Mechanisms of cadmium mitigation in tomato plants under orthophosphate and polyphosphate fertilization regimes.

Cadmium (Cd) is one of the most toxic elements in soil, affecting morphological, physiological, and biochemical processes in plants. Mineral plant nutrition was tested as an effective approach to mitigate Cd stress in several crop species. In this regard, the present study aimed to elucidate how different phosphorus (P) fertilization regimes can improve some bio-physiological processes in tomato plants exposed to Cd stress. In a hydroponic experiment, the impact of two phosphorus fertilizer forms (Polyphosphate (poly-P): condensed P-form with 100% polymerization rate and orthophosphate (ortho-P): from orthophosphoric acid) on the photosynthetic activity, plant growth, and nutrient uptake was assessed under three levels of Cd stress (0, 12, and 25 µM of CdCl2). The obtained results confirmed the negative effects of Cd stress on the chlorophyll content and the efficiency of the photosynthesis machinery. The application of poly-P fertilizer significantly improved the chlorophyll stability index (82%) under medium Cd stress (Cd12), as compared to the ortho-P form (55%). The analysis of the chlorophyll α fluorescence transient curve revealed that the amplitude of Cd effect on the different steps of electron transfer between PSII and PSI was significantly reduced under the poly-P fertilization regime compared to ortho-P, especially under Cd12. The evaluation of the RE0/RC parameter showed that the electron flux reducing end electron acceptors at the PSI acceptor side per reaction center was significantly improved in the poly-P treatment by 42% under Cd12 compared to the ortho-P treatment. Moreover, the use of poly-P fertilizer enhanced iron uptake and its stoichiometric homeostasis in the shoot tissue which maintained an adequate absorption of iron under Cd stress conditions. Findings from this study revealed for the first time that inorganic polyphosphate fertilizers can reduce Cd toxicity in tomato plants by enhancing photosynthesis activity, nutrient uptake, plant growth, and biomass accumulation despite the high level of cadmium accumulation in shoot tissues.

The use of mouthwash containing trimetaphosphate as an adjunct therapy to fluoridated toothpaste reduces enamel demineralization.

INTRODUCTION: The decline in dental caries has been attributed to the widespread use of fluoride (F). Two forms of presentation are fluoridated toothpaste (FT) and mouthwash (MW), widely used by the population. MATERIALS AND METHODS: This study aimed to evaluate in vitro the effects of combining FT and MW, whether supplemented with sodium trimetaphosphate (TMP) or not, on dental enamel demineralization. Bovine enamel blocks (n = 60) were selected based on initial surface hardness (SHi) and divided into 5 experimental groups (n = 12 each): I) Placebo Toothpaste (without F/TMP); II) 1100 ppm F Toothpaste (FT); III) 1100F associated with a MW at 100 ppm F (FT + MW 100F); IV) 1100F associated with a MW at 225 ppm F (FT + MW 250F); and V) 1100F associated with a MW at 100 ppm F supplemented with 0.4 % TMP (FT + MW 100F-TMP). The blocks were treated twice a day, undergoing 5 pH cycles over 7 days. Thus, the percentage change in surface hardness (%SH), integrated subsurface hardness loss (ΔKHN), and the concentration of F, phosphorus (P), and calcium (Ca) in the enamel were determined. The data were submitted to ANOVA and Student-Newman-Keuls test (p < 0.001). RESULTS: The 1100F group was statistically inferior to the groups associated with MW for %SH, ΔKHN, and the concentration of P and Ca in the enamel (p < 0.001). Blocks treated with FT + MW 225F and FT + MW 100F-TMP showed significantly lower %SH compared to the other groups (p < 0.001). The FT + MW 100F - TMP group exhibited the lowest depth mineral loss (ΔKHN), and higher concentration de P in enamel (p < 0.001). CONCLUSION: The adjunct use of MW with FT produces a greater protective effect in inhibiting enamel demineralization, and the supplementation of TMP to the MW with 100F provides a superior effect compared to MW with 225F. CLINICAL SIGNIFICANCE: This combination of treatments could be regarded as one of several alternative fluoride supplements for subjects at elevated risk of caries.

Optimizing phosphorus removal for municipal wastewater post-treatment with Chlorella vulgaris.

