Amorphous Calcium Carbonate Nanoparticles Research Articles

Amorphous CaCO3-bioreactor for tumor microenvironment regulation to reinforce tumor chemoimmunotherapy

Developing efficient nanomedicines to modulate the immunosuppressive tumor microenvironment (TME) is a highly sought-after method for improving the adverse clinical outcomes of immunotherapy. In this study, we designed a novel tumor immune stimulatory nanomedicine to regulate the immunosuppressive TME in multiple directions. This nanomedicine is liposome-modified amorphous calcium carbonate (ACC) nanoparticles (referred to as CEL/ACC-LP NPs) loaded with chemotherapeutic medication celastrol (CEL). CEL/ACC-LP NPs explosively release calcium ions (Ca2+) and CEL in the tumor acidic microenvironment, and rapidly deplete H+ to alleviate the tumor acidic microenvironment, exhibiting intrinsic immune regulatory activity in synergistic tumor chemoimmunotherapy. CEL induces reactive oxygen species (ROS) to promote Ca2+ influx, exacerbating cellular Ca2+ overload. In addition, CEL/ACC-LP NPs can cause unique immunogenic cell death (ICD) effects. Therefore, the multi-directional regulation of tumor cells through simple nano preparation will offer a potent approach to improve the efficacy of chemoimmunotherapy.

Apoptosis and Paraptosis Induced by Disulfiram-Loaded Ca2+/Cu2+ Dual-Ions Nano Trap for Breast Cancer Treatment.

Regarded as one of the hallmarks of tumorigenesis and tumor progression, the evasion of apoptotic cell death would also account for treatment resistance or failure during cancer therapy. In this study, a Ca2+/Cu2+ dual-ion "nano trap" to effectively avoid cell apoptosis evasion by synchronously inducing paraptosis together with apoptosis was successfully designed and fabricated for breast cancer treatment. In brief, disulfiram (DSF)-loaded amorphous calcium carbonate nanoparticles (NPs) were fabricated via a gas diffusion method. Further on, the Cu2+-tannic acid metal phenolic network was embedded onto the NPs surface by self-assembling, followed by mDSPE-PEG/lipid capping to form the DSF-loaded Ca2+/Cu2+ dual-ions "nano trap". The as-prepared nanotrap would undergo acid-triggered biodegradation upon being endocytosed by tumor cells within the lysosome for Ca2+, Cu2+, and DSF releasing simultaneously. The released Ca2+ could cause mitochondrial calcium overload and lead to hydrogen peroxide (H2O2) overexpression. Meanwhile, Ca2+/reactive oxygen species-associated mitochondrial dysfunction would lead to paraptosis cell death. Most importantly, cell paraptosis could be further induced and strengthened by the toxic dithiocarbamate (DTC)-copper complexes formed by the Cu2+ combined with the DTC, the metabolic products of DSF. Additionally, the released Cu2+ will be reduced by intracellular glutathione to generate Cu+, which can catalyze the H2O2 to produce a toxic hydroxyl radical by a Cu+-mediated Fenton-like reaction for inducing cell apoptosis. Both in vitro cellular assays and in vivo antitumor evaluations confirmed the cancer therapeutic efficiency by the dual ion nano trap. This study can broaden the cognition scope of dual-ion-mediated paraptosis together with apoptosis via a multifunctional nanoplatform.

Size and phase preservation of amorphous calcium carbonate nanoparticles in aqueous media using different types of lignin for contrast-enhanced ultrasound imaging

