Inhibition of autophagy reduces the rate of fluoride-induced LS8 apoptosis via regulating ATG5 and ATG7.

Excessive fluoride affects ameloblast differentiation and tooth development. The fate of fluorinated ameloblasts is determined by multiple signaling pathways in response to a range of stimuli. Both autophagy and apoptosis are involved in the regulation of dental fluorosis as well as in protein synthesis and enamel mineralization. Emerging evidence suggests that autophagy and apoptosis are interconnected and that their interaction greatly influences cell death. However, the effect of autophagy on apoptosis in fluoride-treated ameloblasts is unclear. Here, we employed an in vitro cellular model of fluorosis in mouse ameloblast-like LS8 cells and induced autophagy using sodium fluoride (NaF). Our findings suggest that NaF treatment induces autophagy in LS8 cells, and ATG5 and ATG7 are important molecules involved in this process. We also showed that NaF treatment reduced cell viability in Atg5/7 siRNA and autophagy inhibitor-treated LS8 cells. More importantly, NaF-induced apoptosis can be reversed by inhibiting early stage of autophagy. In conclusion, our study shows that autophagy is closely related to dental fluorosis, and inhibition of autophagy, especially ATG5/7, reduces fluoride-induced cell death and apoptosis.

MAP kinase phosphatase MKP-1 regulates p-ERK1/2 signaling pathway with fluoride treatment.

Dental fluorosis is characterized by hypomineralization of tooth enamel caused by ingestion of excessive fluoride during enamel formation. Excess fluoride could have effects on the ERK signaling, which is essential for the ameloblasts differentiation and tooth development. MAP kinase phosphatase-1 (MKP-1) plays a critical role in regulating ERK related kinases. However, the role of MKP-1 in ameloblast and the mechanisms of MKP-1/ERK signaling in the pathogenesis of dental fluorosis are incompletely understood. Here, we adopted an in vitro fluorosis cell model using murine ameloblasts-like LS8 cells by employing sodium fluoride (NaF) as inducer. Using this system, we demonstrated that fluoride exposure led to an inhibition of p-MEK and p-ERK1/2 with a subsequent increase in MKP-1 expression in a dose-dependent manner. We further identified, under high dose fluoride, MKP-1 acted as a negative regulator of the fluoride-induced p-ERK1/2 signaling, leading to downregulation of CREB, c-myc, and Elk-1. Our results identify a novel MKP-1/ERK signaling mechanism that regulates dental fluorosis and provide a framework for studying the molecular mechanisms of intervention and fluorosis remodeling under normal and pathological conditions. MKP-1 inhibitors may prove to be a benefit therapeutic strategy for dental fluorosis treatment.

Promoting Immune Efficacy of the Oral Helicobacter pylori Vaccine by HP55/PBCA Nanoparticles against the Gastrointestinal Environment.

The immunogenicity of oral subunit vaccines is poor partly as a result of the harsh milieu of the gastrointestinal (GI) tract. For some pathogens that restrictedly inhabit the GI tract, a vaccine that works in situ may provide more potent protection than vaccines that operate parenterally. Yet, no appropriate delivery system is available for oral subunit vaccines. In this study, we designed HP55/poly( n-butylcyanoacrylate) (PBCA) nanoparticles (NPs) to carry Helicobacter pylori ( H. pylori) subunit vaccine CCF for oral administration in a prophylactic mice model. These NPs, which are synthesized using an interfacial polymerization method, protected the CCF antigen not only from the acidic pH in simulated gastric fluid (SGF, pH 1.2) but also from the proteolysis in simulated intestinal fluid (SIF, pH 7.4). Oral vaccination of mice with HP55/PBCA-CCF NPs promoted the production of serum antigen-specific antibodies, mucosal secretory IgA, and proinflammatory cytokines. Moreover, a Th1/Th17 response and augmented lymphocytes were found in the gastric tissue of HP55/PBCA-CCF NP-immunized mice, which might eventually limit H. pylori colonization. Collectively, these results indicate that HP55/PBCA NPs are promising carriers against the severe situation of the GI tract and thereby may be further utilized for other orally administrated vaccines or drugs.

A Novel Anisotropic Hydrogel with Integrated Self-Deformation and Controllable Shape Memory Effect.