The microalgal species Chlorella vulgaris was cultivated in batch conditions to identify the optimum set of initial conditions for the best biomass growth rate, phosphate removal, polyphosphate accumulation, and protein productivity. To study the effect of phosphorus deficiency caused stress, the microalgal biomass was exposed to phosphorus deficiency conditions for periods varying between 1 and 10 days and inoculated at different initial biomass and phosphate concentrations. A 10-day period of phosphate deficiency, supported by low initial biomass concentration (∼0.25 mg DW L-1), increased the phosphate removal by 62-175% when compared to the reference conditions. A 10-day period of biomass P-deficiency also boosted the polyphosphate accumulation and protein productivity, increasing them up to 40 and 46.8 times, respectively, if compared to reference conditions. At the same time, optimization algorithm model results suggested one-day biomass P-starvation with low initial biomass concentration as the optimum combination to achieve the highest performance while the initial phosphate concentration had less impact. The initial conditions suggested by the optimization model were validated in a sequencing batch photobioreactor, giving 101.7 and 138.0% more phosphate removal and polyphosphate accumulation, compared to the reference conditions. The obtained results present microalgae exposure to phosphorus stress as a supplementary tool for wastewater post-treatment targeted on rapid phosphorus removal.

Self-healing hydrogel based on polyphosphate-conjugated pectin with hemostatic property for wound healing applications.

Self-healing hydrogels have important application in hemostasis and wound repairing. In this research, pectin based self-healing hydrogel was fabricated with conjugated polyphosphate for hemostatic and wound healing applications. The hydrogel formed without any stimulus and hydrogel kept its biocompatibility; at the same time, the hydrogel degraded completely by enzyme and in vivo. The polyphosphate conjugated hydrogel also showed self-healing property and sustained release performance with strong coagulation characteristic. More importantly, the in vivo experiment revealed that the polyphosphate conjugated hydrogel reduced the blood loss and hemostasis time in hemorrhage model; meanwhile, the hydrogel accelerated the wound repairing rate of the open wound by preventing bacterial invasion. Altogether, the PolyP conjugated hemostatic pectin-based hydrogel is a good candidate as wound dressing material applied in clinic or open wound repairing.

Chickpea plant responses to polyphosphate fertiliser forms and drip fertigation frequencies: effect on photosynthetic performance and phenotypic traits.

Photosynthesis is the main biophysiological process that governs plant growth and development. Under nutrient deficiency in crops and soils, many photosynthetic reactions can be disturbed. We compared two polyphosphates (Poly-A and Poly-B) and an orthophosphate fertiliser (Ortho-P) to an unfertilised treatment under three drip fertigation frequencies. Results showed that the electron transport chain between PSII and PSI was significantly enhanced in fertigated chickpea plants compared with the control treatment. The polyphosphate fertiliser (Poly-A) enhanced the number of electron acceptors of the photosynthetic linear electron transport chain compared with the other fertiliser forms. Furthermore, the time for reaching the maximum intensity F m was shortened in the fertilised chickpea plant indicating that the rate of light trapping and electron transport was enhanced under phosphorus drip fertigation. Also, the energy needed to close all reaction centres was decreased with P fertigated treatments, as revealed by the electron acceptor pool size of PSII (Sm/tFmax ). However, no significant effects of fertiliser forms or fertigation frequencies were observed on the energetic demand for reaction centres closure. Plants grown under polyphosphate fertigation absorbed significantly more phosphorus. Positive correlations between phosphorus uptake, photosynthetic yield, chickpea podding dynamic, and grain yield showed the beneficial effects of adequate phosphorus nutrition on chickpea growth and productivity.

The therapeutic potential of inorganic polyphosphate: A versatile physiological polymer to control coronavirus disease (COVID-19).

