Study on osteoinductive activity of biotin film by low-energy electron beam deposition.

Biotin film was prepared by low-energy electron beam deposition (LEBD). The molecular structure, chemical composition and micromorphology of the biotin film were investigated by 1HNMR, FTIR, XPS, AFM and SEM. The results showed the molecular structure of a monolayer of biotin film is fully consistent with the molecular structure of the initial biotin powders. The contact angle test showed that the biotin film exhibit good hydrophilicity. The release kinetics of biotin film was tested by UV-Vis method. It was found that the film was almost completely released in about two weeks. The cell viability of MC3T3-E1 cells on the surface of the biotin film was attaining 100.54 ± 1.7% (P < 0.05), showing excellent biocompatibility and biosafety. Titanium implant with surface of biotin film was implanted into the femoral head of rabbits as experimental group. The animals were euthanized after four weeks. Compared with the control group, mature lamellar bone formation was observed with dense trabecular bone, and the expression of Coll-I, Runx2 and BMP-2 was better. The results showed that the repair effect of bone defect in the experimental group was excellent.

Osteogenic potential evaluation of biotin combined with magnesium-doped hydroxyapatite sustained-release film.

Biotin is one of the water-soluble B-complex group of vitamins. Recent studies have found that the relative protein expression of BMP2, BSP and OPG in MC3T3-E1 cells is prominent after 14 days of co-culture with biotin film, especially for BMP2. It is also found that the rapid degradation of biotin film in vivo limits its application value. In this work, magnesium-doped hydroxyapatite (MgHA) film can form a porous network structure as a biological sustained-release film. Therefore, the multilayer (MgHA|biotin|MgHA|biotin) film was prepared by pulsed laser assisted electron beam deposition technique. The morphology, structure and properties of biotin film and multilayer film were analyzed and characterized. Also, the osteogenic effect of biotin film and multilayer film was evaluated after implantation into the femoral bone marrow cavity of SD rats. The results of micro-CT scan and 3D reconstruction showed that there were a large number of trabecular bones around the multilayer film, which was superior to biotin film in osteogenesis. Hematoxylin-eosin staining showed cancellous bone structure and intact bone marrow structure around the multilayer film, and the newly formed bone became lamellar. Masson-trichromatic staining revealed abundant osteoid and braided bone formation around the multilayer film. In conclusion, MgHA sustained release film can realize the continuous release of bioactive drugs, which provides a new route to accelerate the repair of bone defects.

Permeation of Nanoparticles into Pulmonary Surfactant Monolayer: In Situ X-ray Standing Wave Studies.

High-resolution X-ray techniques were applied to examine the effects of gold nanoparticles (size <5 nm) on natural pulmonary surfactant and pure DPPC monolayers preliminarily formed on water subphase in a Langmuir trough. Hydrophobic and hydrophilic nanoparticles were delivered from nanoaerosol using electrodeposition method. Grazing incidence diffraction, X-ray reflectivity, and X-ray standing wave measurements allow to monitor the changes in molecular organization of lipid monolayer and to locate the position of gold nanoparticles. X-ray experiments were performed over a period of 9-14 h. The obtained results evidenced that, on a long time scale, the deposition of nanoparticles, even at low doses, can induce pronounced alterations in lipid monolayer. The presented data can help to elucidate the mechanism of pulmonary translocation of inhaled nanoparticles that is of special interest for biomedical investigations of potential risk of nanoaerosols for human health.

Features of electron beam deposition of polymer coatings with the prolonged release of the drug component.

The first part of the paper provides a comprehensive analysis of the features of electron beam formation of polymer coatings with the prolonged release of the drug compound using ciprofloxacin and clotrimazole as an example. The influence features of the low-energy electron beam on the molecular structure of medicinal chemical preparations have been established. The impossibility of producing the coatings based on medicinal compounds with a complex molecular structure (vancomycin, micafungin, etc.) by a low-energy electron beam has been justified. The second part of the paper introduces a fundamentally new vacuum method for the formation of the composite coatings based on antibiotics and antifungal drugs, accompanied by the prolonged release of the drug component. This method allows the formation of composite coatings based on medicinal compounds with a complex molecular structure. It is effective for modifying implants to prevent the risk of implant-associated infectious complications which are the result of the occurrence of mixed biofilms. The method can be used to form composite layers based on topical antitumor drugs for cancer control.

