Pilot clinical study to evaluate the efficacy of a professionally delivered high fluoride varnish on erosive tooth wear in an in-situ model.

OBJECTIVES: The aim for this pilot study was to investigate the effect of a sodium fluoride varnish on step height measured by a profilometer from human enamel worn by healthy volunteers with a novel in situ/ex vivo erosion design. METHOD: Healthy volunteers aged 18-70 years wore a palatal splint containing 8 human enamel samples and underwent two 3-day treatment periods for 6 h a day with a varnish containing sodium fluoride at 22,600 ppm and the control with the same ingredients but without fluoride. Each splint contained 4 polished and 4 unpolished samples. The interventions were applied to the surface of the enamel samples in randomised order, removed after 6 h, then immersed ex-vivo in 1 %, pH 2.7 citric acid for 2 min, repeated 4 times a day, over 2 days. Measurements of enamel were assessed blindly by microhardness on day 2 and by non-contact laser profilometry on day 3 for the two treatments. RESULTS: 24 volunteers, 2 males and 22 females aged 27-54 years, were screened and recruited. The delta microhardness, from polished samples removed at the end of day 2, for the control and fluoride treatment was 95.7 (22.9) kgf/mm2 and 123.7 (28.9) kgf/mm2, respectively (p < .005). The mean (SD) step height for the control polished enamel surfaces was 3.67 (2.07) µm and for the fluoride varnish was 1.79 (1.01) µm (p < .0005). The control unpolished enamel surfaces had a mean 2.09 (1.53) µm and the fluoride varnish was 2.11 (1.53) µm but no statistical difference was detected. CONCLUSIONS: The results from this pilot study, utilizing an in-situ model where enamel was exposed to acid over the course of 2 days, demonstrated that a high fluoride varnish containing sodium fluoride at 22,600 ppm prevented erosive wear compared to a control on the polished enamel surfaces. CLINICAL SIGNIFICANCE: Intra-oral study demonstrated that a high fluoride varnish containing sodium fluoride at 22,600 ppm reduced erosive tooth wear.

Temperature Rise Associated with Adiabatic Shear Band: Causality Clarified.

One of the most important issues related to adiabatic shear failure is the correlation among temperature elevation, adiabatic shear band (ASB) formation and the loss of load capacity of the material. Our experimental results show direct evidence that ASB forms several microseconds after stress collapse and temperature rise reaches its maximum about 30 µs after ASB formation. This observation indicates that temperature rise cannot be the cause of ASB. Rather, it might be the result of adiabatic shear localization. As such, the traditional well-accepted thermal-softening mechanism of ASB needs to be reconsidered.

Peptide-Mediated Biomimetic Regrowth of Human Enamel In Situ.

Mimicking the dynamics of mineral loss and gain involved in dental caries formation can help us evaluate and compare the mineralization efficacy of different treatment agents used in enamel remineralization. Here, we offer an abridged study design outlining the preparation of tooth samples, creation of artificial dental lesions, application of a peptide, and characterization of the regrown enamel-like mineral layer.

Peptide-Based Bioinspired Approach to Regrowing Multilayered Aprismatic Enamel.

The gradual discovery of functional domains in native enamel matrix proteins has enabled the design of smart bioinspired peptides for tooth enamel mimetics and repair. In this study, we expanded upon the concept of biomineralization to design smaller amelogenin-inspired peptides with conserved functional domains for clinical translation. The synthetic peptides displayed a characteristic nanostructured scaffold reminiscent of 'nanospheres' seen in the enamel matrix and effectively controlled apatite nucleation in vitro resulting in the formation of smaller crystallites. Following application of the peptides to sectioned human molar teeth, a robust, oriented, synthetic aprismatic enamel was observed after 7 days of incubation in situ. There was a two-fold increase in the hardness and modulus of the regrown enamel-like apatite layers and an increase in the attachment of the tooth-regrown layer interface compared to control samples. Repeated peptide applications generated multiple enamel-like hydroxyapatite (HAP) layers of limited thickness produced by epitaxial growth in which c-axis oriented nanorods evolved on the surface of native enamel. We conclude that peptide analogues with active domains can effectively regulate the orientation of regenerated HAP layers to influence functional response. Moreover, this enamel biofabrication approach demonstrates the peptide-mediated growth of multiple microscale HAP arrays of organized microarchitecture with potential for enamel repair.

Efficacy of amelogenin-chitosan hydrogel in biomimetic repair of human enamel in pH-cycling systems.