Rationale: The pandemic caused by the novel coronavirus SARS-CoV-2 is advancing rapidly. In particular, the number of severe courses of the disease is still dramatically high. An efficient drug therapy that helps to improve significantly the fatal combination of damages in the airway epithelia, in the extensive pulmonary microvascularization and finally multiorgan failure, is missing. The physiological, inorganic polymer, polyphosphate (polyP) is a molecule which could prevent the initial phase of the virus life cycle, the attachment of the virus to the target cells, and improve the epithelial integrity as well as the mucus barrier. Results: Surprisingly, polyP matches perfectly with the cationic groove on the RBD. Subsequent binding studies disclosed that polyP, with a physiological chain length of 40 phosphate residues, abolishes the binding propensity of the RBD to the ACE2 receptor. In addition to this first mode of action of polyP, this polymer causes in epithelial cells an increased gene expression of the major mucins in the airways, of MUC5AC and MUC1, as well as a subsequent glycoprotein production. MUC5AC forms a gel-like mucus layer trapping inhaled particles which are then transported out of the airways, while MUC1 constitutes the periciliary liquid layer and supports ciliary beating. As a third mode of action, polyP undergoes enzymatic hydrolysis of the anhydride bonds in the airway system by alkaline phosphatase, releasing metabolic energy. Conclusions: This review summarizes the state of the art of the biotherapeutic potential of the polymer polyP and the findings from basic research and outlines future biomedical applications.

Effect of fluoride, casein phosphopeptide-amorphous calcium phosphate and sodium trimetaphosphate combination treatment on the remineralization of caries lesions: An in vitro study.

OBJECTIVE: To evaluate the effects of combination of treatments with fluoridated toothpastes supplemented with sodium trimetaphosphate (TMP) and casein phosphopeptide-amorphous calcium phosphate (MI Paste Plus®), on the remineralization of dental enamel. DESIGN: Enamel blocks with artificial caries were randomly allocated into six groups (n = 12), according to the toothpastes: 1) without F-TMP-MI Paste Plus® (Placebo); 2) 1100 ppm F (1100 F), 3) MI Paste Plus®, 4) 1100 F + MI Paste Plus® (1100 F-MI Paste Plus®), 5) 1100 F + 3% TMP (1100 F-TMP) and 6) 1100 F-TMP + MI Paste Plus® (1100 F-TMP-MI Paste Plus®). Blocks were treated 2×/day with slurries of toothpastes (1 min). Furthermore, groups 4 and 6 received the application of MI Paste Plus® for 3 min. After pH cycling, the percentage of surface hardness recovery (%SHR); integrated loss of subsurface hardness (ΔKHN); profile analysis and lesion depth subsurface through polarized light microscopy (PLM), confocal laser scanning microscopy (LSCM), scanning electron microscopy (SEM), fluoride (F), calcium (Ca), phosphorus (P) concentrations in the enamel were determined. The data were analyzed by ANOVA (1-criterion) and Student-Newman-Keuls test (p < 0.001). RESULTS: 1100 F-TMP-MI Paste Plus® group showed the best results of %SHR, ΔKHN and PLM (p < 0.001). F concentration was similar between the 1100 F, 1100 F-MI Paste Plus®, and 1100 F-TMP-MI Paste Plus® groups (p > 0.001). 1100 F-TMP-MI Paste Plus® group showed the highest concentration of Ca and P in the enamel (p < 0.001). CONCLUSION: The association of 1100 F-TMP and MI Paste Plus® led to a significant increase in the remineralization of initial carious lesions.

Self-Healing Properties of Bioinspired Amorphous CaCO3/Polyphosphate-Supplemented Cement.

There is a strong interest in cement additives that are able to prevent or mitigate the adverse effects of cracks in concrete that cause corrosion of the reinforcement. Inorganic polyphosphate (polyP), a natural polymer that is synthesized by bacteria, even those on cement/concrete, can increase the resistance of concrete to progressive damage from micro-cracking. Here we use a novel bioinspired strategy based on polyP-stabilized amorphous calcium carbonate (ACC) to give this material self-healing properties. Portland cement was supplemented with ACC nanoparticles which were stabilized with 10% (w/w) Na-polyP. Embedding these particles in the hydrated cement resulted in the formation of calcite crystals after a hardening time of 10 days, which were not seen in controls, indicating that the particles dissolve and then transform into calcite. While there was no significant repair in the controls without ACC, almost complete closure of the cracks was observed after a 10 days healing period in the ACC-supplemented samples. Nanoindentation measurements on the self-healed crack surfaces showed a similar or slightly higher elasticity at a lower hardness compared to non-cracked surfaces. Our results demonstrate that bioinspired approaches, like the use of polyP-stabilized ACC shown here, can significantly improve the repair capacity of Portland cement.

