Antibacterial, cytotoxic and mechanical properties of a orthodontic cement with phosphate nano-sized and phosphorylated chitosan: An in vitro study.

OBJECTIVES: Evaluate, in vitro, the effect of incorporating nano-sized sodium trimetaphosphate (TMPnano) and phosphorylated chitosan (Chi-Ph) into resin-modified glass ionomer cement (RMGIC) used for orthodontic bracket cementation, on mechanical, fluoride release, antimicrobial and cytotoxic properties. METHODS: RMGIC was combined with Chi-Ph (0.25%/0.5%) and/or TMPnano (14%). The diametral compressive/tensile strength (DCS/TS), surface hardness (SH) and degree of conversion (%DC) were determined. For fluoride (F) release, samples were immersed in des/remineralizing solutions. Antimicrobial/antibiofilm activity was evaluated by the agar diffusion test and biofilm metabolism (XTT). Cytotoxicity in fibroblasts was assessed with the resazurin method. RESULTS: After 24 h, the RMGIC-14%TMPnano group showed a lower TS value (p < 0.001); after 7 days the RMGIC-14%TMPnano-0.25%Chi-Ph group showed the highest value (p < 0.001). For DCS, the RMGIC group (24 h) showed the highest value (p < 0.001); after 7 days, the highest value was observed for the RMGIC-14%TMPnano-0.25%Chi-Ph (p < 0.001). RMGIC-14%TMPnano, RMGIC-14%TMPnano-0.25%Chi-Ph, RMGIC-14%TMPnano-0.5%Chi-Ph showed higher and similar release of F (p > 0.001). In the SH, the RMGIC-0.25%Chi-Ph; RMGIC-0.5%Chi-Ph; RMGIC-14%TMPnano-0.5%Chi-Ph groups showed similar results after 7 days (p > 0.001). The RMGIC-14%TMPnano-0.25%Chi-Ph group showed a better effect on microbial/antibiofilm growth, and the highest efficacy on cell viability (p < 0.001). After 72 h, only the RMGIC-14%TMPnano-0.25%Chi-Ph group showed cell viability (p < 0.001). CONCLUSION: The RMGIC-14%TMPnano-0.25%Chi-Ph did not alter the physical-mechanical properties, was not toxic to fibroblasts and reduced the viability and metabolism of S. mutans. CLINICAL RELEVANCE: The addition of phosphorylated chitosan and organic phosphate to RMGIC could provide an antibiofilm and remineralizing effect on the tooth enamel of orthodontic patients, who are prone to a high cariogenic challenge due to fluctuations in oral pH and progression of carious lesions.

Elevated extracellular inorganic phosphate inhibits ecto-phosphatase activity in breast cancer cells: Regulation by hydrogen peroxide.

For cells to obtain inorganic phosphate, ectoenzymes in the plasma membrane, which contain a catalytic site facing the extracellular environment, hydrolyze phosphorylated molecules. In this study, we show that increased Pi levels in the extracellular environment promote a decrease in ecto-phosphatase activity, which is associated with Pi-induced oxidative stress. High levels of Pi inhibit ecto-phosphatase because Pi generates H2 O2 . Ecto-phosphatase activity is inhibited by H2 O2 , and this inhibition is selective for phospho-tyrosine hydrolysis. Additionally, it is shown that the mechanism of inhibition of ecto-phosphatase activity involves lipid peroxidation. In addition, the inhibition of ecto-phosphatase activity by H2 O2 is irreversible. These findings have new implications for understanding ecto-phosphatase regulation in the tumor microenvironment. H2 O2 stimulated by high Pi inhibits ecto-phosphatase activity to prevent excessive accumulation of extracellular Pi, functioning as a regulatory mechanism of Pi variations in the tumor microenvironment.

Effect of phosphate on arsenic species uptake in plants under hydroponic conditions.

