Dissolution Of Hydroxyapatite Research Articles

Phosphate Recovery Mechanism from Low P-Containing Wastewaters via CaP Crystallization Using Apatite as Seed: Seed Adsorption, Surface-Induced Crystallization, or Ion Clusters Aggregation?

Low P-containing wastewaters (LPWs) exhibit huge P recovery potential, considering their larger volume. P recovery via CaP crystallization using apatite as seed is documented as being potentially well suited for LPWs. However, its responsible mechanisms remain a subject for debate. Taking hydroxyapatite (HAP) as the seed of LPWs, this paper conducted HAP adsorption/dissolution experiments, titration experiments, and P recovery experiments to distinguish the primary responsible mechanism. Results showed that it was HAP dissolution, not P adsorption, that occurred when the initial P concentration was no higher than 5 mg/L, ruling out adsorption mechanism of P recovery from LPWs using HAP as the seed. Significant OH− consumption and rapid P recovery occurred simultaneously within the first 60 s in titration experiments, suggesting CaP crystallization should be responsible for P recovery. Moreover, the continuous increase in P recovery efficiency with seed dosages observed in P recovery experiments seemed to follow well the mechanism of pre-nucleation ion clusters (PNCs) aggregation. During PNCs aggregation, P aggregates with Ca2+ quickly, generating CaP PNCs; then, CaP PNCs aggregate with seed particles, followed by CaP PNCs fusion, and ultimately transform into fines attached to the seed surface. PNCs’ aggregation mechanism was further supported by a comparison of seed SEM images before and after P recovery, since denser and smaller rod-shaped fines were observed on the seed surface after P recovery. This study suggests that PNCs’ aggregation is the dominant mechanism responsible for the recovery of P from LPWs via CaP crystallization using HAP as the seed.

Immobilization of hexavalent uranium U(VI) by hydroxyapatite under oxic conditions

The immobilization of hexavalent uranium U(VI) by hydroxyapatite (HAP) was investigated under oxic conditions as a function of pH and surface loading ([U(VI)]0/[HAP]0) to evaluate the applicability of HAP as migration barriers or waste forms in nuclear waste repositories. This study focused on the retention mechanisms, which were sought by integrating the solid-phase characterization (e.g., X-ray diffraction, scanning electron microscopy with energy dispersive spectroscopy, and U LIII-edge X-ray absorption spectroscopy) with the solution-phase analysis of dissolved Ca, P, and U. At acidic pH, due to the increasing solubility of HAP, U(VI) retention is mainly controlled by the formation of a uranyl phosphate following HAP dissolution. Importantly, this phosphate is in the form of a solid solution with Na meta-autunite being the principal component, which considerably expands the stable domain of uranyl phosphates compared to the pure phases (e.g., chernikovite, Na meta-autunite, and meta-autunite). At neutral to basic pH, however, the uranyl phosphate formation does not prevail till [U(VI)]0/[HAP]0 = 25 mg/g, below which the formation of U(VI)-phosphate ternary surface complexes mainly accounts for U(VI) retention. At basic pH, the surface substitution of PO43− for CO32− and the dominance of uranyl carbonate species make surface complexation less favorable under this pH condition. In this study, the initially present U(VI) is near completely immobilized (> 99.99 %) by HAP, with the greatest retention noted at circumneutral pH, thus demonstrating the effectiveness of HAP for the containment of U(VI) under ambient geochemical conditions.

Tumor therapy by targeting extracellular hydroxyapatite using novel drugs: A paradigm shift.