Lidocaine tripotassium phosphate complex laden microemulsion for prolonged local anaesthesia: In vitro and in vivo studies.

Lidocaine is widely used as a local anaesthetic in the clinical practice to manage pre- and post-operative pain, skin burns, etc. However, the short duration of action (< 2 h) of marketed dosage forms limit their ability to meet clinical needs. Herein, we prepared a lidocaine-tPP(tri potassium phosphate)-complex loaded microemulsion to achieve greater penetration, followed by destabilization of microemulsion in the skin layer to precipitate oil-complex to produce a depot effect in the skin for prolonging the effects of anaesthesia. The lidocaine-tPP-complex-microemulsion was compared with lidocaine base loaded microemulsion, marketed ointment USP and lidocaine HCl. The pseudo ternary phase diagrams at three Smix ratios (1:2, 1:3 and 1:4; Pluronic F127: PEG 400) were constructed using Capmul MCM C8 EP as oil phase. The Smix at 1:4 ratio showed large microemulsion area in comparison to 1:2 and 1:6 ratio. The lidocaine base (LD-1:4-ME10O45SM and LD-1:4-ME20O45SM) and lidocaine-tPP-complex (LDC-1:4-ME10O45SM and LDC-1:4-ME20O45SM) loaded microemulsion batches (1:4 ratio) were thermodynamically stable. The ex vivo diffusion study showed sustained release up to 12 h with microemulsion batches, in comparison to lidocaine HCl (4 h) and ointment base (7 h). The selected LDC-1:4-ME20O45SM batch was non-irritating on the rabbit skin. In drug retention studies, LD-1:4-ME20O45SM and LDC-1:4-ME20O45SM batches showed 2.68- and 3.93-fold greater lidocaine retention in comparison to ointment USP. The radiant heat tail-flick test showed prolong local anaesthesia using LDC-1:4-ME20O45SM in comparison to ointment USP. The findings suggest that lidocaine-tPP-complex loaded microemulsion could be a potential strategy for providing prolong local anaesthesia.

Calcium Polyphosphate Nanoparticles Act as an Effective Inorganic Phosphate Source during Osteogenic Differentiation of Human Mesenchymal Stem Cells.

The ability of bone-marrow-derived mesenchymal stem/stromal cells (BM-MSCs) to differentiate into osteoblasts makes them the ideal candidate for cell-based therapies targeting bone-diseases. Polyphosphate (polyP) is increasingly being studied as a potential inorganic source of phosphate for extracellular matrix mineralisation. The aim of this study is to investigate whether polyP can effectively be used as a phosphate source during the in vitro osteogenic differentiation of human BM-MSCs. Human BM-MSCs are cultivated under osteogenic conditions for 28 days with phosphate provided in the form of organic ß-glycerolphosphate (BGP) or calcium-polyP nanoparticles (polyP-NP). Mineralisation is demonstrated using Alizarin red staining, cellular ATP content, and free phosphate levels are measured in both the cells and the medium. The effects of BGP or polyP-NP on alkaline phosphatase (ALP) activity and gene expression of a range of osteogenic-related markers are also assessed. PolyP-NP supplementation displays comparable effects to the classical BGP-containing osteogenic media in terms of mineralisation, ALP activity and expression of osteogenesis-associated genes. This study shows that polyP-NP act as an effective source of phosphate during mineralisation of BM-MSC. These results open new possibilities with BM-MSC-based approaches for bone repair to be achieved through doping of conventional biomaterials with polyP-NP.

Impact of a Novel Cryoprotectant Blend on the Sensory Quality of Frozen Lobster (Homarus americanus).

