Synthesis and Characterization of Bioactive Glass and Zinc Oxide Nanoparticles with Enamel Remineralization and Antimicrobial Capabilities.

BACKGROUND: We investigated the effect of bioactive glass and zinc oxide nanoparticles on enamel remineralization, as well as their antimicrobial effect on cariogenic microbes. This is the first study that investigated the properties of bioactive glass and zinc oxide nanoparticles with mixed materials. METHODS: Fluoride gel (F), bioactive glass microparticles (µB), bioactive glass nanoparticles (nB), zinc oxide nanoparticles (Z), and a mixed suspension of nB and Z (nBZ) were prepared and characterized by scanning and transmission electron microscopy, zeta potential measurement, X-ray diffraction, and acid buffering capacity testing. Further, we performed a remineralization cycle test of 28 days, and nanoindentation testing was carried out during the immersion period, and then the enamel surfaces were examined using scanning electron microscopy. Additionally, the antimicrobial effects of the sample suspensions were evaluated by measuring their minimum microbicidal concentrations against various cariogenic microbes. RESULTS: Our results revealed that nB had a near-circular shape with an amorphous structure and a considerably large specific surface area due to nanoparticulation. Additionally, nB possessed a rapid acid buffering capacity that was comparable to that of µB. In the remineralization test, faster recovery of mechanical properties was observed on the enamel surface immersed in samples containing bioactive glass nanoparticles (nB and nBZ). After remineralization, demineralized enamel immersed in any of the samples showed a rough and porous surface structure covered with mineralized structures. Furthermore, nBZ exhibited a broad antimicrobial spectrum. CONCLUSIONS: These results demonstrated that bioactive glass and zinc oxide nanoparticles have superior demineralization-suppressing and remineralization-promoting effects.

Regaining enamel color quality using enamel matrix derivative.

This study aimed to demonstrate and compare the accuracy of tooth shade selection due to the remineralized enamel crystal with enamel matrix derivative (EMD) in vitro. Etched enamel slices were immersed in four types of mineralization buffers for 16 h. Sodium fluoride (NaF) was added to final concentrations of 1-100 ppm with the mineralization buffer that demonstrated the highest mineralization efficiency. EMD was added to the mineralization buffer containing NaF to see if it has any remineralization capacities. The remineralized enamel crystal was analyzed by SEM and XRD. The tooth shade was evaluated by CIE L*a*b*. The results showed that, without NaF, plate-like nanocrystals were formed on the enamel surface, but with NaF, needle-like nanocrystals were formed. By adding EMD, a layer of well-compacted hydroxyapatite crystals was successfully precipitated onto the natural enamel surface. No significant differences were observed in the L* value of the mineralization surface pre-etching and after mineralization buffer containing NaF and EMD. A new method has been developed to recover the color quality of enamel, as well as to mineralize the tooth enamel by constructing hydroxyapatite crystals with mineralization buffers containing NaF and EMD on the etched tooth surface.

Human dentin materials for minimally invasive bone regeneration: Animal studies and clinical cases.

OBJECTIVES: Bone, platelet concentrate, and tooth-derived dentin/cementum have been used as autologous materials in regenerative medicine Dentin materials were first recycled in 2002 for bone regeneration in humans, although bone autografts were noted in the 19th century, and auto-platelet concentrates were developed in 1998. Dentin/cementum-based material therapy has been applied as an innovative technique for minimally invasive bone surgery, while bone autografts are associated with donor site morbidity. METHODS: In October 2021, PubMed, Google Scholar, Scopus, and the Cochrane Library databases from 1980 to 2020 were screened. RESULTS: The demineralized dentin/cementum matrix (DDM) had better performance in bone induction and bone regeneration than mineralized dentin. CONCLUSIONS: Unlike cell culture therapy, DDM is a matrix-based therapy that includes growth factors. A matrix-based system is a realistic and acceptable treatment, even in developing countries. The aim of this review was to summarize the evidence related to both animal studies and human clinical cases using human dentin materials with a patch of cementum, especially DDM.

Effect of photofunctionalization on titanium bone-implant integration in ovariectomized rats.

