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The acidic nature of malignant tumors leads to increased drug sequestration and the evasion of apoptotic damage, which is further exacerbated by abnormal lysosomes in tumor cells. In this study, a "lysosomal bomb" will be constructed using a type of acid-neutralized amorphous calcium carbonate (ACC) to encapsulate the sonosensitizer protoporphyrin IX (PpIX), and then coated with homologous tumor cell membranes to increase water solubility and homologous targeting. The PpIX-ACC@CMs designed in this paper are popcorn-like structures, which can not only neutralize the tumor's acidic microenvironment to balance the pH value and release excess Ca 2+ , but also cause lysosomal dysfunction and achieve drug lysosomal escape to increase drug accumulation. Additionally, the CO 2 gas nucleus produced by the acid reaction of ACC can increase the ultrasonic cavitation effect to amplify the sonodynamic therapy (SDT) effect. In vitro and in vivo experiments demonstrated that PpIX-ACC@CMs, serving as a "lysosomal bomb," successfully localized to lysosomes of tumor cells and exhibited lysosomal escape ability through its acid reaction ability, achieving excellent SDT efficacy under ultrasound stimulation. Furthermore, exogenous Ca 2+ overload also increased the likelihood of tumor calcification, which could contribute to in vivo tumor inhibition and facilitate CT medical imaging to monitor treatment efficacy., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Transient receptor potential vanilloid 1 (TRPV1) pathway can result in the disruption of intracellular calcium ion (Ca 2+ ) homeostasis, but how to effectively control the process of Ca 2+ overload needs to be addressed for tumor treatment. Herein, we developed a mild photothermal activation of TRPV1 pathway strategy for enhanced Ca 2+ overload therapy of hepatocellular carcinoma, in which the multifunctional nanoparticle (CNQ) loading with CaCO 3 , a novel polycyclic conjugated small molecule (2Br-NDIA) and quercetin (Qu) as the core for the efficient mild photothermal therapy (PTT). The photothermal property of CNQ can specifically activate the TRPV1 channel, which leads to mitochondrial Ca 2+ overloading and the disruption of intracellular Ca 2+ homeostasis. This CNQ-mediated activation of the TRPV1 channel by mild PTT provides an emerging paradigm for enhancing mitochondrial Ca 2+ overload therapy and dismantling tumor self-defense, safely expanding cancer therapies for highly refractory tumors., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Magnesium phosphate cement (MPC) has been evaluated as a novel bone substitute owing to its favorable biocompatibility, plasticity, and osteogenic potential. However, the low porosity of MPC prevents growth factors and osteoblasts from fully growing into the material, thereby limiting its clinical use. In this study, different concentrations (0-5%) of calcium carbonate and citric acid (CA) were used as foaming agents to prepare porous MPC. The MPC containing 3% CaCO 3 /CA exhibited the best physicochemical properties, including greater porosity, improved injectability, extended setting time, and decreased hydration temperature. The proliferation and adhesion of cells on 3%CaCO 3 /CA-MPC were higher than those on MPC alone. To explore its osteogenesis in vivo, 3% CaCO 3 /CA-MPC and Bio-Oss® bone powder were implanted into periodontal bone defects in rats for 4 weeks and 12 weeks, respectively. Micro-CT and histological analysis demonstrated the improved bone regeneration of 3%CaCO 3 /CA-MPC compared to the blank group (P < 0.05); it had slightly lower bone regeneration than the Bio-Oss® group but no statistical difference. The results indicated that porous MPC foamed with calcium carbonate and CA improved its physicochemical properties and enhanced its biocompatibility, making it a promising material for bone regeneration., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The effect of microbial phytase and limestone particle size (LmPS) was assessed in Lohmann Tradition laying hens from 31 to 35 wk of age. Seventy-two hens were used in a completely randomized trial according to a 2 × 2 factorial arrangement with 2 levels of phytase/basal available P (aP); 0 FTU/kg with 0.30% aP or 300 FTU/kg with 0.15% aP, and 2 limestone particle sizes; fine particles (FL, <0.5 mm) or a mix (MIX) of 75% coarse limestone (CL, 2-4 mm) and 25% FL. Diets contained equivalent levels of Ca (3.5%), phytic P (PP; 0.18%), and aP (0.30%) considering the P equivalency of phytase. Thus, dietary treatments were FL0 and MIX0 without phytase, and FL300 and MIX300 with 300 FTU/kg phytase. Performance were recorded daily and eggshell quality (eggshell weight proportion, weight, thickness, and breaking strength) was measured weekly. At the end of the trial, bone parameters (tibia breaking strength, elasticity, and ash) and the apparent precaecal digestibility (APCD) of P and Ca were determined. No differences were observed between treatments in feed intake, FCR and bone parameters. Addition of MIX increased the eggshell proportion, weight and thickness in groups receiving no phytase (+6.5, +6.9, and +4.5%, respectively) while no effect was observed in groups receiving phytase (Phytase × LmPS, P < 0.05). In hens receiving FL, the APCD of P was lower in diets supplemented with phytase (-14 percentage points; Phytase × LmPS, P < 0.001). A higher phytate disappearance was observed in hens fed diets with phytase in combination with MIX (Phytase × LmPS, P = 0.005). Phytase and MIX together increased the APCD of Ca by 7.3 percentage points (Phytase × LmPS, P < 0.001). In conclusion, addition of CL could limit the formation of Ca-phytate complex thus improving the response of the birds to phytase compared to FL., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

The growing biomedical challenges impose the continuous development of novel platforms. Ensuring the biocompatibility of drug delivery and implantable biomedical devices is an essential requirement. Calcium carbonate (CaCO3) in the form of vaterite nanoparticles is a promising platform, which has demonstrated distinctive optical and biochemical properties, including high porosity and metastability. In this study, the biocompatibility of differently shaped CaCO 3 vaterite particles (toroids, ellipsoids, and spheroids) are evaluated by bacterial toxicity mode-of-action with a whole-cell biosensor. Different Escherichia coli (E. coli) strains were used in the bioluminescent assay, including cytotoxicity, genotoxicity and quorum-sensing. Firstly, both scanning electron microscopy (SEM) and fluorescence microscopy characterizations were conducted. Bacterial cell death and aggregates were observed only in the highest tested concentration of the vaterite particles, especially in toroids 15-25 µm. After, the bioluminescent bacterial panel was exposed to the vaterite particles, and their bioluminescent signal reflected their toxicity mode-of-action. The vaterite particles resulted in an induction factor (IF > 1) on the bacterial panel, which was higher after exposure to the toroids (1.557 ≤ IF ≤ 2.271) and ellipsoids particles (1.712 ≤ IF ≤ 2.018), as compared to the spheroids particles (1.134 ≤ IF ≤ 1.494), in all the tested bacterial strains. Furthermore, the vaterite particles did not affect the viability of the bacterial cells. The bacterial monitoring demonstrated the biofriendly nature of especially spheroids vaterite nanoparticles., Competing Interests: Declaration of Competing Interest The authors declared no potential conflicts of interest concerning the research, authorship, and/or publication of this article., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Artesunate (AS), the first-line treatment of malaria with a satisfactory safety profile, has been repurposed as a potential anticancer candidate as it mainly generates reactive oxygen species (ROS) through its intrinsic endoperoxide bridge reacting with ferrous-based catalysts to suppress cancer cell growth. However, further clinical translation of AS is hindered by the attenuated anticancer efficacy due to insufficient ROS generation. Herein, we rationally integrated hydrophobic-modified AS (hAS) with biomimetic polydopamine (PDA) and biomineral calcium carbonate to fabricate high AS-loaded nanomedicine (Ca-PDA/hAS@PEG) for cancer chemo-photothermal therapy, which exerted anticancer effects in the following ways: (1) the heat was generated when PDA was irradiated by near-infrared (NIR) light for photothermal therapy. Meanwhile, the increased temperature accelerated the production of ROS from hAS, thus enhancing the anticancer efficacy of hAS-based chemotherapy; (2) hAS-mediated chemotherapy boosted the cancer inhibition effect of photothermal therapy by arousing the intracellular ROS levels in the presence of endogenous ferrous ions and sensitizing cancer cells to thermal ablation; (3) the integration of calcium carbonate into the nanoparticle facilitated the pH-responsive drug release for precise treatment. Such hybrid nanoparticles exhibited a combinational antitumor effect of photothermal therapy and chemotherapy in vivo with no systemic toxicity. Taken together, our work presents a facile strategy to improve the anticancer efficacy of AS by combining chemical modification and photothermal therapy-assisted endoperoxide bridge cleavage, which may offer opportunities to pave the way for clinical translation of AS-based nanomedicines. STATEMENT OF SIGNIFICANCE: The clinical translation of artesunate (AS) is hindered by the attenuated anticancer efficacy due to insufficient ROS generation. Herein, we rationally integrated hydrophobic-modified AS (hAS) with biomimetic polydopamine (PDA) and biomineral calcium carbonate to fabricate high AS-loaded nanomedicine (Ca-PDA/hAS@PEG) for improved cancer chemo-photothermal therapy. The heat generated from PDA in response to near-infrared light irradiation could locally ablate tumor as well as accelerate the production of ROS by hAS, thus enhancing the anticancer efficacy of hAS-based chemotherapy. On the other hand, hAS-based chemotherapy amplified the intracellular oxidative stress, sensitizing cancer cells to thermal ablation. Our work presents a facile strategy to improve the anticancer efficacy of AS by combining chemical modification and photothermal therapy-assisted endoperoxide bridge cleavage., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2022. Published by Elsevier Ltd.)

