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.

Surface characteristics and in vitro biocompatibility of titanium preserved in a vitamin C-containing saline storage solution.

The purpose of this study is to explore a storage solution for titanium implants and investigate its osteogenic properties. The commercial pure titanium (cp-Ti) surface and double-etched (SLA) titanium surface specimens were preserved in air, saline, 10 mM Vitamin C (VitC)-containing saline and 100 mM VitC-containing saline storage solutions for 2 weeks. The surface microtopography of titanium was observed by scanning electron microscopy (SEM), the surface elemental compositions of the specimens were analyzed by Raman and X-ray photoelectron spectroscopy (XPS), and water contact angle and surface roughness of the specimens were tested. The protein adsorption capacity of two titanium surfaces after storage in different media was examined by BCA kit. The MC3T3-E1 osteoblasts were cultured on two titanium surfaces after storage in different media, and the proliferation, adhesion and osteogenic differentiation activity of osteoblasts were detected by CCK-8, laser confocal microscope (CLSM) and Western blot. The SEM results indicated that the titanium surfaces of the air group were relatively clean while scattered sodium chloride or VitC crystals were seen on the titanium surfaces of the other three groups. There were no significant differences in the micromorphology of the titanium surfaces among the four groups. Raman spectroscopy detected VitC crystals on the titanium surfaces of two experimental groups. The XPS, water contact angle and surface roughness results suggested that cp-Ti and SLA-Ti stored in 0.9% NaCl and two VitC-containing saline storage solutions possessed less carbon contamination and higher surface hydrophilicity. Moreover, the protein adsorption potentials of cp-Ti and SLA-Ti surfaces were significantly improved under preservation in two VitC-containing saline storage solutions. The results of in vitro study showed that the preservation of two titanium surfaces in 100 mM VitC-containing saline storage solution upregulated the cell adhesion, proliferation, osteogenic related protein expressions of MC3T3-E1 osteoblasts. In conclusion, preservation of cp-Ti and SLA-Ti in 100 mM VitC-containing saline storage solution could effectively reduce carbon contamination and enhance surface hydrophilicity, which was conducive to osteogenic differentiation of osteoblasts.

The potential influence of high uric acid exposure on surface and corrosion susceptibility of pure titanium.

This study investigated the corrosion susceptibility of pure titanium under uric acid exposure for 7 days based on surface analysis. The prepared pure titanium specimens, exposed to different concentrations of uric acid, were examined for surface microstructure, surface element composition and surface wettability using scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and static contact angle measurement, respectively. The corrosion behaviors of titanium specimens were measured by open-circuit potential (OCP), electrochemical impedance spectroscopy (EIS) and potentiodynamic polarization. The titanium ion release from the prepared specimens, which were immersed in Hank's balanced salt solution (HBSS) containing different amount of uric acid, was measured by inductively coupled plasma atomic emission spectrometry (ICP-AES). More irregular pitting holes were observed on titanium surfaces exposed to a high concentration of uric acid, and XPS analyses revealed that the amount of titanium dioxide (TiO2) decreased. Titanium surfaces pre-treated with high uric acid became more hydrophobic. Furthermore, the results of OCP and potentiodynamic polarization tests showed increased corrosion susceptibility of titanium samples, while EIS data indicated more active corrosion behavior of titanium materials. The high concentration of uric acid also induced titanium ion release. High concentration of uric acid negatively influenced the surface characteristics and corrosion properties of titanium materials, which destroyed the titanium oxide film barrier. High uric acid exposure increased corrosion susceptibility of pure titanium specimens and accelerated titanium ion release. Graphical abstract.

Targeting NF-κB c-Rel in regulatory T cells to treat corneal transplantation rejection.

The relevance of Tregs in the induction of tolerance against corneal allografts has been well established. Although it is well known that the conversion of Tregs into effector-like cells contributes to the loss of corneal immune privilege, the underlying mechanism is still not fully understood. Using heterologous penetrating keratoplasty model, we found that Tregs from corneal allograft rejected mice (inflam-Tregs) exhibit impaired function and characteristics of effector T cells. Further study showed that the expression of NF-κB c-Rel, a key mediator of effector T cell function, was significantly increased in inflam-Tregs. Mechanistic study revealed that elevated NF-κB c-Rel level in inflam-Tregs impaired Treg function through the promotion of inflammatory cytokine production and glycolysis. More importantly, we demonstrated that targeting NF-κB c-Rel was able to improve the immune suppressive function of inflam-Tregs in vitro and enhance the potential of them to suppress corneal transplantation rejection. Therefore, our current study identified NF-κB c-Rel as a key mediator of the conversion of Tregs into effector-like cells when under inflammatory environment.