It has been shown that tumor microenvironment (TME) hydroxyapatite (HAP) is typically associated with many malignancies and plays a role in tumor progression and growth. Additionally, acidosis in the TME has been reported to play a key role in selecting for a more aggressive tumor phenotype, drug resistance and desensitization to immunotherapy for many types of cancers. TME-HAP is an attractive target for tumor detection and treatment development since HAP is generally absent from normal soft tissue. We provide strong evidence that dissolution of hydroxyapatite (HAP) within the tumor microenvironment (TME-HAP) using a novel therapeutic can be used to kill cancer cells both in vitro and in vivo with minimal adverse effects. We developed an injectable cation exchange nano particulate sulfonated polystyrene solution (NSPS) that we engineered to dissolve TME-HAP, inducing localized acute alkalosis and inhibition of tumor growth and glucose metabolism. This was evaluated in cell culture using 4T1, MDA-MB-231 triple negative breast cancer cells, MCF10 normal breast cells, and H292 lung cancer cells, and in vivo using orthotopic mouse models of cancer that contained detectable microenvironment HAP including breast (MMTV-Neu, 4T1, and MDA-MB-231), prostate (PC3) and colon (HCA7) cancer using 18 F-NaF for HAP and 18 F-FDG for glucose metabolism with PET imaging. On the other hand, H292 lung tumor cells that lacked detectable microenvironment HAP and MCF10a normal breast cells that do not produce HAP served as negative controls. Tumor microenvironment pH levels following injection of NSPS were evaluated via Chemical Exchange Saturation (CEST) MRI and via ex vivo methods. Within 24 h of adding the small concentration of 1X of NSPS (~7 μM), we observed significant tumor cell death (~ 10%, p < 0.05) in 4T1 and MDA-MB-231 cell cultures that contain HAP but ⟨2% in H292 and MCF10a cells that lack detectable HAP and in controls. Using CEST MRI, we found extracellular pH (pHe) in the 4T1 breast tumors, located in the mammary fat pad, to increase by nearly 10% from baseline before gradually receding back to baseline during the first hour post NSPS administration. in the tumors that contained TME-HAP in mouse models, MMTV-Neu, 4T1, and MDA-MB-231, PC3, and HCA7, there was a significant reduction (p<0.05) in 18 F-Na Fuptake post NSPS treatment as expected; 18 F- uptake in the tumor = 3.8 ± 0.5 %ID/g (percent of the injected dose per gram) at baseline compared to 1.8 ±0.5 %ID/g following one-time treatment with 100 mg/kg NSPS. Of similar importance, is that 18 F-FDG uptake in the tumors was reduced by more than 75% compared to baseline within 24 h of treatment with one-time NSPS which persisted for at least one week. Additionally, tumor growth was significantly slower (p < 0.05) in the mice treated with one-time NSPS. Toxicity showed no evidence of any adverse effects, a finding attributed to the absence of HAP in normal soft tissue and to our therapeutic NSPS having limited penetration to access HAP within skeletal bone. Dissolution of TME-HAP using our novel NSPS has the potential to provide a new treatment paradigm to enhance the management of cancer patients with poor prognosis.

Epigallocatechin gallate-immobilized antimicrobial resin with rechargeable fluorinated synergistic composite for enhanced caries control

ObjectivesGiven the global prevalence of dental caries, impacting 2.5 billion individuals, the development of sophisticated prevention filling materials is crucial. Streptococcus mutans, the principal caries-causing strain, produces acids that demineralize teeth and initiate dental caries. To address this issue, we aimed to develop a synergistic resin-based composite for enhancing caries control. MethodsThe synergistic resin composite incorporates fluorinated kaolinite and silanized Al2O3 nanoparticle fillers into an epigallocatechin gallate (EGCG) immobilized urethane-modified epoxy acrylate (U-EA) resin matrix, referred to the as-prepared resin composite. The EGCG-modified TPGDA/U-EA network was synthesized by preparing methacrylate-functionalized isocyanate (HI), reacting it with EGCG to form HI-EGCG, and then incorporating HI-EGCG into the TPGDA/U-EA matrix. The lamellar space within the kaolinite layer was expanded through the intercalation of acrylamide into kaolinite, enhancing its capability to adsorb and release fluoride ions (F–). The layered structure of acrylamide/ kaolinite in the U-EA resin composite acts as a F– reservoir. ResultsThe physico-mechanical properties of the as-prepared resin composites are comparable to those of commercial products, exhibiting lower polymerization shrinkage, substantial F– release and recharge and favorable diametral tensile strength. The immobilized EGCG in the composite exhibits potent antimicrobial properties, effectively reducing the biofilm biomass. Furthermore, the synergistic effect of EGCG and fluorinated kaolinite efficiently counteracts acid-induced hydroxyapatite dissolution, thereby suppressing demineralization and promoting enamel remineralization. SignificanceOur innovative EGCG and fluoride synergistic composite provides enhanced antimicrobial properties, durable anti-demineralization, and tooth remineralization effects, positioning it as a promising solution for effective caries control and long-term dental maintenance.