Amelogenin-chitosan (CS-AMEL) hydrogel has shown great potential for the prevention, restoration, and treatment of defective enamel. As a step prior to clinical trials, this study aimed to examine the efficacy of CS-AMEL hydrogel in biomimetic repair of human enamel with erosive or caries-like lesions in pH-cycling systems. Two models for enamel defects, erosion and early caries, were addressed in this study. Two pH-cycling systems were designed to simulate the daily cariogenic challenge as well as the nocturnal pH conditions in the oral cavity. After pH cycling and treatment with CS-AMEL hydrogel, a synthetic layer composed of oriented apatite crystals was formed on the eroded enamel surface. CS-AMEL repaired the artificial incipient caries by re-growing oriented crystals and reducing the depth of the lesions by up to 70% in the pH-cycling systems. The results clearly demonstrate that the CS-AMEL hydrogel is effective at the restoration of erosive and carious lesions under pH-cycling conditions.

Matrix metalloproteinase-20 mediates dental enamel biomineralization by preventing protein occlusion inside apatite crystals.

Reconstruction of enamel-like materials is a central topic of research in dentistry and material sciences. The importance of precise proteolytic mechanisms in amelogenesis to form a hard tissue with more than 95% mineral content has already been reported. A mutation in the Matrix Metalloproteinase-20 (MMP-20) gene results in hypomineralized enamel that is thin, disorganized and breaks from the underlying dentin. We hypothesized that the absence of MMP-20 during amelogenesis results in the occlusion of amelogenin in the enamel hydroxyapatite crystals. We used spectroscopy and electron microscopy techniques to qualitatively and quantitatively analyze occluded proteins within the isolated enamel crystals from MMP-20 null and Wild type (WT) mice. Our results showed that the isolated enamel crystals of MMP-20 null mice had more organic macromolecules occluded inside them than enamel crystals from the WT. The crystal lattice arrangements of MMP-20 null enamel crystals analyzed by High Resolution Transmission Electron Microscopy (HRTEM) were found to be significantly different from those of the WT. Raman studies indicated that the crystallinity of the MMP-20 null enamel crystals was lower than that of the WT. In conclusion, we present a novel functional mechanism of MMP-20, specifically prevention of unwanted organic material entrapped in the forming enamel crystals, which occurs as the result of precise amelogenin cleavage. MMP-20 action guides the growth morphology of the forming hydroxyapatite crystals and enhances their crystallinity. Elucidating such molecular mechanisms can be applied in the design of novel biomaterials for future clinical applications in dental restoration or repair.

Assembly of Layered Monetite-Chitosan Nanocomposite and Its Transition to Organized Hydroxyapatite.

Bioinspired synthesis of hierarchically structured calcium phosphate (CaP) material is a highly promising strategy for developing improved bone substitute materials. However, synthesis of CaP materials with outstanding mechanical properties still remains an ongoing challenge. Inspired by the formation of lamellar structure in nacre, we designed an organic matrix composed of chitosan and cis-butenediolic acid (maleic acid, MAc) that could assemble into a layered complex and further guide the mineralization of monetite crystals, resulting in the formation of organized and parallel arrays of monetite platelets with a brick-and-mortar structure. Using the layered monetite-chitosan composite as a precursor, we were able to synthesize hydroxyapatite (HAp) with multiscale hierarchically ordered structure via a topotactic phase transformation process. On the nanoscale, needlelike HAp crystallites assembled into organized bundles that aligned to form highly oriented plates on the microscale. On the large-scale level, these plates with different crystal orientations were stacked together to form a layered structure. The organized structures and composite feature yielded CaP materials with improved mechanical properties close to those of bone. Our study introduces a biomimetic approach that may be practical for the design of advanced, mechanically robust materials for biomedical applications.

Amelogenin Affects Brushite Crystal Morphology and Promotes Its Phase Transformation to Monetite.

Amelogenin protein is involved in organized apatite crystallization during enamel formation. Brushite (CaHPO4·2H2O), one of the precursors of hydroxyapatite mineralization in vitro, has been used for fabrication of biomaterials for hard tissue repair. In order to explore its potential application in biomimetic material synthesis, we studied the influence of the enamel protein amelogenin on brushite morphology and phase transformation to monetite. Our results show that amelogenin can adsorb onto the surface of brushite, leading to the formation of layered morphology on the (010) face. Amelogenin promoted the phase transformation of brushite into monetite (CaHPO4) in the dry state, presumably by interacting with crystalline water layers in brushite unit cells. Changes to the crystal morphology mediated by amelogenin continued even after the phase transformation from brushite to monetite, leading to the formation of organized platelets with an interlocked structure. This effect of amelogenin on brushite morphology and the phase transformation to monetite could provide a new approach to developing biomimetic materials.