Role of the Hippo-YAP/NF-κB signaling pathway crosstalk in regulating biological behaviors of macrophages under titanium ion exposure.

The presence of metal ions, such as titanium (Ti) ions, is toxic to adjacent tissues of implants. Indeed, Ti ions may induce an inflammatory response through the NF-κB pathway, thus causing damage to soft and hard tissues. The involvement of Yes-associated protein (YAP), a key factor of the Hippo pathway, in an immuno-inflammatory response has been confirmed, whereas its role in Ti ion-mediated inflammation has not been elucidated. Therefore, this study aimed to investigate the role of signal crosstalk between the Hippo/YAP and NF-κB signaling pathways in the pro-inflammatory effect of Ti ions on macrophages. In our work, RAW264.7 cells were cocultured with Ti ions. The migration capacity of macrophages under Ti ion exposure was measured by transwell assay. Western blot analysis was used to detect the expressions of related proteins. Polymerase chain reaction was used to evaluate the expression of pro-inflammatory cytokines. The nucleus translocation of YAP and P65 was visualized and analyzed via immunofluorescence staining. The results showed that the migration of macrophages was promoted under Ti ion exposure. Ten parts per million Ti ions induced nuclear expression of YAP and activated the NF-κB pathway, which finally upregulated the expression of pro-inflammatory cytokines in macrophages. Moreover, the inhibition of the NF-κB pathway rescued the reduction of YAP expression under Ti ion exposure. Most importantly, the overexpression of YAP exacerbated the inflammatory response mediated by Ti ions through the NF-κB pathway. In summary, this study explored the mechanism of Hippo-YAP/NF-κB pathway crosstalk involved in the regulation of macrophage behaviors under Ti ion exposure.

NAD+ precursors protect corneal endothelial cells from UVB-induced apoptosis.

Excessive exposure of the eye to ultraviolet B light (UVB) leads to corneal edema and opacification because of the apoptosis of the corneal endothelium. Our previous study found that nicotinamide (NIC), the precursor of nicotinamide adenine dinucleotide (NAD), could inhibit the endothelial-mesenchymal transition and accelerate healing the wound to the corneal endothelium in the rabbit. Here we hypothesize that NIC may possess the capacity to protect the cornea from UVB-induced endothelial apoptosis. Therefore, a mouse model and a cultured cell model were used to examine the effect of NAD+ precursors, including NIC, nicotinamide mononucleotide (NMN), and NAD, on the UVB-induced apoptosis of corneal endothelial cells (CECs). The results showed that UVB irradiation caused apparent corneal edema and cell apoptosis in mice, accompanied by reduced levels of NAD+ and its key biosynthesis enzyme, nicotinamide phosphoribosyltransferase (NAMPT), in the corneal endothelium. However, the subconjunctival injection of NIC, NMN, or NAD+ effectively prevented UVB-induced tissue damage and endothelial cell apoptosis in the mouse cornea. Moreover, pretreatment using NIC, NMN, and NAD+ increased the survival rate and inhibited the apoptosis of cultured human CECs irradiated by UVB. Mechanistically, pretreatment using nicotinamide (NIC) recovered the AKT activation level and decreased the BAX/BCL-2 ratio. In addition, the capacity of NIC to protect CECs was fully reversed in the presence of the AKT inhibitor LY294002. Therefore, we conclude that NAD+ precursors can effectively prevent the apoptosis of the corneal endothelium through reactivating AKT signaling; this represents a potential therapeutic approach for preventing UVB-induced corneal damage.

The proinflammatory cytokines IL-1ß and TNF-α modulate corneal epithelial wound healing through p16Ink4a suppressing STAT3 activity.

The proinflammatory cytokines interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) are involved in the corneal inflammatory response and wound healing following corneal injuries. However, the mechanism by which proinflammatory cytokines modulate corneal epithelial wound healing remains unclear. In this study, we found that IL-1ß or TNF-α was transiently elevated during corneal epithelial wound healing in mice. After corneal epithelial debridement, persistent treatment with IL-1ß or TNF-α restrained the level of phosphorylated signal transducer and activator of transcription 3 (p-STAT3) and boosted the level of cell cycle inhibitor p16Ink4a , resulting in impaired corneal epithelial repair. When p16Ink4a was deleted, the p-STAT3 level in corneal epithelium was enhanced and corneal epithelial wound healing was clearly accelerated. In diabetic mice, IL-1ß, TNF-α, and p16Ink4a appeared a sustained and strong expression in the corneal epithelium, and p16Ink4a knockdown partially reverted the defective diabetic corneal epithelial repair. Furthermore, immunoprecipitation proved that p16Ink4a interacted with p-STAT3 and thus possibly suppressed the STAT3 activity. Our findings revealed a novel mechanism that the proinflammatory cytokines modulate corneal epithelial wound healing via the p16Ink4a -STAT3 signaling.

