feasibility of a biphasic calcium phosphate cement (CPC)-calcium sulfate hemihydrate (CSH) biomaterial containing simvastatin (Sim) and collagenase (Col) for VPT. METHODS: Combinations of varying CPC and CSH concentrations were analyzed for their handling properties and setting times, with their structures observed through scanning electron microscopy-energy dispersive X-ray spectrometry (SEM-EDS). Drug release patterns of simvastatin and collagenase combined with CPC-CSH (CPC-CSH-Sim-Col) were also analyzed, followed by biocompatibility and bioactivity tests on human dental pulp stem cells (hDPSCs) and in vivo animal study in canine models; the in vivo results were obtained through microcomputed tomography and histological analysis. RESULTS: The results revealed that 70 wt% CPC (CPC7) with 30 wt% CSH (CSH3) exhibited optimal setting time and porous structure for clinical use. The cell viability and cytotoxicity analysis demonstrated that CPC7-CSH3 with or without simvastatin or collagenase did not injure hDPSCs. In vivo, the CPC7-CSH3-Sim-Col induced dentin bridge formation. SIGNIFICANCE: CPC7-CSH3-Sim-Col in this study has great potential as a VPT biomaterial to enhance the dentin bridge formation.

Characterization of Phosphorus Species in Human Dentin by Solid-State NMR.

The rat has been considered as an appropriate animal model for the study of the mineralization process in humans. In this work, we found that the phosphorus species in human dentin characterized by solid-state NMR spectroscopy consist mainly of orthophosphate and hydrogen phosphate. Some orthophosphates are found in a disordered phase, where the phosphate ions are hydrogen-bonded to structural water, some present a stoichiometric apatite structure, and some a hydroxyl-depleted apatite structure. The results of this study are largely the same as those previously obtained for rat dentin. However, the relative amounts of the various phosphorus species in human and rat dentin are dramatically different. In particular, stoichiometric apatite is more abundant in human dentin than in rat dentin, whereas the converse is true for disordered-phase orthophosphates. Furthermore, spatial proximity among all phosphorus species in human dentin is identical within experimental error, in contrast to what observed for rat dentin. Although it is not clear how these spectroscopic data could relate to the hierarchical structure or the mechanical properties of teeth, our data reveal that the molecular structures of human and rat dentin at different growth stages are not exactly the same.

Nanocrystalline calcium sulfate/hydroxyapatite biphasic compound as a TGF-ß1/VEGF reservoir for vital pulp therapy.

OBJECTIVES: Vital pulp therapy aims to treat reversible pulpal injuries via protective dentinogenesis and to preserve more tooth structure. Mineral trioxide aggregate (MTA)-based capping materials demonstrate prolonged setting time increases the risk of pulpal infection during multi-visit treatment. Their non-degradable property occupies pulp space and limits dentin-pulp regeneration. This study reports an inorganic degradable biomaterial that presents a short initial setting time and acts as a growth factor reservoir to promote reparative dentinogenesis. METHODS: We synthesize nanocrystalline calcium sulfate hemihydrate (nCS), hydroxyapatite (HAp) and calcium sulfate hemihydrate (CS) as a reservoir to which transforming growth factor-beta 1 (TGF-ß1) and vascular endothelial growth factor (VEGF) are added (denoted as nCS/HAp/CS/TGF-ß1/VEGF). In vitro biocompatibility and mineralization (the activity and expression of alkaline phosphatase, ALP) were evaluated. Rat animal model was created to test in vivo efficacy. RESULTS: Cultured human dental pulp cells (HDPCs) showed that nCS/HAp/CS/TGF-ß1/VEGF cement has excellent biocompatibility and the potential to elevate the activity and expression of ALP. The in vivo efficacy (rat animal model) indicates protective dentin by micro-computed tomography (µ-CT) measurements and histological analyses. The 3D µ-CT non-destructive analysis also determines volume changes during pulpotomy, suggesting that the degraded space of the nCS/HAp/CS/TGF-ß1/VEGF cement is repaired by the formation of dentin-pulp tissue. SIGNIFICANCE: These findings demonstrate that nCS/HAp/CS cement acts as a potent reservoir for the sustained release of growth factors, and that nCS/HAp/CS/TGF-ß1/VEGF cement has a high potential to form the reparative dentinogenesis in vivo.

Development of calcium phosphate/sulfate biphasic cement for vital pulp therapy.

