Advanced Mg, Zn, Sr, Si Multi-Substituted Hydroxyapatites for Bone Regeneration.

PURPOSE: Compositional tailoring is gaining more attention in the development of advanced biomimetic nanomaterials. In this study, we aimed to prepare advanced multi-substituted hydroxyapatites (ms-HAPs), which show similarity with the inorganic phase of bones and might have therapeutic potential for bone regeneration. MATERIALS: Novel nano hydroxyapatites substituted simultaneously with divalent cations: Mg2+ (1.5%), Zn2+ (0.2%), Sr2+ (5% and 10%), and Si (0.2%) as orthosilicate (SiO4 4-) were designed and successfully synthesized for the first time. METHODS: The ms-HAPs were obtained via a wet-chemistry precipitation route without the use of surfactants, which is a safe and ecologically friendly method. The composition of synthesized materials was determined by inductively coupled plasma optical emission spectrometry (ICP-OES). The materials were characterized by X-ray powder diffraction (XRD), FT-IR and FT-Raman spectroscopy, BET measurements and by imaging techniques using high-resolution TEM (HR-TEM), FE-SEM coupled with EDX, and atomic force microscopy (AFM). The ion release was measured in water and in simulated body fluid (SBF). RESULTS: Characterization methods confirmed the presence of the unique phase of pure stoichiometric HAP structure and high compositional purity of all synthesized nanomaterials. The doping elements influenced the crystallite size, the crystallinity, lattice parameters, morphology, particle size and shape, specific surface area, and porosity. Results showed a decrease in both nanoparticle size and crystallinity degree, coupled with an increase in specific surface area of these advanced ms-HAP materials, in comparison with pure stoichiometric HAP. The release of biologically important ions was confirmed in different liquid media, both in static and simulated dynamic conditions. CONCLUSION: The incorporation of the four substituting elements into the HAP structure is demonstrated. Synthesized nanostructured ms-HAP materials might inherit the in vivo effects of substituting functional elements and properties of hydroxyapatite for bone healing and regeneration. Results revealed a rational tailoring approach for the design of a next generation of bioactive ms-HAPs as promising candidates for bone regeneration.

A New High Hydrostatic Pressure Process to Assure the Microbial Safety of Human Milk While Preserving the Biological Activity of Its Main Components.

Background: The main process used to pasteurize human milk is the low-temperature, long-time Holder method. More recently, the high-temperature, short-time method has been investigated. Both processes lead to the appropriate inactivation of vegetative bacterial forms but are ineffective against bacterial spores. Research Aims/Questions: We aimed to accomplish two main objectives: inactivation of all pathogens, including spores; and preservation of the activity of milk components. Design/Methods: Recently, a novel high-hydrostatic pressure process has been developed by HPBioTECH. Using the same raw human milk samples, we compared the effects of this method with those of the Holder method on vegetative and spore forms of pathogens and on bioactive components (lipase activity, immunoproteins). Results: Two main microbial strains were selected: Staphylococcus aureus (as a reference for vegetative forms) and Bacillus cereus (as a reference for spores). Use of the high-hydrostatic pressure process led to microbial decontamination of 6 log for both S. aureus and B. cereus. Additionally, the bioactivity of the main components of human milk was preserved, with activities of lipase, α-lactalbumin, casein, lysozyme, lactoferrin, and sIgA of ~80, 96-99, 98-100, 95-100, 93-97, and 63-64%, respectively. Conclusions: Use of this novel high-hydrostatic pressure process to generate microbiologically safe human milk may provide important benefits for preterm infants, including improved assimilation of human milk (leading increased weight gain) and improved resistance to infections. Because 10% of all human milk collected is contaminated by B. cereus, use of this method will also prevent waste.

High hydrostatic pressure treatment for the inactivation of Staphylococcus aureus in human blood plasma.