Osteoporosis is considered a risk factor for osseointegration during implant treatment. Photofunctionalization of titanium has been shown to improve bone-based cell adhesion, proliferation, and functional expression, increasing the bone-implant contact rate and bone-implant integration strength. This study aimed to test the hypothesis that photofunctionalization is effective for implant fixation using an osteoporosis rat model. In the biomechanical push-in test, the bone-implant integration strength of the photofunctionalization treatment group was 1.53 times that of the control group (p<0.05). These values implied that photofunctionalization restored the ovariectomy-induced low bone-implant integration strength to normal states. In the micro-CT analysis, the BV/TV of the photofunctionalization treatment group was 1.32 times that of the control group (p<0.05). These values implied that photofunctionalization restored the ovariectomy-induced low peri-implant bone formation to normal states. These results indicate that photofunctionalization treatment increased peri-implant bone formation and bone-implant integration strength in ovariectomized rats.

Surface analysis of titanium disks with strontium coating.

Peri-implantitis is one of the most common complications after dental implant placement. Researchers have demonstrated that the peri-implantitis tends to occur around dental implants with a rough surface rather than those with a smooth surface. We aimed to investigate the ability of a smooth titanium (Ti) surface containing strontium (Sr) to enhance bone formation as a result of strontium's capacity to support osteoblast proliferation and differentiation. A thin titanium oxide film was formed on an as-mirror polished Ti surface by dipping in 5% sodium hypochlorite (NaOCl) solution for 24 h, followed by thermal treatment at 350°C. The Ti surface was then treated with 1% strontium nitrate (Sr(NO3)2) solution and turned in spin coater. The surface morphology, chemical composition, and release of strontium ions (Sr2+) were evaluated. The results demonstrate that strontium in the form of Sr2+ was successfully doped into the titanium dioxide (TiO2) film by this simple chemical treatment.

Chemical interaction of 4-META with enamel in resin-enamel bonds.

X-ray photoelectron spectroscopy (XPS) is used to analyze 4-META resin and enamel that are debonded at an adhesive interface. The XPS spectra showed two chemical states for Ca: one resulted from Ca of hydroxyapatite and the other, an unknown chemical state, suggested that Ca was chemically bonded with 4-META. We postulate that for a chemical reaction of 4-META and hydroxyapatite, the chemical structure of carboxyl groups will resemble that of calcium phthalate. Hence, calcium phthalate was used as a reference material. Additionally, the spectra obtained from the adhesive interface and the mixture of calcium phthalate with hydroxyapatite particles were compared using peak deconvolution analysis. XPS analysis revealed that the chemical bond of 4-META with enamel resembled the chemical state of Ca in calcium phthalate. Consequently, we suggest that Ca of the enamel and the carboxyl group of 4-META were chelate-bonded at the interface.

In vitro evaluation of NaOCl-mediated functionalization of biologically aged titanium surfaces.

The aim of this study was to evaluate the NaOCl-mediated biofunctionalization of titanium surfaces. Titanium disks stored for 2 weeks were immersed in 5% NaOCl solution for 24 h. A disk immersed in distilled water for 24 h was used as a control. X-ray photoelectron spectrometer assay of the titanium surface after NaOCl treatment demonstrated that organic contaminants containing carbon and nitrogen were removed and the number of hydroxyl groups increased. The NaOCl treatment substantially converted the titanium surface to superhydrophilic status (θ<5°), which resulted in an increased number of attached cells and enhanced cell spreading on the NaOCl-treated surfaces. These results indicate that biofunctionalization of the biologically degraded titanium surfaces can be achieved by chemical surface treatment with 5% NaOCl. The mechanism for desorption of strongly adsorbed organic molecules with polar groups such as amino and aldehyde groups from titanium surfaces by ClO- was elucidated.

Protective Effects of GIC and S-PRG Filler Restoratives on Demineralization of Bovine Enamel in Lactic Acid Solution.

This study was aimed at investigating the protective effects of glass ionomer cement (GIC) and surface pre-reacted glass ionomer (S-PRG) fillers used as dental restorative materials on demineralization of bovine enamel. GlasIonomer FX ULTRA (FXU), Fuji IX GP Extra (FIXE), CAREDYNE RESTORE (CDR) were used as GICs. PRG Barrier Coat (BC) was used as the S-PRG filler. They were incubated in a lactic acid solution (pH = 4.0) for six days at a temperature of 37 °C. The mineral was etched from the enamel surface, and a large number of Ca and P ions were detected in solution. The Al, F, Na, Sr, and Sr ions were released in GICs and S-RPG fillers. The Zn ion was released only in CDR and the B ion was released only in BC. The presence of apparent enamel prism peripheries was observed after six days of treatment for the group containing only enamel blocks. pH values for the FXU, FIXE, CDR, BC, and enamel block groups after six days were 6.5, 6.6, 6.7, 5.9, and 5.1, respectively. Therefore, the observed pH neutralization effect suppressed progression of caries due to the release of several ions from the restoratives.