Unusual features of the c-ring of F1FO ATP synthases.

Membrane integral ATP synthases produce adenosine triphosphate, the universal "energy currency" of most organisms. However, important details of proton driven energy conversion are still unknown. We present the first high-resolution structure (2.3 Å) of the in meso crystallized c-ring of 14 subunits from spinach chloroplasts. The structure reveals molecular mechanisms of intersubunit contacts in the c14-ring, and it shows additional electron densities inside the c-ring which form circles parallel to the membrane plane. Similar densities were found in all known high-resolution structures of c-rings of F1FO ATP synthases from archaea and bacteria to eukaryotes. The densities might originate from isoprenoid quinones (such as coenzyme Q in mitochondria and plastoquinone in chloroplasts) that is consistent with differential UV-Vis spectroscopy of the c-ring samples, unusually large distance between polar/apolar interfaces inside the c-ring and universality among different species. Although additional experiments are required to verify this hypothesis, coenzyme Q and its analogues known as electron carriers of bioenergetic chains may be universal cofactors of ATP synthases, stabilizing c-ring and prevent ion leakage through it.

The Enhancement of Metal-Binding Properties in Hemoglobin: The Role of Mild Damaging Factors.

X-ray studies revealed the considerable enhancement of metal-binding properties in human hemoglobin under exposure to mild damaging factors (in the presence of 0.09 M urea or upon heating for 30 min at 50 °C). Changes in the element composition of the hemoglobin monolayer, formed on the water subphase in the Langmuir trough, have been monitored in real time by the total external reflection X-ray fluorescence measurements. X-ray absorption spectroscopy has been applied to study the local environment of zinc ions bound on hemoglobin molecules. According to these data, each zinc ion is coordinated by four ligands, two of which are cysteine and histidine. The oxidative stress has been found to accelerate extensively the enhancement of metal-binding ability in protein. A two-stage mechanism has been proposed as a possible explanation of the observed phenomenon: First, in the presence of the mild damaging agents, protein molecules can undergo a transition from the native conformation to a more labile intermediate state that increases the accessibility of amino acid residues (in particular cysteine). At the second stage, oxidation of cysteine and the subsequent activation of cysteine SH groups can affect markedly the protein-metal interaction. The presented investigations provide a deeper insight into the pathogenesis of metabolic disorders that excessive concentrations of the endogenic toxicants might trigger in an organism.

Grazing-incidence small-angle X-ray scattering study of correlated lateral density fluctuations in W/Si multilayers.

A structural characterization of W/Si multilayers using X-ray reflectivity (XRR), scanning transmission electron microscopy (STEM) and grazing-incidence small-angle X-ray scattering (GISAXS) is presented. STEM images revealed lateral, periodic density fluctuations in the Si layers, which were further analysed using GISAXS. Characteristic parameters of the fluctuations such as average distance between neighbouring fluctuations, average size and lateral distribution of their position were obtained by fitting numerical simulations to the measured scattering images, and these parameters are in good agreement with the STEM observations. For the numerical simulations the density fluctuations were approximated as a set of spheroids distributed inside the Si layers as a 3D paracrystal (a lattice of spheroids with short-range ordering but lacking any long-range order). From GISAXS, the density of the material inside the density fluctuations is calculated to be 2.07 g cm-3 which is 89% of the bulk value of the deposited layer (2.33 g cm-3).

[Biocompatible antibacterial polymer coatings with ciprofloxacin extended release].

A procedure for application of thin composite coats based on polymers (polyurethane or polyethylene) and ciprofloxacin in active gaseous phase generated by electron beam dispersion under vacuum was developed. The coats were shown to be highly dispersed systems whose components preserved the initial molecular structure and had antibacterial activity. The composite coats based on polyurethane were more stable to ciprofloxacin leaching in isotonic sodium chloride solution.