Although shape memory polymers have been highlighted widely and developed rapidly, it is still a challenging task to realize complex temporary shapes automatically in practical applications. Herein, a novel shape memory hydrogel with the ability of self-deformation is presented. Through constructing an anisotropic poly(acrylic acid)-polyacrylamide (PAAc-PAAm) structure, the obtained hydrogel exhibits stable self-deformation behavior in response to pH stimulus, and the shapes that formed automatically can be fixed by the coordination between carboxylic groups and Fe3+ ; therefore, self-deformation and shape memory behaviors are integrated in one system. Moreover, the magnitude of auto-deformation and shape memory could be adjusted with the concentration of corresponding ions, leading to programmable shape memory and shape recovery processes.

Structural elucidation and targeted valorization of untractable lignin from pre-hydrolysis liquor of xylose production via a simple and robust separation approach.

Effective separation of lignin macromolecules from the xylose pre-hydrolysates (XPH) during the xylose production, thus optimizing the separation and purification process of xylose, is of great significance for reducing the production costs, achieving the high value-added utilization of lignin and increasing the industrial revenue. In this study, a simple and robust method (pH adjustment) for the separation of lignin from XPH was proposed and systematically compared with the conventional acid-promoted lignin precipitation method. The results showed that the lignin removal ratio (up to 60.34 %) of this simple method was higher than that of the conventional method, and the proposed method eliminated the necessity of heating and specialized equipment, which greatly reduced the separation cost. Meanwhile, this simple method does not destroy the components in XPH (especially xylose), ensuring the yield of the target product. On the other hand, the obtained lignin was nano-scale with less condensed structures, which also possessed small molecular weights with narrow distribution, excellent antioxidant activity (8-14 times higher than commercial antioxidants) and UV protection properties. In conclusion, the proposed simple separation method could effectively separate lignin from XPH at low cost, and the obtained lignin had potential commercial applications, which would further enhance the overall profitability of industrial production.

The association of healthy eating index with periodontitis in National Health and Nutrition Examination Study 2011-2012.

Aim: Periodontitis is a chronic inflammatory disorder caused by periodontopathic bacteria that causes inflammation of the supporting tissues around teeth. Previous studies have found that daily dietary nutritional intake can influence the development of periodontal disease. However, research on the Healthy Eating Index's involvement in periodontitis is limited. The purpose of this study was to look at the link between the Healthy Eating Index and periodontitis. Methods and design: We examined data from the National Health and Nutrition Examination Study (NHANES), a nationally representative survey that was performed in 2-year cycles from 2011 to 2012. As part of our investigation, we used multivariate logistic regression models to investigate the independent relationship between the Healthy Eating Index and periodontitis. We used odds ratios (OR) with 95% confidence intervals to assess the significance of the connection (95% CI). Results: Individuals with a lower total healthy eating index were more likely to have periodontitis. A higher healthy diet index was associated with a lower prevalence of periodontitis (OR = 0.69; 95%CI: 0.49-0.97), according to adjusted multivariate regression models. The restricted cubic spline (RCS) analysis revealed that the non-linear relationship between HEI-2015 and periodontitis was statistically significant and that high HEI-2015 reduced periodontitis prevalence. Conclusion: The study's findings revealed that dietary structure was linked to the prevalence of periodontitis. Patients with a higher Healthy Eating Index were less likely to have periodontitis. There is a need for future prospective longitudinal studies to confirm causality.

Clinical application study of immediate implantation without bone grafting in maxillary molars: a clinical study with one-year follow up.