Dissolution of Nb-doped hydroxyapatite prepared via low-temperature mechanochemical method: Spectroscopy studies

Calcium phosphate glass ceramics with nominal hydroxyapatite stoichiometry doped with niobium were synthesized using simple as well as low-temperature mechanochemical method and then in the form of compressed pellet were submitted to the static dissolution process in distilled water for one month. The results of structural analysis, performed mainly on the base of spectroscopic methods such as: infrared absorption spectroscopy, energy-dispersive X-ray spectroscopy, photoelectron spectroscopy as well as soft and tender X-ray absorption spectroscopies, indicate that during the dissolution process the presence of niobium reduced the dynamics of the calcium phosphate new layer formation, without a significant impact on the morphology and stoichiometry of this layer. Moreover, the Nb addition favored the formation of hydroxyapatite and Ca-deficient hydroxyapatite, both during the synthesis and dissolution processes, and facilitated the incorporation of the CO3-2 groups into the ceramic matrix, leading to the formation of B-type carbonated apatite.

Film-Forming Polymers for Inhibition of Hydroxyapatite Dissolution: A Screening Study

The aim of this study was to evaluate the effect of film-forming polymer solutions of different concentrations and pH values, either associated or not with sodium fluoride (F; 225 ppm F⁻), when applied during the initial stage of salivary pellicle formation, to prevent the dissolution of hydroxyapatite (HA), which was determined by the pH-stat method. Polyacrylic acid (PA), chitosan, sodium linear polyphosphate (LPP), polyvinyl methyl ether/maleic anhydride (PVM/MA), and propylene glycol alginate (PGA) were tested in three concentrations (lower, medium, and higher), two pH values (native or adjusted), and either associated or not with F. Distilled water, F, and stannous ion+fluoride (Sn/F; 225 ppm F⁻ and 800 ppm Sn²⁺, as SnCl₂) solutions were the controls, totalizing 63 groups. HA crystals were pretreated with human saliva for 1 min to allow pellicle formation, then immersed in the experimental solutions (1 min), and exposed to saliva for another 28 min. Subsequently, they were added to a 0.3% citric acid solution (pH = 3.8), connected to a pH-stat system that added aliquots of 28 μL 0.1 N HCl for a total reaction time of 5 min. Data were analyzed with one-way ANOVA and Tukey’s tests (α = 0.05). For PA alone, the concentrations of 0.1% (native pH), 0.06%, and 0.08% (both pH adjusted) showed significantly lower HA dissolution than the negative control. PA concentrations of 0.1% and 0.08%, of both pH values, improved the effect of F against HA dissolution to a near-identical value as Sn/F. All solutions containing chitosan and LPP significantly reduced HA dissolution in comparison with the control. For chitosan, the concentration of 0.5% (in both pH values) improved the effect of F. LPP at 0.5% (native pH) and all associations of LPP with F outperformed the effect of F. Some PVM/MA solutions significantly reduced HA dissolution but PVM/MA could not improve the protection of F. PGA was incapable of reducing HA dissolution or improving F effect. It was concluded that chitosan, LPP, and some PA and PVM/MA solutions used alone were capable of reducing HA dissolution. Only PA, chitosan, and LPP were able to enhance fluoride protection, but for PA and chitosan, this was influenced by the polymer concentration.

Effects of Calcium and Phosphate on Dissolution of Enamel, Dentin and Hydroxyapatite in Citric Acid

The aim was to evaluate the effect of dissolved calcium and phosphate on dissolution rate of enamel, dentin and compressed hydroxyapatite (HA) in citric acid solution as a function of pH. At pH 2.5, dissolution rate of enamel increased significantly by 6% in 20 mmol/L added calcium but, otherwise, dissolution rates of neither enamel, dentin nor HA were significantly affected by 10 or 20 mmol/L calcium. However, enamel dissolution rate was reduced by &gt; 50 mmol/L calcium. At pH 3.25 and 4.0, 10-20 mmol/L calcium inhibited dissolution of enamel by 29-100% and HA by 65-75% but did not affect dentin dissolution. Phosphate (10 or 20 mmol/L) did not inhibit dissolution of enamel, dentin or HA at any pH, but there were increases in dissolution rate of all three substrates at pH 2.5 and, in one test with dentine (at 20 mmol/L phosphate), at pH 3.25. The results suggest that calcium addition to soft drinks and other acidic products such as medications may reduce erosivity against enamel, provided that pH is not too low; that phosphate would not reduce erosivity against enamel; and that neither calcium nor phosphate at these concentrations would reduce erosivity against dentin.&#x0D; Accepted for publication: December 12, 2022&#x0D; Published online: February 21, 2023