Frozen storage of lobster meat (Homarus americanus) can result in undesirable quality changes that decrease consumer acceptability of these products. Current seafood industry methods use cryoprotective agents that contain phosphates including sodium tripolyphosphates (STPP). However, recent evidence suggests that cryoprotective mixtures that combine different carbohydrates and STPP can have equal or even greater cryoprotective properties compared to using STPP alone. The objective of this study was to compare the overall consumer acceptability of lobster meat stored for 6 months in different blends of these cryoprotective solutions. One hundred and seven panelists were recruited to score the acceptability of the lobster samples using nine-point hedonic scales. A check-all-that-apply (CATA) question containing 27 literature-informed, sensory descriptors was also used to identify terms frequently used to describe lobster meat. Analysis of variance analysis, indicated a significant increase for overall liking (22.1%, P < 0.0001), liking of flavor (23.6%, P < 0.0001) and texture (15.6%, P = 0.000) scores for samples stored in a novel carbohydrate blend plus sodium chloride (NaCl) and STPP compared to the water control. Subsequent penalty analysis revealed that overall liking scores were most positively associated with the attributes tender, sweet, moist and soft. Moreover, the attributes with the highest positive mean impact were more frequently used to describe lobster samples stored in solutions containing NaCl and the novel carbohydrate blend, as well as NaCl and STPP (Lobster-3 and Lobster-5 samples, respectively). PRACTICAL APPLICATION: The positive impact on the sensory quality of this novel blend of cryoprotective compounds (carbohydrates and NaCl) is proof of concept that this mixture is comparable, if not better than preservatives currently used by the seafood industry. Given the necessary regulatory approval and industry acceptance, lobster processors may consider this novel blend as a suitable alternative to freeze lobster products for up to 6 months.

Suppressive effects of collismycin C on polyphosphate-mediated vascular inflammatory responses.

Human endothelial cells-derived polyphosphate (PolyP) is one of the pro-inflammatory mediators as suggested by the previous reports. 2,2'-bipyridine containing natural products are generally accepted to have antimicrobial, cytotoxic and anti-inflammatory properties. This study was undertaken to investigate whether a 2,2'-bipyridine containing natural product, collismycins C, can modulate PolyP-mediated inflammatory responses in human umbilical vein endothelial cells (HUVECs) and in mice. After HUVECs or mice were activated with PolyP, cells or mice were post-treated with collismycins C. The anti-inflammatory activities of collismycins C were determined by measuring permeability, leukocytes adhesion and migration, and activation of pro-inflammatory proteins in PolyP-activated HUVECs and mice. In addition, the beneficial effects of collismycins C on survival rate in PolyP-injected mice. Collismycins C inhibits PolyP-mediated barrier disruption, the expressions of cell adhesion molecules, and leukocyte to HUVEC adhesion/migration. Interestingly, PolyP-induced NF-κB activation and the productions of TNF-α and IL-6 were inhibited by collismycins C in HUVECs. These anti-inflammatory functions of collismycins C were confirmed in PolyP injected mice. In conclusion, based on the anti-inflammatory effects of collismycins C in PolyP-mediated septic response, collismycins C have therapeutic potential for various systemic inflammatory diseases.

Global effect of the lack of inorganic polyphosphate in the extremophilic archaeon Sulfolobus solfataricus: A proteomic approach.

Inorganic polyphosphates (polyP) are present in all living cells and several important functions have been described for them. They are involved in the response to stress conditions, such as nutrient depletion, oxidative stress and toxic metals amongst others. A recombinant strain of Sulfolobus solfataricus unable to accumulate polyP was designed by the overexpression of its endogenous ppx gene. The overall impact of the lack of polyP on this S. solfataricus polyP (-) strain was analyzed by using quantitative proteomics (isotope-coded protein label, ICPL). Stress-related proteins, such as peroxiredoxins and heat shock proteins, proteins involved in metabolism and several others were produced at higher levels in the ppx expression strain. The polyP deficient strain showed an increased copper sensitivity and an earlier transcriptional up-regulation of copA gene coding for the P-type copper-exporting ATPase. This implies a complementary function of both copper resistance systems. These results strongly suggests that the lack of polyP makes this hyperthermophilic archaeon more sensitive to toxic conditions, such as an exposure to metals or other harmful stimuli, emphasizing the importance of this inorganic phosphate polymers in the adaptations to live in the environmental conditions in which thermoacidophilic archaea thrive. SIGNIFICANCE: Inorganic polyphosphate (polyP) are ubiquitous molecules with many functions in living organisms. Few studies related to these polymers have been made in archaea. The construction of a polyP deficient recombinant strain of Sulfolobus solfataricus allowed the study of the global changes in the proteome of this thermoacidophilic archaeon in the absence of polyP compared with the wild type strain. The results obtained using quantitative proteomics suggest an important participation of polyP in the oxidative stress response of the cells and as having a possible metabolic role in the cell, as previously described in bacteria. The polyP deficient strain also showed an increased copper sensitivity and an earlier transcriptional up-regulation of copA, implying a complementary role of both copper resistance systems.

