Enhanced osteogenic and antibacterial properties of titanium implant surface modified with Zn-incorporated nanowires: Preclinical in vitro and in vivo investigations.

OBJECTIVE: This study aimed to synthesize zinc-incorporated nanowires structure modified titanium implant surface (Zn-NW-Ti) and explore its superior osteogenic and antibacterial properties in vitro and in vivo. MATERIALS AND METHODS: Zn-NW-Ti was synthesized via displacement reactions between zinc sulfate solutions and the titanium (Ti) surface, which was pretreated by hydrofluoric acid etching and hyperthermal alkalinization. The physicochemical properties of the Zn-NW-Ti surface were examined. Moreover, the biological effects of Zn-NW-Ti on MC3T3-E1 cells and its antibacterial property against oral pathogenic bacteria (Staphylococcus aureus, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans) compared with sandblasted and acid-etched Ti (SLA-Ti) and nanowires modified Ti (NW-Ti) surface were assessed. Zn-NW-Ti and SLA-Ti modified implants were inserted into the anterior extraction socket of the rabbit mandible with or without exposure to the mixed bacterial solution (S. aureus, P. gingivalis, and A. actinomycetemcomitans) to investigate the osteointegration and antibacterial performance via radiographic and histomorphometric analysis. RESULTS: The Zn-NW-Ti surface was successfully prepared. The resultant titanium surface appeared as a nanowires structure with hydrophilicity, from which zinc ions were released in an effective concentration range. The Zn-NW-Ti surface performed better in facilitating the adhesion, proliferation, and differentiation of MC3T3-E1 cells while inhibiting the colonization of bacteria compared with SLA-Ti and NW-Ti surface. The Zn-NW-Ti implant exhibited enhanced osseointegration in vivo, which was attributed to increased osteogenic activity and reduced bacterial-induced inflammation compared with the SLA-Ti implant. CONCLUSIONS: The Zn-incorporated nanowires structure modified titanium implant surface exhibited improvements in osteogenic and antibacterial properties, which optimized osteointegration in comparison with SLA titanium implant surface.

Effect of titanium ions on the Hippo/YAP signaling pathway in regulating biological behaviors of MC3T3-E1 osteoblasts.

Titanium (Ti) and its corresponding alloys have been widely applied in dental and orthopedic implants. Owing to abrasion and corrosion of implants in the unfavorable electrolytic aqueous environment of the host body, Ti ions could be released from implants and accumulated in local tissues. Recent studies have found that excessive Ti ions were toxic to osteoblasts in adjacent bone tissues and subsequently influenced long-term effects on implant prostheses. However, the potential molecular mechanisms underlying the damage to osteoblasts induced by Ti ions remained unclear. Hippo signaling has been confirmed to be involved in organ size and tissue regeneration in many organs, while its roles in osteoblasts differentiation and bone repair remained elusive. Therefore, we hypothesize that YAP, a regulator of Hippo pathway, inhibited osteoblast growth, skeletal development and bone repair, as well as excessive Ti ions promoted the progression of YAP activation. This study aimed to explore the role of Hippo/YAP signaling pathway in the biotoxicity effect of Ti ions on osteoblast behaviors. Here, we confirmed that 10 ppm Ti ions, a minimum concentration gradient previously reported that was capable of suppressing osteoblasts growth, induced nuclear expression of YAP in osteoblasts in our study. Furthermore, 10 ppm Ti ion-induced YAP activation was found to downregulate osteogenic differentiation of MC3T3-E1 cells. Most importantly, the hypothesis we proposed that knockdown of YAP did reverse the inhibitory effect of 10 ppm Ti ions on osteogenesis has been verified. Taken together, our work provides insights into the mechanism of which YAP is involved in regulating osteoblast behaviors under the effect of Ti ions, which may help to develop therapeutic applications for Ti implant failures and peri-implantitis.

PLGA film/Titanium nanotubues as a sustained growth factor releasing system for dental implants.