The marble bury test is a commonly applied behavioral test often used to screen pharmaceuticals for treatment of compulsivity or anxiety disorders and to better understand the underlying neurobiology of these conditions using rodent models. We explore the use of the marble bury test in a repeated fashion over longer time intervals to assess how it may be used in a more chronic context. Assuming that marble bury scores represent a neurobiological phenotype of an individual, we hypothesize that the measurement of this phenotype is repeatable over time in a healthy animal. We performed four trials over the course of 12 weeks, using three overlapping scores for marble burying: 100%, >50%, and >0% buried. Despite intertrial differences in the mean number of marbles buried, we found significant repeatability scores across 12 weeks for two measurements: marbles buried 100% and marbles buried >50%. Exploration of pairwise repeatabilities between the trials revealed highest repeatability during shorter time intervals. Although the effect of time is difficult to disentangle from possible effects of body size/development on the scores, we hypothesize that a combination of these two factors together influence repeatability of scores and should be explored further. Understanding the patterns in repeatability in marble burying scores can inform experimental design to use this test over longer time intervals for chronic conditions or long-term interventions. Additionally, these data indicate that a pre-test, post-test design is possible, which can reduce the number of animals needed for research applications while maintaining statistical power., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Calcium carbonate is used as bone-filling material due to its good biocompatibility, bioactivity, and bioabsorbability, but the prevalence of infectious complications associated with calcium carbonate has created a persisting challenge in the treatment of bone defect. Therefore, this greatly necessitate the need to endow calcium carbonate with antibacterial properties. In this study, calcium carbonate powders loaded with silver nanoparticles (Ag-CaCO 3 ) were prepared in attempt to serve as a novel antibacterial inorganic filler material. This objective was achieved using ultrasonic spray-pyrolysis (USSP) route to produce Ag-CaCO 3 with 1, 5 and 10 mol% silver. The size of silver nanoparticles on CaCO 3 microspheres could be regulated by adjusting silver concentration to facilitate effective release of Ag + ions. This was demonstrated in Ag-CaCO 3 (1), where the lowest silver content at 1 mol% achieved the highest Ag + ions release over 28 days. This in turn gave rise to effective antibacterial efficiency against Staphylococcus aureus and Escherichia coli. Furthermore, CaCO 3 (1) could also support osteoblast-like cells (MG-63) at a cell viability of 80%. Overall, this work extends the capabilities in employing USSP to produce inorganic filler materials with sustained antibacterial properties, bringing one step closer to the development of antibacterial products., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The development of heterogeneous drug delivery systems leads to innovative strategies for targeted therapy of common pathologies, such as cancer, immunological and neurological disorders. Nowadays, it is possible to choose among a great variety of nanoparticles on the basis of the needs they have to satisfy. However, a candidate for the treatment of cardiovascular pathologies is still missing. In this context, a targeted therapy implies the conceptualization of nanoparticles that take active part in the treatment of vascular pathologies. The aim of this work was to provide a method to produce multi-layered calcium carbonate (CaCO 3 ) nanoparticles encapsulating a model protein, bovine serum albumin, with model antibodies on their surface. CaCO 3 nanoparticles were produced by the combination of complex coacervation and mineralization and were engineered using layer-by-layer technique with a polysaccharide, dextran sulfate, and a homo-poly-amino acid, poly-L-arginine. Morphology, biocompatibility, cellular uptake, influence on cell expression of the inflammatory marker matrix metalloproteinase-9, and hemocompatibility of the nanoparticles were studied. The presence of the dextran/poly-L-arginine layers did not negatively affect the nanoparticle overall characteristics and they did not trigger proinflammatory response in vitro. Taking together all the obtained results, we consider the proposed CaCO 3 nanoparticles as a promising tool in cardiovascular field., (Copyright © 2021. Published by Elsevier B.V.)

We investigated effects of rearing cage type and dietary limestone particle size (LPS) on egg production, egg weight, eggshell, and bone quality in laying hens. The pullets were reared in conventional (CON; 20 chicks/cage, 270 cm 2 /chick) or furnished (FUR; 30 chicks/cage; 636 cm 2 /chick) cages and fed 3 LPS (fine, <0.595 mm; medium, 0.595 to <1.68 mm; and 1:1 mixture of F and M wt/wt) to 16 wk of age (woa). Pullets were transitioned to laying furnished cages and retained rearing treatment combination identities (n = 5, 20 hens/cage). Hens had free access to common commercial layer diet and water through to 40 woa. Eggs were recorded daily for calculation of hen day egg production (HDEP). Subsamples of eggs laid on the first day of 24, 28, 32, 36, and 40 woa were used for eggshell quality analyses. Two hens per cage were sacrificed on the last day of 24 and 40 woa for femur and tibia quality assessments. There was no interaction (P > 0.05) between rearing cage type and dietary LPS on response variables. At 19 and 20 woa, HDEP was higher (P < 0.01) for FUR than CON reared hens but was similar (P > 0.05) afterward. At 40 woa, FUR reared hens had higher (P < 0.05) body weight (BW), egg weight (EW), eggshell thickness, and eggshell weight and tended (P < 0.10) to have higher femur and tibia mineral density (BMD) and mineral content (BMC) than CON reared hens. Rearing dietary LPS had no effect (P > 0.05) on HDEP, BW, EW, and eggshell quality. Although, rearing dietary LPS did not affect (P > 0.05) femur and tibia BMD and BMC; at 24 woa, hens reared on medium LPS tended to have higher femur BMD (0.17 vs. 0.14 g/cm 2 ; P = 0.079) and BMC (0.99 vs.0.78 g; P = 0.088) than hens reared on fine LPS. In conclusion, hens reared in furnished cages had better eggshell quality but had marginal effects on femur and tibia quality, whereas rearing dietary LPS had no effects on eggshell and bone attributes in hens., (Copyright © 2020. Published by Elsevier Inc.)

The hypothesis was tested that an increased digestion of coarse compared with fine limestone can alleviate the negative effects of a low dietary Ca/P ratio on the growth performance and characteristics of tibia strength (CTS) in broilers. A total of 1,152 Ross 308 broiler chickens received a standard commercial starter feed from day 0 to 13. From day 14 onward, birds received 1 of 12 diets containing 1 of 6 Ca/P ratios (0.50, 0.75, 1.00, 1.25, 1.50, and 1.75) and 1 of 2 limestone particle sizes (<500 [fine] and 500 to 2,000 [coarse] μm) in a study with a 6 × 2 factorial arrangement of treatments. Total P content was fixed at 5.5 g/kg for all treatment diets. Each treatment was replicated 6 times with 16 birds per replicate pen. On day 20 and 21, twelve birds per pen were randomly selected from 4 of the 6 replicate pens for tibia analysis and digesta collection from different gut segments. The apparent Ca digestibility was higher for fine than coarse limestone in the jejunum (P = 0.043). However, this difference in Ca digestibility disappeared for the low, whereas it remained for the high Ca/P ratios in the proximal (P interaction = 0.067) and distal (P interaction = 0.052) ileum. In addition, coarse limestone improved apparent P digestibility in the proximal and distal ileum (P < 0.001) but not in the jejunum (P = 0.305). Regardless of limestone particle size, reducing dietary Ca/P ratio linearly improved apparent Ca and P digestibility in the proximal and distal ileum (P < 0.001). Moreover, decreasing dietary Ca/P ratio linearly (P < 0.001) and quadratically (P < 0.046) reduced the CTS. Reducing dietary Ca/P ratio linearly (P < 0.003) and quadratically (P ≤ 0.006) decreased body weight gain and increased feed conversion ratio. For both fine and coarse limestone, the optimal Ca/P ratio was 1.00 to 1.25 to optimize apparent Ca and P digestibility while maintaining growth performance and CTS. Reducing Ca/P ratio from 1.75 to 1.00 improved distal ileal Ca and P apparent digestibility from 36.6 to 53.7% and 48.0 to 58.3%, respectively. In conclusion, coarse limestone is equally digestible with fine limestone at a low Ca/P ratio but is less digestible at a high Ca/P ratio, and the optimal Ca/P ratio in the diet is 1.00 to 1.25 for both fine and coarse limestone., (Copyright © 2020. Published by Elsevier Inc.)