In situ effect of fluoride toothpaste supplemented with nano-sized sodium trimetaphosphate on enamel demineralization prevention and biofilm composition.

OBJECTIVE: To evaluate the effect of a fluoride toothpaste containing nano-sized sodium trimetaphosphate (TMPnano) on enamel demineralization in situ and composition of the biofilm. DESIGN: This crossover double-blind study consisted of four phases (seven days each) and 12 volunteers who wore oral appliances containing four enamel bovine blocks. The cariogenic challenge was performed by 30% sucrose solution (6x/day). The toothpaste treatments (3x/day) were as follows: no F/TMP/TMPnano (Placebo), 1100 ppm F (1100F), 1100F plus 3% micrometric or nano-sized TMP (1100F/TMP; 1100F/TMPnano). Percentage of surface hardness loss (%SH), and integrated loss of subsurface hardness (ΔKHN), as well as enamel calcium (Ca), phosphorus (P), and fluoride (F) were determined. Moreover, biofilm formed on the blocks were analyzed for F, Ca, P, and insoluble extracellular polysaccharide (EPS) concentrations. Data were analyzed using one-way ANOVA, repeated measures followed by Fisher LSD test (p < 0.001). RESULTS: 1100F/TMPnano promoted the lowest %SH and ΔKHN among all groups (p < 0.001). Regarding the F concentrations in the enamel and in the biofilm, there were no significant differences between 1100 F and 1100 F/TMPnano, but significantly increased enamel Ca concentrations (p < 0.001). 1100F/TMPnano showed lower values of EPS concentration when compared with 1100F (∼80%) (p < 0.001). CONCLUSION: 1100F/TMPnano promoted a greater protective effect against enamel demineralization and significantly affected the composition of biofilm formed in situ when compared to 1100F toothpaste.

Use of sodium trimetaphosphate in the inhibition of dentin matrix metalloproteinases and as a remineralizing agent.

OBJECTIVES: Because of its ability to act as an antiproteolytic agent, the effect of sodium trimetaphosphate (STMP) against specific enzymes extracted from sound dentin and its performance under acidic challenge on demineralized dentin were investigated. METHODS: The antiproteolytic potential of STMP (0.5%, 1.0%, and 1.5%) was assessed in triplicate by zymography. For the evaluation of remineralization activity, 50 bovine-root dentin specimens were selected and randomly divided into 5 groups (n=10). Three areas were determined for each specimen: 1) control (no treatment); 2) demineralized (artificial caries-like challenge); 3) treated (demineralized and subjected to pH-cycling for 7days, and treated for 10min with 1.5% STMP, 1.5% STMP+calcium hydroxide (Ca[OH]2), 1.5% STMP+sodium fluoride (NaF), NaF, or deionized H2O). The dentin specimens were analyzed for superficial hardness (SH) and cross-sectional hardness (CSH) at different depths (10, 30, 50, 70, 90, 110, and 220µm) using a Knoop penetrator (10g/10s). Statistical analyses were performed with analysis of variance (ANOVA) and Tukey tests (p<0.05). RESULTS: The zymographic analysis showed that 1.5% STMP promoted complete inhibition of gelatinolytic activity. Therefore, 1.5% STMP was investigated in association with supplemented calcium or fluoride; a combination of 1.5% STMP and Ca(OH)2 significantly increased the mechanical properties of the treated dentin. CONCLUSION: 1.5% STMP serves as an antiproteolytic agent against matrix metalloproteinases extracted from human dentin. Furthermore, when supplemented with Ca(OH)2, 1.5% STMP may potentially induce remineralization. CLINICAL SIGNIFICANCE: STMP can be introduced as a novel strategy that combines enzymatic inhibition and remineralizing potential, which can serve to strengthen dentin and improve stability. STMP may have potential in the treatment of demineralized dentin lesions, especially when supplemented with calcium.