Ti-based implants sometimes fail to integrate with surrounding bone tissue due to insufficiency of new bone formation and surface bonding. To overcome this problem, this research focused on establishing a sustained bone growth factor delivery system by applying anodized TiO2 nanotube arrays and PLGA film on the titanium implant surface. TiO2 nanotube arrays were made by anodic oxidation method, and were then filled with rhBMP2 by vacuum freeze-drying. Next, PLGA was deposition on the surface of this material. The designed system was characterized, pharmacokinetic release rate of rhBMP2 was determined. Adhesion, proliferation, and differentiation activity of osteoblasts cultured on the new surfaces and traditional titanium surfaced were compared. SEM showed that a surface of TiO2 nanotube arrays were successfully generated. PLGA membranes of 50 nm, 250 nm, 800 nm thickness were successfully deposited on the surfaces of TiO2 nanotube layers by using 1%, 3%, 10% PLGA solutions. PLGA film of 250 nm thickness showed ideally controlled release of rhBMP2, lasting for 4 weeks. Furthermore, 250 nm thickness PLGA film improved osteoblast adhesion, proliferation, and levels of alkaline phosphatase. In conclusion, the PLGA film / TiO2 nanotube growth factor delivery system can effectively sustain the release of rhBMP-2, and promote proliferation and differentiation of MC3T3-E1 osteoblasts.

Effectiveness of resin-based materials against erosive and abrasive enamel wear.

OBJECTIVE: The objective of this study was to test the effectiveness of resin-based materials against erosive enamel wear under erosive and abrasive challenges by orange juice and tooth brushing. METHODS: Fifty enamel specimens from third molars were assigned to five groups: ICON resin infiltration with no etching (ICON-NE), ICON resin infiltration with 15 % HCl etching (ICON-AE), Seal & Protect sealant (S&P), Tetric EvoFlow (TEF), and control. Erosive lesions were first created on enamel, then treated with resin-based materials. Erosive and abrasive challenges by orange juice and tooth brushing were repeated after treatments. Erosive wear of the treated areas was measured with 3D scanning microscopy, and data were analyzed using ANOVA and paired t tests. RESULTS: Treatments with ICON, S&P, and TEF created a protective material coating of 4.5 ± 1.9 µm, 44.3 ± 8.1 µm, and 84.6 ± 15.7 µm in thickness on the lesion surfaces, respectively. After 15 cycles of erosive and abrasive challenges, enamel or material losses were 21.9 ± 2.3 µm for control, 24.5 ± 4.0 µm for ICON-NE, 24.6 ± 7.4 µm for ICON-AE, 11.2 ± 4.1 µm for S&P, and 3.9 ± 1.9 µm for TEF, respectively. The protective coatings were completely lost in the ICON infiltration groups but remained intact in both the S&P and TEF groups after erosive and abrasive challenges. CONCLUSION: In contrast to a resin sealant and a flowable composite, ICON infiltration resin was not effective in protecting enamel surfaces from erosive and abrasive wear. CLINICAL RELEVANCE: Sealant and flowable composite resin may create protective coatings on eroded enamel surfaces and prevent further tissue loss.

Early loading of splinted implants in the posterior mandible: a prospective multicentre case series.

AIM: To evaluate the 12-months clinical and radiological outcomes with the OsseoSpeed(™) TX implant using an early loading protocol in patients with missing teeth in the posterior mandible. MATERIAL AND METHODS: Forty-five subjects, with Kennedy class I or II edentulism in the mandible, were enrolled at three centres in China. Two or three implants were placed in one edentulous region using a one-stage procedure. Patients received a screw-retained splinted fixed permanent restoration in one edentulous region 6-8 weeks after surgery. Follow-up took place at 6 and 12 months after loading. Marginal bone level alteration, implant survival and clinical findings were assessed using descriptive statistics. The data were analysed on a patient level, implying that the mean overall implants by patient was used as the statistical unit. The data from the three centres were pooled in the statistical analyses. RESULTS: A total of 107 implants were inserted in 45 patients. Twelve months after loading, the implant survival rate was 100%, with a mean (± std) marginal bone gain of 0.08 ± 0.411 mm and healthy soft tissue status. CONCLUSIONS: Early loading of splinted OsseoSpeed(™) TX implants was an effective and safe treatment for partial edentulism of the posterior mandible. CLINICAL TRIAL REGISTRATION NUMBER ON CLINICALTRIALS.GOV: NCT01346683.

Reciprocal interaction between dental alloy biocorrosion and Streptococcus mutans virulent gene expression.