Monothioarsenate (MTA) is a newly discovered arsenic (As) compound that can be formed under reduced sulfur conditions, mainly in paddy soil pore waters. It is structurally similar to arsenate As(V) and inorganic phosphate (Pi), which is taken up through phosphate transporters. Due to the similarity between As(V) and Pi, As(V) enters into plants instead of Pi. The important role played by phytochelatin (PC), glutathione (GSH), and the PC-vacuolar transporters ABCC1 and ABCC2 under As stress in plants is well known. However, the plant uptake and mechanisms surrounding MTA still have not been completely addressed. This investigation was divided in two stages: first, several hydroponic assays were set up to establish the sensibility-tolerance of wild-type Arabidopsis thaliana (accession Columbia-0, Col-0). Then Col-0 was used as a control plant to evaluate the effects of As(V) or MTA in (PC)-deficient mutant (cad1-3), glutathione biosynthesis mutant (cad2), and PC transport (abcc1-2). The inhibitory concentration (IC50) root length was calculated for both As species. According to the results, both arsenic species (As(V) and MTA) exhibited high toxicity for the genotypes evaluated. This could mean that these mechanisms play a constitutive role in MTA detoxification. Second, for the Pi-MTA and As(V)-Pi competition assays, a series of experiments on hydroponic seedlings of A. thaliana were carried out using Col-0 and a pht1;1. The plants were grown under increasing Pi concentrations (10 µM, 0.1 mM, or 1 mM) at 10 µM As(V) or 50 µM MTA. The total As concentration in the roots was significantly lower in plants exposed to MTA, there being less As content in the pht1;1 mutant at the lowest Pi concentrations tested compared with the As(V)/Pi treatments. In addition, a higher rate of As translocation from the roots to the shoots under MTA was observed in comparison to the As(V)-treatments.

Effect of experimental and commercial artificial saliva formulations on the activity and viability of microcosm biofilm and on enamel demineralization for irradiated patients with head and neck cancer (HNC).

The effect of different artificial saliva formulations on biofilm activity and viability, and on enamel demineralization for head and neck cancer (HNC) patients was evaluated. Irradiated enamel samples were treated (1 min) with BioXtra® or with experimental formulations containing carboxymethylcellulose plus inorganic constituents alone (AS) or containing 0.1 mg mL-1 CaneCPI-5 (AS + Cane), 1.0 mg mL-1 hemoglobin (AS + Hb) or combination of both (AS + Cane + Hb). Phosphate-buffered-saline and chlorhexidine (0.12%) were negative and positive control, respectively. Biofilm was produced from the saliva of five male HNC patients, under 0.2% sucrose exposure for 5 days, and daily treated with the formulations (1 min). No significant effects were observed for the different experimental treatments. BioXtra® significantly reduced lactobacilli, demonstrating antibacterial potential for this group. Chlorhexidine was an effective treatment to significantly reduce all parameters, being an important antimicrobial and anticaries agent. Future in vitro studies must be performed using a new approach for the design of the experimental formulations.

Biofilm accumulation and sucrose rinse modulate calcium and fluoride bioavailability in the saliva of children with early childhood caries.

This study aimed at investigating the combined effect of biofilm accumulation and 20% sucrose rinse on the modulation of calcium (Ca2+), phosphate (Pi), and fluoride (F-) bioavailability in the saliva of children with early childhood caries (ECC). Fifty-six preschoolers of both genders were evaluated according to caries experience and activity: caries-free (CF, n = 28) and with ECC (n = 28) and then, submitted to biofilm intervention (biofilm accumulation). In each situation, saliva samples were collected before and five minutes after a 20% sucrose rinse to determine the concentrations of Ca2+, Pi, and F-. Calcium concentration was significantly lower in the biofilm accumulation situation compared to the situation of biofilm mechanical control (p ≤ 0.01), except for CF children after sucrose rinse. Biofilm accumulation increased salivary calcium concentration in children with ECC after sucrose rinse (p = 0.04), whereas mechanical biofilm control reduced it in both groups (p = 0.000). Phosphate concentration was influenced by mechanical control of biofilm in CF children (p = 0.03). The fluoride bioavailability was reduced by sucrose rinse and biofilm accumulation in CF and ECC children (p ≤ 0.002). In conclusion, the combined effect of biofilm accumulation and sucrose rinse modifies the bioavailability of calcium and fluoride in the saliva of children with early childhood caries.

Reducing the negative impact of ceftriaxone and doxycycline in aqueous solutions using ferrihydrite/plant-based composites: mechanism pathway.