For the past 30years, pressure inactivation of microorganisms has been developed in biosciences, in particular for foods and more recently for biological products, including pharmaceutical ones. In many past studies, the effect of high hydrostatic pressure (HHP) processes on pathogens focused mainly on the effect of an increase of the pressure value. To assure the safety of pharmaceutical products containing fragile therapeutic components, development of new decontamination processes at the lowest pressure value is needed to maintain their therapeutic properties. The aim of this study was therefore to evaluate the impact of the process parameters characterizing high-pressure treatments [such as the pressurization rate (PR) and the application mode (AM)] on the inactivation of pathogens, in particular to determine how these parameters values could help decrease the pressure value necessary to reach the same inactivation level. The effect of these physical parameters was evaluated on the inactivation of Staphylococcus aureus ATCC 6538 which is an opportunistic pathogen of important relevance in the medical, pharmaceutical and food domains. Human blood plasma was chosen as the suspension medium because of its physiological importance in the transfusion field. It was shown that the optimization of all the selected parameters could lead to a high inactivation level (≈5log(10) decrease of the initial bacterial load) at a pressure level as low as 200MPa, underlining some synergistic effects among these parameters. Complete inactivation of the initial bacterial population was achieved for the following conditions: PR=50MPas(-1), AM=5×2min, T≈-5°C and P=300MPa.

Optical and magnetic characterisation of Co3+ and Ni3+ in LaAlO3: interplay between the spin state and Jahn-Teller effect.

The coordination, the electronic structures and the spin of the ground state of Ni(3+) (3d(7)) and Co(3+) (3d(6)) introduced as impurities in LaAlO(3) are investigated through optical spectroscopy and magnetic measurements. The unusual trivalent valence state in both transition-metal ions was stabilised via a sol-gel process followed by high oxygen pressure treatments. We show that the crystal-field strength at the nearly O(h) transition-metal site in LaAlO(3) locates Ni(3+) and Co(3+) near the spin state crossover, yielding a low-spin ground state in both cases. We analyse how the interplay between the Jahn-Teller (JT) effect and the spin state affects the magnetic moment of the ion and its temperature dependence. The optical spectra reveal a JT effect associated with a low-spin ground state in Ni(3+) and with a thermally populated high-spin low-lying first excited state in Co(3+). The corresponding JT distortions are derived from structural correlations. We conclude that the JT effect is unable to stabilise the intermediate spin state in Co(3+). A low-spin ground state in thermal equilibrium with a high-spin low-lying first excited state is detected in diluted Co(3+)-doped LaAlO(3). These results are compared with those obtained in the parent pure compounds LaNiO(3) and LaCoO(3).

High hydrostatic pressure and biology: a brief history.

Pressure as a thermodynamical parameter was successively introduced in physics, hydrometallurgy, geochemistry, and biology. In all cases, the main objective was to recreate a natural phenomenon (gas or liquid compressibility, synthesis or crystal growth of minerals, survival of deep sea microorganisms…). The introduction of high hydrostatic pressure (HHP) in Biology was an important scientific feature over the last hundred years. This paper describes the different steps that have led to the spreading of pressure in biology and the opening of new frontiers either in basic and applied researches due to the specific characteristics of the pressure parameter. Because of the low energy conveyed by this parameter, leading to the preservation of most organoleptic properties of foods, and its ability to inactivate many pathogens, the use of HHP began to spread at the end of the twentieth century into the food industry, in particular for the development of pathogen inactivation processes. Today, even if this field is still the first application domain for HHP, more and more research works have shown that this parameter could be of great interest in health and medicine sciences.

Development of high hydrostatic pressure in biosciences: pressure effect on biological structures and potential applications in biotechnologies.

Compared to temperature, the development of pressure as a tool in the research field has emerged only recently (at the end of the XIXth century). Following several developments in Physics and Chemistry during the first half of the XXth century (in particular the synthesis of diamond in 1953-1954), high pressures were applied in Food Science, especially in Japan. The main objective was then to achieve the decontamination of foods while preserving their organoleptic properties. Now, a new step is engaged: the biological applications of high pressures, from food to pharmaceuticals and biomedical applications. This paper will focus on three main points: (i) a brief presentation of the pressure parameter and its characteristics, (ii) a description of the pressure effects on biological constituents from simple to more complex structures and (iii) a review of the different domains for which the application of high pressures is able to initiate potential developments in Biotechnologies.