Corrosion of dental alloys is a major concern in dental restorations. Streptococcus mutans reduces the pH in oral cavity and induces demineralization of the enamel as well as corrosion of restorative dental materials. The rough surfaces of dental alloys induced by corrosion enhance the subsequent accumulation of plaque. In this study, the corrosion process of nickel-chromium (Ni-Cr) and cobalt-chromium (Co-Cr) alloys in a nutrient-rich medium containing S. mutans was studied using inductively coupled plasma atomic emission spectrometry (ICP-AES), X-ray photoelectron spectroscopy (XPS) and electrochemical corrosion test. Our results showed that the release of Ni and Co ions increased, particularly after incubation for 3 days. The electrochemical corrosion results showed a significant decrease in the corrosion resistance (Rp) value after the alloys were immersed in the media containing S. mutans for 3 days. Correspondingly, XPS revealed a reduction in the relative dominance of Ni, Co, and Cr in the surface oxides after the alloys were immersed in the S. mutans culture. After removal of the biofilm, the pre-corroded alloys were re-incubated in S. mutans medium, and the expressions of genes associated with the adhesion and acidogenesis of S. mutans, including gtfBCD, gbpB, fif and ldh, were evaluated by detecting the mRNA levels using real-time reverse transcription polymerase chain reaction (RT-PCR). We found that the gtfBCD, gbpB, ftf and Idh expression of S. mutans were noticeably increased after incubation with pre-corroded alloys for 24 h. This study demonstrated that S. mutans enhanced the corrosion behavior of the dental alloys, on the other hand, the presence of corroded alloy surfaces up-regulated the virulent gene expression in S. mutans. Compared with smooth surfaces, the rough corroded surfaces of dental alloys accelerated the bacteria-adhesion and corrosion process by changing the virulence gene expression of S. mutans.

Effects of lipopolysaccharides on the corrosion behavior of Ni-Cr and Co-Cr alloys.

STATEMENT OF PROBLEM: Lipopolysaccharides (LPS) are constituents of gingival crevicular fluid and may affect the base metal alloys used in metal ceramic crowns. The role of LPS in base metal alloys is currently unknown. PURPOSE: The purpose of this in vitro study was to evaluate the effects of gram-negative bacterial LPS on the electrochemical behavior of Ni-Cr and Co-Cr alloys. MATERIAL AND METHODS: Alloy specimens were divided into 4 groups according to Escherichia coli LPS concentration (0, 0.15, 15, and 150 µg/mL) in acidic saliva (pH 5). Open circuit potential (OCP) and potentiodynamic polarization behavior were examined using a computer-controlled potentiostat. Metal ions released from the 2 alloys were measured by immersion in LPS-free solution and 150 µg/mL LPS solution and analyzed by inductively coupled plasma atomic emission spectrometry (ICP-AES). Data were evaluated using 1-way ANOVA (α=.05). RESULTS: Compared with control groups, medium LPS concentration (15 µg/mL) accelerated Ni-Cr alloy corrosion (P<.05), whereas high LPS concentration (150 µg/mL) accelerated Co-Cr alloy corrosion (P<.05), as determined by OCP, corrosion current density, and polarization resistance parameters. After immersion in high LPS concentrations (150 µg/mL), a slight increase in Ni ion release (P >.05) was observed for the Ni-Cr alloy, while a more significant Co ion release (P<.05) was observed for the Co-Cr alloy. CONCLUSIONS: LPS negatively affected the electrochemical behavior of both the Ni-Cr and Co-Cr alloys.

Corrosion behavior of pure titanium in the presence of Actinomyces naeslundii.

It is well known that some microorganisms affect the corrosion of dental metal. Oral bacteria such as Actinomyces naeslundii may alter the corrosion behavior and stability of titanium. In this study, the corrosion behavior of titanium was studied in a nutrient-rich medium both in the presence and the absence of A. naeslundii using scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and electrochemical impedance spectroscopy (EIS). A. naeslundii was able to colonize the surface of titanium and then form a dense biofilm. The SEM images revealed the occurrence of micropitting corrosion on the metal surface after removal of the biofilm. The electrochemical corrosion results from EIS showed a significant decrease in the corrosion resistant (R(p)) value after immersing the metal in A. naeslundii culture for 3 days. Correspondingly, XPS revealed a reduction in the relative levels of titanium and oxygen and an obvious reduction of dominant titanium dioxide (TiO2) in the surface oxides after immersion of the metal in A. naeslundii culture. These results suggest that the metabolites produced by A. naeslundii can weaken the integrity and stability of the protective TiO2 in the surface oxides, which in turn decreases the corrosion resistance of titanium, resulting in increased corrosion of titanium immersed in A. naeslundii solution as a function of time.