Our aim was to evaluate the clinical effect and feasibility of immediate implant placement combined with flap surgery with no bone grafting and non-submerged healing in the maxillary molar area. Thirty-five patients with failed single teeth in the molar area were selected. After minimally invasive extraction of the tooth, the flaps were elevated, and an implant inserted immediately; thereafter a healing abutment was connected. The mucoperiosteal flaps were sutured around the abutment without tension, and a permanent repair was performed six months later. During the study period, the implant survival rate, cone-beam computed tomography (CBCT) data, torque value, and the results of a subjective satisfaction survey conducted with a visual analogue scale (VAS), were evaluated to assess the procedure's therapeutic effect and feasibility. All 35 teeth were successfully implanted immediately after flap surgery. The mean (SD) torque value was 42.79 (5.70) N∙cm at the time of placement. During the six-month follow up and after one year of restoration, all 35 teeth showed no loosening, shedding of implants and restoration, or inflammation around the site. The mean (SD) value of the bony space around the implant immediately after the operation was 2.47 (0.56) mm. The bony spaces were filled with new bone after six months postoperatively. The mean (SD) VAS for satisfaction was 8.71 (1.05). Immediate implant placement combined with flap surgery without bone grafting and non-submerged healing is feasible for the maxillary molar area and produces a satisfactory clinical effect.

The association of healthy eating index with periodontitis in NHANES 2013-2014.

Background: Periodontal disease is very common worldwide and is one of the main causes of tooth loss in adults. Periodontal disease is characterized by chronic inflammation that can destroy adjacent alveolar bone and lead to a loss of periodontal ligaments. Although previous studies have found that a daily diet can influence the development of periodontal disease (e.g., a diet low in carbohydrates and rich in vitamins C and D and fiber can have a protective effect). Periodontal disease may present as gingivitis or periodontitis. However, studies on the role of healthy eating index in periodontitis are lacking. The purpose of this study was to assess the association between healthy eating index and periodontitis. Methods: We analyzed data collected from participants in the National Health and Nutrition Examination Survey (NHANES), a nationally representative survey conducted in 2-year cycles from 2013 to 2014. As part of our analysis, we developed multivariate logistic regression models to examine the independent association between the healthy eating index and periodontitis. We evaluated the significance of association using odds ratios (OR) with 95% confidence intervals (95%CI). Results: Individuals with a lower total healthy eating index had a higher prevalence of periodontitis. Adjusted multivariate regression models showed that a higher healthy diet index was associated with a lower prevalence of periodontitis (OR = 0.69, 95% CI: 0.55-0.86, P < 0.05). Conclusion: The results of the study showed that dietary structure was associated with the prevalence of periodontitis. Patients with a higher healthy eating index had a lower prevalence of periodontitis. These findings will need to be confirmed by longitudinal, prospective studies in the future.

Dietary magnesium intake is protective in patients with periodontitis.

Background: Periodontitis is a chronic inflammatory disease of the oral cavity characterized by inflammation of the periodontal tissue and resorption of the alveolar bone, which has a high incidence and is the main cause of tooth loss in adults. In addition to its role in promoting osteogenesis, magnesium also has a role in regulating the inflammatory response, both systemically and locally. There is growing evidence that magnesium is an important factor in maintaining the normal functioning of the body's immune system. Hypomagnesaemia can lead to a variety of chronic inflammatory diseases throughout the body, including periodontitis. Two-thirds of the US population suffers from magnesium deficiency. The connection between dietary magnesium and periodontitis is unknown. As a result, we set out to investigate the link between dietary magnesium intake and periodontitis. Methods: In this study, we collected data from the National Health and Nutrition Examination Survey (NHANES) database from 2013 to 2014. Through 24-h dietary recalls, information about food consumption was collected. We examined the association between the dietary magnesium and periodontitis using multivariable logistic regression model. Based on odds ratios (OR) and 95% confidence intervals (CIs), a strong association was detected. Results: Multivariable logistic regression analysis showed that the OR for periodontitis comparing the highest to the lowest quintile of dietary magnesium intake was 0.69 (95% CIs = 0.52~0.92). The restricted cubic spline (RCS) analysis showed that the non-linear association between dietary magnesium and periodontitis was statistically significant and that dietary magnesium supplementation reduced the prevalence of periodontitis. Conclusion: Dietary magnesium intake is associated with the prevalence of periodontitis. Dietary magnesium deficiency increases the prevalence of periodontitis.

Studies of poly(ethylene glycol) modification of HM-3 polypeptides.