Stereolithographic Surgical Guide with a Combination of Tooth and Bone Support: Accuracy of Guided Implant Surgery in Distal Extension Situation.

A distal free-end situation could result in insufficient stability of the surgical guide, and could reduce accuracy of the static guided implant surgery (sGIS). The purpose of this study was to investigate the accuracy of sGIS using a combination tooth-and-bone supported stereolithographic (SLA) surgical guide in distal extension situation. Thirty dentists, each placed three implants at the Federal Dentaire Internationale (FDI) teeth positions #46, #47 (a distal extension situation), and #36 (a single tooth gap) via the surgical guide on a model fixed to a manikin. Pre- and post-operative computed tomography (CT) images of the models were superimposed, and the positional and angular deviations of the implants were measured with metrology software. An analysis of variance (ANOVA) test was performed to evaluate the intergroup differences. No significant differences were found for all the positional and angular deviations among the three implant sites, except the bucco-lingual deviation at the implant platform in the #47 position (0.43 ± 0.19 mm) that was significantly larger than the #46 (0.21 ± 0.14 mm) and #36 (0.24 ± 0.25 mm) positions (p < 0.0001). Within the limits of this study, we conclude that, in distal extension situation of missing mandibular molars, adding a bone-supported strut in the distal part of the surgical guide can be beneficial to the accuracy of the sGIS.

Inhibition of enamel demineralization by an ion-releasing tooth-coating material.

PURPOSE: To evaluate the inhibitory effect of a surface pre-reacted glass-ionomer (S-PRG) filler-containing tooth-coating material on enamel demineralization. The outer surface of the S-PRG filler is in a state in which ions are readily released. METHODS: Human enamel blocks were incubated in lactic acid solution (pH 4.0) with and without a disk (n=6) made of the cured tooth-coating material. Test solutions were changed every 24 hours and incubation was continued for 4 days. The pH and amount of fluoride released were measured with an electrode and ion meter, respectively. The concentrations of ions (aluminum, boron, calcium, phosphorus, silicon, sodium, and strontium) were measured by inductively coupled plasma atomic emission spectroscopy. The surface of the enamel block was observed with a scanning electron microscope. RESULTS: Enamel demineralization was not observed in an enamel block incubated with a disk of the tooth-coating material. Ions released from S-PRG filler had an acid buffering action in the low pH lactic acid solution. However, in the enamel block-only solution showing high levels of calcium ion release, the degree of demineralization was correlated with morphological changes of the enamel surface. CLINICAL SIGNIFICANCE: Due to the buffering effects of the pre-reacted glass-ionomer surface by ion release, the S-PRG filler-containing tooth-coating material inhibited enamel demineralization by neutralizing the acidic environment at an early time point.

Designing an antibacterial acrylic resin using the cosolvent method -Effect of ethanol on the optical and mechanical properties of a cold-cure acrylic resin.

Antimicrobial cetylpyridinium chloride (CPC) has low miscibility with acrylic resin monomer but can be homogeneously mixed using ethanol as a cosolvent. This study investigated the effects of ethanol addition on the properties of a cold-cure acrylic resin. Ethanol was an excellent cosolvent for CPC and methyl methacrylate monomer (MMA), but the cured resin exhibited a strong change in coloration to yellow (ΔE*ab>8) and a drastically reduced bending strength (from 97 to 25 MPa) and elastic modulus (from 2.7 to 0.6 GPa) when equal volumes of ethanol and monomer were used together, possibly due to the solvation and deactivation of radicals by ethanol. However, these unfavorable effects diminished when the ethanol/MMA ratio was reduced to 0.25, and became smaller when each specimen was depressurized and excess ethanol was removed. Thus, it may be possible to develop a molecularly uniform antibacterial acrylic resin with acceptable color and strength using this simple technique.

Effect of gold deposition onto titanium on the adsorption of alkanethiols as the protein linker functionalizing the metal surface.