6-Bromoindirubin-3'-oxime Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells and Facilitates Bone Regeneration in a Mouse Periodontitis Model.

Effective bone tissue engineering is important to overcome the unmet clinical challenges of periodontal tissue regeneration. Successful bone tissue engineering comprises three key factors: stem cells, growth factors, and scaffolds. 6-Bromoindirubin-3'-oxime (BIO) is an inhibitor of glycogen synthase kinase-3 (GSK-3) that can activate the Wnt signaling pathway by enhancing ß-catenin activity. In this study, the effects of BIO on the proliferation, migration, and osteogenic differentiation of periodontal ligament stem cells (PDLSCs) were investigated. Poly(lactic-co-glycolic acid) (PLGA) and hyaluronic acid (HA) emerged as promising biomaterials; thus, we developed a novel HA hydrogel embedded with BIO-encapsulated PLGA microspheres and injected the formulation into the gingival sulcus of mice with experimental periodontitis. The release speed of this system was fast in the first week and followed a sustained release phase until week 4. In vivo experiments showed that this PLGA-BIO-HA hydrogel system can inhibit periodontal inflammation, promote bone regeneration, and induce the expression of bone-forming markers alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), and osteocalcin (OCN) in a mouse periodontitis model. Therefore, this PLGA-BIO-HA hydrogel system provides a promising therapeutic strategy for periodontal bone regeneration.

Biocorrosion of pure and SLA titanium surfaces in the presence of Porphyromonas gingivalis and its effects on osteoblast behavior.

Objective: The study aims to investigate the biocorrosion behavior of Porphyromonas gingivalis on pure and SLA titanium surfaces and its effects on surface characteristics and osteoblast behavior. Methods: Pure and SLA titanium specimens were immersed in culture medium with P. gingivalis and incubated for 7 days. P. gingivalis colonization on the pure and SLA titanium surfaces was observed by scanning electron microscopy (SEM). The pure and SLA titanium surface characteristics were analyzed via X-ray photoelectron spectroscopy (XPS), surface roughness and surface wettability. The corrosion behaviors of pure and SLA titanium specimens were evaluated by electrochemical corrosion test. The osteoblast behavior of MC3T3-E1 cells on the pure and SLA titanium surfaces after P. gingivalis colonization was investigated by cell adhesion and western blot assays. Results: P. gingivalis colonized on the pure and SLA titanium surfaces was observed by SEM. The XPS analysis demonstrated reductions in the relative levels of titanium and oxygen and obvious reductions of dominant titanium dioxide (TiO2) on both titanium surfaces after immersing the metal in P. gingivalis culture. In addition, their roughness and wettability were changed. Correspondingly, the electrochemical corrosion test results revealed significant decreases in the corrosion resistance and increases in the corrosion rate of the pure and SLA titanium specimens after immersion in P. gingivalis culture. The results of the in vitro study showed that the pre-corroded pure and SLA titanium surfaces by P. gingivalis exhibited lower osteocompatibility and down-regulated the adhesion, spreading and osteogenic differentiation abilities of MC3T3-E1 cells. Conclusions: P. gingivalis was able to colonize on the pure and SLA titanium surfaces and weaken their surface properties, especially a decrease in the protective TiO2 film, which induced the biocorrosion and further negatively affected the osteoblast behavior.

Elimination of Cr(VI) from chromium slag with poplar lignin by electrochemical treatment in sulfuric acid solution.

In this paper, we proposed a novel method to eliminate nocuous Cr(VI) from chromium slag with poplar lignin by electrochemical treatment in sulfuric acid solution. In this electrochemical process, self-made Ti/SnO2-Sb anode and graphite cathode were applied, and the oxidative degradation of lignin proceeded simultaneously with the reduction of Cr(VI) in one pot. The influences of pivotal factors on electrocatalytic redox efficiency were investigated, such as chromium slag concentration, lignin concentration, current density, sulfuric acid concentration, and reaction time. The results showed that the elimination rate of Cr(VI) in chromium slag was 97.16 ± 1.13% and the total yield of lignin degradation products reached 93.78 g/kg lignin under the optimal conditions. X-ray photoelectron spectroscopy (XPS), energy dispersive X-ray spectroscopy (EDS), and UV-visible spectrophotometer studies confirmed that most of the Cr(VI) ions were reduced to Cr(III) ions with the aid of lignin, and a small amount of Cr(VI) ions were adsorbed by lignin residue. Importantly, this method provides a typical example of "waste control by waste", which is treating waste chromium slag with waste lignin that can be an effective way to eliminate Cr(VI).