Effect of fluoride toothpaste with nano-sized trimetaphosphate on enamel demineralization: An in vitro study.

OBJECTIVE: This study evaluated the effect of toothpastes containing 1100ppm F associated or not with micrometric or nano-sized sodium trimetaphosphate (TMP) on enamel demineralization in vitro, using a pH cycling model. DESIGN: Bovine enamel blocks (4mm×4mm, n=96) were randomly allocated into eight groups (n=12), according to the test toothpastes: Placebo (without fluoride or TMP); 1100ppm F (1100F); 1100F plus micrometric TMP at concentrations of 1%, 3% or 6%; and 1100F plus nanosized TMP at 1%, 3% or 6%. Blocks were treated 2×/day with slurries of toothpastes and submitted to a pH cycling regimen for five days. Next, final surface hardness (SHf), integrated hardness loss (IHL), differential profile of integrated hardness loss (ΔIHL) and enamel fluoride (F) concentrations were determined. Data were analyzed by ANOVA and Student-Newman-Keuls' test (p<0.05). RESULTS: The use of 1100F/3%TMPnano led to SHf 30% higher (p<0.001) and IHL∼80% lower (p<0.001) when compared to 1100F. This toothpaste also resulted in ∼64% reduction of mineral loss (ΔIHL) when compared to 1100F. Moreover, the addition of nano-sized TMP promoted increases in enamel F uptake of 90%, 160% and 100%, respectively for the concentrations of 1%, 3% and 6%, when compared to 1100F (p<0.001). CONCLUSION: The addition of nano-sized TMP at 3% to a conventional toothpaste significantly decreased enamel demineralization when compared to its counterparts without TMP or supplemented with micrometric TMP.

Fluoride and calcium concentrations in the biofilm fluid after use of fluoridated dentifrices supplemented with polyphosphate salts.

OBJECTIVES: The present study evaluated fluoride (F) and calcium (Ca) concentrations in the biofilm fluid formed in situ under cariogenic challenge after using F dentifrices supplemented or not with sodium trimetaphosphate (TMP) or calcium glycerophosphate (CaGP). METHODS: Volunteers (n = 12) were randomly divided into 5 groups according to the toothpastes used: placebo (without F, CaGP or TMP), 1100 ppm F (1100F) and low-fluoride dentifrice (LFD, 550 ppm F) with no supplementation (550F) or supplemented with 1 % TMP (550F-TMP) or 0.25 % CaGP (550F-CaGP). In each phase, volunteers wore palatal appliances containing 4 bovine enamel blocks. Cariogenic challenge was performed with 30 % sucrose solution, 6 times/day. On the morning of the eigth day, biofilm samples were collected 12 h and 1 h after brushing and cariogenic challenge. F and Ca analyses in the biofilm fluid were performed with the inverted electrode after buffering with TISAB III and using the Arsenazo III method, respectively. Data were submitted to two-way ANOVA (repeated measures) and Student-Newman-Keuls test (p < 0.05). RESULTS: A dose-response relationship was verified between F concentrations in the dentifrices and in the biofilm fluid. Significant differences were observed among placebo, 550F, and 1100F only 1 h after brushing, without statistical differences among 550F, 550F-TMP, and 550F-CaGP. No defined trend was observed among the groups regarding Ca concentrations, with the highest values seen for placebo and 550F-CaGP. CONCLUSION: The anticaries effect of LFDs supplemented with CaGP or TMP cannot be related to an increased availability of F and Ca in the biofilm fluid. CLINICAL SIGNIFICANCE: The better performance of LFDs containing CaGP or TMP shown in previous studies should be attributed to their ability to interact with tooth enamel and with the biofilm, rather to their effect on the biofilm fluid.

Inorganic polyphosphates enhances nucleus pulposus tissue formation in vitro.