CERAMIC MATERIALS OF A GIVEN PHASE COMPOSITION BASED ON HYDROXYAPATITE POWDER TREATED IN SOLUTIONS OF AMMONIUM, SODIUM AND POTASSIUM DIHYDROPHOSPHATES

Powders of hydroxyapatite (HA) Ca10(PO4)6(OH)2 treated in 0.25M aqueous solutions of ammonium NH4H2PO4, sodium NaH2PO4 and potassium KH2PO4 dihydrophosphates for 1 hour under stirring were used for ceramics production. According to the XRD data, there was no changes in the phase composition of the powders after such treatment. After firing in the range of 900 – 1100 ?С, the phase composition of ceramics based on HA Ca10(PO4)6(OH)2 powder treated with an aqueous solution of NH4H2PO4 included ?-tricalcium phosphate ?-Ca3(PO4)2 and HA Ca10(PO4)6(OH)2; The phase composition of ceramics based on HA Ca10(PO4)6(OH)2 powder treated with NaH2PO4 aqueous solution included sodium-substituted tricalcium phosphate Ca10Na(PO4)7 and HA Ca10(PO4)6(OH)2; the phase composition of ceramics based on HA Ca10(PO4)6(OH)2 powder treated with an aqueous solution of KH2PO4 included potassium-substituted tricalcium phosphate Ca10K(PO4)7 and HA Ca10(PO4)6(OH)2. The formation of biphasic ceramics occurred due to a decrease in the molar ratio Ca/P of HA powder after treatment in aqueous solutions of ammonium NH4H2PO4, sodium NaH2PO4 and potassium KH2PO4 dihydrophosphates. HA-particle surface adsorption of cations and anions from the solution, slight dissolution of HA in solutions of dihydrophosphates with acidic pH, as well as ion exchange of cations and anions of HA for cations and anions from solutions, were possible processes providing change in the ratio of cations and anions in the HA powder. The resulting ceramic composites contain biocompatible phases, and can be recommended for the creation of bone implants.

Recycling of phosphate tailings for an efficient hydroxyapatite-based adsorbent to immobilize heavy metal cations.

Hydroxyapatite (HAP) is a promising adsorbent for immobilizing heavy metals in soil and water. However, the preparation and modification of HAP from pure chemicals increases its cost and limits its large-scale practical application. In this study, a hydroxyapatite-based adsorbent (HAPPT) was prepared from phosphate tailing produced in the phosphorus industry to sequester Pb, Cd and Zn from solution. The results showed that HAPPT was composed of HAP and MgO, with a surface area of 27.74 m2/g. The kinetics studies showed that most Pb and Cd were removed from the initial solution in 4h and the adsorption of Zn increased with increasing contact time. Metals presented higher adsorption capacities at 35°C than that at 25°C. The adsorption isotherms showed that HAPPT presented high adsorption capacities for Pb, Cd and Zn in mono-metal solutions. The adsorption capacity of Cd at pH 6 was higher than that at pH 3, but the adsorption for Pb and Zn was similar at both pHs. HAPPT has selectivity for Pb in Pb-Cd-Zn multi-metals solution, and competitive adsorption reduced the adsorption quantity by 53%, 93% and 79% for Pb, Cd and Zn, respectively. The combined results of TEM-EDS, XRD and XPS showed that Pb was immobilized by forming phosphates due to the dissolution of HAP, whereas Cd and Zn were immobilized by forming hydroxide precipitates resulting from the function of MgO in HAPPT. The results of this study provided an efficient adsorbent for the removal of heavy metals in solution and provided a new perspective on the recycling of phosphate tailings in the phosphorus industry.

Phosphorus transformation behavior and phosphorus cycling genes expression in food waste composting with hydroxyapatite enhanced by phosphate-solubilizing bacteria

This study aimed to explore the effect of phosphate-solubilizing bacteria (PSB) Bacillus inoculation in the cooling stage on hydroxyapatite dissolution, phosphorus (P) forms transformation, and bacterial P cycling genes in food waste composting with hydroxyapatite. Results indicated that PSB inoculation promoted the dissolution of hydroxyapatite, increased P availability of compost by 8.1% and decreased the ratio of organic P to inorganic P by 10.2% based on sequential fractionation and 31P nuclear magnetic resonance spectroscopy. Illumina sequencing indicated Bacillus relative abundance after inoculation increased up to one time higher than control after the cooling stage. Network analysis and metabolic function of bacterial community analysis suggested inorganic P solubilizing genes of Bacillus and organic P mineralization genes of other genera were improved after inoculation in the core module. Therefore, bioaugmentation of PSB in the cooling stage may be a potential way to improve P bioavailability of bone and food waste in composting.