Surface analysis and corrosion behavior of pure titanium under fluoride exposure.

STATEMENT OF PROBLEM: The corrosive effects of oral fluoride products on titanium have been reported, and chronic fluorosis, which causes hyperfluoemia, is one of the world's health problems. Nevertheless, the relationship between high serum fluoride and corrosion on the titanium surface, which might have adverse effects on titanium implant osseointegration, has not been elucidated. PURPOSE: The purpose of this in vitro study was to investigate the corrosion behavior of pure titanium exposed to high serum fluoride with different pH values based on surface analysis. MATERIAL AND METHODS: Pure titanium specimens, exposed to different electrolytes with 0.04 and 0.4 ppm NaF at pH 7.3 and 5.0 values, were examined for surface microstructure by using scanning electron microscopy (SEM) and for surface element composition with X-ray photoelectron spectroscopy (XPS). The corrosion behavior and metal ion release of specimens immersed in the Hanks' balanced salt solution (HBSS) containing 0.04 and 0.4 ppm serum fluoride concentrations (NaF) at 7.3 and 5.0 pH values were measured by electrochemical impedance spectroscopy (EIS) and inductively coupled plasma atomic emission spectrometry (ICP-AES). RESULTS: Pitting holes were observed on pure titanium surfaces exposed to high serum fluoride. The surfaces became rougher with the increase of serum fluoride concentration, especially under acidic conditions. XPS analysis revealed a reduction of dominant titanium dioxide (TiO2) on the pure titanium surface under serum fluoride exposure, corresponding to an increase in the relative level of F. EIS data showed an active corrosion behavior of pure titanium exposed to high serum fluoride and gradually decreased corrosion resistance with increasing concentration of serum fluoride, which was more severe under acidic conditions. The release of titanium ions was also induced by high serum fluoride and acidic conditions. CONCLUSIONS: High serum fluoride had a negative influence on the corrosion behavior of pure titanium. The titanium oxide film barrier could be broken down in the fluoride ions condition, and the corrosion resistance of pure titanium decreased with the increasing concentration of serum fluoride. The increased corrosion susceptibility of pure titanium accelerated the release of titanium ions after exposure to high serum fluoride; this was more pronounced in an acidic environment.

Nicotinamide inhibits corneal endothelial mesenchymal transition and accelerates wound healing.

Corneal endothelial cells (CECs) maintain the clarity of the cornea through the barrier and pump function. Ex vivo culture or injury may cause corneal endothelial-mesenchymal transition (EnMT) and lead to loss of function. In this study, we explored the effects of nicotinamide (NIC) on the wound healing of rabbit corneal endothelium and the proliferation, migration, and EnMT of cultured human CEC lines. The animal results showed that corneal clarity was rapidly recovered within seven days through topical application of NIC in the rabbits with mechanical injury of the corneal endothelium, while the control corneas remained edematous and cloudy. Whole-mounted corneal staining found the expressions of Na+/K+-ATPase, aquaporin-1, and zonula occludens-1 were mainly localized to the boundaries of regenerated endothelium in NIC-treated eyes, in contrast to the scattered staining in vehicle-treated eyes. Interestingly, we found that NIC application inhibited the expression of typical EnMT marker alpha-smooth muscle actin, which appeared in the rabbit corneal endothelial wound healing. In vitro, NIC promoted the proliferation, but not the migration, of cultured human CECs. Moreover, NIC effectively inhibited transforming growth factor beta-1-induced corneal EnMT and decreased the levels of EnMT regulators snail and slug. Therefore, our study indicates that NIC enhances corneal endothelial wound healing through the promotion of proliferation and the inhibition of EnMT, which may provide a potential pharmaceutical agent for treating corneal endothelial dysfunction.

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.

The colonization and divergence patterns of Brandt's vole (Lasiopodomys brandtii) populations reveal evidence of genetic surfing.