Hypothesis: Calcium carbonate (CaCO3) nanoparticles could have great potential for contrast-enhanced ultrasound imaging (CEUS) due to their gas-generating properties and sensitivity to physiological conditions. However, the use of nano CaCO3 for biomedical applications requires the assistance of stabilizers to control the size and avoid the fast dissolution/recrystallization of the particles when exposed to aqueous conditions.Experiments: Herein, we report the stabilization of nano CaCO3 using lignin, and synthesized core–shell amorphous CaCO3-lignin nanoparticles (LigCC NPs) with a diameter below 100 nm. We have then investigated the echogenicity of the LigCC NPs by monitoring the consequent generation of contrast in vitro for 90 min in linear and non-linear B-mode imaging.Findings: This research explores how lignin type and structure affect stabilization efficiency, lignin structuration around CaCO3 cores, and particle echogenicity. Interestingly, by employing lignin as the stabilizer, it becomes possible to maintain the echogenic properties of CaCO3, whereas the use of lipid coatings prevents the production of signal generation in ultrasound imaging. This work opens new avenue for CEUS imaging of the vascular and extravascular space using CaCO3, as it highlights the potential to generate contrast for extended durations at physiological pH by utilizing the amorphous phase of CaCO3.

Producing amorphous calcium carbonate using waste Ca-rich solution generated through the bio-leaching of slag

This study presents a new method for treating waste Ca-rich solutions (WCS), from which amorphous calcium carbonate (ACC) nanoparticles, an industrial product in high demand, are produced. The distinctive feature of this method is the use of tannic acid (TA) as a surfactant, employing a CO2 bubbling method under ambient conditions. By modulating the concentration of TA, the morphology and crystallization of the generated CaCO3 nanoparticles can be effectively controlled. Experimental results reveal that with increasing TA dosage, the crystallinity of CaCO3 decreases until an amorphous phase is obtained. The crystallization of ACC in solution was completely inhibited when TA was added at a concentration of more than 1%. Therefore, the proposed method provides a new strategy for the efficient preparation of ACC from WCS.

Dynamic Growth of Macroscopically Structured Supramolecular Hydrogels through Orchestrated Reaction-Diffusion.

Living organisms are capable of dynamically changing their structures for adaptive functions through sophisticated reaction-diffusion processes. Here we show how active supramolecular hydrogels with programmable lifetimes and macroscopic structures can be created by relying on a simple reaction-diffusion strategy. Two hydrogel precursors (poly(acrylic acid) PAA/CaCl2 and Na2 CO3 ) diffuse from different locations and generate amorphous calcium carbonate (ACC) nanoparticles at the diffusional fronts, leading to the formation of hydrogel structures driven by electrostatic interactions between PAA and ACC nanoparticles. Interestingly, the formed hydrogels are capable of autonomously disintegrating over time because of a delayed influx of electrostatic-interaction inhibitors (NaCl). The hydrogel growth process is well explained by a reaction-diffusion model which offers a theoretical means to program the dynamic growth of structured hydrogels. Furthermore, we demonstrate a conceptual access to dynamic information storage in soft materials using the developed reaction-diffusion strategy. This work may serve as a starting point for the development of life-like materials with adaptive structures and functionalities.

Dynamic Growth of Macroscopically Structured Supramolecular Hydrogels through Orchestrated Reaction‐Diffusion

AbstractLiving organisms are capable of dynamically changing their structures for adaptive functions through sophisticated reaction‐diffusion processes. Here we show how active supramolecular hydrogels with programmable lifetimes and macroscopic structures can be created by relying on a simple reaction‐diffusion strategy. Two hydrogel precursors (poly(acrylic acid) PAA/CaCl2 and Na2CO3) diffuse from different locations and generate amorphous calcium carbonate (ACC) nanoparticles at the diffusional fronts, leading to the formation of hydrogel structures driven by electrostatic interactions between PAA and ACC nanoparticles. Interestingly, the formed hydrogels are capable of autonomously disintegrating over time because of a delayed influx of electrostatic‐interaction inhibitors (NaCl). The hydrogel growth process is well explained by a reaction‐diffusion model which offers a theoretical means to program the dynamic growth of structured hydrogels. Furthermore, we demonstrate a conceptual access to dynamic information storage in soft materials using the developed reaction‐diffusion strategy. This work may serve as a starting point for the development of life‐like materials with adaptive structures and functionalities.

A Comparative Study of Ultrasmall Calcium Carbonate Nanoparticles for Targeting and Imaging Atherosclerotic Plaque.