A new method to enhance the biofunctionalization of Ti materials was developed by the immersion of Au-sputtered titanium plates into a solution of alkanethiol with a carboxy group end (HS-C11-COOH). X-ray photoelectron spectroscopy (XPS) analysis showed that the gold content increased with the Au sputtering time and reached over 98 at% after 100 s. The content of the characteristic alkanethiol elements (S, C, and O) in the immersed Ti specimens increased with the Au sputtering time, which indicates that the binding of alkanethiol molecules is enhanced by the Au modification on the Ti surface. Qualitative analysis of XPS (Au-S, COO(-), and OH(-) species) and FTIR spectra (peaks assignable to an aliphatic carboxylic acid) for the sample after immersion also supported this. A strong positive correlation between the S and Au content confirms that Au sputtering is an effective method to control the alkanethiol treatment of Ti substrates.

Structural requirements essential for elastin coacervation: favorable spatial arrangements of valine ridges on the three-dimensional structure of elastin-derived polypeptide (VPGVG)n.

The elastin precursor tropoelastin possesses a number of polymeric peptides with repeating 3-9 mer sequences. One of these is the pentapeptide Val-Pro-Gly-Val-Gly (VPGVG) present in almost all animal species, and its polymer (VPGVG)n coacervates just as does tropoelastin. In the present study, in order to explore the structural requirements essential for coacervation, (VPGVG)n and its shortened repeat analogs (VPGV)n, (VPG)n, and (PGVG)n were synthesized and their structural properties were investigated. In our turbidity measurements, (VPGVG)n demonstrated complete reversible coacervation in agreement with previous findings. The Gly(5) -deleted polymer (VPGV)n also achieved self-association, though the onset of self-association occurred at a lower temperature. However, the dissociation of (VPGV)n upon temperature lowering was found to occur in a three-step process; the Val(i) (4) -Val(i+1) (1) structure arising in the VPGV polypeptide appeared to perturb the dissociation. No self-association was observed for (VPG)n or (PGVG)n repeats. Spectroscopic measurements by CD, FT-IR, and (1) H-NMR showed that the (VPGV)n and (VPG)n both assumed ordered structures similar to that of (VPGVG)n. These results demonstrated that VPGVG is a structural element essential to achieving the ß-spiral structure required for self-association followed by coacervation, probably due to the ideal spatial arrangement of the hydrophobic Val residues.

Proteome analyses of human macrophages exposed to low cytotoxic IC90 copper (2+) ions.

Dental noble alloys often contain copper (Cu). Eluted metal ions sometimes irritate oral tissues. The most eluted ions are Cu ions. The purpose of this study was to investigate the effects of low cytotoxic (IC90, 100 µmol/L) Cu ions on macrophages by proteome analyses consisting of two-dimensional (2D) electrophoresis and matrix-assisted laser desorption/ionization -time of flight (MALDI-TOF) mass spectrometry. The analyses revealed that stimulation with IC90 Cu ions for 1 day caused the macrophage to significantly increase five specific protein spots. Mascot peptide mass finger-print matching suggested that four of them were attributed to 70 kDa heat shock protein 1A/1B (HSP70). HSP70 expression was verified by expressions of corresponding HSPA1A and HSPA1B mRNAs of the macrophage in quantitative real-time PCR analyses. It was concluded that by producing abundant HSP70, the macrophage protected itself against intracellularly intruding cytotoxic Cu ions that might un-fold and crosslink cellular proteins.

Viscoelastic behavior of commercially available tissue conditioners under compression.

Three commercially available tissue conditioners -Tissue conditioner II (STII), Tissue conditioner (GTC), and Tissue Care (TTC)- were tested using a rheometer to evaluate their viscoelasticity under compression at a constant distortion (10%). Immediately after fabrication, STII showed the highest hardness and compressive modulus of elasticity, followed by TTC and GTC. TTC showed the highest relaxation rate, followed by GTC and STII. STII and GTC showed equivalent relaxation times, while TTC showed the shortest relaxation time. The modulus of viscosity was highest in STII, followed by GTC and TTC. The immersion of each sample in water resulted in increases in hardness, compressive modulus of elasticity, relaxation time, and modulus of viscosity and decreases in relaxation rates. The three products have different viscoelastic properties. Further, the results indicate that STII and GTC exhibit better performance when used as functional impression materials, while TTC exhibits better performance when used as a tissue conditioner.

Viscoelastic adlayers of collagen and lysozyme studied using quartz crystal microbalance with dissipation monitoring.