Effects of rearing cage type and dietary limestone particle size (LPS) on growth, apparent retention (AR) of nutrients, and bone quality were investigated. The treatments were arranged in a 2 × 3 factorial with cage (conventional, CON and furnished, FUR) and LPS (fine, < 0.595 mm, F; medium, 0.595 to < 1.68 mm, M; and 1:1 mixture of F and M wt/wt; FM). A total of 900-day-old Lohmann LSL-Lite chicks were placed in CON (20 chicks/cage) and FUR (30 chicks/cage) based on BW. The diets were formulated according to breeder's nutrient specifications for starter, grower, and developer phases. At the end of 4, 12, and 16 wk of age (woa), 2 pullets/cage were euthanized for samples. At 12 and 16 woa, 1 pullet/cage was transferred to metabolism cages for AR measurements. There was no interaction (P > 0.05) between cage type and LPS on response variables. At 4 woa, body (P = 0.002) and bone (P < 0.05) weight was higher for CON than FUR pullets, but this was reversed (P < 0.01) at 16 woa. Pullets fed M LPS had higher (P < 0.05) AR of Ca, whole body mineral density (BoMD), and whole body mineral content (BoMC) than pullets fed F LPS. However, pullets fed F LPS had higher (P < 0.05) femur bone mineral density (BMD) and tended (P = 0.059) to have higher tibia bone breaking strength (BBS) than pullets fed M LPS at 16 woa. Pullets reared in CON cages had higher (P < 0.05) AR of Ca than FUR pullets. At 4 woa, CON pullets had lower (P < 0.05) femur and tibia BMD but higher tibia (93 vs. 83 N P = 0.012) BBS than FUR pullets. However, at 16 woa, FUR pullets had higher (P < 0.05) BoMD, BoMC, and tibia BBS than CON pullets. In conclusions, cage type and dietary LPS had independent effects on Ca utilization and skeletal development. Despite poor Ca retention, FUR caged pullets showed improved bone quality at 16 woa. Finer LPS improved femur mineral density suggesting coarser LPS had limited effects on pullet bone quality., (Copyright © 2020. Published by Elsevier Inc.)

Calcium carbonate nanoparticles (CaCO 3 NPs) derived from CO 2 are promising materials for different industrial applications. It is imperative to understand their toxicological profile in biological systems as the human and environmental exposures to CaCO 3 NPs increases with growing production. Here, we analyse the cytotoxicity of CaCO 3 NPs synthesized from a CaO slurry on two cell lines, and in vivo on zebrafish (Danio Rerio). Our results demonstrate the CaCO 3 NPs in vitro safety as they do not cause cell death or genotoxicity. Moreover, zebrafish treated with CaCO 3 NPs develop without any abnormalities, confirming the safety and biocompatibility of this nanomaterial., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

Polymeric coatings with positive surface charge offer potential antimicrobial activity, which they owe to a simple electrostatic attraction with negatively charged bacterial walls and membranes. We describe synthesis and characterization of poly(2-aminoethyl methacrylate) and its copolymers with methyl methacrylate and butyl acrylate, as potential binders for antimicrobial solvent-cast paints. TiO 2 and CaCO 3 mineral particles were employed as model pigments/fillers, as they are used in most real-life paint formulations. Electrokinetic (ζ) potential and antimicrobial activity of thin films made of the (co)polymers in the absence and presence of TiO 2 and CaCO 3 nanopowders were assessed using streaming current measurements and microbial growth inhibition tests, respectively. Independently of the structure of the monomers used for the synthesis, the films showed positive ζ-potential values (up to +95 mV) in the pH range 3.5-8.0. The presence of mineral particles at 50% dry weight of the films did not affect significantly the ζ(pH) curves. The films made of the mixed dispersions remained positively charged and inhibited growth of both Gram-negative (E. coli) and Gram-positive (S. aureus) bacteria, as well as yeast (C. albicans). The mixed polymeric-mineral films described in this study seem to be promising potential candidates for designing antimicrobial coatings aimed to prevent spreading of bacterial infections., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The present work reports the very first hydrothermal synthesis of 100% triclinic phase pure aragonite (A1) with microdumbbell microstructural architecture and Au Nanoparticle-decorated (AuNP-decorated) aragonites (A2, A3 and A4) with spherical, pentagonal/hexagonal and agglomerated AuNP-decorated microdumbbells having triclinic aragonite phase as the major and cubic AuNPs as the minor phase. Even in dark the AuNP-decorated aragonites (especially A2) show efficacies as high 90% against gram-negative e.g., Pseudomonas putida (P. putida) bacteria. Further the AuNP-decorated aragonites (A3) show anti-biofilm capability of as high as about 20% against P. putida. Most importantly the AuNP-decorated aragonites (A3) offer anti-cancer efficacy of as high as 53% while those of A1, A2, and A4 are e.g., 26%, 46% and 37%, respectively. For the very first time, based on detailed investigations, the mechanisms behind such advance antibiofilm and anticancer activities are linked to the generation of excess labile toxic reactive oxygen species (ROS). Thus, these materials show enormous potential as futuristic, multi-functional biomaterials for anti-bacterial, anti-biofilm and anti-cancer applications., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Carbonate apatites are sought as a bone substitute due to their biocompatibility and excellent resorbability. The present study deals with Cowrie's shell derived powder (CSDP) as natural biomaterial for bone regenerative medicine. Structural and physicochemical analysis showed that Cowrie's shells, presenting brick and mortar microstructures, were mainly composed of aragonite crystals, which were converted into poorly crystalline B-type carbonate apatite once soaked, at 37 °C, in simulated body fluid for 7 days, reflecting bioactive features. Cytotoxic assays showed that CSDP boosted human stem cell proliferation over the study time compared to nacre derived powder (NDP), used as positive control. Human stem cells adopted a flattened morphology and established physical contact with CSDP, signature of a good biocompatibility. Thus, these results suggested that CSDP presents a great interest for bone regenerative medicine, and could be a useful and versatile carrier/scaffold for bone tissue engineering or a raw material for 3D printed orthopedic devices., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Indocyanine green (ICG) is an efficient photosensitizer that can facilitate producing cytotoxic reactive oxygen species (ROS). At the same time, ICG also has characteristic absorption of near-infrared light and thus can induce a strong photothermal effect. Both of these important features of ICG may be applied for noninvasive light-induced tumor ablation. On the other hand, ICG is lack of stability in blood circulation and susceptible to aggregation or premature clearance from the body. These issues need to be effectively addressed before antitumor application of ICG becomes possible. Herein, a nanocomposite consisting of calcium carbonate modified magnetic polydopamine (PDA) nanoparticles and loaded with ICG, namely Fe 3 O 4 @PDA@CaCO 3 /ICG (FPCI) NPs, was developed to integrate the photothermal capability of PDA with the photodynamic capability of ICG. Particularly, calcium carbonate not only entrapped ICG in the form of stable aggregate to evade blood clearance, but also facilitated controlled release of ICG in response to acidic tumor microenvironment via self-decomposition. With the aid of magnetic guidance, this multifunctional therapeutic agent makes it possible to achieve the combination of photothermal (PTT) and photodynamic therapies (PDT) against tumors, which was demonstrated by this proof-of-concept study based on in vitro and in vivo tumor models. STATEMENT OF SIGNIFICANCE: Currently, there is an ongoing trend of realizing precise and targeted tumor therapy using functional nanocomplexes. Magnetic particles, which can be manipulated by a magnetic field, have attracted increasing attention for tumor therapy. This submitted work demonstrated that calcium carbonate nanoshell was precipitated onto magnetic nanocores mediated by polydopamine. Moreover, indocyanine green (ICG), as a potent photosensitizer, was embedded in this nanocomplex and protected by the calcium carbonate nanoshell, resulting in high drug loading efficiency and enhanced drug stability on the carrier. This new nanocomposite was demonstrated to achieve controlled and pH-responsive release of ICG in tumor environment. This work explored the relationship between the physiochemical properties of the nanocomplex and their potential biomedical applications, aiming to inspire the development of analogous nanoplatforms featured with calcium carbonate blocks., (Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