The adsorption of ceftriaxone (CET) and doxycycline (DOX) from aqueous solution using ferrihydrite/plant-based composites (silica rice husk) to reduce their negative impact on the ecosystem was adequately studied. On the other hand, phosphate and humic acid are often found in water and soil; in view of this, their effects on the adsorption of CET and DOX were investigated. The results showed that the removal of ceftriaxone decreased with an increase in pH, while that of doxycycline did not. Ferrihydrite with 10% silica rice husk (Fh-10%SRH) has the highest maximum adsorption capacity of 139 and 178 mg g-1 for CET and DOX, respectively, at room temperature based on Liu's adsorption isotherm. This implies that the presence of silica rice husk increases CET and DOX uptake due to an increase in the pore volume of FH-10%SRH. The results showed that phosphate had a significant inhibition role on CET adsorption and minor on DOX, whereas humic acid salt affected neither case. Increase in temperature up to 333 K favored the adsorption of both contaminants. The proposed adsorption mechanisms of ceftriaxone are electrostatic interaction, n-π interaction, and hydrogen bond, while that of DOX entails n-π interaction and hydrogen bond.

Extracellular Inorganic Phosphate-Induced Release of Reactive Oxygen Species: Roles in Physiological Processes and Disease Development.

Inorganic phosphate (Pi) is an essential nutrient for living organisms and is maintained in equilibrium in the range of 0.8-1.4 mM Pi. Pi is a source of organic constituents for DNA, RNA, and phospholipids and is essential for ATP formation mainly through energy metabolism or cellular signalling modulators. In mitochondria isolated from the brain, liver, and heart, Pi has been shown to induce mitochondrial reactive oxygen species (ROS) release. Therefore, the purpose of this review article was to gather relevant experimental records of the production of Pi-induced reactive species, mainly ROS, to examine their essential roles in physiological processes, such as the development of bone and cartilage and the development of diseases, such as cardiovascular disease, diabetes, muscle atrophy, and male reproductive system impairment. Interestingly, in the presence of different antioxidants or inhibitors of cytoplasmic and mitochondrial Pi transporters, Pi-induced ROS production can be reversed and may be a possible pharmacological target.

Vitamin D, zinc and glutamine: Synergistic action with OncoTherad immunomodulator in interferon signaling and COVID­19 (Review).

Coronavirus disease 2019 (COVID­19), caused by severe acute respiratory syndrome coronavirus 2 (SARS­CoV­2), was identified in December, 2019 in Wuhan, China. Since then, it has continued to spread rapidly in numerous countries, while the search for effective therapeutic options persists. Coronaviruses, including SARS­CoV­2, are known to suppress and evade the antiviral responses of the host organism mediated by interferon (IFN), a family of cytokines that plays an important role in antiviral defenses associated with innate immunity, and has been used therapeutically for chronic viral diseases and cancer. On the other hand, OncoTherad, a safe and effective immunotherapeutic agent in the treatment of non­muscle invasive bladder cancer (NMIBC), increases IFN signaling and has been shown to be a promising therapeutic approach for COVID­19 in a case report that described the rapid recovery of a 78­year­old patient with NMIBC with comorbidities. The present review discusses the possible synergistic action of OncoTherad with vitamin D, zinc and glutamine, nutrients that have been shown to facilitate immune responses mediated by IFN signaling, as well as the potential of this combination as a therapeutic option for COVID­19.

Identification of Z nucleotides as an ancient signal for two-component system activation in bacteria.

Two-component systems (TCSs) in bacteria are molecular circuits that allow the perception of and response to diverse stimuli. These signaling circuits rely on phosphoryl-group transfers between transmitter and receiver domains of sensor kinase and response regulator proteins, and regulate several cellular processes in response to internal or external cues. Phosphorylation, and thereby activation, of response regulators has been demonstrated to occur by their cognate histidine kinases but also by low molecular weight phosphodonors such as acetyl phosphate and carbamoyl phosphate. Here, we present data indicating that the intermediates of the de novo syntheses of purines and histidine, 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranosyl 5'-monophosphate (ZMP) and/or 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranosyl 5'-triphosphate (ZTP), activate the response regulator UvrY, by promoting its autophosphorylation at the conserved aspartate at position 54. Moreover, these Z nucleotides are shown to also activate the nonrelated response regulators ArcA, CpxR, RcsB, and PhoQ. We propose that ZMP and/or ZTP act as alarmones for a wide range of response regulators in vivo, providing a novel mechanism by which they could impact gene expression in response to metabolic cues.

Evaluation of hematologic, biochemical, and blood gas variables in stored canine packed red blood cells, and the impact of storage time on blood recipients.