Amelogenin and Enamel Biomimetics.

Mature tooth enamel is acellular and does not regenerate itself. Developing technologies that rebuild tooth enamel and preserve tooth structure is therefore of great interest. Considering the importance of amelogenin protein in dental enamel formation, its ability to control apatite mineralization in vitro, and its potential to be applied in fabrication of future bio-inspired dental material this review focuses on two major subjects: amelogenin and enamel biomimetics. We review the most recent findings on amelogenin secondary and tertiary structural properties with a focus on its interactions with different targets including other enamel proteins, apatite mineral, and phospholipids. Following a brief overview of enamel hierarchical structure and its mechanical properties we will present the state-of-the-art strategies in the biomimetic reconstruction of human enamel.

Amelogenin-chitosan matrix for human enamel regrowth: effects of viscosity and supersaturation degree.

We recently reported an amelogenin-chitosan (CS-AMEL) hydrogel as a promising biomimetic material for future in situ human enamel regrowth. To further optimize the necessary conditions for clinical applicability of CS-AMEL hydrogel, herein we studied the effects of viscosity and supersaturation degree on the size and orientation of synthetic crystals by means of scanning electron microscopy (SEM) and X-ray diffraction (XRD). Raising the hydrogel viscosity by increasing chitosan concentration from 1% to 2% (w/v) improved the orientation of the crystals, while a higher supersaturation (σ(HAp) >10.06, [Ca(2+)] >5 mM) resulted in the formation of random crystals with larger sizes and irregular structures. We conclude that optimal conditions to produce organized enamel-like crystals in a CS-AMEL hydrogel are: 2% (w/v) chitosan, 2.5 mM calcium, and 1.5 mM phosphate (degree of supersaturation = 8.23) and 200 µg/ml of amelogenin.

Development of amelogenin-chitosan hydrogel for in vitro enamel regrowth with a dense interface.

Biomimetic enamel reconstruction is a significant topic in material science and dentistry as a novel approach for the treatment of dental caries or erosion. Amelogenin has been proven to be a critical protein for controlling the organized growth of apatite crystals. In this paper, we present a detailed protocol for superficial enamel reconstruction by using a novel amelogenin-chitosan hydrogel. Compared to other conventional treatments, such as topical fluoride and mouthwash, this method not only has the potential to prevent the development of dental caries but also promotes significant and durable enamel restoration. The organized enamel-like microstructure regulated by amelogenin assemblies can significantly improve the mechanical properties of etched enamel, while the dense enamel-restoration interface formed by an in situ regrowth of apatite crystals can improve the effectiveness and durability of restorations. Furthermore, chitosan hydrogel is easy to use and can suppress bacterial infection, which is the major risk factor for the occurrence of dental caries. Therefore, this biocompatible and biodegradable amelogenin-chitosan hydrogel shows promise as a biomaterial for the prevention, restoration, and treatment of defective enamel.

An amelogenin-chitosan matrix promotes assembly of an enamel-like layer with a dense interface.

Biomimetic reconstruction of tooth enamel is a significant topic of study in materials science and dentistry as a novel approach to the prevention, restoration, and treatment of defective enamel. We have developed a new amelogenin-containing chitosan hydrogel for enamel reconstruction that works through amelogenin supramolecular assembly, stabilizing Ca-P clusters and guiding their arrangement into linear chains. These amelogenin Ca-P composite chains further fuse with enamel crystals and eventually evolve into enamel-like co-aligned crystals, anchored to the natural enamel substrate through a cluster growth process. A dense interface between the newly grown layer and natural enamel was formed and the enamel-like layer improved the hardness and elastic modulus compared with etched enamel. We anticipate that this chitosan hydrogel will provide effective protection against secondary caries because of its pH-responsive and antimicrobial properties. Our studies introduce an amelogenin-containing chitosan hydrogel as a promising biomaterial for enamel repair and demonstrate the potential of applying protein-directed assembly to biomimetic reconstruction of complex biomaterials.

Macromolecules on nano-outlets responding to electric field and pH for dual-mode drug delivery.

Macromolecules responsive to both electric field and pH are modified on the outlets of mesoporous silica nanospheres to form a novel dual-mode drug delivery system. Dual drug delivery modes are indicated by the release patterns obtained by simulating the body's gastric and intestinal fluid ex vivo. In a pH 1.4 release medium (simulated gastric fluid, SGF) without an alternating electric field, only 8.5% of the total payload is found to leach in 8 h, whereas 50.3% and 60.2% of the total payload are released in the same time period by elevating the pH of the release medium or applying an alternating electric field. The two stimuli signals can work independently or corporately to regulate the release kinetics to form a dual-mode drug delivery mechanism, which makes it more flexible for use in certain complicated situations.