OBJECTIVES: Bioactive calcium phosphate cement (CPC) has been used widely to repair bone defects because of its excellent biocompatibility and bioactivity. However, the poor handling properties, low initial mechanical strength, and long setting time of CPC limit its application in vital pulp therapy (VPT). The aim of this study was to synthesize biphasic calcium phosphate/sulfate cements and evaluate the feasibility of applying these cements in VPT. METHODS: The physical, chemical, and mechanical properties of CPC were improved by mixing the cement with various amounts of α-calcium sulfate hemihydrate (CSH). The hydration products and crystalline phases of the materials were characterized using scanning electron microscopy and X-ray diffraction analysis. In addition, the physical properties, such as the setting time, compressive strength, viscosity, and pH were determined. Water-soluble tetrazolium salt-1 and lactase dehydrogenase were used to evaluate cell viability and cytotoxicity. RESULTS: The developed CPC (CPC/CSH cement), which contains 50wt% CSH cement, exhibited no obvious temperature increase or pH change during setting when it was used as a paste. The initial setting time of the CPC/CSH biphasic cement was substantially shorter than that of CPC, and the initial mechanical strength was 23.7±5.6MPa. The CPC/CSH cement exhibited higher viscosity than CPC and, thus, featured acceptable handling properties. X-ray diffraction analysis revealed that the relative peak intensity for hydroxyapatite increased, and the intensity for calcium sulfate dehydrate decreased as the amount of CPC was increased. The cell viability and cytotoxicity test results indicated that the CPC/CSH cement did not harm dental pulp cells. SIGNIFICANCE: The developed CPC/CSH biphasic cement exhibits substantial potential for application in VPT.

Development of in vitro tooth staining model and usage of catalysts to elevate the effectiveness of tooth bleaching.

OBJECTIVES: Whole blood or tea was frequently used to stain the teeth for measuring the effectiveness of different bleaching materials. However, the components of blood or tea cannot be quantitatively determined and variability might exist among different brands of tea. The purpose of this study was to develop a reproducible in vitro tooth-staining model to simulate the intrinsic discoloration of teeth and evaluate the ability of two catalysts to enhance the bleaching activity of H(2)O(2). METHODS: Rhodamine B, Orange II, Fe(III) phthalocyanine, and tea were used to stain the tooth specimens for 4-72 h and subsequently bleached by H(2)O(2) for 4-72 h. The process was photographed using a digital stereoscopic microscope and a digital camera. The image was transformed to get L*, a*, b* values of CIE Lab system with image processing software. The catalytic ability of light irradiation plus addition of Fe/Sodium-Y or Mn/Sodium-Y for specimens stained by Orange II was evaluated in test tubes and in extracted tooth model. RESULTS: The color of specimens stained by Rhodamine B could not be sufficiently recovered after bleaching by H(2)O(2). In addition, the reaction of Fe(III) phthalocyanine with H(2)O(2) in test tubes was too fast to be monitored. Light activation plus use of Fe/Sodium-Y or Mn/Sodium-Y could significantly accelerate the bleaching efficiency of H(2)O(2). SIGNIFICANCE: Orange II was the most appropriate dye for tooth staining among the dyes used in this study. Addition of Fe/Sodium-Y or Mn/Sodium-Y plus light irradiation could elevate the bleaching efficacy of H(2)O(2) for those specimens stained by Orange II.

Effects of different debonding techniques on the debonding forces and failure modes of ceramic brackets in simulated clinical set-ups.

INTRODUCTION: The objectives of this study were to evaluate the effects of different debonding techniques on the in-vitro mean debonding forces and failure modes of ceramic brackets bonded to enamel with clinically simulated setups. METHODS: Three kinds of ceramic brackets (Clarity; 3M Unitek, Monrovia, Calif; Inspire and Inspire Ice; Ormco, Orange, Calif) were bonded to extracted premolars with the same bonding system. Thirty ceramic brackets, 10 of each type, were removed by hand; 60 ceramic brackets, 20 of each type, were tested on a universal testing machine with the pliers according to the manufacturers' recommendations. To simulate clinical debonding conditions, specially designed setups were used to debond the ceramic brackets. Debonding forces and failure modes were investigated. Fractographic evaluations were performed by using scanning electron microscopy. RESULTS: Most brackets failed at the bracket-adhesive interface. Cohesive bracket fractures were noted in all 3 types of ceramic brackets (debonded by hand: 70% of Inspire, 20% of Inspire Ice, and 10% of Clarity; debonded by machine: 75% of Inspire, 30% of Inspire Ice, and 25% of Clarity). The cohesive ceramic fractures of the Clarity brackets were located at the junction between the wings and the body, and at the slot. However, for the Inspire and the Inspire Ice brackets, the cohesive ceramic fractures were located at the occlusal aspect of the base. The mean debonding forces of Inspire, Inspire Ice, and Clarity brackets were 25.72 +/- 11.98, 17.92 +/- 5.03, and 76.89 +/- 23.47 N, respectively. No enamel damage was found after the brackets were removed. CONCLUSIONS: The results of the failure modes showed that the new designs with a ball reduction band in the Inspire Ice bracket and the vertical debonding slot in the Clarity bracket significantly reduced the risk of ceramic bracket fracture during debonding. The force required to debond the Inspire Ice bracket was significantly lower than that of the Inspire bracket.