BACKGROUND: Canine packed red blood cells (pRBCs) can be stored under refrigeration for several days; however, cellular metabolism remains active inside the units, thus producing substances that affect their quality. OBJECTIVES: We aimed to evaluate hematologic, biochemical, and blood gas variable alterations that occur in canine pRBCs during storage, and their effects on recipient clinicopathologic parameters. METHODS: The study was conducted in two phases. In phase I, 15 pRBC units containing CPDA-1 were stored for 28 days; samples were collected weekly from the units of days 0 to 28 to measure the packed cell volume (PCV), pH, partial pressure carbon dioxide (PCO2 ), partial pressure oxygen (PO2 ), concentrations of lactate and potassium, and the percent hemolysis. In phase II, another 22 canine pRBC units stored for different time periods (maximum of 21 days) were transfused, and the recipients were evaluated before and after transfusion for changes in clinical parameters (heart rate, respiratory rate, systolic arterial pressure, and rectal temperature) and hematologic variables (PCV, lactate and potassium concentrations, pH, PCO2 , the ratio of arterial oxygen partial pressure to fractional inspired oxygen [PO2 /FiO2 ] ratio, oxygen saturation [SaO2 ], base excess, and bicarbonate [HCO3 ]). RESULTS: In the pRBC units, the PCV increased from 70% to 78.33%, the lactate concentration increased 627%, the potassium concentration increased 183%, the percent hemolysis reached 0.69%, and the pH decreased 9% after 28 days. However, the dogs who received transfusions were not negatively affected. There was a significant increase in PCVs, and a significant decrease in heart rates. CONCLUSION: Canine pRBCs undergo hematologic, blood gas, and biochemical alterations during storage; however, the transfusion of pRBCs stored for up to 21 days increased PCVs without causing harm to the dogs.

Effect of transient thermal shocks on alcoholic fermentation performance.

Stuck and sluggish fermentations are among the main problems in winemaking industry leading to important economic losses. Several factors have been described as causes of stuck and sluggish fermentations, being exposure to extreme temperatures barely studied. The objective of this study was to identify thermal conditions leading to stuck and sluggish fermentations, focusing on the impact of an abrupt and transient decrease/increase of temperature on fermentation performance and yeast viability/vitality. Different strains of Saccharomyces cerevisiae, SBB11, T73, and PDM were evaluated in synthetic grape must fermentations. Cold shocks (9 °C and 1.5 °C for 16 h) carried out on different days during the fermentation process were unable to alter fermentation performance. Conversely, shock temperatures higher than 32 °C, applied in early stages of the process, lead to sluggish fermentation showing a delay directly related to the temperature increase. Fermentation delay was associated with a decrease in cell vitality. The impact of the heat shock on fermentation performance was different depending on the strain evaluated and nitrogen supplementation (with or without diammonium phosphate addition). None of the conditions evaluated produced a stuck fermentation and importantly, in all cases must nutrition improved fermentation performance after a heat shock.

Nanopartículas de clorexidina-hexametafosfato na endodontia: síntese, caracterização, avaliação biológica e físico-química da incorporação aos cimentos endodônticos / Chlorhexidine hexametaphosphate nanoparticles in endodontics: synthesis, characterization, biological and physicochemical evaluation of incorporation to endodontic sealers