An effective gold nanotubes electrode for amperometric biosensor.

A sensitive and effective amperometric glucose biosensor based on gold nanotubes electrode (GNTE) was investigated. Gold nanotubes (GNTs), which were prepared by electroless plating of the metal within the pores of nanoporous polycarbonate (PC) track-etched membranes, were filled into a hollow teflon cylinder to construct a GNTE. Glucose oxidase (GOD) was immobilized on the electrode via glutaraldehyde cross-linkage method. The electrochemical properties were investigated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The km value of the immobilized glucose oxidase on GNTE was 0.47 mM. The biosensor showed a linear range from 0.4 to 11 mM with excellent sensitivity of 8.77 microA cm(-2) mM(-1) and fast response time within 5 s.

Synthesis of h-AIN nanowires via carbothermal reduction and nitridation method using acetylene black.

Using acetylene black as an assistant reagent in the ammonia atmosphere, one dimensional (1D) hexagonal single crystalline aluminum nitride (AIN) nanowires were successfully synthesized via carbothermal reduction and nitridation method. With smooth surface and uniform diameter, the wool-like product of AIN nanowires grow along [001] direction and are 80-120 nm in diameter and several tens micrometers in length. According to thermodynamic and kinetic analysis, AI2O3 initially was reduced to a volatile suboxide gas Al2O at 1800 degrees C. Then gaseous Al2O was transported by N2/NH3 mixtures to about 1200 degrees C, where 1D AIN nanowires formed by the reaction of Al2O gas, graphite and NH3. Due to high surface area, small particle size and amorphous structure, acetylene black maybe possess more active sites and more effective contact area, which benefits the first step reaction. The electrostatic polar charge model and crystallographic characteristics are employed to explain the growth mechanism of AIN nanowires. To our knowledge, acetylene black is the first time to be used for synthesizing 1D AIN nanostructures via carbothermal reduction and nitridation method, which would be an efficient, economical assistant reagent for fabricating nitride nanostructures.

Dipolar molecules as impellers achieving electric-field-stimulated release.

Here we report the design of a new external electric field-controlled release system using functional dipolar molecules as nanoimpellers. The dipolar molecule 4-(3-cyanophenyl)butylene, which can reorient in response to external electric fields with different frequencies because of its strong inherent dipole moment, was synthesized and grafted onto the inner surfaces of mesopores. Under an alternating electric field, the swinging flexible molecular chains consequently push guest molecules out of the pore voids. This innovative approach to controlled release may provide important application opportunities in tumor treatment with a number of advantages in terms of local release with targetability, external remote control, and the nonelectrochemical nature of the process.

Investigation of layer-by-layer assembled heparin and chitosan multilayer films via electrochemical spectroscopy.

The layer-by-layer (LbL) assembly as a simple and effective method has been extensively used to prepare polyelectrolyte films but the buildup mechanism is expected to be further clarified. In this work, the structure and formation mechanism of LbL-assembled heparin/chitosan multilayer composite films were characterized by electrochemical system, scanning electron microscope and atom force microscope. The results revealed that the film grew linearly in the first 10 bilayers based on measured linear increase of film resistance with number of layers, while the film grew exponentially in the later 10 bilayers based on measured nonlinear increase of film resistance. The charge-transfer resistance increased in an oscillatory way or a linear way at different growing stages, which was discussed with their formation mechanism and the interfacial structure on electrode. A buildup mode of the LbL film was suggested based on the structural and electrochemical characters.

Ultrasonic-irradiation-assisted oriented assembly of ordered monetite nanosheets stacking.

Bioactive monetite (anhydrous calcium hydrogen phosphate, CaHPO(4)) with orderly layered structure assembled by nanosheets has been successfully synthesized by a sonochemical-assisted method in the presence of cetyltrimethylammonium bromide (CTAB). The thicknesses of the nanosheets are 100-200 nm, and the lateral sizes are about 2 microm. Because of the strong affinity with the phosphate ions as well as the (200) faces of the crystals, CTAB molecules can make the formation and stabilization of monetite nanosheets with (200) exposed face. Ultrasonic irradiation makes the transition from disordered state to oriented state before the oriented assembly of monetite nanosheets. The ultrasonic irradiation provides enough external work to make the assemble process possible in thermodynamics. The drastic flow stirred by the supersonic jet in the solution accomplishes the transition and successive oriented assembly of nanosheets in dynamics. This study would offer a simple method to design and synthesize oriented-assembled materials.