In vitro cellular biocompatibility and in vivo degradation behavior of calcium phosphate-coated ZK60 magnesium alloy

Calcium phosphate (Ca-P) surface coating is a simple but effective way to enhance both corrosion resistance and biocompatibility of ZK60 magnesium alloy. However, cell compatibility on different Ca-P layers coated on ZK60 alloy has seldom been investigated. In this study, the effects of type, morphology and corrosion protection of several Ca-P coatings formed at pH 6.5, 7.8 and 10.2 on cell behavior were examined by using an osteoblastic cell line MC3T3-E1. Furthermore, in vivo behavior in rabbits of the alloy coated with the optimum Ca-P layer was also studied. It was found that the surface factors governed the cell morphology and density. The coating morphology plays a dominant role in these surface factors. The sample coated at pH 7.8 showed the best cellular biocompatibility, suggesting that the hydroxyapatite (HAp) layer formed at pH 7.8 was the optimum coating. In rabbits, this optimum coating enhanced remarkably the corrosion resistance of the alloy. During implantation, the outermost crystals of the HAp coating were shortened and thinned due to the dissolution of HAp caused by the body fluid of the rabbits. It is indicated that ZK60 alloy coated at pH 7.8 can be applied as a biodegradable implant.

Phytate Intake, Health and Disease: "Let Thy Food Be Thy Medicine and Medicine Be Thy Food".

Phytate (myo-inositol hexakisphosphate or InsP6) is the main phosphorus reservoir that is present in almost all wholegrains, legumes, and oilseeds. It is a major component of the Mediterranean and Dietary Approaches to Stop Hypertension (DASH) diets. Phytate is recognized as a nutraceutical and is classified by the Food and Drug Administration (FDA) as Generally Recognized As Safe (GRAS). Phytate has been shown to be effective in treating or preventing certain diseases. Phytate has been shown to inhibit calcium salt crystallization and, therefore, to reduce vascular calcifications, calcium renal calculi and soft tissue calcifications. Moreover, the adsorption of phytate to the crystal faces can inhibit hydroxyapatite dissolution and bone resorption, thereby playing a role in the treatment/prevention of bone mass loss. Phytate has a potent antioxidation and anti-inflammatory action. It is capable of inhibiting lipid peroxidation through iron chelation, reducing iron-related free radical generation. As this has the effect of mitigating neuronal damage and loss, phytate shows promise in the treatment/prevention of neurodegenerative disease. It is reported that phytate improves lipid and carbohydrate metabolism, increases adiponectin, decreases leptin and reduces protein glycation, which is linked with macrovascular and microvascular diabetes complications. In this review, we summarize the benefits of phytate intake as seen in in vitro, animal model, epidemiological and clinical trials, and we also identify questions to answer in the future.

Enamel Remineralization and Crystallization after Fluoride Iontophoresis

Early caries lesions consist of noncavitated subsurface demineralization caused by the dissolution of hydroxyapatite from the surface to the subsurface area of the enamel. Such lesions cannot be remineralized effectively by the conventional treatment. Thus, there is a need for a noninvasive technique capable of delivering the remineralizing agent to subsurface sites. For this purpose, fluoride iontophoresis (IP) using weak currents has been investigated with some conflicting results and no information on the crystal structure and composition. Because enamel remineralization involves the role of fluid from dentin, the presence of enamel fluid is necessary to determine the repair associated with the physiological condition. This study aimed to investigate structural and compositional characteristics, including the remineralizing effect of 5% sodium fluoride (NaF) IP with different polarities, cathodal iontophoresis (CIP), and anodal iontophoresis (AIP) for the treatment of natural enamel caries under simulated pulpal pressure. A bulk measurement of the crystal structure inside the lesion was first determined using calcium (Ca) K-edge X-ray absorption spectroscopy. IP with both polarities significantly promoted subsurface remineralization. The CIP generated a significant increase in the Ca/phosphorus ratio, and fluoride at the surface lesion significantly correlated with higher mineral density (MD) and more strengthening crystal structure of the lesion volume, while the lesion’s MD and other impurities at the lesion surface, mostly the carbonate ions, affected the significant increase in MD with the unchanged structure of the lesion volume after AIP. The CIP of NaF is an ideal method for rapid enamel remineralization and recrystallization of fluoroapatite/fluorohydroxyapatite.