To meet the progressive requirements for bone regeneration purpose, injectable hydrogels have attracted increasing attention in tissue regeneration and local drug delivery applications. In this study, we report a facile method to prepare injectable and degradable polysaccharide-based hydrogels doubly integrated with hydroxyapatite (HAp) nanoparticles and calcium carbonate microspheres (CMs) under physiological condition. The mechanism of cross-linking is attributed to the Schiff-base reaction between amino and aldehyde groups of carboxymethyl chitosan (CMCS) and oxidized alginate (OAlg), respectively. Synchronously, tetracycline hydrochloride (TH) loaded CMs were fabricated by the precipitation reaction with an average diameter of 6.62 μm. To enhance bioactive and mechanical properties, nano-HAp and CMs containing TH were encapsulated into the polysaccharide-based hydrogel to form injectable gel scaffolds for imitation of bone niche. The gelation time, morphology, mechanical properties, swelling ratio and in vitro degradation of the gel scaffolds could be controlled by varying HAp and CMs contents. Moreover, the composite gel scaffolds had good sustained drug release and antibacterial properties, as confirmed by drugs release calculation and antibacterial evaluation. In addition, the gel scaffolds were found to be self-healing due to dynamic equilibrium of the Schiff-base linkages. These results suggested that the prepared composite gel scaffolds hold great potential for drug delivery and regeneration of irregular bone defects., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objectives: From a pharmaceutical point of view, we see a need to develop stable preclinical test systems to identify and investigate effects of potentized remedies as used in Anthroposophic Medicine and Homeopathy. We evaluated a plant bioassay regarding its capacity to distinguish homeopathic remedies from placebo, applied as sucrose pillules., Methods: Pea seed (Pisum sativum L) was soaked for 24 hours in water with dissolved homeopathic or placebo pillules, or in water only. Shoot length was measured 14 days after planting and treatment groups were compared by analysis of variance (ANOVA). The stability of the system was validated by systematic negative control experiments., Results: The system is suitable to test a common application form - sucrose pillules - of a potentized preparation without influence of the pharmaceutical carrier substance. A screening of 13 potentized preparations revealed Calcium carbonicum to affect pea shoot growth (p < 0.05). Three independent series of main experiments were performed with potentized Calcium carbonicum to assess reproducibility. Meta-analysis of all data revealed significant effects of Calcium carbonicum 12c and 30c on pea shoot growth (p < 0.05), which were however dependent on the date of experiment and/or the experimental series., Conclusions: Potentized Calcium carbonicum, applied as sucrose pillules, influenced pea shoot growth in the assay investigated. However, due to the small effect size and due to the modulation of the effects by still unknown external factors, further optimization of this bioassay is necessary to be used in pharmaceutical quality control or in investigating the biological or pharmaceutical mode of action of potentized preparations., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Sorafenib is a first-line drug for hepatocellular carcinoma (HCC). Autophagy has been shown to facilitate sorafenib resistance. miR-375 has been shown to be an inhibitor of autophagy. In this study, miR-375 and sorafenib were co-loaded into calcium carbonate nanoparticles with lipid coating (miR-375/Sf-LCC NPs). The nanoparticles had high loading efficiency and were ∼50 nm in diameter. Besides, the NPs could increase the stability and residence time of both drugs. Moreover, we demonstrated that autophagy was activated in HCC cells by sorafenib but not by miR-375/Sf-LCC NPs. In vitro, miR-375/Sf-LCC NPs exhibited pH-dependent drug release and potent cytotoxicity. In vivo, miR-375/Sf-LCC NPs increased miR-375 and sorafenib uptake in tumor (2 folds compared with Lipofectamine 2000-miR-375 and 2-5 folds compared with free sorafenib). Furthermore, miR-375/Sf-LCC NPs showed greatly enhanced therapeutic efficacy in an HCC xenograft model. These findings suggest that miR-375/Sf-LCC NPs may be a promising agent for the HCC therapy., Statement of Significance: Hepatocellular carcinoma (HCC) is the most common primary liver tumor and the third leading cause of cancer mortality globally. In this manuscript, miR-375 and sorafenib were co-loaded into calcium carbonate nanoparticles with lipid coating (miR-375/Sf-LCC NPs) to treat HCC. We demonstrated that miR-375/Sf-LCC NPs can deliver sorafenib and miR-375 into HCC cells and tumor tissues, increase drug retention time in tumor, significantly inhibit autophagy and produce enhanced anti-tumor effect., (Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