Disc degeneration is associated with low back pain for which currently there is no optimal therapy so there is a great need to identify new treatment approaches. Inorganic polyphosphates (polyP) are linear polymers of orthophosphate units varying in chain length and present in many cell types. As polyP has anabolic effects on chondrocytes, we hypothesized that polyP treatment would enhance matrix accumulation by nucleus pulposus (NP) cells. NP cells isolated from bovine caudal discs were grown in 3D culture under normoxic or in select experiments under hypoxic conditions, in the presence or absence of various concentrations and sizes of polyP. Gene expression was determined using RT-PCR. Matrix accumulation was quantified by measuring proteoglycan and collagen contents. DAPI fluorescence shift was used to stain for polyP in tissue. DAPI staining showed polyP present predominantly in the pericellular region of in vitro formed tissue. PolyP treatment enhanced matrix accumulation in a concentration and chain length dependant manner. NP cells exposed to polyP-22 (22 phosphate units length) showed an increase in gene expression of aggrecan, Collagen II, Sox 9, and MMP-13 which was maintained for the 14 days of culture. This suggests that polyP may enhance NP tissue formation in vitro by upregulating the expression of matrix genes. As polyP enhances proteoglycan accumulation even under hypoxic conditions, this raises the possibility that polyP may be a novel treatment to induce NP regeneration. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:41-50, 2017.

A randomised clinical in situ study to evaluate the effects of novel low abrasivity anti-sensitivity dentifrices on dentine wear.

OBJECTIVES: To compare the abrasive wear on human dentine in an in situ model associated with use of an experimental low abrasivity anti-sensitivity dentifrice containing 1% alumina and 5% sodium tripolyphosphate (STP) with an experimental ultra-low abrasivity non-alumina 5% STP dentifrice, a higher abrasivity daily-use whitening dentifrice, and water as controls. METHODS: This was a single-centre, single-blind, randomised, split-mouth, four-treatment, two-period, crossover in situ study in 29 healthy subjects. Subjects wore bilateral lower buccal appliances, each fitted with four dentine specimens. Study treatments were applied ex vivo (three times daily). Dentine loss was measured by non-contact profilometry after 5, 10 and 15days' treatment. RESULTS: All 29 subjects were included in the efficacy analysis. Significantly less dentine loss was associated with brushing with the low and ultra-low abrasivity dentifrices than with the higher abrasivity dentifrice at all timepoints (p<0.01). Brushing with ultra-low abrasivity dentifrice or water resulted in statistically significantly less dentine loss compared with brushing with the low abrasivity dentifrice at all timepoints (p<0.05). Dentine loss after brushing with ultra-low abrasivity dentifrice was not significantly different from brushing with water. CONCLUSIONS: The degree of dentine loss observed in this in situ model reflected the abrasivity of the study dentifrices. Brushing with low or ultra-low abrasivity STP-containing anti-sensitivity dentifrices resulted in significantly less dentine loss (equating to dentine wear) than with a higher abrasivity daily-use whitening dentifrice.

Remineralizing Potential of a Low Fluoride Toothpaste with Sodium Trimetaphosphate: An in situ Study.

OBJECTIVE: To evaluate the effect of a low-fluoride (F) toothpaste supplemented with sodium trimetaphosphate (TMP) on enamel remineralization in situ. DESIGN: Bovine enamel blocks were selected on the basis of their surface hardness (SH) after caries-like lesions had been induced, and randomly divided into 4 treatment groups, according to the toothpastes used: without F or TMP (placebo); 500 ppm F; 500 ppm F plus 1% TMP; and 1,100 ppm F. The study design was blinded and crossover and performed in 4 phases of 3 days each. Eleven subjects used palatal appliances containing 4 bovine enamel blocks which were treated 3 times per day during 1 min each time, with natural slurries of saliva and toothpaste formed in the oral cavity during toothbrushing. After each phase, the percentages of surface (%SHR) and subsurface hardness recovery (%ΔKHNR) were calculated. F, calcium (Ca), and phosphorus (Pi) contents in enamel were also determined. Data were analyzed by 1-way, repeated-measures ANOVA, followed by the Student-Newman-Keuls test (p < 0.05). RESULTS: Toothpaste with 500 ppm F + TMP and 1,100 ppm F showed similar %SHR and %ΔKHNR as well as enamel F, Ca, and Pi concentrations. CONCLUSION: The addition of TMP to a low-fluoride toothpaste promoted a similar remineralizing capacity to that of a standard (1,100 ppm F) toothpaste in situ.