Hydroxyapatite Modified Silica Aerogel Nanoparticles: In Vitro Cell Migration Analysis

The ability of silica aerogel nanoparticles (SA-Np) to improve the stability of hydroxyapatite (HA) was investigated. Using the sol-gel method, the HA was incorporated into SA-Np at a weight ratio of 0.5 of HA to SiO2 (HA-SA-Np). The efficacy of HA-SA-Np, SA-Np, and HA on the in vitro migration of normal human dermal fibroblast cells (HSF1184) was compared. The cell migration was measured at 0, 6, and 24 hours after scratching using ImageJ and an inverted optical microscope. To ascertain the resorbability of HA-SA-Np, the phosphate and silicic acid concentrations in media treated for 2, 5, and 7 days were examined. The high dissolution of HA could be reduced by incorporating the HA into the silica nanosphere. The HA-SA-Np significantly stimulated cell migration and increased closure with increasing treatment time. It was caused by the release of silicic acid, which aided in healing cells. It also demonstrates the ability of HA-SA-Np to be resorbed and eventually increase the adhesion and migration of normal human fibroblast cells. As a result, the potential application of HA-SA-Np as an alternative biomaterial was confirmed.

Free and Encapsulated Phosphate-Solubilizing Bacteria for the Enhanced Dissolution of Swine Wastewater-Derived Struvite—An Attractive Approach for Green Phosphorus Fertilizer

Struvite and hydroxyapatite are byproducts of phosphorus removal from wastewater that can be used as phosphate fertilizers. Due to their low water solubility, especially in alkaline soils, their use is currently limited. The use of phosphate-solubilizing bacteria to enhance the dissolution of struvite and hydroxyapatite could be an attractive solution for expanding their use, but literature reports on this are limited. In this study, Arthrobacter sp. (TBRC 5201), Azotobacter vinelandii (TBRC 7231), and Bacillus megaterium (TBRC 1396) were evaluated for their ability to dissolve struvite and hydroxyapatite on agar media with struvite or hydroxyapatite as the sole source of phosphorus. Only B. megaterium (TBRC 1396) was able to use struvite and hydroxyapatite for growth. After 14 d of incubation in liquid medium, B. megaterium (TBRC 1396) dissolved phosphorus from struvite up to 835.45 ± 11.76 mg P/l compared with 196.08 ± 3.92 mg P/l in a control without cells, whereas the dissolution of hydroxyapatite by B. megaterium was minimal. B. megaterium (TBRC 1396) was also capable of dissolving phosphorus from swine wastewater-derived struvite. Both free cells and alginate-encapsulated cells of B. megaterium (TBRC 1396) were able to rapidly dissolve phosphorus from swine wastewater-derived struvite, resulting in soluble phosphorus concentrations that reached 400 mg P/l within 2 days, compared with those without cells that required 12 days. In conclusion, the application of struvite with phosphate-solubilizing bacteria is a promising tool for green sustainable agriculture.

Remediation of chromium, zinc, arsenic, lead and antimony contaminated acidic mine soil based on Phanerochaete chrysosporium induced phosphate precipitation

Microbial induced phosphate precipitation (MIPP) is an advanced bioremediation technology to reduce the mobility and bioavailability of heavy metals (HMs), but the high level of HMs would inhibit the growth of phosphate solubilizing microbes. This study proposed a new combination system for the remediation of multiple HMs contaminated acidic mine soil, which included hydroxyapatite (HAP) and Phanerochaete chrysosporium (P. chrysosporium, PC) that had high phosphate solubilizing ability and HMs tolerance. Experimental data suggested that in HAP/PC treatment after 35 d of remediation, labile Cr, Zn and As could be transformed into the stable fraction with the maximum immobilization efficiencies increased by 53.01 %, 22.43 %, and 35.65 %, respectively. The secretion of organic acids by P. chrysosporium was proved to promote the dissolution of HAP. Besides, the pH value, available phosphorus (AP) and organic matter (OM) increased in treated soil than in original soil, which also indicated the related dissolution-precipitation mechanism of HMs immobilization. Additionally, characterization results revealed that adsorption and ion exchange also played an important role in the remediation process. The overall results suggested that applying P. chrysosporium coupled with HAP could be considered as an efficient strategy for the remediation of multiple HMs contaminated mine soil and laid a foundation for the future exploration of soil microenvironment response during the remediation process.