The elaboration of biocompatible and biodegradable carriers for photosensitizer targeted delivery is one of the most promising approaches in a modern photodynamic therapy (PDT). This approach is aimed at reducing sides effects connected with incidental toxicity in healthy tissue whilst also enhancing drug accumulation in the tumour area. In the present work, Photosens-loaded calcium carbonate (CaCO3) submicron particles in vaterite modification are proposed as a novel platform for anticancer PDT. Fast penetration of the carriers (0.9±0.2μm in diameter) containing 0.12% (w/w) of the photosensitizer into NIH3T3/EGFP cells is demonstrated. The captured particles provide the dye localization inside the cell increasing its local concentration, compared with "free" Photosens solution which is uniformly distributed throughout the cell. The effect of photosensitizer encapsulation into vaterite submicron particles on cell viability under laser irradiation (670nm, 19mW/cm(2), 10min) is discussed in the work. As determined by a viability assay, the encapsulation renders Photosens more phototoxic. By this means, CaCO3 carriers allow improvement of the photosensitizer effectiveness supposing, therefore, the reduction of therapeutic dose. Summation of these effects with the simplicity, upscalability and cheapness of fabrication, biocompatibility and high payload ability of the vaterite particles hold out the prospect of a novel PDT platform., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In a previous study, robust calcium carbonate composite ceramics (CC/PG) were prepared by using phosphate-based glass (PG) as an additive, which showed good cell response. In the present study the in vivo response of porous CC/PG was compared to that of porous biphasic calcium phosphate ceramics (BCP), using a rabbit femoral critical-size grafting model. The materials degradation and bone formation processes were evaluated by general observation, X-ray radiography, micro-computed tomography, and histological examination. The results demonstrated excellent biocompatibility and osteoconductivity, and progressive degradation of CC/PG and BCP. Although the in vitro degradation rate of CC/PG was distinctly faster than that of BCP, at 4week post-implantation, the bone generation and material degradation of CC/PG were less than those of BCP. Nevertheless, at postoperative week 8, the increment of bone formation and material degradation of CC/PG was pronouncedly larger than that of BCP. These results show that CC/PG is a potential resorbable bone graft aside from the traditional synthetic ones., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Biomimetic synthesis of calcium carbonate with various polymorphs, sizes and morphologies by using organic substrates has become an interesting topic for the last years. Calcium carbonate has been synthesized by the reaction of Na2CO3 and CaCl2 in the presence of extract components of croaker gill. The products were characterized by powder X-ray diffraction (PXRD) and Fourier transform infrared (FT-IR) spectrum, and particle morphologies were observed by scanning electron microscope (SEM). The results show that at lower concentration yellow croaker gill extract has no effect on calcium carbonate crystal polymorph. Calcite was obtained only. But the morphologies of calcite particle change with the increase of the concentration. The corners of the particle change from angular to curved. However, with the further increase of the concentration of yellow croaker gill extract, the calcium carbonate obtained is a mixture of calcite and vaterite. The vaterite component in the mixture rises with increasing concentration of extract solution, indicating that the proteins from the yellow croaker gill during growth play a crucial role in stabilizing and directing the crystal growth., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Unlabelled: Iron deficiency is a yield-limiting factor with major implications for crop production, especially in soils with high CaCO3. Because stems are essential for the delivery of nutrients to the shoots, the aim of this work was to study the effects of Fe deficiency on the stem proteome of Medicago truncatula. Two-dimensional electrophoresis separation of stem protein extracts resolved 276 consistent spots in the whole experiment. Iron deficiency in absence or presence of CaCO3 caused significant changes in relative abundance in 10 and 31 spots, respectively, and 80% of them were identified by mass spectrometry. Overall results indicate that Fe deficiency by itself has a mild effect on the stem proteome, whereas Fe deficiency in the presence of CaCO3 has a stronger impact and causes changes in a larger number of proteins, including increases in stress and protein metabolism related proteins not observed in the absence of CaCO3. Both treatments resulted in increases in cell wall related proteins, which were more intense in the presence of CaCO3. The increases induced by Fe-deficiency in the lignin per protein ratio and changes in the lignin monomer composition, assessed by pyrolysis-gas chromatography-mass spectrometry and microscopy, respectively, further support the existence of cell wall alterations., Biological Significance: In spite of being essential for the delivery of nutrients to the shoots, our knowledge of stem responses to nutrient deficiencies is very limited. The present work applies 2-DE techniques to unravel the response of this understudied tissue to Fe deficiency. Proteomics data, complemented with mineral, lignin and microscopy analyses, indicate that stems respond to Fe deficiency by increasing stress and defense related proteins, probably in response of mineral and osmotic unbalances, and eliciting significant changes in cell wall composition. The changes observed are likely to ultimately affect solute transport and distribution to the leaves., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Although chitin, poly(N-acetylglucosamine), possesses considerable potential as a biomaterial, it has not been as thoroughly studied as its derivative chitosan. In this study, the potential of chitin gel beads has been evaluated for surface modification via vinyl polymer grafting. Grafting behavior of two well-established vinyl monomers, namely 2-hydroxyethylmethacrylate (HEMA) and 4-vinylpyridine (4-VP) were investigated using cerium (IV) ammonium nitrate as the redox initiator with the aim of obtaining chemically functionalized more hydrophilic chitin surfaces. The intractable nature of chitin, which is one of its primary drawbacks as a grafting substrate was overcome by applying a CaCO3 treatment during bead preparation. The maximum grafting percentage of poly(HEMA) onto chitin bead without CaCO3 treatment was found to be 65%, while the value for CaCO3 treated chitin beads was 515%. The maximum grafting yield of poly(4-VP) on to CaCO3 treated chitin powder was 380% at optimum conditions. The grafting system was extensively characterized before and after grafting by FT-IR, SEM, C-13 NMR and XRD analyses. Significant improvement on the swelling capacities of chitin based gel beads in aqueous acidic, basic and neutral media was obtained. An account of the pros and cons of the system has been presented., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The robust calcium carbonate composite ceramics (CC/PG) can be acquired by fast sintering calcium carbonate at a low temperature (650 °C) using a biocompatible, degradable phosphate-based glass (PG) as sintering agent. In the present study, the in vitro degradation and cell response of CC/PG were assessed and compared with 4 synthetic bone substitute materials, calcium carbonate ceramic (CC), PG, hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP) ceramics. The degradation rates in decreasing order were as follows: PG, CC, CC/PG, β-TCP, and HA. The proliferation of rat bone mesenchymal stem cells (rMSCs) cultured on the CC/PG was comparable with that on CC and PG, but inferior to HA and β-TCP. The alkaline phosphatase (ALP) activity of rMSCs on CC/PG was lower than PG, comparable with β-TCP, but higher than HA. The rMSCs on CC/PG and PG had enhanced gene expression in specific osteogenic markers, respectively. Compared to HA and β-TCP, the rMSCs on the CC/PG expressed relatively lower level of collagen I and runt-related transcription factor 2, but showed more considerable expression of osteopontin. Although CC, PG, HA, and β-TCP possessed impressive performances in some specific aspects, they faced extant intrinsic drawbacks in either degradation rate or mechanical strength. Based on considerable compressive strength, moderate degradation rate, good cell response, and being free of obvious shortcoming, the CC/PG is promising as another choice for bone substitute materials., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Lipase extracted from Rhizopus oryzae was immobilized in alginate gel beads. The effects of the immobilization conditions, such as, alginate concentration, CaCl2 concentration and amount of initial enzyme on retained activity (specific activity ratio of entrapped active lipase to free lipase) were investigated. The optimal conditions for lipase entrapment were determined: 2% (w/v) alginate concentration, 100mM CaCl2 and enzyme ratio of 2000IU/mL.In such conditions, immobilized lipase by inclusion in alginate showed a highest stability and activity, on olive oil hydrolysis reaction where it could be reused for 10 cycles. After 15min of hydrolysis reaction, the mass composition of monoolein, diolein and triolein were about 78%, 10% and 12%. Hydrolysis' products purification by column chromatography lead to a successful separation of reaction compounds and provide a pure fraction of monoolein which is considered as the widest used emulsifier in food and pharmaceutical industries., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The cellular effects of calcium carbonate (CaCO₃) nanoparticles were evaluated. Three kinds of CaCO₃ nanoparticles were employed in our examinations. One of the types of CaCO₃ nanoparticles was highly soluble. And solubility of another type of CaCO₃ nanoparticle was lower. A stable CaCO₃ nanoparticle medium dispersion was prepared and applied to human lung carcinoma A549 cells and human keratinocyte HaCaT cells. Then, mitochondrial activity, cell membrane damage, colony formation ability, DNA injury, induction of oxidative stress, and apoptosis were evaluated. Although the influences of CaCO₃ nanoparticles on mitochondrial activity and cell membrane damage were small, "soluble" CaCO₃ nanoparticles exerted some cellular influences. Soluble CaCO₃ nanoparticles also induced a cell morphological change. Colony formation was inhibited by CaCO₃ nanoparticle exposure. In particular, soluble CaCO₃ nanoparticles completely inhibited colony formation. The influence on intracellular the reactive oxygen species (ROS) level was small. Soluble CaCO₃ nanoparticles caused an increase in C/EBP-homologous protein (CHOP) expression and the activation of caspase-3. Moreover, CaCO₃ exposure increased intracellular the Ca²⁺ level and activated calpain. These results suggest that cellular the influences of CaCO₃ nanoparticles are mainly caused by intracellular calcium release and subsequently disrupt the effect of calcium signaling. In conclusion, there is possibility that soluble CaCO₃ nanoparticles induce cellular influences such as a cell morphological change. Cellular influence of CaCO₃ nanoparticles is caused by intracellular calcium release. If inhaled CaCO₃ nanoparticles have the potential to influence cellular events. However, the effect might be not severe because calcium is omnipresent element in cell., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Sulcona, a Siddha proprietary medicine used for the treatment of burns, has been in practice for more than 50 years. This medicine has been successfully used on several burned patients with an excellent recovery and safety record. In this manuscript, we investigate some of its pharmacological and safety profiles. Treatment of cells with Sulcona induced a statistically significant increase in population doubling compared to concurrent controls in proliferating human lymphocytes as well as in Balb/c 3T3 cells, suggesting that it stimulates cell proliferation. Sulcona exhibited some antibacterial activity against Pseudomonas aeruginosa, Salmonella typhi and Staphylococcus aureus. Carrageenan-induced rat paw edema testing suggested that Sulcona has some anti-inflammatory properties. Patch testing showed that Sulcona has mild anesthetic effects. The above properties suggest Sulcona's pharmacological properties aidin treatment of burns. Sulcona did not show any skin irritation or sensitization or mutagenic potential suggesting that it is safe for use. Further work is necessary to elucidate its exact mechanisms of action., (Copyright © 2013 Elsevier Ltd and ISBI. All rights reserved.)

There is little consensus as to the most appropriate methodology for the measurement of gastrointestinal pH in chickens. An experiment was conducted to establish the optimum sampling method for the determination of broiler digesta pH in birds fed differing levels of dietary calcium. Ross 308 broilers (n = 60) were fed 1 of 2 experimental diets, one containing 0.8% monocalcium phosphate and 2% limestone and one containing 0.4% monocalcium phosphate and 1% limestone. Four factors were investigated to determine the most appropriate method of measuring broiler gastrointestinal digesta pH: removal from the tract, prolonged air exposure, altering the temperature of the assay, and controlling the water content of the digesta. The conditions were assessed at bird ages from 7 to 42 d posthatch. Dietary Ca content had no significant effect on in situ pH, but it contributed toward variance in ex situ pH of both gizzard and duodenum digesta. Digesta pH read higher when the digesta was removed from the tract, but the amount of time the digesta was exposed to air did not affect the reading. Digesta pH read higher when measured at room temperature than when measured at 41°C; temperature made the strongest unique contribution to explaining variance in duodenum pH, and the second strongest contribution to explaining variance in gizzard pH, after diet. When water was added to the digesta, before pH determination, the pH of the digesta read higher (P < 0.001) than when measured in situ. The method that resulted in pH readings that were most representative of bird gastrointestinal environment was insertion of a pH probe directly into the gut lumen posteuthanasia, because measurement ex situ likely encourages dissociation of carbonic acid, the major buffer in the gastrointestinal tract, which causes pH to read to be higher than when measured in situ. This study shows that the method of pH measurement needs careful consideration to ensure the validity of the result.

In the present paper, the impact of freshwater (ARC21 and LS0519) and marine (PCC8806) Synechococcus cyanobacteria on calcium carbonate (CaCO3) precipitation has been examined in respect of the formation rates and morphology of crystals. Acid-base potentiometric titrations were employed to study surface functional groups, while CaCO3 experiments have been carried out in presence and absence of cells at low to near-equilibrium conditions in respect to CaCO3. During these experiments, the pH values have been monitored, Ca and alkalinity were measured and precipitates have been investigated by Raman spectroscopy and Atomic Force and Scanning Electron microscopy. Our results showed that the Synechococcus strains exhibited different surface reactivity with total concentration of surface functional groups of 0.342 and 0.350 mMg(-1) of dry bact. for freshwater strains, and 0.662 mMg(-1) of dry bact. for the marine strain, which are on the same order of magnitude as that reported for bacterial cell surfaces. The marine strain showed the highest CaCO3 formation rate with Ca(2+) removal of 18 mMg(-1) dry bact. compared to 6-7 mMg(-1) dry bact. for freshwater strains. The morphological diversity in crystals has been linked to presence of specific functional groups. The linking cell surface properties to crystal morphologies and precipitation rates propose that bacterial surfaces may modulate CaCO3 formation. Results of this work should allow better understanding of biominiralization in marine and freshwater systems as they define the precipiatation rates in typical range of pH necessary for estimation of CaCO3 formation by cyanobacterial communities., (Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.)

