Insights into the In Vitro Formation of Apatite from Mg-Stabilized Amorphous Calcium Carbonate

A protein-free formation of bone-like apatite from amorphous precursors through ball-milling is reported. Mg²⁺ ions are crucial to achieve full amorphization of CaCO₃. Mg²⁺ incorporation generates defects which strongly retard a recrystallization of ball-milled Mg-doped amorphous calcium carbonate (BM-aMCC), which promotes the growth of osteoblastic and endothelial cells in simulated body fluid and has no effect on endothelial cell gene expression. Ex situ snapshots of the processes revealed the reaction mechanisms. For low Mg contents (<30%) a two phase system consisting of Mg-doped amorphous calcium carbonate (ACC) and calcite “impurities” was formed. For high (>40%) Mg²⁺ contents, BM-aMCC follows a different crystallization path via magnesian calcite and monohydrocalcite to aragonite. While pure ACC crystallizes rapidly to calcite in aqueous media, Mg-doped ACC forms in the presence of phosphate ions bone-like hydroxycarbonate apatite (dahllite), a carbonate apatite with carbonate substitution in both type A (OH⁻) and type B (PO₄³⁻) sites, which grows on calcite “impurities” via heterogeneous nucleation. This process produces an endotoxin-free material and makes BM-aMCC an excellent “ion storage buffer” that promotes cell growth by stimulating cell viability and metabolism with promising applications in the treatment of bone defects and bone degenerative diseases.     

Enhancing the Selectivity and Transparency of the Electron Contact in Silicon Heterojunction Solar Cells by Phosphorus Catalytic Doping

An intrinsic hydrogenated amorphous silicon (a-Si:H(i)) film and a doped silicon film are usually combined in the heterojunction contacts of silicon heterojunction (SHJ) solar cells. In this work, a post-doping process called catalytic doping (Cat-doping) on a-Si:H(i) is performed on the electron selective side of SHJ solar cells, which enables a device architecture that eliminates the additional deposition of the doped silicon layer. Thus, a single phosphorus Cat-doping layer combines the functions of two other layers by enabling excellent interface passivation and high carrier selectivity. The overall thinner layer on the window side results in higher spectral response at short wavelengths, leading to an improved short-circuit current density of 40.31 mA cm⁻² and an efficiency of 23.65% (certified). The cell efficiency is currently limited by sputter damage from the subsequent transparent conductive oxide fabrication and low carrier activation in the a-Si:H(i) with Cat-doping. Numerical device simulations show that the a-Si:H(i) with Cat-doping can provide sufficient field effect passivation even at lower active carrier concentrations compared to the as-deposited doped layer, due to the lower defect density.     

Protein function microarrays based on self-immobilizing and self-labeling fusion proteins

Protein microarrays are an attractive approach for the high-throughput analysis of protein function, but their impact on proteomics has been limited by the technical difficulties associated with their generation. Here we demonstrate that fusion proteins of O6-alkylguanine-DNA alkyltransferase (AGT) can be used for the simple and reliable generation of protein microarrays for the analysis of protein function. Important features of the approach are the selectivity of the covalent immobilization; this allows for direct immobilization of proteins out of cell extracts, and the option both to label and to immobilize AGT fusion proteins, which allows for direct screening for protein-protein interactions between different AGT fusion proteins. In addition to the identification of protein-protein interactions, AGT-based protein microarrays can be used for the characterization of small molecule-protein interactions or post-translational modifications. The potential of the approach was demonstrated by investigating the post-translational modification of acyl carrier protein (ACP) from E. coli by different phosphopantetheine transferases (PPTases), yielding insights into the role of selected ACP amino acids in the ACP-PPTase interaction.     

Investigation of sodium sulfate phase transitions in a porous material using humidity- and temperature-controlled X-ray diffraction

Crystals growing in confined spaces can generate stress and are a major cause of damage in porous materials. To investigate such deleterious processes, appropriate in situ techniques are required. This paper describes the use of X-ray diffractometry under controlled conditions of temperature and relative humidity (RH-XRD) for the direct observation of phase transition reactions in a porous substrate. An improved environmental chamber without temperature gradients is presented and applied to the investigation of phase transformations in the system Na2SO4 + H2O. This salt is generally considered as particularly damaging and frequently used in accelerated weathering tests. It is demonstrated that RH-XRD can be successfully applied for the direct observation of several relevant phase transitions in glass frits used as porous substrates. The conversion of Na2SO4(III) to Na2SO4(V) and the hydration of Na2SO4(V) both proceed fairly rapidly as true solid-state reactions without deliquescence of the educt phases. In contrast, crystallization from solution is kinetically hindered as there is a strong tendency of aqueous Na2SO4 to form supersaturated solutions also in narrow pores. The important implications of this behavior of the salt are also briefly discussed in the paper.     

Quantitative Evaluation of Cellular Uptake and Trafficking of Plain and Polyethylene Glycol‐Coated Gold Nanoparticles

This study addresses the cellular uptake and intracellular trafficking of 15‐nm gold nanoparticles (NPs), either plain (i.e., stabilized with citrate) or coated with polyethylene glycol (PEG), exposed to human alveolar epithelial cells (A549) at the air–liquid interface for 1, 4, and 24 h. Quantitative analysis by stereology on transmission electron microscopy images reveals a significant, nonrandom intracellular distribution for both NP types. No particles are observed in the nucleus, mitochondria, endoplasmic reticulum, or golgi. The cytosol is not a preferred cellular compartment for both NP types, although significantly more PEG‐coated than citrate‐stabilized NPs are present there. The preferred particle localizations are vesicles of different sizes (<150, 150–1000, >1000 nm). This is observed for both NP types and indicates a predominant uptake by endocytosis. Subsequent inhibition of caveolin‐ and clathrin‐mediated endocytosis by methyl‐β‐cyclodextrin (MβCD) results in a significant reduction of intracellular NPs. The inhibition, however, is more pronounced for PEG‐coated than citrate‐stabilized NPs. The latter are mostly found in larger vesicles; therefore, they are potentially taken up by macropinocytosis, which is not inhibited by MβCD. With prolonged exposure times, both NPs are preferentially localized in larger‐sized intracellular vesicles such as lysosomes, thus indicating intracellular particle trafficking. This quantitative evaluation reveals that NP surface coatings modulate endocytotic uptake pathways and cellular NP trafficking. Other nonendocytotic entry mechanisms are found to be involved as well, as indicated by localization of a minority of PEG‐coated NPs in the cytosol.