POS0478 TOPICAL CALCIUM CHELATORS FOR TARGETING CUTANEOUS CALCIFICATION: PRECLINICAL EVIDENCE OF EFFICACY

BackgroundCutis calcinosis, defined as sub-epidermal deposition of calcium salts, is a painful, disabling, disfiguring, problem in 20-40% of patients with systemic sclerosis. Its pathogenesis remains poorly understood and currently there is no effective disease-modifying pharmacotherapy. One potential strategy is topical application of calcium chelating agents, able to penetrate the epidermal barrier and dissolve subcutaneous calcinotic deposits in situ. To date, the only compound reported for this application has been sodium thiosulfate (STS), with often-contradictory results.ObjectivesTo test the hypothesis, in pre-clinical studies, that polycarboxylic acids can induce calcium dissolution without skin toxicity, with the long-term aim of developing an effective topical treatment for cutis calcinosis.MethodsWe compared the metal ion-chelating agents citric acid (CA) and ethylenediaminetetraacetic acid (EDTA) - polycarboxylic acids with well-characterised chelation profiles – to STS for their ability to chelate calcium, without inducing cytotoxicity or inflammation (pro-inflammatory cytokine expression and release), using in vitro 2D (keratinocyte [HaCaT]; fibroblast ([HCA2]) and recombinant human epidermal (RHE) models. The resultant data was subsequently used to predict therapeutic concentrations for assessment in a validated skin irritation model (SkinEthicTM; Episkin SA) and to assay maximal percutaneous absorption. At relative dermal concentrations, the dissolution performance of each chelator was further assessed using two different models of calcinosis: 1) pharmaceutical dissolution of a hydroxyapatite (HAp) tablet (1) and; 2) dissolution of a calcified extracellular matrix laid down by mineralising SaOS2 in vitro monolayer culture (2).ResultsIncubation with CA, EDTA and STS induced cytotoxicity in both in vitro cell lines studied at concentrations of &gt;10 mM; only EDTA (10 mM) resulted in inflammatory cytokine release (IL8) from cells at these higher concentrations (cf positive control, Lipopolysaccharide 10 mg/mL). When applied topically to RHE models as near-saturated solutions, none of the chelators were categorised as skin irritants. Due to differences in their relative aqueous solubility, higher concentrations of CA (1600 mM) and STS (1200 mM) could be delivered through the RHE model than EDTA (200 mM). Using a simple linear regression model, the rate of compound absorption was: CA, 0.43 ± 0.05; STS, 0.26 ± 0.03 and; EDTA, 0.05 ± 0.01 g/L/hr. At each time-point, the cumulative concentration of compound in the receptor media was CA &gt; STS &gt; EDTA. Incubation with chelators had no effect on the integrity of the RHE by standard histology. Based on the rate of percutaneous absorption, the dissolution performance of each chelator was tested at relative dermal concentrations for phosphate dissolution (nmoles) of HAp (CA, 9.61 ±0.97; EDTA, 5.38 ± 0.28; 3.78 ± 0.58) and in the calcified in vitro model (Figure 1; CA, 3285 ± 105, STS, 947 ± 95, EDTA, 1174 ± 89), showing the superiority of CA in both model systems.Figure 1.Dissolution of a calcified extracellular matrix by citric acidConclusionOverall, this study highlights the promise of polycarboxylic acids, particularly CA, to target subcutaneous calcification, which are neither toxic nor inflammatory to the skin. Specifically, we have identified CA as a potentially more efficacious alternative to STS for the topical treatment of cutis calcinosis.

Diluted Acetic Acid Softened Intermuscular Bones from Silver Carp (Hypophthalmichthys molitrix) by Dissolving Hydroxyapatite and Collagen.