Atherosclerosis is a complex disease that can lead to life-threatening events, such as myocardial infarction and ischemic stroke. Despite the severity of this disease, diagnosing plaque vulnerability remains challenging due to the lack of effective diagnostic tools. Conventional diagnostic protocols lack specificity and fail to predict the type of atherosclerotic lesion and the risk of plaque rupture. To address this issue, technologies are emerging, such as noninvasive medical imaging of atherosclerotic plaque with customized nanotechnological solutions. Modulating the biological interactions and contrast of nanoparticles in various imaging techniques, including magnetic resonance imaging, is possible through the careful design of their physicochemical properties. However, few examples of comparative studies between nanoparticles targeting different hallmarks of atherosclerosis exist to provide information about the plaque development stage. Our work demonstrates that Gd (III)-doped amorphous calcium carbonate nanoparticles are an effective tool for these comparative studies due to their high magnetic resonance contrast and physicochemical properties. In an animal model of atherosclerosis, we compare the imaging performance of three types of nanoparticles: bare amorphous calcium carbonate and those functionalized with the ligands alendronate (for microcalcification targeting) and trimannose (for inflammation targeting). Our study provides useful insights into ligand-mediated targeted imaging of atherosclerosis through a combination of in vivo imaging, ex vivo tissue analysis, and in vitro targeting experiments.

Amorphous biomineral-reinforced hydrogels with dramatically enhanced toughness for strain sensing

Ionic conductive hydrogels are promising candidates for flexible wearable strain sensors and artificial skin. However, achieving high mechanical and sensing performance concurrently remains challenging. Herein, a novel biomineral-reinforced hydrogel composed of polyacrylamide (PAM) and highly stable amorphous calcium carbonate (ACC) is reported. Benefiting from the dual ionic doping strategy (Mg2+ and PO43−), ACC nanoparticles in hybrid hydrogels show a super stable amorphous nature. The resulting mineral hydrogel displays a high stretchability (>1150% strain), a dramatically enhanced fracture toughness (9.57±1.28 vs. 0.91±0.12 kJ m−2), and a desirable linear strain sensitivity. Moreover, the novel mineral hydrogel exhibits high biocompatibility and flame retardance, making it an appealing candidate for wearable device applications.

Protection of Historical Mortars through Treatment with Suspensions of Nanoparticles

Mortars, which are very important elements for the integrity of historic monuments, consist mainly of calcium carbonate and silicates in different proportions. Chemical dissolution due to exposure in open air is very important for the degradation of mortars. Inorganic nanoparticles with chemical and crystallographic affinity with mortar components are expected to be effective structure stabilizers and agents offering resistance to chemical dissolution. In the present work, we have developed and applied suspensions of amorphous calcium carbonate (ACC), silicon oxide (am-SiO2) and composite nanoparticles by the precipitation of ACC on am-SiO2 and vice versa. The application of suspensions of the synthesized nanoparticles on three different historical mortars of Roman times (1st century AD), retarded their dissolution rate in solutions undersaturated with respect to calcite, in acid pH (6.50, 25 °C). All three test historic mortars, treated with suspensions of the nanoparticles prepared, showed high resistance towards dissolution at pH 6.50. The ability of the nanoparticles’ suspension to consolidate the damaged mortar was the key factor in deciding the corresponding effectiveness in the retardation of the rate of dissolution. The combination of ACC with am-SiO2 nanoparticles showed high efficiency for protection from the dissolution of calcite rich mortars.

Bioprocess inspired formation of calcite mesocrystals by cation-mediated particle attachment mechanism.

Calcite mesocrystals were proposed, and have been widely reported, to form in the presence of polymer additives via oriented assembly of nanoparticles. However, the formation mechanism and the role of polymer additives remain elusive. Here, inspired by the biomineralization process of sea urchin spine comprising magnesium calcite mesocrystals, we show that calcite mesocrystals could also be obtained via attachment of amorphous calcium carbonate (ACC) nanoparticles in the presence of inorganic zinc ions. Moreover, we demonstrate that zinc ions can induce the formation of temporarily stabilized amorphous nanoparticles of less than 20nm at a significantly lower calcium carbonate concentration as compared to pure solution, which is energetically beneficial for the attachment and occlusion during calcite growth. The cation-mediated particle attachment crystallization significantly improves our understanding of mesocrystal formation mechanisms in biomineralization and offers new opportunities to bioprocess inspired inorganic ions regulated materials fabrication.

