Influence of surface pre-treatment with mechanical polishing, chemical, electrochemical and ion sputter etching on the surface properties, corrosion resistance and MG-63 cell colonization of commercially pure titanium.

The impact of four pre-treatment techniques on the surface morphology and chemistry, residual stress, mechanical properties, corrosion resistance in a physiological saline solution and cell colonization of commercially pure titanium is examined in detail. Mechanical polishing, electrochemical etching, chemical etching in Kroll's reagent, and ion sputter etching with argon ions were applied. Surface morphologies reflect the nature of surface layer removal. Significant roughening of the surface and a characteristic microtopology become apparent as a result of the sensitivity of chemical and ion sputter etching to the grain orientation. The hardness in the near surface region was controlled by the amount of residual stress. Etching of the stressed surface layer led to a reduction in residual stress and surface hardness. A compact passivation layer composed of TiO, TiO2 and Ti2O3 native oxides imparted high corrosion resistance to the surface after mechanical polishing, chemical and electrochemical etching. The ion sputter etched surface showed substantially reduced corrosion resistance, where the corrosion process was controlled by electron transfer. The specific topology affected the adhesion of the cell to the surface rather than the cell area coverage. The cell area coverage increased with the corrosion stability of the surface.

Silicalite-1 Layers as a Biocompatible Nano- and Micro-Structured Coating: An In Vitro Study on MG-63 Cells.

Silicalite-1 is a purely siliceous form of zeolite, which does not contain potentially harmful aluminum in its structure as opposed to ZSM-5 aluminosilicate types of zeolite. This paper reports on a study of a silicalite-1 film, deposited on a silicon Si(100) substrate, as a potential anti-corrosive and biocompatible coating for orthopaedic implants. Silicalite-1 film was prepared in situ on the surface of Si(100) wafers using a reaction mixture of tetrapropyl-ammonium hydroxide (TPAOH), tetraethyl-orthosilicate (TEOS), and diH2O. The physico-chemical properties of the obtained surface were characterized by means of X-ray photoelectron spectroscopy, water contact angle measurement, atomic force microscopy, and scanning electron microscopy. The biocompatibility was assessed by interaction with the MG-63 cell line (human osteosarcoma) in terms of cell adhesion, morphology, proliferation, and viability. The synthesized silicalite-1 film consisted of two layers (b- and a, b-oriented crystals) creating a combination of micro- and nano-scale surface morphology suitable for cell growth. Despite its hydrophobicity, the silicalite-1 film increased the number of initially adhered human osteoblast-like MG-63 cells and the proliferation rate of these cells. The silicalite-1 film also improved the cell viability in comparison with the reference Si(100) substrate. It is therefore a promising candidate for coating of orthopaedic implants.

Properties of titanium-alloyed DLC layers for medical applications.

DLC-type layers offer a good potential for application in medicine, due to their excellent tribological properties, chemical resistance, and bio-inert character. The presented study has verified the possibility of alloying DLC layers with titanium, with coatings containing three levels of titanium concentration prepared. Titanium was present on the surface mainly in the form of oxides. Its increasing concentration led to increased presence of titanium carbide as well. The behavior of the studied systems was stable during exposure in a physiological saline solution. Electrochemical impedance spectra practically did not change with time. Alloying, however, changed the electrochemical behavior of coated systems in a significant way: from inert surface mediating only exchange reactions of the environment in the case of unalloyed DLC layers to a response corresponding rather to a passive surface in the case of alloyed specimens. The effect of DLC layers alloying with titanium was tested by the interaction with a simulated body fluid, during which precipitation of a compound containing calcium and phosphorus--basic components of the bone apatite--occurred on all doped specimens, in contrast to pure DLC. The results of the specimens' surface colonization with cells test proved the positive effect of titanium in the case of specimens with a medium and highest content of this element.

Photosynthesis in dynamic light: systems biology of unconventional chlorophyll fluorescence transients in Synechocystis sp. PCC 6803.