Intermuscular bones (IBs) pose physical hazards that threaten consumer health and food safety. This study aimed to investigate the mechanism of softening IBs from silver carp with diluted acetic acid. IBs (separated from muscle) and fillets (without removing IBs) were treated with diluted acetic acid. Analyses of sensory attributes and the hardness of treated IBs indicated that diluted acetic acid (<10 mmol/L) could soften IBs effectively. Additionally, 0.5 mmol/L acetic acid softened IBs within fillets without significantly affecting the texture and flavor of fillets. Analyses of microstructure, minerals (calcium and phosphorus) and collagen content, and the Fourier transform infrared (FTIR) spectra of IBs indicated that acetic acid broke connections (formed by collagen that shared hydroxyl groups) between collagen molecules, and between collagen and hydroxyapatite (HAP), thus inducing the dissolution of collagen and HAP. The dissolution of HAP contributed more to IBs softening than collagen.

Lead immobilization in simulated polluted soil by Douglas fir biochar-supported phosphate

This study compared the lead (Pb2+) immobilization efficacy of biochar-supported phosphate to conventional in-situ heavy metal immobilization methods (with lime, neat biochar and phosphate). The biochar-supported phosphate was obtained by treating Douglas fir biochar (BC) with anhydrous calcium chloride and potassium dihydrogen phosphate. The amount of Pb2+ immobilized was determined by comparing the concentration of ammonium nitrate extractable Pb2+ lead from lead-spiked soil (without amendment) to that of a 30 d incubation with (a) lead-spiked soil plus 5% (wt./wt.) biochar supported-phosphate, (b) lead-spiked soil plus 5% (wt./wt.) untreated Douglas fir biochar, (c) lead-spiked soil plus 5% (w/w) lime and (d) lead-spiked soil plus 5% (wt./wt.) potassium dihydrogen phosphate. The control (lead-spiked soil without amendment) produced the largest quantity (96.08 ± 9.22 mg L−1) of NH4NO3-extractable Pb2+, while lead-spiked soil treated with 5% (wt./wt.) biochar-supported phosphate resulted in the lowest quantity of NH4NO3 extractable Pb2+ (0.3 ± 0.2 mg L−1). The mechanism for immobilization of Pb2+ by BP occurs at pH < 7 through dissolution of hydroxyapatite embedded in BP during modification, followed by precipitation of insoluble Pb10(PO4)6(OH)2. The residual lead fraction in the lead-spiked soil increased by 20.9% following amendment with BP. These results indicate that biochar-supported phosphate is a candidate to reduce lead mobility (bioavailability) in polluted soil. This amendment may lower Pb2+ uptake into plants while minimizing the potential for water contamination due to Pb2+mobility.

Hydroxyapatite as a scavenger of reactive radiolysis species in graphene liquid cells for in situ electron microscopy

Liquid cell electron microscopy is an imaging technique allowing for the investigation of the interaction of liquids and solids at nanoscopic length scales. Such in situ observations are increasingly in-demand in an array of fields, from biological sciences to medicine to batteries. Graphene liquid cells (GLCs), in particular, have generated a great interest as a low-scattering window material with the potential for increasing the quality of both imaging and spectroscopy. However, preserving the stability of the liquid and of the sample in the GLC remains a considerable challenge. In the present work we encapsulate water and hydroxyapatite (HAP), a pH-sensitive biological material, in GLCs to observe the interactions between the graphene, HAP, and the electron beam. HAP was chosen for several reasons. One is its ubiquity in biological specimens such as bones and teeth, and the second is the presence of phosphate ions in common buffer solutions. Finally, there is its sensitivity to changes in pH, which result from beam-induced hydrolysis in liquid cells. A dynamic process of dissolution and recrystallization of HAP was observed, which correlated with the production of H+ ions by the beam during imaging. In addition, a large increase in the stability of the GLC under irradiation was noted. Specifically, no stable hydrogen bubbles were detected under the electron fluxes routinely exceeding 170 e− Å−2 s−1. With the measured threshold dose for the bubble formation in pure water equaling 9 e− Å−2 s−1, it was concluded that the presence of HAP increases the resistance of water against radiolysis in the GLC by more than an order of magnitude. These results confirm the possibility of using biological materials, such as HAP, as stabilizers in liquid cell electron microscopy. They outline a potential route for stabilization of specimens in liquid cells through the addition of a scavenger of reactive species generated by the beam-induced hydrolysis of water. These improvements are essential for enhancing both the resolution of imaging and the available imaging time, as well as avoiding the beam-induced artifacts.