Evidence for calcifying nanoparticles in gingival crevicular fluid and dental calculus in periodontitis.

BACKGROUND: Calcifying nanoparticles (CNPs), also known as nanobacteria, can produce carbonate apatite on their cell walls and initiate pathologic calcification. The objective of this study was to determine whether CNPs are present in the gingival crevicular fluid (GCF) from subjects with periodontal disease and whether they can induce the pathologic calcification of primary cultured human gingival epithelial cells. METHODS: GCF and dental calculus samples were collected from 10 subjects with gingivitis and 10 subjects with chronic periodontitis. CNPs in GCF and calculus filtrates were detected with nanocapture enzyme-linked immunosorbent assay kits. The CNPs in cultures of dental calculus filtrates were also identified using immunofluorescence staining, transmission electron microscopy (TEM), and chemical analysis. Pathologic changes in the CNP-treated gingival epithelial cells were observed with TEM, alizarin red staining, and disk-scanning confocal microscopy. RESULTS: CNPs were found in GCF samples from two subjects with chronic periodontitis. Based on chemical analysis, the surface-associated material from CNPs isolated and cultured from calculus has a composition similar to dental calculus. The pathologic calcification of CNP-treated gingival epithelial cells was also observed. CONCLUSIONS: Self-replicating calcifying nanoparticles can be cultured and identified from dental calculus. This raises the issue of whether CNPs contribute to the pathogenesis of periodontitis.

[The effect of bacteria reaction time on corrosion properties of Ni-Cr alloys pretreated with different proteins].

PURPOSE: To evaluate the corrosion properties of absorbed protein on the surface of NiCr alloys, and provide experimental base for corrosion resistance of dental casting alloys. METHODS: NiCr alloy specimens were divided into 3 groups: one group was exposed to the artificial saliva(control group), and the other 2 groups were exposed to the artificial saliva with 1% bovine serum albumin(BSA), or 0.22% lysozyme(LSZ). Group of BSA and group of LSZ were the experimental group. Specimens in 3 groups were cultured in solution of Streptococcus mutans for 12 h, 24 h, 36 h and 48h, and investigated with electrochemical impedance spectroscopy measurement(EIS) and potentiodynamic polarization measurement(POT) to determine the corrosion resistance of the alloys. The data was analyzed with SPSS 17.0 software package. RESULTS: The results indicated that the corrosion resistance of both BSA group and LSZ group were higher than that of the control group (P<0.05) and LSZ group was superior to BSA group cultured in the solution of Streptococcus mutans for 12 h. When cultured for 24 h, the corrosion resistance of BSA group and LSZ group had no significant difference (P>0.05), but was still higher than that of the control group. After 36 h culture time, the control group and the BSA group had no statistical difference in corrosion resistance （P>0.05), while the LSZ group had the poorest corrosion resistance. When the culture time extended to 48 h, the control group had a better corrosion resistance compared with the BAS group and the LSZ group(P<0.05), but BSA group had displayed lower corrosion properties than LSZ group. The potentiodynamic polarization curve and electrochemical impedance spectroscopy had similar results. CONCLUSIONS: The adhesion of BSA and LSZ on the surface of the NiCr alloys in the early time could effectively inhibit the corrosive effect of Streptococcus mutans. The LSZ had better effect than BSA. With the continuing role of bacteria and the consumption of the absorb protein, the corrosion resistance of NiCr alloys toward Streptococcus mutans becomes lower than the alloys without absorb protein, which demonstrated that the adhesion of protein would change the surface structure of NiCr alloys and BSA had a greater effect.

Direct conversion of cellulose to 1-(furan-2-yl)-2-hydroxyethanone in zinc chloride solution under microwave irradiation.

Conversion of cellulose to 1-(furan-2-yl)-2-hydroxyethanone has been demonstrated in concentrated zinc chloride solution under microwave irradiation. Compared with the conventional oil-bath heating mode, microwave irradiation significantly reduced the reaction time and increased the yield of 1-(furan-2-yl)-2-hydroxyethanone. A typical degradation reaction with cellulose produced 1-(furan-2-yl)-2-hydroxyethanone in 12.0% molar yield in ZnCl(2) solution (ZnCl(2)-H(2)O ratio=2.25:1, w/w) with microwave irradiation at 600 W for 5 minutes at 135°C.