Bactericidal effects of diode laser on Streptococcus mutans after irradiation through different thickness of dentin.

BACKGROUND AND OBJECTIVES: A reliable method to eradicate the bacteria of residual carious dentin has not yet been developed. The aim of this study was to evaluate the antibacterial effect of a diode laser on Streptococcus mutans through different thickness (500, 1,000, and 2,000 microm) of human dentin. The thermal effect of laser irradiation was also investigated. STUDY DESIGN/MATERIALS AND METHODS: Dentin specimens were inoculated with 2 microl of S. mutans on one side and irradiated by a diode laser on the other side with a power output ranging from 0.5 to 7 W. The laser tip was swept with the whole irradiation area of 7 mm x 3 mm at a speed of about 10 mm/second with a total irradiation time of 30 seconds. Cooling with distilled water (30 ml/minute) was applied simultaneously during laser irradiation. After laser irradiation, the bacteria was removed from the dentin surfaces and cultured for 48 hours at 37 degrees C anaerobically to assess the colony forming units (CFU) per ml. The morphology of the lased bacteria and the temperature rise during laser irradiation were observed by scanning electron microscope (SEM) and measured by thermocouple, respectively. RESULTS: The results revealed that 7 W of laser power could kill 97.7% of CFU through 500 microm thickness of dentin. However, the bactericidal efficiency was significantly reduced as the dentin thickness was increased. The morphological changes of lased bacteria ranged from less affected such as loss of their wall bands and existence of minicells to more severely degenerated, such as disintegration and fusion of cells with pores on the cell wall. Only the dentin specimens with a thickness of 500 microm exhibited a temperature rise greater than 5.5 degrees C after receiving 5 or 7 W of laser irradiation. CONCLUSIONS: A diode laser can eliminate the Streptococcus mutans of the residual carious dentin without inducing high pulpal temperature rise when the remaining dentin thickness is greater than 1 mm.

Structural changes of Er:YAG laser-irradiated human dentin.

OBJECTIVE: The aim of this study was to investigate the phase, compositional, and morphological changes of Er:YAG laser-irradiated dentin. BACKGROUND DATA: To date, nothing comprehensive has been reported about the phase and compositional changes of human dentin after Er:YAG laser irradiation. METHODS: The human dentin was irradiated by Er:YAG laser with irradiation energies from 300 mJ/pulse-10 pps-10 sec to 700 mJ/pulse-10 pps-10 sec with and without water spray. After irradiation, the specimens were analyzed by means of x-ray diffractometer (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). RESULTS: After exposure to Er:YAG laser, dentin showed no phase changes on the x-ray diffractometer. In Fourier transform infrared analysis, two absorption bands at 2200 and 2015 cm(-1) could be traced on dentin treated by Er:YAG laser with irradiation energies beyond 300 mJ/pulse-10 pps-10 sec and without water spray. The OH(-) band disappeared at 1630 cm(-1). Scanning electron micrographs revealed that laser energy of 500 mJ/pulse-10 pps-10 sec was sufficient to prompt melting and recrystallization of dentin crystals. CONCLUSIONS: Our study demonstrated that Er:YAG laser irradiation on dentin with water spray would not significantly change the structure and composition. Therefore, water cooling is important for reducing the thermal effect of Er:YAG laser.

Ultrastructural changes of human dentin after irradiation by Nd:YAG laser.

BACKGROUND AND OBJECTIVE: The use of Nd:YAG laser has been proposed for endodontic treatment. However, its ability to reduce dentin permeability, which is important for the success of root canal treatment, remains controversial. STUDY DESIGN/MATERIALS AND METHODS: Nd:YAG laser irradiation was performed in pulsed mode on human dentin. The parameters were: pulse energy (100 mJ), rate (10 pps), and total irradiation time (4 seconds). The crystalline phases, electron diffraction patterns, morphology, and microstructure of specimens after laser irradiation were observed by dark-field emission transmission electron microscope (TEM). RESULTS: Three ultrastructural zones could be delineated in the dentin: (1) an outer zone with an ordered columnar structure composed of hydroxyapatite and beta-tricalcium phosphate, (2) an intermediate zone composed of an amorphous substance (about 40-70 nm in diameter), and (3) an inner zone of well-crystallized hydroxyapatite grains. These three zones were free of pores or voids. CONCLUSIONS: Our study demonstrated that laser-irradiation might be used to reduce dentin permeability.