Este estudo avaliou as influências da incorporação de nanopartículas (NPs) de clorexidina-hexametafosfato em diversas propriedades dos cimentos endodônticos: AH Plus, MTA Fillapex e Pulp Canal Sealer (PCS). As NPs foram sintetizadas através de uma suspensão contendo 5 mM de hexametafosfato de sódio (HMP) e 4 mM de digluconato de clorexidina (CHX) a 20%, caracterizadas por microscopia eletrônica de varredura (MEV), espalhamento dinâmico de luz (DLS), potencial zeta, microscopia de força atômica (AFM) e espectroscopia de energia dispersiva de raios-X (EDS). Os cimentos endodônticos sofreram incorporação, em peso, de 2% e 5% das NPs. A ação antimicrobiana das amostras foi avaliada pelo teste de difusão em ágar e contato direto (DCT) frente a Enterococcus faecalis. O DCT foi conduzido em 3 tempos de armazenamento: cimentos frescos (T0), após 7 dias (T7) e após 30 dias (T30), e nos tempos de contato: 10 e 60 min. A citotoxicidade foi avaliada conforme as normas ISO 10993-12 através de extratos preparados em meio de cultura Dulbecco's modified Eagle's medium. As interações dos fibroblastos da linhagem MRC-5 com as diluições seriadas (1:1 a 1:8) dos extratos foram analisadas através do ensaio de quantificação do metabolismo mitocondrial pelo tetrazollium (MTT). O escoamento, radiopacidade, solubilidade e tempo de presa das amostras foi avaliado segundo a norma ISO 6876:2012. A solubilidade foi avaliada após 24 horas e 7 dias de imersão das amostras e o pH aferido após: 3h, 24h, 48h, 72h e 7 dias de armazenamento. Os resultados obtidos foram submetidos à análise estatística por análise de variância one-way e teste de Tukey, com nível de significância de 5%. A análise por MEV demonstrou aglomeração das NPs CHX-HMP, o diâmetro médio efetivo aferido por DLS foi de 169,39 nm e o potencial zeta foi de -10,18 mV. As NPs foram mensuradas por AFM, apresentando: 22,99 a 52,75 nm. Os elementos químicos: C, N, O, Na, P, Cl, identificados por EDS, comprovam a interação entre o digluconato de CHX e HMP. A ação antimicrobiana das NPs confirmou-se preliminarmente no teste de difusão em ágar e o DCT demonstrou o aumento da ação antimicrobiana dos cimentos endodônticos após incorporação das NPs, principalmente no MTA Fillapex, eliminando totalmente as cepas após 30 dias. A citotoxicidade dos cimentos analisados não foi potencializada pelas NPs. Houve redução do escoamento após a incorporação das NPs, no entanto sem infringir a norma ISO 6876:2012. A incorporação de NPs não alterou a radiopacidade das amostras, mas o MTA Fillapex não atingiu a radiopacidade mínima preconizada. As amostras incorporadas apresentaram redução da solubilidade após 24h de imersão, no entanto o MTA Fillapex em todas as amostras excedeu os valores preconizados pela ISO 6876:2012. Quanto ao pH, todas as amostras apresentaram decréscimo do valor com a progressão do tempo de imersão. O tempo de presa do AH Plus aumentou após a incorporação das NPs e o MTA Fillapex não atingiu a presa em nenhuma das condições testadas. Com os resultados obtidos conclui-se que a incorporação das NPs pode beneficiar o desempenho antimicrobiano dos cimentos endodônticos.(AU)

This study evaluated the influence of the incorporation of chlorhexidine hexametaphosphate NPs in various properties of the following endodontic sealers: AH Plus, MTA Fillapex and Pulp Canal Sealer (PCS). The NPs were synthesized using a suspension containing 5 mM sodium hexametaphosphate (HMP) and 4 mM 20% chlorhexidine digluconate (CHX), and characterized by scanning electron microscopy (SEM), dynamic light scattering (DLS), zeta potential, atomic force microscopy (AFM), energy-dispersive X-ray spectroscopy (EDS). Endodontic sealers were incorporated by weight of 2% and 5% of NPs. The antimicrobial activity of the samples was evaluated by agar diffusion test and direct contact (DCT) against E. faecalis. DCT was conducted in 3 storage times: fresh sealers (T0), after 7 days (T7) and after 30 days (T30) storage, and two contact times with E. faecalis suspension for 10 and 60 minutes. The cytotoxicity of the samples was evaluated according to ISO 10993-12 standards, through extracts prepared in Eagle modified by Dulbecco culture medium. The interactions of MRC-5 fibroblasts cell culture with the extracts serial dilutions (1: 1 to 1: 8) were analyzed by tetrazolium mitochondrial metabolism quantification assay (MTT). Samples flow, radiopacity, solubility and setting time were evaluated according to ISO 6876: 2012. Solubility was evaluated after 24 hours and 7 days of sample immersion and the pH measured after: 3h, 24h, 48h, 72h and 7 days of storage. The results were submitted to statistical analysis by one-way analysis of variance test and Tukey test, with a significance level of 5%. SEM analysis showed a tendency for CHX-HMP NPs to cluster, the effective mean diameter measured by DLS was 169,39 nm and the zeta potential: -10.18 mV. The NPs were individually measured by AFM, showing: 22.99 to 52.75 nm. The chemical elements: C, N, O, Na, P, Cl, identified by EDS, prove the interaction between CHX digluconate and HMP. The antimicrobial action of NPs was preliminarily confirmed in the agar diffusion test and the DCT demonstrated an increase in the antimicrobial action of endodontic sealers after incorporation of NPs, mainly in the MTA Fillapex, totally eliminating the strains after 30 days. There was reduction of flow after NPs incorporation, however without violating the ISO 6876: 2012 standard. The incorporation of NPs did not change the radiopacity of the samples, but MTA Fillapex did not reach the minimum radiopacity recommended. The incorporated samples showed reduced solubility after 24 hours of immersion, however the MTA Fillapex in all samples exceeded the values recommended by ISO 6876:2012.. Regarding the pH, all samples showed a decrease in the value as the immersion time progressed. AH Plus setting time increased after incorporation of NPs and MTA Fillapex did not reached setting under none of the conditions tested. With the obtained results it can be concluded that the incorporation of NPs can benefit the antimicrobial performance of endodontic sealers.(AU)