Synthesis of stable calcium carbonate nanoparticles for pH-responsive controlled drug release

The construction of controlled release for drugs is a promising strategy in disease treatment, which can improve the bioavailability of utilized drugs and reduce the side effects. Herein, a simple and green strategy was developed to prepare stable amorphous calcium carbonate (ACC) using polyvinyl pyrrolidone (PVP) as stabilizer. The crystallization behavior of ACC nanoparticles in methanol–water system and the effect of PVP content on ACC was investigated. Most importantly, the obtained ACC-PVP exhibited high drug loading efficiency and pH responsive release properties for anticancer drugs 5-Fluorouracil (5-FU) and curcumin (CUR) in phosphate buffer solution and simulated gastric juice. All evidences demonstrate that smart mesoporous ACC-PVP nanoparticles provide a potential clinical application platform for cancer therapy.

Crystallization pathways, fabrics and the capture of climate proxies in speleothems: Examples from the tropics

The quality of climate proxy data from speleothem archives depends to varying degrees on crystallization processes, which result in diverse fabrics. Here, we document shifts in calcite growth mechanisms, from ion-by-ion to nanoparticle/nanocrystal attachment, in stalagmites from the tropical island of Atiu (South Pacific). Changes in solution stoichiometry and organic matter content result in the development of two columnar fabrics that are common elsewhere in settings characterized by seasonal contrast. A porous columnar fabric, characterized by intracrystalline micro and nanoporosity grows via a non-classical pathway through amorphous calcium carbonate (ACC) nanoparticles (∼2–4 nm in diameter) and calcite nanocrystal attachment. Despite subsequent transformation of nanoparticles/crystals into a large calcite crystal, the porous columnar fabric appears to preserve a δ18O signal that faithfully reflects that of the parent fluid via quasi-equilibrium fractionation. Furthermore, the porous fabric shows random and fuzzy lateral distributions of Sr, another hydrological proxy, yet this element's incorporation follows equilibrium partitioning. The chemical properties of compact columnar fabrics, which appear to grow by classical ion-by-ion attachment, may not directly reflect those of the original depositional environment because of degassing, the presence of growth inhibitors (such as Na) and very early diagenetic modifications. Columnar porous calcite fabrics that formed through non-classical pathways in other settings may faithfully record the original properties of the parent drip water, whereas compact fabrics that formed through classical pathways elsewhere may not. It is concluded that the study of fabrics at the nano-scale is a necessary complement to speleothem research to identify the influence of crystallization pathways on the accuracy of proxy data.

Eco-friendly processes for the synthesis of amorphous calcium carbonate nanoparticles in ethanol and their stabilisation in aqueous media

New understandings in the amorphous calcium carbonate nanoparticle synthesis lead to a final mass concentration increase by a factor of 3.5. The stabilisation in aqueous media is achieved by a 2-minute scalable process using bio-sourced stabilisers.

Pressure-induced crystallization and densification of amorphized calcium carbonate hexahydrate controlled by interfacial water

Amorphous calcium carbonate (ACC) is widely known as a metastable precursor in the formation of crystalline calcium carbonate biominerals. However, the exact role of water during the crystallization of ACC remains elusive. Here, a novel ACC with high specific surface area and nanopores is synthesized by solvent-induced dehydration and amorphization of crystalline calcium carbonate hexahydrate (ikaite), denoted as I-ACC. Comparing I-ACC and typical spherical ACC (S-ACC) nanoparticles, it reveals that the crystallization pathways of ACC under heating or pressure are not dictated by the total amount of water in ACC as reported, but rather the interfacial water that is released from ACC bulk and adsorbed on the surface of the particles. We show that the crystallization pathways of I-ACC to calcite single crystal with high specific surface area or vaterite can be easily controlled by tuning the release of water during heating. In addition, densely packed pure vaterite can be obtained via pressured-induced transformation of I-ACC at room temperature, which is otherwise difficult to form using S-ACC. These insights contribute to the understanding of the biological control of mineral formation via amorphous precursors and offer new opportunities to bioprocess inspired fabrication of strong bulk material at room temperature.