Implant-associated infection is a serious problem in orthopaedic surgery. One of the most effective ways is to introduce a controlled antibiotics delivery system into the bone filling materials, achieving sustained release of antibiotics in the local sites of bone defects. In the present work, mesoporous carbonated hydroxyapatite microspheres (MCHMs) loaded with gentamicin have been fabricated according to the following stages: (i) the preparation of the MCHMs by hydrothermal method using calcium carbonate microspheres as sacrificial templates, and (ii) loading gentamicin into the MCHMs. The MCHMs exhibit the 3D hierarchical nanostructures constructed by nanoplates as building blocks with mesopores and macropores, which make them have the higher drug loading efficiency of 70-75% than the conventional hydroxyapatite particles (HAPs) of 20-25%. The gentamicin-loaded MCHMs display the sustained drug release property, and the controlled release of gentamicin can minimize significantly bacterial adhesion and prevent biofilm formation against S. epidermidis. The biocompatibility tests by using human bone marrow stromal cells (hBMSCs) as cell models indicate that the gentamicin-loaded MCHMs have as excellent biocompatibility as the HAPs, and the dose of the released gentamicin from the MCHMs has no toxic effects on the hBMSCs. Hence, the gentamicin-loaded MCHMs can be served as a simple, non-toxic and controlled drug delivery system to treat bone infections., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Objectives: To evaluate the influence of desensitising paste containing 8% arginine and calcium carbonate (Ar-Ca) on biofilm formation on dentine., Methods: Dentine discs were cut from extracted third molars and divided into the following three groups: no treatment, pumice treatment and Ar-Ca treatment. Surface topography and roughness were examined using scanning electron microscopy (SEM) and non-contact 3D surface profiler. After sterilisation, samples were incubated with Streptococcus mutans (S. mutans) for 4 h, 24 h and 72 h. Bacterial adhesion and biofilm formation were analysed using SEM, whereas MTT and lactic acid production assays were used to analyse the metabolic activity of S. mutans., Results: After polishing with either pumice or Ar-Ca, the surfaces of the samples became smoother than in the control group. The Ra values of the three experimental groups decreased significantly to 0.43 μm, 0.3 μm and 0.26 μm, respectively. Compared to the control group, fewer bacteria adhered to the dentine surface in the Ar-Ca group, while biofilm thickness decreased significantly for both groups after incubating for 24 h and 72 h. MTT and lactic acid production levers also showed a significant reduction in the Ar-Ca group., Conclusions: Ar-Ca appears to present antibiofilm efficacy and may provide a promising approach to combat bacterial infection in hypersensitive dentinal lesions., Clinical Significance: As a clinical application of desensitising polishing paste, the paste containing 8% arginine and calcium carbonate could also inhibit the biofilm formation effectively., (Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.)

Objective: The mode of action of an arginine mouthwash using the Pro-Argin™ Mouthwash Technology, containing 0.8% arginine, PVM/MA copolymer, pyrophosphates and 0.05% sodium fluoride, has been proposed and confirmed as occlusion using a variety of in vitro techniques., Methods: Quantitative and qualitative laboratory techniques were employed to investigate the mode of action of the new arginine mouthwash. Confocal laser scanning microscopy (CSLM) and atomic force microscopy (AFM) investigated a hydrated layer on dentine surface. Electron spectroscopy for chemical analysis (ESCA), secondary ion mass spectroscopy (SIMS) and near-infrared spectroscopy (NIR) provided information about its chemical nature., Results: CLSM was used to observe the formation of a hydrated layer on exposed dentine tubules upon application of the arginine mouthwash. Fluorescence studies confirmed penetration of the hydrated layer in the inner walls of the dentinal tubules. The AFM investigation confirmed the affinity of the arginine mouthwash for the dentine surface, supporting its adhesive nature. NIR showed the deposition of arginine after several mouthwash applications, and ESCA/SIMS detected the presence of phosphate groups and organic acid groups, indicating the deposition of copolymer and pyrophosphates along with arginine., Conclusion: The studies presented in this paper support occlusion of the dentine surface upon the deposition of an arginine-rich layer together with copolymer and phosphate ions from an alcohol-free mouthwash containing 0.8% arginine, PVM/MA copolymer, pyrophosphates and 0.05% sodium fluoride., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Close to the bone mineral phase, the calcic bioceramics, such as hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP), are commonly used as substitutes or filling materials in bone surgery. Besides, calcium carbonate (CaCO3) is also used for their excellent biocompatibility and bioactivity. However, the problem with the animal-origin aragonite demands the new technique to synthesize pure calcite capable of forming 3D bone implant. This study aims to manufacture and evaluate a highly-pure synthetic crystalline calcite with good cytocompatibility regarding to the osteoblasts, comparing to that of HA and β-TCP. After the manufacture of macroporous bioceramic scaffolds with the identical internal architecture, their cytocompatibility is studied through MC3T3-E1 osteoblasts with the tests of cell viability, proliferation, vitality, etc. The results confirmed that the studied process is able to form a macroporous material with a controlled internal architecture, and this synthesized calcite is non-cytotoxic and facilitate the cell proliferation. Indeed requiring further improvement, the studied calcite is definitely an interesting alternative not only to coralline aragonite but also to calcium phosphate ceramics, particularly in bone sites with the large bone remodelling., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Antibacterial activity of photocatalytic substrates is primarily induced by ultraviolet light irradiation. Visible light-responsive photocatalysts were recently discovered, offering greater opportunity to use photocatalysts as disinfectants in our living environment. The development of antibacterial photocatalysts, however, has mainly focused on titanium oxide (TiO(2))-related materials with antibacterial properties not comparable with conventional chemical disinfectants. This study demonstrated that a core-shell structured In(2)O(3)@CaIn(2)O(4) substrate has superior visible light-induced bactericidal properties, as compared with several commercially available and laboratory-prepared visible light-responsive photocatalysts. The high performance is enhanced by more easily photoexcited electron transfer between the interfaces of In(2)O(3) and CaIn(2)O(4) to minimize the electron-hole recombination during photocatalysis. Additionally, when compared with TiO(2)-based photocatalysts, In(2)O(3)@CaIn(2)O(4) treatments did not induce significant cell death and tissue damage, implying a superior biocompatibility. These findings suggest that In(2)O(3)@CaIn(2)O(4) may have potential application in the development of a safer and highly bactericidal photocatalyst., From the Clinical Editor: A photocatalytic susbstrate is described that functions in visible light, possesses bactericidal properties and better biocompatibility than the standard TiO(2) based methods., (Copyright © 2012 Elsevier Inc. All rights reserved.)

A metal-arsenic polluted soil from sulphide-mine waste was treated, in all possible combinations, with two different amounts of marble sludge (98% CaCO3), compost (41% organic carbon), and Byferrox (70% Fe). Lixiviate and pore water from each treated and untreated soil were analysed, and lettuce-seed bioassays were performed. None of the treatments decreased the electrical conductivity of lixiviates or the concentrations of all pollutants found in both solutions. Marble sludge and compost increased the pH values and decreased the zinc, cadmium, copper, and lead concentrations in both solutions while increasing the arsenic concentrations in the lixiviates. Byferrox did not alter the physicochemical parameters or the concentrations of zinc, cadmium, copper, or lead in either solution but significantly decreased the arsenic concentrations in pore water. Compared with the Byferrox treatment, the mixture of marble sludge and Byferrox decreased redox potential values, increasing the arsenic concentrations in both solutions and the electrical conductivity of the pore water. All lixiviates were highly phytotoxic and seeds did not germinate. Pore-water phytotoxicity was related to electrical conductivity values and heavy-metal concentrations. The combination of marble sludge and compost was most effective at diminishing toxicity in lettuce. The soils treated with Byferrox, alone or mixed with marble sludge or compost, were the most phytotoxic., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The present work underlined the negative effects of increasing CaCO(3) concentrations (5, 10 and 20 mM) both on the chicory root growth and the arbuscular mycorrhizal fungus (AMF) Glomus irregulare development in monoxenic system. CaCO(3) was found to reduce drastically the main stages of G. irregulare life cycle (spore germination, germinative hyphae elongation, root colonization, extraradical hyphae development and sporulation) but not to inhibit it completely. The root colonization drop was confirmed by the decrease in the arbuscular mycorrhizal fungal marker C16:1ω5 amounts in the mycorrhizal chicory roots grown in the presence of CaCO(3). Oxidative damage evaluated by lipid peroxidation increase measured by (i) malondialdehyde (MDA) production and (ii) the antioxidant enzyme peroxidase (POD) activities, was highlighted in chicory roots grown in the presence of CaCO(3). However, MDA formation was significantly higher in non-mycorrhizal roots as compared to mycorrhizal ones. This study pointed out the ability of arbuscular mycorrhizal symbiosis to enhance plant tolerance to high levels of CaCO(3) by preventing lipid peroxidation and so less cell membrane damage., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