Isolation and characterization of a human cementocyte-like cell line, HCY-23.

Cementum is the mineralized tissue covering the tooth root that functions in tooth attachment and post-eruptive adjustment of tooth position. It has been reported to be highly similar to bone in several respects but remains poorly understood in terms of development and regeneration. Here, we investigate whether cementocytes, the residing cells in cellular cementum, have the potential to be protagonist in cementum homeostasis, responding to endocrine signals and directing local cementum metabolism. Cells from healthy erupted human teeth were isolated using sequential collagenase/EDTA digestions, and maintained in standard cell culture conditions. A cementocyte-like cell line was cloned (HCY-23, for human cementocyte clone 23), which presented a cementocyte compatible gene expression signature, including the expression of dentin matrix protein 1 ( DMP1 ), sclerostin ( SOST ), and E11/gp38/podoplanin ( E11 ). In contrast, these cells did not express the odontoblast/dentin marker dentin sialoprotein ( DSPP ). HCY-23 cells produced mineral-like nodules in vitro under differentiation conditions, and were highly responsive to inorganic phosphate (Pi). Within the limits of the present study, it can be concluded that cementocytes are phosphate-responsive cells, and have the potential do play a key role in periodontal homeostasis and regeneration.

Evaluation of an antibacterial orthodontic adhesive incorporated with niobium-based bioglass: an in situ study.

This in situ study aimed to evaluate the antibacterial and anti-demineralization effects of an experimental orthodontic adhesive containing triazine and niobium phosphate bioglass (TAT) around brackets bonded to enamel surfaces. Sixteen volunteers were selected to use intra-oral devices with six metallic brackets bonded to enamel blocks. The experimental orthodontic adhesives were composed by 75% BisGMA and 25% TEGDMA containing 0% TAT and 20% TAT. Transbond XT adhesive (TXT) was used as a control group. Ten volunteers, mean age of 29 years, were included in the study. The six blocks of each volunteer were detached from the appliance after 7 and 14 days to evaluate mineral loss and bacterial growth including total bacteria, total Streptococci, Streptococci mutans, and Lactobacilli. Statistical analysis was performed using GLM model - univariate analysis of variance for microhardness and 2-way ANOVA for bacterial growth (p<0.05). The 20% TAT adhesive caused no difference between distances from bracket and the sound zone at 10-µm deep after 7 and 14 days. After 14 days, higher mineral loss was shown around brackets at 10- to 30-µm deep for TXT and 0% TAT adhesives compared to 20% TAT. S. mutans growth was inhibited by 20% TAT adhesive at 14 days. Adhesive with 20% TAT showed lower S. mutans and total Streptococci growth than 0% TAT and TXT adhesives. The findings of this study show that the adhesive incorporated by triazine and niobium phosphate bioglass had an anti-demineralization effect while inhibiting S. mutans and total Streptococci growth. The use of this product may inhibit mineral loss of enamel, preventing the formation of white spot lesions.

A novel application of nano eggshell/titanium dioxide composite on occluding dentine tubules: an in vitro study.