In this study the effects of molecular structure of proteins on their adsorption behaviour and viscoelasticity are investigated using a QCM-D method. The adsorption measurements show that spherical lysozyme is rapidly adsorbed on the gold surface as a stiff monolayer, as indicated by a sharp drop in the oscillation frequency (Deltaf) of the sensor, and by a very small increase in the energy dissipation of the adlayer (DeltaD). Fibrous calfskin collagen (CSC) is, however, adsorbed on the same surface rather slowly in two steps to form a thick and soft multilayer (large Deltaf and DeltaD) at pH 3 in salt-free conditions. The two-step adsorption is not observed for stiffly aggregated CSC. This study clearly demonstrates that the polymer structure strongly affects not only how adsorption develops but also the viscoelastic properties of the adlayer.

Microscopic observations and inflammatory cytokine productions of human macrophage phagocytising submicron titanium particles.

This study was performed to microscopically observe and measure inflammatory cytokine production by human macrophages phagocytosing submicron titanium (Ti) particles. Observations with secondary electron microscopy (SEM), SEM/electron probe microanalysis (EPMA) and transmission electron microscopy (TEM) indicated that macrophages [phorbol-12-myristate-13-acetate (PMA)-differentiated THP-1 cells] at 24 h in culture actively phagocytosed and accumulated submicron Ti particles in intracellular phagosomes, in which refinement of Ti particles occurred. The macrophages were also cultured for 24 h in four media with and without submicron Ti particles and lipopolysaccharide (LPS; components of bacteria). Whilst neither stimulus reduced cell viability, submicron Ti particles and LPS activation independently and synergistically caused the macrophages to produce three inflammatory cytokines (TNF-alpha, IL-1beta and IL-6) at high levels in the culture supernatants. The inflammatory and osteolysis conditions caused by macrophages phagocytosing submicron Ti particles would be worsened by challenge with LPS in patients wearing Ti prostheses.

Gene expression analyses of human macrophage phagocytizing sub-micro titanium particles by allergy DNA chip (Genopal).

The purpose of this study was to examine gene expressions of macrophage phagocytizing sub-micro Ti particles by a DNA chip. Human monocytic cell line THP-1 was differentiated into macrophages by culturing for two days in medium supplemented with 200 nM phorbol ester (PMA). The macrophages were then cultured in four media: medium without PMA (control); medium with suspended sub-micro Ti particles (0.5 wt%); medium with 1.0 microg/ml lipopolysaccharide (LPS); and medium with LPS and Ti particles. After 6 hours' culture, total RNA were extracted and gene expressions were evaluated by DNA allergy chip with 205 allergy and inflammation related gene spots. We found that phagocytosis of sub-micro Ti particles and LPS independently and synergistically up-regulated 17 inflammation-related genes more than two-fold. The extensive expressions of four genes (CCL1, IL1B, IL6 and IL8) were further confirmed by real-time quantitative PCR. It turned out that dual stimulation of LPS and Ti particles most up-regulated three genes (IL1B, IL6 and IL8), followed by LPS while Ti particles moderately but least increased, suggesting that phagocytosis of sub-micro Ti particles induces moderate inflammation with its degree less than LPS, but phagocytosis of sub-micro Ti particles has the potential to worsen inflammation caused by LPS-stimulated macrophages.

Preparation and in vivo evaluation of apatite/collagen packed composite by alternate immersion method and Newton press.

Further development of bio-compatible, bio-absorbable, and osteo-conductive bio-materials is desired for bone grafts in dental and medical clinics. One candidate material might be a high-density apatite/collagen composite, which cures relatively large bone defects. To produce such a composite, we freeze-dried type I collagen solution, cross-linked the formed sponge by 2 wt % glutaraldehyde, immersed the insoluble sponge in CaCl(2) and Na(2)HPO(4) solutions alternately five times, and compacted the sponge by Newton press at 5000 kgf. For comparison, cross-linked collagen without alternate immersion was also pressed. SEM/EPMA, XRD, and FTIR analyses clarified that alternate immersion successfully coated the collagen sponge with hydroxyapatite. Packed apatite/collagen composite and collagen disks 6 mm in diameter and 0.5 mm in height were implanted in the subperiostea of rabbit tibiae for 2, 4, 8, and 12 weeks to assess bio-compatibility, bio-absorbability, and osteo-conductivity. Histological observations showed that the packed apatite/collagen composite was biocompatible, osteo-conductive for up to 8 weeks, and largely bio-absorbed at 12 weeks, while the packed collagen sponge caused an undesirable foreign body reaction, which worsened with the implantation period. The overall findings suggest that this packed apatite/collagen composite might be used as a new bio-absorbable bone graft material.