An RGD modified endostatin-derived synthetic peptide, named HM-3, is a polypeptide angiogenesis inhibitor previously synthesized in our laboratory. Its robust inhibitory effects on endothelial cell migration and tumor growth have been demonstrated by in vivo and in vitro activity assays. The RGD integrin recognition sequence enables the selective binding of HM-3 and its specific targeting to tumor cells that express high levels of integrin. However, the drug has relatively short half-life in vivo, thus requiring administration twice a day to achieve its optimal in vivo antitumor efficacy. In the current study designed to prolong HM-3 half-life, we used methoxy-poly(ethylene glycol)-aldehyde (mPEG-ALD) to specifically modify its N terminus and optimized the reaction condition via monitoring the modification by reverse-phase high-performance liquid chromatograph (RP-HPLC) under varying stoichiometric ratios (n(mPEG10k-ALD):n(HM-3)), reaction times, and pH values. The maximal modification rate was achieved in a reaction when substrates mPEG10k-ALD and HM-3 were mixed at the molar ratio of 2:1 in a pH 6 phosphate buffer after 4 h incubation at room temperature. The reaction product of this optimal reaction was purified to 96% purity by RP-HPLC. Compared with HM-3, the newly modified PEG(10k)-HM-3 was shown to be more active in the inhibition of angiogenesis in the chorioallantoic membrane of chick embryos (CAM), its rate of in vitro degradation in serum was markedly reduced, and its in vivo half-life was prolonged by 5.86-fold relative to unmodified HM-3 after intravenous injection into male SD rats.

Oral Helicobacter pylori vaccine-encapsulated acid-resistant HP55/PLGA nanoparticles promote immune protection.

Oral vaccination, is notoriously weak or nonimmunogenic. One of the major reasons is the inefficient antigen uptake caused by enzymolysis and hydrolysis in the gastrointestinal tract. In this study, acid-resistant HP55/PLGA nanoparticle was developed as an oral delivery system to protect H. pylori recombinant antigen CCF against the complex gastrointestinal environment. These ∼200nm particles controlled the release of antigen in the acidic environment (pH⩽5.5). Immunized mice with HP55/PLGA-CCF nanoparticles induced high levels of urease-specific antibodies and memory T cell responses. A month after H. pylori challenge, 43% of mice were completely protected. The protection was highly associated with the Th1/Th17-bias immune response, which had been recognized as an optimal immunity against H. pylori infection. In addition, a mass of T-cells were observed in the lamina propria of mice immunized with CCF, especially in the HP55/PLGA-CCF nanoparticles administered recipients, and contributed to the development of postimmunization gastritis. These results indicate that oral immunization with acid-resistant HP55/PLGA nanoparticles encapsulating vaccine antigens represent a promising strategy for antigen protection, slow-release and targeting, and thus prevented gastrointestinal infection.

Selective toxicity of antimicrobial peptide S-thanatin on bacteria.

S-thanatin, an analog of thanatin, was synthesized by substituting the 15th amino acid of threonine with serine, which showed a broad antimicrobial activity against bacteria. We reported earlier that membrane phospholipid was found to be the target for S-thanatin with different mechanism from other antimicrobial peptides. In this study, we have performed its structural characterization by circular dichroism (CD) spectroscopy. The CD analysis showed that S-thanatin retained its overall conformation beta-sheet in aqueous buffer, beta-turn in 50% trifluoroethanol (TFE) and beta-hairpin in 0.4 mM POPC-LUVs. In hemolysis assay, S-thanatin exhibited low hemolytic activity and bacteria selectivity. We investigated the effect of the presence of 33 mol percent cholesterol on the interactions of the antimicrobial peptide S-thanatin with phosphatidylcholine (PC) model membrane systems. The results showed that S-thanatin was more potent at disrupting cholesterol-free bacterial than cholesterol-containing eukaryotic membranes. Thus, in all respects, fluorescence dye leakage experiments indicated that cholesterol inhibited the S-thanatin-induced permeabilization of PC vesicles. Finally, flow cytometry was used to monitor changes in bacterial cell membrane potential and cell membrane integrity, with specific fluorescent dyes DiBAC(4)(3) and PI. Adding the respiratory poison CCCP seemed to prevent peptide-induced membrane damage, which suggested that S-thanatin acted at the metabolic level on respiratory chain. These findings might explain why S-thanatin was selective toxicity towards bacteria, but low toxicity towards erythrocytes. It might be due to three factors at least: electrostatic interaction (namely anionic phospholipids); cholesterol; respiratory chain.