To synthesize Nano eggshell-titanium-dioxide (EB@TiO2) biocomposite and to evaluate its effectiveness in occluding opened dentine tubules. EB@TiO2 was synthesized and characterized using X-ray diffraction (XRD), and Transmission Electron Microscope (TEM). Sixteen simulated bovine dentine discs were prepared and randomly assigned into four groups according to the following treatment (n = 4): Group 1: No treatment; Group 2: eggshell powder; Group 3: EB@TiO2; Group 4: Sensodyne. These were then agitated in a solution of 1g powder and 40mL water for 3hours. Thereafter, each dentine discs from the respective groups were post-treated for 5 min with 2wt% citric acid to test their acid resistant characteristics. Scanning Electron Microscope (SEM) was used to observe the effectiveness of occluded dentine pre-and post-treatment. The cytotoxicity of the synthesized EB@TiO2 was tested using NIH 3T3 assay. ANOVA was used to evaluate the mean values of the occluded area ratio and the data of MTS assay. This was followed by a multi-comparison test with Bonferroni correction (α = .05). The XRD confirmed that EB@TiO2 was successfully modified through ball-milling. The TEM revealed the presence of both spherical and irregular particle shape powders. The SEM result showed that EB@TiO2 could effectively occlude open dentine tubules. Equally, the result demonstrated that EB@TiO2 exhibited the highest acid resistant stability post-treatment. NIH 3T3 assay identified that EB@TiO2 had little effect on the NIH 3T3 cell line even at the highest concentration of 100µg/ml. This study suggests that the application of EB@TiO2 effectively occluded dentine tubules and the occlusion showed a high acid resistant stability.

A novel application of nano eggshell/titanium dioxide composite on occluding dentine tubules: an in vitro study

Abstract To synthesize Nano eggshell-titanium-dioxide (EB@TiO2) biocomposite and to evaluate its effectiveness in occluding opened dentine tubules. EB@TiO2 was synthesized and characterized using X-ray diffraction (XRD), and Transmission Electron Microscope (TEM). Sixteen simulated bovine dentine discs were prepared and randomly assigned into four groups according to the following treatment (n = 4): Group 1: No treatment; Group 2: eggshell powder; Group 3: EB@TiO2; Group 4: Sensodyne. These were then agitated in a solution of 1g powder and 40mL water for 3hours. Thereafter, each dentine discs from the respective groups were post-treated for 5 min with 2wt% citric acid to test their acid resistant characteristics. Scanning Electron Microscope (SEM) was used to observe the effectiveness of occluded dentine pre-and post-treatment. The cytotoxicity of the synthesized EB@TiO2 was tested using NIH 3T3 assay. ANOVA was used to evaluate the mean values of the occluded area ratio and the data of MTS assay. This was followed by a multi-comparison test with Bonferroni correction (α = .05). The XRD confirmed that EB@TiO2 was successfully modified through ball-milling. The TEM revealed the presence of both spherical and irregular particle shape powders. The SEM result showed that EB@TiO2 could effectively occlude open dentine tubules. Equally, the result demonstrated that EB@TiO2 exhibited the highest acid resistant stability post-treatment. NIH 3T3 assay identified that EB@TiO2 had little effect on the NIH 3T3 cell line even at the highest concentration of 100µg/ml. This study suggests that the application of EB@TiO2 effectively occluded dentine tubules and the occlusion showed a high acid resistant stability.

Evaluation of an antibacterial orthodontic adhesive incorporated with niobium-based bioglass: an in situ study

Abstract This in situ study aimed to evaluate the antibacterial and anti-demineralization effects of an experimental orthodontic adhesive containing triazine and niobium phosphate bioglass (TAT) around brackets bonded to enamel surfaces. Sixteen volunteers were selected to use intra-oral devices with six metallic brackets bonded to enamel blocks. The experimental orthodontic adhesives were composed by 75% BisGMA and 25% TEGDMA containing 0% TAT and 20% TAT. Transbond XT adhesive (TXT) was used as a control group. Ten volunteers, mean age of 29 years, were included in the study. The six blocks of each volunteer were detached from the appliance after 7 and 14 days to evaluate mineral loss and bacterial growth including total bacteria, total Streptococci, Streptococci mutans, and Lactobacilli. Statistical analysis was performed using GLM model - univariate analysis of variance for microhardness and 2-way ANOVA for bacterial growth (p<0.05). The 20% TAT adhesive caused no difference between distances from bracket and the sound zone at 10-µm deep after 7 and 14 days. After 14 days, higher mineral loss was shown around brackets at 10- to 30-µm deep for TXT and 0% TAT adhesives compared to 20% TAT. S. mutans growth was inhibited by 20% TAT adhesive at 14 days. Adhesive with 20% TAT showed lower S. mutans and total Streptococci growth than 0% TAT and TXT adhesives. The findings of this study show that the adhesive incorporated by triazine and niobium phosphate bioglass had an anti-demineralization effect while inhibiting S. mutans and total Streptococci growth. The use of this product may inhibit mineral loss of enamel, preventing the formation of white spot lesions.

