Neutrophil Extracellular Traps Initiate Gallstone Formation.

The presence of gallstones (cholelithiasis) is a highly prevalent and severe disease and one of the leading causes of hospital admissions worldwide. Due to its substantial health impact, we investigated the biological mechanisms that lead to the formation and growth of gallstones. We show that gallstone assembly essentially requires neutrophil extracellular traps (NETs). We found consistent evidence for the presence of NETs in human and murine gallstones and describe an immune-mediated process requiring activation of the innate immune system for the formation and growth of gallstones. Targeting NET formation via inhibition of peptidyl arginine deiminase type 4 or abrogation of reactive oxygen species (ROS) production, as well as damping of neutrophils by metoprolol, effectively inhibit gallstone formation in vivo. Our results show that after the physicochemical process of crystal formation, NETs foster their assembly into larger aggregates and finally gallstones. These insights provide a feasible therapeutic concept to prevent cholelithiasis in patients at risk.

Calcium carbonate mineralization mediated by in vitro cultured mantle cells from Pinctada fucata.

Formation of the molluscan shell is believed to be an extracellular event mediated by matrix proteins. We report calcium carbonate mineralization mediated by Pinctada fucata mantle cells. Crystals only appeared when mantle cells were present in the crystallization solution. These crystals were piled up in highly ordered units and showed the typical characteristics of biomineralization products. A thin organic framework was observed after dissolving the crystals in EDTA. Some crystals had etched surfaces with a much smoother appearance than other parts. Mantle cells were observed to be attached to some of these smooth surfaces. These results suggest that mantle cells may be directly involved in the nucleation and remodeling process of calcium carbonate mineralization. Our result demonstrate the practicability of studying the mantle cell mechanism of biomineralization and contribute to the overall understanding of the shell formation process.

Different calcium sources affect the products and sites of mineralized Cr(VI) by microbially induced carbonate precipitation.

Microbially induced carbonate precipitation (MICP) is a common biomineralization method, which is often used for remediation of heavy metal pollution such as hexavalent chromium (Cr(VI)) in recent years. Calcium sources are essential for the MICP process. This study investigated the potential of MICP technology for Cr(VI) remediation under the influence of three calcium sources (CaCl2, Ca(CH3COO)2, Ca(C6H11O7)2). The results indicated that CaCl2 was the most efficient in the mineralization of Cr(VI), and Ca(C6H11O7)2 could significantly promote Cr(VI) reduction. The addition of different calcium sources all promoted the urease activity of Sporosarcina saromensis W5, in which the CaCl2 group showed higher urease activity at the same Ca2+ concentration. Besides, with CaCl2, Ca(CH3COO)2 and Ca(C6H11O7)2 treatments, the final fraction of Cr species (Cr(VI), reduced Cr(III) and organic Cr(III)-complexes) were mainly converted to the carbonate-bound, cytoplasm and cell membrane state, respectively. Furthermore, the characterization results revealed that three calcium sources could co-precipitate with Cr species to produce Ca10Cr6O24(CO3), and calcite and vaterite were present in the CaCl2 and Ca(CH3COO)2 groups, while only calcite was present in the Ca(C6H11O7)2 group. Overall, this study contributes to the optimization of MICP-mediated remediation of heavy metal contaminated soil. CaCl2 was the more suitable calcium source than the other two for the application of MICP technology in the Cr(VI) reduction and mineralization.

Advances in the identification of calcium carbonate urinary crystals.

The examination of the urinary sediment of a 64-year-old woman showed the presence of three different types of crystals, all with unusual morphology, which could not be identified with bright field microscopy, polarized light, and the knowledge of urine pH (7.5). The use of microscopic infrared spectroscopy, Raman spectroscopy and energy dispersive X-ray spectroscopy led to the identification of the three types of crystals as calcite, vaterite and aragonite, which are all variants of calcium carbonate crystals. This paper confirms the complex morphology and nature that urinary crystals may at times have and the utility of advanced infrared spectroscopy techniques for their identification.

Composite Magnetite and Protein Containing CaCO3 Crystals. External Manipulation and Vaterite → Calcite Recrystallization-Mediated Release Performance.

Biocompatibility and high loading capacity of mesoporous CaCO3 vaterite crystals give an option to utilize the polycrystals for a wide range of (bio)applications. Formation and transformations of calcium carbonate polymorphs have been studied for decades, aimed at both basic and applied research interests. Here, composite multilayer-coated calcium carbonate polycrystals containing Fe3O4 magnetite nanoparticles and model protein lysozyme are fabricated. The structure of the composite polycrystals and vaterite → calcite recrystallization kinetics are studied. The recrystallization results in release of both loaded protein and Fe3O4 nanoparticles (magnetic manipulation is thus lost). Fe3O4 nanoparticles enhance the recrystallization that can be induced by reduction of the local pH with citric acid and reduction of the polycrystal crystallinity. Oppositely, the layer-by-layer assembled poly(allylamine hydrochloride)/poly(sodium styrenesulfonate) polyelectrolyte coating significantly inhibits the vaterite → calcite recrystallization (from hours to days) most likely due to suppression of the ion exchange giving an option to easily tune the release kinetics for a wide time scale, for example, for prolonged release. Moreover, the recrystallization of the coated crystals results in formulation of multilayer capsules keeping the feature of external manipulation. This study can help to design multifunctional microstructures with tailor-made characteristics for loading and controlled release as well as for external manipulation.