Investigation of ultrasonication and/or mechanical stirring on the reactive crystallization of calcium carbonate at lower temperature and higher supersaturation condition

Calcium carbonate (CaCO3, CC) distributes diversely among the abiotic and biogenic minerals/biominerals. It has six phases (three anhydrous crystalline, two hydrates and one amorphous). Phase selectivity of the synthetic CC is of considerable interest and determines its physicochemical properties and destinated applications.In the present study the reactive crystallization process of CC has been subjected to the treatment of different disturbances, i.e., mechanical stirring (ms), ultrasonication (us), and composed ms + us at lower temperature (0 ± 1℃) and higher supersaturation condition (initial concentration of Ca2+ is 0.1 M), in comparison with the quiescent condition. In order to gain insight on the role of these disturbances in controlling the crystallization process, nature, crystal habits of precipitated CC and their consecutive phase transformation, samples were collected at different time intervals (30 s, 1 min, 5 min, 10 min, 20 min, 45 min and 60 min) for investigation. Results reveals that these disturbing ways play a significant role on the CC crystallization process and can stabilize the amorphous calcium carbonate (ACC) phase for prolonged periods of time (1–60 min) in aqueous solution, which will be a powerful way for large-scale fabrication of pure ACC nanoparticles (about 100 nm) without addition of any chemicals. Longer disturbing time tends to transform the pure and spherical ACC nanoparticles competitively into spherical vaterite particles and rhombohedral calcite particles through the self-assembly of nanoparticles aggregation or Ostwald ripening mechanism, respectively. The investigation also suggested that the reactive crystallization of CC with the three disturbances undergoes a non-classical crystallization pathway to crystallize CC particles, in comparison with the quiescent reference condition. To the best of our knowledge, the report on reactive crystallization of CC crystallites as a function of different disturbances is first of its kind in the literature.

Intragastric amorphous calcium carbonate consumption triggered generation of in situ hydrogel piece for sustained drug release.

Sustained-release systems in solid forms are widely accepted except for those patients with dysphagia such as the elderly and infants. Herein, we have developed a sustained-release oral drug solution by dispersing polyaspartic acid stabilized amorphous calcium carbonate (ACC) nanoparticles (NPs) (<100nm) in drug-containing sodium alginate (SA) solution (2%). It can be supposed that ACC NPs would consume in the gastrointestinal tract acid environment and provide calcium ions as the crosslinker for SA polymer solution to gain in situ hydrogels. The pre-gel solution system appeared a distinct reversible shear-thin characteristic with high viscosity and stability during storage. Whereas, it can regain fine flowability for convenient oral administration after slightly shaking. In this study, ACC NPs consumption triggered solution-gel transition of this drug-containing pre-gel solution system was demonstrated in simulated gastric solution and in vivo, with the distinct sustained drug release profiles lasting for 10h. In addition, pharmacokinetics studies in SD rats showed that the Tmax (up to 1.79±0.24h) and AUC0-12 (46.37±2.71μg/mL·h) of ACC/SA in situ hydrogels treated group were significantly higher (p<0.05) than that of free drug solution. Overall, these results highlighted a novel promising platform for oral administration of drugs, reduced dosing frequency, and high potential compliance for patients with dysphagia.