Inactivation of Escherichia coli and Listeria innocua in kiwifruit and pineapple juices by high hydrostatic pressure.

Escherichia coli and Listeria innocua in kiwifruit and pineapple juices were exposed to high hydrostatic pressure (HHP) at 300 MPa for 5 min. Both bacteria showed equal resistance to HHP. Using low (0 degrees C) or sub-zero (-10 degrees C) temperatures instead of room temperature (20 degrees C) during pressurization did not change the effectiveness of HHP treatment on both bacteria in studied juices. Pulse pressure treatment (multiple pulses for a total holding time of 5 min at 300 MPa) instead of continuous (single pulse) treatment had no significant (p>0.05) effect on the microbial inactivation in kiwifruit juice; however, in pineapple juice pulse treatment, especially after 5 pulses, increased the inactivation significantly (p<0.05) for both bacteria. Following storage of pressure-treated (350 MPa, 20 degrees C for 60 s x 5 pulses) juices at 4, 20 and 37 degrees C up to 3 weeks, the level of microbial inactivation further increased and no injury recovery of the bacteria were detected. This work has shown that HHP treatment can be used to inactivate E. coli and L. innocua in kiwifruit and pineapple juices at lower pressure values at room temperature than the conditions used in commercial applications (>400 MPa). However, storage period and temperature should carefully be optimized to increase the safety of HHP treated fruit juices.

The stenlying effect of high hydrostatic pressure on thermally and hydrolytically labile nanosized carriers.

PURPOSE: To investigate whether high hydrostatic pressure (HHP) treatment allows the sterilization of thermosensitive polymer nanoparticle suspensions without jeopardizing their physicochemical integrity. METHODS: Application of HHP was explored on a wide variety of thermosensitive poly(cyanoacrylate) nanoparticles, varying by their type (nanospheres or nanocapsules), by their preparation method (nanoprecipitation or emulsion/solvent evaporation), as well as by their surface characteristics. Physicochemical characterization before and after pressurization included turbidimetry, size measurement, zeta potential, scanning electron microscopy and infrared analysis. A sterility test also conducted according to pharmacopoeial requirements on an importantly contaminated nanoparticle suspension. RESULTS: Poly(cyanoacrylate) nanoparticles appeared to be extremely baroresistant. Continuous or oscillatory HHP treatment up to 500 MPa during 30 min induced generally neither physical, nor chemical damage. However, precautions should be taken when surface modifiers are adsorbed onto nanoparticles, as a layer destabilization may occur. Finally, this process allowed the successful inactivation of vegetative bacteria, yeast, and fungi. CONCLUSIONS: This work proposes HHP as a new method for polymer drug carriers sterilization, taking into account that further exploration in this area is needed to propose novel protocols for spores inactivation.

Effects of high hydrostatic pressure on several sensitive therapeutic molecules and a soft nanodispersed drug delivery system.

PURPOSE: According to the development in the last decade of industrial processes using high hydrostatic pressure (HHP) for preservation of several commercial food products, novel sterilization or decontamination processes for pharmaceutical products could be conceivable. The aim of this work is to evaluate the effects of HHP on the integrity of insulin and heparin solutions, suspension of monoclonal antibodies and Spherulites. METHODS: High performance liquid chromatography, thin layer chromatography, capillary electrophoresis assays, ELISA tests, laser granulometry and spectrophotometry analyses have been performed to compare HHP treated drugs (in a domain of pressure and temperature ranging respectively from 20 up to 500 MPa and from 20 degrees C up to 37 degrees C) vs. untreated ones. RESULTS: No difference has been detected except for monoclonal antibodies that are altered above 500 MPa. CONCLUSIONS: The structure integrity of sensitive molecule due to the small energy involved by HHP and the development of industrial plants (intended for the decontamination of food products) confer to this technology the potential of a new method for sterilization of fragile drugs and an original alternative to aseptic processes and sterilizing filtration.