Isolation and characterization of a human cementocyte-like cell line, HCY-23

Abstract Cementum is the mineralized tissue covering the tooth root that functions in tooth attachment and post-eruptive adjustment of tooth position. It has been reported to be highly similar to bone in several respects but remains poorly understood in terms of development and regeneration. Here, we investigate whether cementocytes, the residing cells in cellular cementum, have the potential to be protagonist in cementum homeostasis, responding to endocrine signals and directing local cementum metabolism. Cells from healthy erupted human teeth were isolated using sequential collagenase/EDTA digestions, and maintained in standard cell culture conditions. A cementocyte-like cell line was cloned (HCY-23, for human cementocyte clone 23), which presented a cementocyte compatible gene expression signature, including the expression of dentin matrix protein 1 ( DMP1 ), sclerostin ( SOST ), and E11/gp38/podoplanin ( E11 ). In contrast, these cells did not express the odontoblast/dentin marker dentin sialoprotein ( DSPP ). HCY-23 cells produced mineral-like nodules in vitro under differentiation conditions, and were highly responsive to inorganic phosphate (Pi). Within the limits of the present study, it can be concluded that cementocytes are phosphate-responsive cells, and have the potential do play a key role in periodontal homeostasis and regeneration.

Uremia Impacts VE-Cadherin and ZO-1 Expression in Human Endothelial Cell-to-Cell Junctions.

Endothelial dysfunction in uremia can result in cell-to-cell junction loss and increased permeability, contributing to cardiovascular diseases (CVD) development. This study evaluated the impact of the uremic milieu on endothelial morphology and cell junction's proteins. We evaluated (i) serum levels of inflammatory biomarkers in a cohort of chronic kidney disease (CKD) patients and the expression of VE-cadherin and Zonula Occludens-1 (ZO-1) junction proteins on endothelial cells (ECs) of arteries removed from CKD patients during renal transplant; (ii) ECs morphology in vitro under different uremic conditions, and (iii) the impact of uremic toxins p-cresyl sulfate (PCS), indoxyl sulfate (IS), and inorganic phosphate (Pi) as well as of total uremic serum on VE-cadherin and ZO-1 gene and protein expression in cultured ECs. We found that the uremic arteries had lost their intact and continuous endothelial morphology, with a reduction in VE-cadherin and ZO-1 expression. In cultured ECs, both VE-cadherin and ZO-1 protein expression decreased, mainly after exposure to Pi and uremic serum groups. VE-cadherin mRNA expression was reduced while ZO-1 was increased after exposure to PCS, IS, Pi, and uremic serum. Our findings show that uremia alters cell-to-cell junctions leading to an increased endothelial damage. This gives a new perspective regarding the pathophysiological role of uremia in intercellular junctions and opens new avenues to improve cardiovascular outcomes in CKD patients.

Optimized 1,3-propanediol production from crude glycerol using mixed cultures in batch and continuous reactors.

The production of 1,3-propanediol from crude glycerol and mixed anaerobic sludge was investigated in batch experiments and continuous reactors. Using a 23 complete factorial design, the effects of the concentration of glycerol (22-30 g L-1), KH2PO4 (1.50-2.00 g L-1), and vitamin B12 (7-8 mg L-1) were examined in batch reactors. As an evaluated response, the highest 1,3-PD yields occurred for high concentrations of vitamin B12 and low levels of KH2PO4, reaching 0.57 g g-1 glycerol consumed. The variable glycerol concentration was not significant in the studied range. In addition, the condition that provided the best 1,3-PD yield was applied to an anaerobic fluidized bed reactor fed with crude glycerol (26.0 g L-1), which was monitored as the hydraulic retention time (HRT) decreased from 36 to 12 h. The greatest 1,3-PD yield, of 0.31 g g-1 glycerol, was obtained with an HRT of 28 h.