This paper demonstrates a general approach for fabrication of lactobionic chitosan microcapsules using layer-by-layer assembly via click chemistry. Chitosan was selectively modified with either azide (CHI-Az) or alkyne (CHI-Alk) groups. The growth of the CHI-Az/CHI-Alk click multilayer was studied experimentally by multilayer assembly on planar supports. Linear buildup of the film was observed. The chitosan click capsules were also analyzed with confocal laser scanning microscopy and transmission electron microscopy. Capsules were found to have regular spherical shapes. In addition, (CHI-Az/CHI-Alk)-coated particles were modified with fluorescein isothiocyanate to ensure that the particles can be easily post-functionalized. Finally, lactobionic acid was conjugated onto the (CHI-Az/CHI-Alk)-coated particles and the lactobionic particles exhibited hepatoma cell (HepG2) targeting behavior., (Copyright © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Silicon-releasable microfiber meshes consisting of silicon-doped vaterite (SiV) particles and poly(lactic acid) (PLA) hybrids were prepared by electrospinning. Due to their flexibility and porosity they formed ideal membranes or scaffolds for guided bone regeneration. In addition, a trace amount of silicon species has been reported to stimulate osteogenic cells to mineralize and enhance bone formation. We propose a new method of preparation of silicon-releasing microfiber meshes by electrospinning. Their structure and hydroxyapatite (HA)-forming abilities in simulated body fluid were examined. In addition, we studied their stimulatory effects on osteoblast-like cells in vitro and bone-forming ability in vivo, with a special emphasis on their ability to release silicon. The meshes consisted of a hybrid of carboxy groups in PLA and amino groups in siloxane, derived from aminopropyltriethoxysilane or calcium ions on the SiV surface. This hybrid exhibited an enhanced ability to form HA. The meshes coated with HA released 0.2-0.7 mg l(-1) silicon species into the culture medium over 7 days. Enhanced proliferation of osteoblast-like cells was observed using the meshes and new bone formed on the meshes when implanted into the calvaria of rabbits. These meshes, therefore, provide an excellent substrate for bone regeneration and exhibit enhanced bone-forming ability under both in vitro and in vivo conditions., (Copyright 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Calcium (Ca) supplements, especially Ca carbonate (CaCO3), are the main alternative sources of dietary Ca and an important part of a treatment regimen for osteoporosis, the most common metabolic bone disorder of aging and menopause. In a female ovariectomized (OVX) rat model for studying postmenopausal osteoporosis, we tested the hypothesis that a small compared with a large particle size of CaCO3 (13.0- vs. 18.5-mum geometric diameter) would result in increased Ca balance and subsequently bone mass and that this would be affected by dietary Ca level. We used 6-mo-old rats that were OVX either at 6 or 3 mo of age as models of early or stable menopausal status, respectively. The rats received semipurified diets that contained either 0.4 or 0.2% dietary Ca provided from CaCO3 of 2 particle sizes. A group of Sham-operated rats with intact ovaries served as control and were fed 0.4% dietary Ca from large particles. Estrogen deficiency as a result of ovariectomy had an adverse effect on bone density, mineral content, and bone mechanical properties (P < 0.001). Reducing dietary Ca from 0.4 to 0.2% resulted in significant adverse effects on bone density and mechanical properties (P < 0.001). The particle size of CaCO3 did not affect total Ca balance, bone dual energy X-ray absorptiometry and peripheral quantitative computed tomography indices, bone ash and Ca content, or the mechanical determinants of bone strength. We conclude that a decrease in particle size of CaCO3 to 70% of that typically found in Ca supplements does not provide a benefit to overall Ca metabolism or bone characteristics and that the amount of Ca consumed is of greater influence in enhancing Ca nutrition and skeletal strength.

A total of 1,152 Lohmann Brown laying hens were used to study the influence of level (3.5 and 4.0%) and source (coded FIN, COA, and OYS) of Ca in the diet on productive performance and egg quality from 58 to 73 wk of age. The FIN diet contained all the Ca carbonate as fine limestone (LIM). In the COA and OYS diets, 40% of the fine LIM was substituted with either coarse LIM or oyster shell. Each treatment was replicated 8 times (24 hens). Productive performance and egg quality traits were recorded every 4 wk, and tibia characteristics and shell quality traits were determined at 73 wk of age. An increase in Ca intake from 4.08 to 4.64 g/hen per day improved egg production (71.2 vs. 74.9%; P < 0.001), egg mass (49.0 vs. 51.4 g; P < 0.05), and feed conversion ratio (2.43 vs. 2.30 kg of feed/kg of egg; P < 0.001). In addition, an increase in Ca intake improved shell weight (9.98 vs. 10.20%; P < 0.05), shell thickness (0.342 vs. 0.351 mm; P < 0.01), and shell density (82.0 vs. 83.8 mg/cm2; P < 0.001). Calcium source had no effect on productive performance, tibia characteristics, or egg quality except for shell density, which was greater for hens fed COA than for hens fed FIN, with hens fed OYS being intermediate (81.9 vs. 84.0 vs. 82.7 mg/cm2, respectively; P < 0.05). It was concluded that Brown egg-laying hens in the late phase of production require more than 3.5% Ca in the diet (4.08 g of Ca/hen per day) and that the substitution of 40% of fine LIM with COA or OYS does not affect productive performance and has little impact on shell quality and tibia characteristics.

Background: Low-energy fractures of the hip, forearm, shoulder, and spine are known consequences of osteoporosis., Objective: We evaluated the effect of 1 y of treatment with calcium and vitamin D on bone mineral density (BMD) and bone markers in patients with a recent low-energy fracture., Design: In a double-blinded design, patients with fracture of the hip (lower-extremity fracture, or LEF) or upper extremity (UEF) were randomly assigned to receive 3000 mg calcium carbonate + 1400 IU cholecalciferol or placebo (200 IU cholecalciferol). BMD of the hip (HBMD) and lumbar spine (LBMD) were evaluated by dual-energy X-ray absorptiometry, and physical performance was assessed by the timed Up & Go test. Serum concentrations of 25-hydroxycholecalciferol, parathyroid hormone (PTH), telepeptide of type I collagen (ICTP), osteocalcin, and N-terminal propeptide of collagen type I were measured., Results: A total of 122 patients were included (84% women; x +/- SD age: 70 +/- 11 y); 68% completed the study. In an intention-to-treat analysis, LBMD increased in the intervention group and decreased in the placebo group, and the difference between the groups was significant after 12 mo: 0.931 +/- 0.211 compared with 0.848 +/- 0.194 (P<0.05). No significant change was shown for HBMD. The effect of treatment was more pronounced in patients aged <70 y. The intervention decreased bone turnover. PTH was significantly lower in the intervention group (P<0.01) for the LEF patients. ICTP and change in LBMD were significantly related to physical performance., Conclusions: A 1-y intervention with calcium and vitamin D reduced bone turnover, significantly increased BMD in patients younger than 70 y, and decreased bone loss in older patients. The effect of treatment was related to physical performance.

Background: Levofloxacin is used in adult patients with cystic fibrosis but its pharmacokinetics is not well characterized in this population. Patients with cystic fibrosis use calcium routinely to prevent osteoporosis. A slower intestinal transit time is common in cystic fibrosis implying that the standard 2-h spacing of minerals and levofloxacin to prevent a chelation interaction may be insufficient. The objectives of this study were to characterize the steady state pharmacokinetics of oral levofloxacin 750 mg with and without 2-h spaced calcium carbonate in patients with cystic fibrosis compared to matched healthy volunteers., Methods: In an open-label, randomized, cross-over study of five patients with cystic fibrosis and five age, sex, race, and serum creatinine matched healthy volunteers received 750 mg of oral levofloxacin alone daily for 5 days and the same dose of levofloxacin with 2-h spaced calcium carbonate supplementation 500 mg po thrice daily with meals in random sequence. Blood was collected for plasma assay of levofloxacin pre-dose, 0.5, 1, 1.5, 2, 4, 8, 12, and 24h after the fifth levofloxacin dose., Results: There was no significant interaction in healthy volunteers, however, when cystic fibrosis patients were given levofloxacin with 2-h spaced calcium, the maximum plasma concentration (Cmax) decreased by 19% and time to Cmax increased by 37% (p<0.05). This difference in peak concentrations resulted in a lack of bioequivalence (Cmax geometric mean ratio 81.6%, 90% confidence intervals: 71.8%, 91.4%) even when levofloxacin and calcium supplements were spaced by the standard 2h administration instruction in patients with cystic fibrosis., Conclusions: These results indicate that multivalent cations such as calcium should be maximally separated from oral levofloxacin administration in adult patients with cystic fibrosis to prevent this drug interaction, thereby better optimizing antibiotic efficacy and decreasing the potential for resistance development.