BACKGROUND: The colonial habit of Brandt's vole (Lasiopodomys brandtii) differs from that of most other species of the genus Microtus. The demographic history of this species and the patterns shaping its current genetic structure remain unknown. Here, we explored patterns of genetic differentiation and infered the demographic history of Brandt's vole populations through analyses of nuclear microsatellite and D-loop sequences. RESULTS: Phylogenetic analyses divided the sampled populations into three main clusters, which represent the southeastern, northeastern and western parts of the total range in Mongolia and China. Molecular data revealed an ancestral area located in the southeast of the extant range, in the Xilinguole District, Inner Mongolia, China, from where Brandt's vole populations began expanding. A gene flow analysis suggested that the most likely colonization route was from the ancestral area and was followed by subsequent northeastward and westward range expansions. We identified decreases in genetic diversity with increasing distance from the founder population within the newly occupied regions (northeastern and western regions), clinal patterns in the allele frequencies, alleles that were rare in the original area that have become common in the newly occupied regions, and higher genetic differentiation in the expanded range compared with the original one. CONCLUSION: Our results indicate that L. brandtii most likely originated from the southeastern part of its current geographic range, and subsequently colonized into the northeastern and western parts by expansion. The genetic patterns among the derived populations and with respect to the original population are consistent with that expected under genetic surfing models, which indicated that genetic drift, rather than gene flow, is the predominant factor underlying the genetic structure of expanding Brandt's vole populations.

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.

Anti-inflammatory effects of the immobilization of SEMA4D on titanium surfaces in an endothelial cell/macrophage indirect coculture model.

In this study, we established a procedure to prepare a Semaphorin4D (SEMA4D)-immobilized titanium surface and explored its effects on macrophage behaviors in an endothelial cell/macrophage indirect coculture model. The SEMA4D-bovine serum albumin complex was immobilized onto a preprocessed poly L-lysine titanium surface through NaOH hydrothermal treatment and self-assembly technology. All titanium specimens were examined for surface microstructure, surface element composition, and surface wettability by field emission scanning electron microscopy, x-ray photoelectron spectroscopy (XPS), and water contact angle measurement, respectively. Subsequently, we constructed an endothelial cell/macrophage indirect coculture model and evaluated the activation of NF-κB signaling pathway and the expression of proinflammatory cytokines (TNFα, IL-6, and IL-1ß) in macrophages. In XPS analysis, the SEMA4D-immobilized titanium surface appeared as a loose porous structure covered with uniform film, which exhibited better hydrophilicity than the control smooth titanium surface. In the indirect coculture model, SEMA4D attenuated the activation of NF-κB signaling pathway of lipopolysaccharide-stimulated THP-1 macrophages, thereby downregulating the expression of proinflammatory cytokines in macrophages. In conclusion, SEMA4D could be immobilized on titanium surfaces through NaOH hydrothermal treatment and self-assembly technology. Meanwhile, SEMA4D immobilization altered the characteristics of the titanium surfaces, which negatively regulated macrophage behaviors in the endothelial cell/macrophage indirect coculture model.

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.

Osteoclast-mediated biocorrosion of pure titanium in an inflammatory microenvironment.

Titanium (Ti) and alloys thereof are commonly utilized in biomedical settings owing to their desirable mechanical properties and good biocompatibility. However, when exposed to biological systems for extended periods of time, Ti still undergoes corrosion. In the present study, we therefore explore the impact of osteoclasts (OC) on the surface characteristics and corrosion of commercially pure Titanium (cpTi) in the context of lipopolysaccharide (LPS)-induced inflammation. We utilized tartrate resistant acidic phosphatase (TRAP) and fluorescence staining to assess OC properties, while scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), optical profilometer, electrochemical impedance spectroscopy (EIS), potentiodynamic polarization tests, and inductively coupled plasma atomic emission spectrometry (ICP-AES) were used to evaluate metal microstructure, surface composition and roughness, electrochemical corrosion properties, and metal ion release. SEM findings demonstrated that the surface of cpTi exhibited micro-pitting as well as the presence of viable OCs. Correspondingly, cpTi that had been exposed to OCs exhibited reduced levels of Ti, oxygen, and oxides within the corroded regions relative to smooth Ti as measured via EDS and XPS. OC exposure was also associated with significant changes in cpTi surface roughness, a significant decrease in corrosion resistance, and a significant increase in the release of Ti ions into the surrounding medium. In summary, these findings indicate that OC culture on the surface of cpTi can directly corrode titanium and lead to the release of Ti ions.