Preparation and characterization of phosphate-stabilized amorphous calcium carbonate nanoparticles and their application in curcumin delivery

Nano-sized amorphous calcium carbonate (ACC) have a great potential for drug transport and drug delivery because of their high drug loading capacity, excellent biocompatibility and biodegradability. However, ACC is the thermodynamically least stable phase of calcium carbonate, and such thermal metastability often transforms it into more stable vaterite or calcite phases, so the inhibition of the phase transformation of ACC is of great significance for its application in drug delivery systems. Herein, phosphate as a morphology controlling additive was introduced to stabilize the metastable ACC. The results showed that the composite nanoparticles (named here the ACCP) with narrow size distribution located at approximately 128 nm (measured by dynamic light scattering) were obtained, when Ca2+ concentration and C/P were 10 mM and 7/3 respectively, prepared at 30 °C for 10 min. The powdered X-ray diffraction (XRD) results showed that phosphate has an excellent ability to stabilize ACC, and even after storage for 60 days at room temperature, the structure of ACCP remains amorphous. The in vitro drug release tests showed that ACCP nanoparticles had a high curcumin (Cur) loading capacity and a sustained drug release property. Moreover, the 1, 1-diphenyl-2-picrylhydrazyl radical (DPPH) radical scavenging activity assay showed that ACCP-encapsulating strategy could effectively prevent the decomposition of Cur. In vitro cytotoxicity tests demonstrated that the resultants have excellent biocompatibility. Therefore, the as-prepared ACCP nanoparticles are promising for drug delivery applications.

Solid-State Phase Transformation and Self-Assembly of Amorphous Nanoparticles into Higher-Order Mineral Structures

Materials science has been informed by nonclassical pathways to crystallization, based on biological processes, about the fabrication of damage-tolerant composite materials. Various biomineralizing taxa, such as stony corals, deposit metastable, magnesium-rich, amorphous calcium carbonate nanoparticles that further assemble and transform into higher-order mineral structures. Here, we examine a similar process in abiogenic conditions using synthetic, amorphous calcium magnesium carbonate nanoparticles. Applying a combination of high-resolution imaging and in situ solid-state nuclear magnetic resonance spectroscopy, we reveal the underlying mechanism of the solid-state phase transformation of these amorphous nanoparticles into crystals under aqueous conditions. These amorphous nanoparticles are covered by a hydration shell of bound water molecules. Fast chemical exchanges occur: the hydrogens present within the nanoparticles exchange with the hydrogens from the surface-bound H2O molecules which, in turn, exchange with the hydrogens of the free H2O molecule of the surrounding aqueous medium. This cascade of chemical exchanges is associated with an enhanced mobility of the ions/molecules that compose the nanoparticles which, in turn, allow for their rearrangement into crystalline domains via solid-state transformation. Concurrently, the starting amorphous nanoparticles aggregate and form ordered mineral structures through crystal growth by particle attachment. Sphere-like aggregates and spindle-shaped structures were, respectively, formed from relatively high or low weights per volume of the same starting amorphous nanoparticles. These results offer promising prospects for exerting control over such a nonclassical pathway to crystallization to design mineral structures that could not be achieved through classical ion-by-ion growth.

Forming amorphous calcium carbonate within hydrogels by enzyme-induced mineralization in the presence of N-(phosphonomethyl)glycine

Amorphous inorganic materials have a great potential in material science. Amorphous calcium carbonate (ACC) is a widely useable system, however, its stabilization often turns out to be difficult and the synthesis is mostly limited to precipitation in solution as nanoparticles. Stable ACC in bulk phases would create new composite materials. Previous work described the enzyme-induced mineralization of hydrogels with crystalline calcium carbonate by entrapping urease into a hydrogel and treating this with an aqueous mineralization solution containing urea und calcium chloride. Here, this method was modified using a variety of crystallization inhibitors attached to the hydrogel matrix or added to the surrounding mineralization solution. It was found that only N-(phosphonomethyl)glycine (PMGly) in solution completely inhibits the crystallization of ACC in the hydrogel matrix. The stability of the homogeneously precipitated ACC could be accounted to the combination of stabilizing effects of the additive and stabilization through confinement. The crystallization could be accelerated at higher temperatures up to 60 °C. Here, a combination of Mg ions and PMGly was required to stabilize ACC in the hydrogel. Variation of these two compounds can be used to control a number of different calcium carbonate morphologies within the hydrogel. While the ACC nanoparticles within the hydrogel are stable over weeks even in water, a calcite layer grows on the surface of the hydrogel, which might be used as self-hardening mechanism of a surface.

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.