Background: In chronic renal failure (CRF), hyperphosphatemia and an elevated calcium-phosphate product are associated with vascular calcification and increased cardiovascular morbidity and mortality. Previous data have demonstrated that 3-month treatment of uremic rats with sevelamer was associated with less nephrocalcinosis compared to calcium carbonate (CaCO3), despite similar control of serum phosphorus, calcium-phosphorus product (Ca x P product), and secondary hyperparathyroidism. There was no evidence of aortic calcification after 3 months of uremia (J Am Soc Nephrol 13:2299-2308, 2002). The present studies explore the influence of sevelamer and CaCO3 on cardiovascular and kidney calcifications in long-term experimental uremia over 6 months., Methods: Normal and 5/6 nephrectomized rats (U) were fed a high phosphorus (HP) diet for 6 months. Two phosphate binders, CaCO3 and sevelamer, were administered and their influence on hyperphosphatemia, secondary hyperparathyroidism, kidney/myocardial/aortic calcification, and renal function was compared., Results: All uremic rats began the study with the same degree of renal failure. Sevelamer was as effective as CaCO3 in reducing serum phosphorus, Ca x P product, and attenuating secondary hyperparathyroidism. Despite similar serum cholesterol levels, rats in the U-HP + sevelamer group had markedly lower calcium deposition in the myocardium and aorta (myocardium, 72 +/- 4 microg/g wet tissue; aorta, 736 +/- 156 microg/g wet tissue) compared to rats in either the U-HP + CaCO3 group (myocardium, 179 +/- 48, P < 0.05; aorta, 1308 +/- 343, P < 0.05) or the U-HP group (myocardium, 98 +/- 10, NS; aorta, 2150 +/- 447, P < 0.05). Dual immunohistochemical analysis for calcium and endothelial cell markers demonstrated that myocardial calcium deposition was intravascular within capillaries. Furthermore, calcium deposition in the kidney of uremic rats treated with sevelamer (582 +/- 111 microg/g wet tissue) was lower than that found in uremic rats treated with CaCO3 (1196 +/- 180 microg/g wet tissue). Sevelamer-treated rats had less deterioration in renal function with an associated lower serum creatinine, higher creatinine clearance, and less proteinuria. There was no difference in overall mortality between the three experimental groups., Conclusion: In long-term experimental CRF, in addition to controlling serum phosphorus and secondary hyperparathyroidism as efficiently as CaCO3, treatment with the phosphate-binder sevelamer attenuates vascular and kidney calcification.

Broilers chickens have limited ability to utilize phytate phosphorus (PP), and the influence of nutrients on the activity and efficacy of intestinal phytase is unclear. Therefore in a 2 x 2 x 2 factorial experiment, male chicks were fed 0 or 0.21 mg/kg 25-hydroxycholecalciferol (25-OH D3), 4 or 9 g/kg Ca supplied from CaCO3 or Ca malate from 14 to 24 d of age (six pens/treatment, eight birds/pen). Source of Ca had no effect on tibia ash, intestinal phytase activity, or apparent ileal PP hydrolysis. Phytase activity (Vmax) within brush border vesicles prepared from small intestinal mucosa was greater in chicks fed 4 vs. 9 g/kg Ca (P < or = 0.05). Similarly, birds fed 4 vs. 9 g/kg Ca were able to hydrolyze an additional 24.36% PP (P < 0.01). Intestinal phytase activity and apparent ileal PP hydrolysis were not affected by 25-OH D3 supplementation, but tibia ash was improved by 2.7% (P < 0.01). A subsequent experiment validated the effect of dietary Ca (4 and 9 g/kg) and elucidated differences between strains (Ross 308 and Hubbard x Peterson) from 8 to 22 d of age (six pens/treatment, eight birds/pen). The strains responded similarly to dietary Ca in terms of intestinal phytase and apparent ileal PP hydrolysis. Intestinal phytase activity was 9% greater in birds fed 4 vs. 9 g/kg Ca (P < 0.05) and apparent ileal PP hydrolysis was 11.9% greater (P < 0.02). In conclusion, a typical dietary Ca concentration (9 g/kg) reduced intestinal phytase activity and apparent ileal PP hydrolysis.

Cooperativity between pepsin and crystallization of calcium carbonate in distilled water was studied. The results show that vaterite was formed under the influence of pepsin and the crystalline product was a composite of vaterite and pepsin. The component of this material was similar to that of nacre. At the same time, the crystallization of calcium carbonate had also an important effect on the secondary structure of the pepsin. The secondary structure of the pepsin was characterized through FT-IR technology. The result indicated that the pure pepsin is composed of 24.38% alpha-helices, 29.91% beta-sheets, 39.32% beta-turns and 6.49% random structures and the pepsin in the CaCO(3)-pepsin solution is composed of 2.09% alpha-helices, 93.304% beta-sheets, 4.592% beta-turns and 0.006% random structure. During the crystallization of the calcium carbonate from the pepsin solution, the secondary structure of the pepsin transformed. These results showed that there was cooperativity between the crystallization of vaterite and the pepsin. The cooperative mechanism is discussed.

The treatment of secondary hyperparathyroidism (HPT) in patients with chronic renal disease has improved markedly in recent years. The skeletal pain, disabling fractures, tendon ruptures, and myriad other symptoms associated with HPT can now be avoided, and the quality of life of patients with end-stage renal disease is improved. Control of hyperphosphatemia, maintenance of normocalcemia, and appropriate dosing of vitamin D analogues can prevent HPT in many cases. Palatable, nutritious diets should be followed; serum calcium, phosphorus, alkaline phosphatase, and parathyroid hormone should be monitored; and treatment regimens should be adjusted accordingly. If prevention fails, and even if severe HPT develops, many of these patients can still be controlled medically with correction of hyperphosphatemia and high doses of intravenous calcitriol. In our experience, only a few patients require surgical parathyroidectomy (usually noncompliant patients or patients whose HPT has been poorly managed from early uremia). The essence to medical management is to correct the two most important pathogenetic factors of HPT, hyperphosphatemia, and calcitriol deficiency. We present the current approach to the management of HPT, with highlights of recent advances.

This study was designed to examine the effect of 500 to 5,000 mg of ascorbic acid on DNA adducts, natural killer (NK) cell activity, programmed cell death, and cell cycle analysis of human peripheral blood leukocytes. According to our hypothesis, if ascorbic acid is a pro-oxidant, doses between 500 and 5,000 mg should enhance DNA adduct formation, decrease immune function, change the cell cycle progression, and increase the rate of apoptosis. Twenty healthy volunteers were divided into four groups and given either placebo or daily doses of 500, 1,000 or 5,000 mg of ascorbic acid for a period of 2 weeks. On days 0, 1, 7, 15, and 21, blood was drawn from them, and the leukocytes were separated and examined for intracellular levels of ascorbic acid, the level of 8-hydroxyguanosine, NK cell activity, cell cycle progression, and apoptosis. Depending on the subjects, between a 0% and a 40% increase in cellular absorption of ascorbic acid was observed when daily doses of 500 mg were used. At doses greater than 500 mg, this cellular absorption was not increased further, and all doses produced equivalent increases in ascorbic acid on days 1 to 15. This increase in cellular concentration of ascorbic acid resulted in no statistically meaningful changes in the level of 8-hydroxyguanosine, increased NK cytotoxic activity, a reduced percentage of cells undergoing apoptosis, and switched cell cycle phases from S and G2/M to G0/G1. After a period of 1 week, with no placebo or vitamin washout, ascorbic acid levels along with functional assays returned to the baseline and became equivalent to placebos. In comparison with baseline values, no change (not more than daily assays variation) was seen in ascorbate concentrations or other assays during oral placebo treatment. We concluded that ascorbic acid is an antioxidant and that doses up to 5,000 mg neither induce mutagenic lesions nor have negative effects on NK cell activity, apoptosis, or cell cycle.

The lipid peroxidative effects of copper sulfate singly (4 mg/l CuSO4.5H2O) and in combination with calcium carbonate (4 mg/l CuSO4.5H2O + 50 mg/l CaCO3) were determined in the liver of the African tilapia Oreochromis mossambicus following exposures of the fish to the chemicals for 24, 48, 72, 96 and 168 h. Lipid peroxidative effects of the treatment with calcium carbonate (50 mg/l CaCO3) and with a known hepatotoxicant, carbon tetrachloride (0.25 ml/l CCl4) were also determined. Fish not exposed to any chemical served as negative controls. The extent of lipid peroxidation was based on hepatic malondialdehyde (MDA) levels as assayed using the thiobarbituric acid reaction test. Results suggested the lipid peroxidative property of the copper salt which was associated with the toxic nature of the heavy metal, although, this effect was not as potent as that of CCl4. Findings also indicated a measure of protection against copper hepatotoxicity provided by the addition of calcium carbonate as a liming agent in the water.