Photosynthetic organisms live in a dynamic environment where light typically fluctuates around a mean level that is slowly drifting during the solar day. We show that the far-from-equilibrium photosynthesis occurring in a rapidly fluctuating light differs vastly from the stationary-flux photosynthesis attained in a constant or slowly drifting light. Photosynthetic organisms in a static or slowly drifting light can be characterized by a steady-state quantum yield of chlorophyll fluorescence emission F' that is changing linearly with small and slow variations of the incident irradiance I+DeltaI(t): F'(I+DeltaI(t)) approximately Fmean '(dF)/(dI).DeltaI(t). In Synechocystis sp. PCC 6803, the linear approximation holds for an extended interval covering largely the static irradiance range experienced by the cyanobacteria in nature. The photosynthetic dynamism and, consequently, the dynamism of the chlorophyll fluorescence emission change dramatically when exposing the organism to a fluctuating irradiance. Harmonically-modulated irradiance I+DeltaI . sin(2pit/T), T approximately 1-25 s induces perpetual, far-from-equilibrium forced oscillations that are strongly non-linear, exhibiting significant hysteresis with multiple fluorescence levels corresponding to a single instantaneous level of the incident irradiance. We propose that, in nature, the far-from-equilibrium dynamic phenomena represent a significant correction to the steady-state photosynthetic activity that is typically investigated in laboratory. Analysis of the forced oscillations by the tools of systems biology suggests that the dynamism of photosynthesis observed in fluctuating light can be explained by a delayed action of regulatory agents.

Negative feedback regulation is responsible for the non-linear modulation of photosynthetic activity in plants and cyanobacteria exposed to a dynamic light environment.

Photosynthetic organisms exposed to a dynamic light environment exhibit complex transients of photosynthetic activities that are strongly dependent on the temporal pattern of the incident irradiance. In a harmonically modulated light of intensity I approximately const.+sin(omegat), chlorophyll fluorescence response consists of a steady-state component, a component modulated with the angular frequency of the irradiance omega and several upper harmonic components (2omega, 3omega and higher). Our earlier reverse engineering analysis suggests that the non-linear response can be caused by a negative feedback regulation of photosynthesis. Here, we present experimental evidence that the negative feedback regulation of the energetic coupling between phycobilisome and Photosystem II (PSII) in the cyanobacterium Synechocystis sp. PCC6803 indeed results in the appearance of upper harmonic modes in the chlorophyll fluorescence emission. Dynamic changes in the coupling of the phycobilisome to PSII are not accompanied by corresponding antiparallel changes in the Photosystem I (PSI) excitation, suggesting a regulation limited to PSII. Strong upper harmonic modes were also found in the kinetics of the non-photochemical quenching (NPQ) of chlorophyll fluorescence, of the P700 redox state and of the CO(2) assimilation in tobacco (Nicotiana tabaccum) exposed to harmonically modulated light. They are ascribed to negative feedback regulation of the reactions of the Calvin-Benson cycle limiting the photosynthetic electron transport. We propose that the observed non-linear response of photosynthesis may also be relevant in a natural light environment that is modulated, e.g., by ocean waves, moving canopy or by varying cloud cover. Under controlled laboratory conditions, the non-linear photosynthetic response provides a new insight into dynamics of the regulatory processes.

Complex metabolic oscillations in plants forced by harmonic irradiance.

Plants exposed to harmonically modulated irradiance, approximately 1 + cos(omegat), exhibit a complex periodic pattern of chlorophyll fluorescence emission that can be deconvoluted into a steady-state component, a component that is modulated with the frequency of the irradiance (omega), and into at least two upper harmonic components (2omega and 3omega). A model is proposed that accounts for the upper harmonics in fluorescence emission by nonlinear negative feedback regulation of photosynthesis. In contrast to simpler linear models, the model predicts that the steady-state fluorescence component will depend on the frequency of light modulation, and that amplitudes of all fluorescence components will exhibit resonance peak(s) when the irradiance frequency is tuned to an internal frequency of a regulatory component. The experiments confirmed that the upper harmonic components appear and exhibit distinct resonant peaks. The frequency of autonomous oscillations observed earlier upon an abrupt increase in CO(2) concentration corresponds to the sharpest of the resonant peaks of the forced oscillations. We propose that the underlying principles are general for a wide spectrum of negative-feedback regulatory mechanisms. The analysis by forced harmonic oscillations will enable us to examine internal dynamics of regulatory processes that have not been accessible to noninvasive fluorescence monitoring to date.