Determinants of biological activity of nanomaterials in innate immunity cells: the role of aspect ratio, environmental contaminants and protein corona

As defined by the European Union, “ ’Nanomaterial’ (NM) means a natural, incidental or manufactured material containing particles, in an unbound state or as an aggregate or agglomerate, where, for 50 % or more of the particles in the number size distribution, one or more external dimensions is in the size range 1 nm-100 nm ” (2011/696/UE). Given their peculiar physico-chemical features, nanostructured materials are largely used in many industrial fields (e.g. cosmetics, electronics, agriculture, biomedical) and their applications have astonishingly increased in the last fifteen years. Nanostructured materials are endowed with very large specific surface area that, besides making them very useful in many industrial processes, renders them very reactive towards the biological systems and, hence, potentially endowed with significant hazard for human health. For these reasons, in recent years, many studies have been focused on the identification of toxic properties of nanostructured materials, investigating, in particular, the mechanisms behind their toxic effects as well as their determinants of toxicity. This thesis investigates two types of nanostructured TiO2 materials, TiO2 nanoparticles (NP), which are yearly produced in tonnage quantities, and TiO2 nanofibres (NF), a relatively novel nanomaterial. Moreover, several preparations of MultiWalled Carbon Nanotubes (MWCNT), another nanomaterial widely present in many products, are also investigated.- Although many in vitro and in vivo studies have characterized the toxic properties of these materials, the identification of their determinants of toxicity is still incomplete. The aim of this thesis is to identify the structural determinants of toxicity, using several in vitro models. Specific fields of investigation have been a) the role of shape and the aspect ratio in the determination of biological effects of TiO2 nanofibres of different length; b) the synergistic effect of LPS and TiO2 NP on the expression of inflammatory markers and the role played therein by TLR-4; c) the role of functionalization and agglomeration in the biological effects of MWCNT. As far as biological effects elicited by TiO2 NF are concerned, the first part of the thesis demonstrates that long TiO2 nanofibres caused frustrated phagocytosis, cytotoxicity, hemolysis, oxidative stress and epithelial barrier perturbation. All these effects were mitigated by fibre shortening through ball-milling. However, short TiO2 NF exhibited enhanced ability to activate acute pro-inflammatory effects in macrophages, an effect dependent on phagocytosis. Therefore, aspect ratio reduction mitigated toxic effects, while enhanced macrophage activation, likely rendering the NF more prone to phagocytosis. These results suggest that, under in vivo conditions, short NF will be associated with acute inflammatory reaction, but will undergo a relatively rapid clearance, while long NF, although associated with a relatively smaller acute activation of innate immunity cells, are not expected to be removed efficiently and, therefore, may be associated to chronic inflammatory responses. As far as the relationship between the effects of TiO2 NP and LPS, investigated in the second part of the thesis, are concerned, TiO2 NP markedly enhanced macrophage activation by LPS through a TLR-4-dependent intracellular pathway. The adsorption of LPS onto the surface of TiO2 NP led to the formation of a specific bio-corona, suggesting that, when bound to TiO2 NP, LPS exerts a much more powerful pro-inflammatory effect. These data suggest that the inflammatory changes observed upon exposure to TiO2 NP may be due, at least in part, to their capability to bind LPS and, possibly, other TLR agonists, thus enhancing their biological activities. Finally, the last part of the thesis demonstrates that surface functionalization of MWCNT with amino or carboxylic groups mitigates the toxic effects of MWCNT in terms of macrophage activation and capability to perturb epithelial barriers. Interestingly, surface chemistry (in particular surface charge) influenced the protein adsorption onto the MWCNT surface, allowing to the formation of different protein coronae and the tendency to form agglomerates of different size. In particular functionalization a) changed the amount and the type of proteins adsorbed to MWCNT and b) enhanced the tendency of MWCNT to form large agglomerates. These data suggest that the different biological behavior of functionalized and pristine MWCNT may be due, at least in part, to the different tendency to form large agglomerates, which is significantly influenced by their different capability to interact with proteins contained in biological fluids. All together, these data demonstrate that the interaction between physico-chemical properties of nanostructured materials and the environment (cells + biological fluids) in which these materials are present is of pivotal importance for the understanding of the biological effects of NM. In particular, bio-persistence and the capability to elicit an effective inflammatory response are attributable to the interaction between NM and macrophages. However, the interaction NM-cells is heavily influenced by the formation at the nano-bio interface of specific bio-coronae that confer a novel biological identity to the nanostructured materials, setting the basis for their specific biological activities.

Refractive index sensing properties of long-period fibre grating with sol-gel derived coatings

Long-period fibre gratings (LPGs) have previously been used to detect quantities such as temperature, strain and refractive index (RI). The responsivity to surrounding refractive index means that, potentially, LPGs could be realised as optical biosensors for applications in biochemical and biomedical application areas. We report here to our best knowledge the first investigation on refractive index sensing properties of LPGs with sol-gel derived titanium and silicon oxide coatings. It is revealed that the RI sensitivity of an LPG is affected by both the thickness and the index value of the coating; the coating with higher index and thickness will enhance the LPG RI sensitivity significantly. The surrounding refractive index induced LPG resonance shift has been evaluated over the LPGs’ most sensitive RI region from 1.42 to 1.44. We have identified that, in this region, the uncoated LPG has an RI sensitivity of (-673.0±0.4)nm/uri (unit of refractive index) while the LPG coated with titanium oxide exhibits a sensitivity as high as (-1067.15±0.04)nm/uri.

Fibre agglomerate transport in a horizontal flow

An experimental and theoretical study of the transport of mineral wool fibre agglomerates in nuclear power plant containment sumps is being performed. A racetrack channel was devised to provide data for the validation of numerical models, which are intended to model the transport of fibre agglomerates. The racetrack channel provides near uniform and steady conditions that lead to either the sedimentation or suspension of the agglomerates. Various experimental techniques were used to determine the velocity conditions and the distribution of the fibre agglomerates in the channel. The fibre agglomerates are modelled as fluid particles in the Eulerian reference frame. Simulations of pure sedimentation of a known mass and volume of agglomerations show that the transport of the fibre agglomerates can be replicated. The suspension of the fibres is also replicated in the simulations; however, the definition of the fibre agglomerate phase is strongly dependent on the selected density and diameter. Detailed information on the morphology of the fibre agglomerates is lacking for the suspension conditions, as the fibre agglomerates may undergo breakage and erosion. Therefore, ongoing work, which is described here, is being pursued to improve the experimental characterisation of the suspended transport of the fibre agglomerates.

Advanced UV inscribed fibre grating structures and applications in optical sensing and laser systems

This thesis presents detailed investigation of UV inscribed fibre grating based devices and novel developments in the applications of such devices in optical sensing and fibre laser systems. The major contribution of this PhD programme includes the systematic study on fabrication, spectral characteristics and applications of different types of UV written in-fibre gratings such as Type I and IA Fibre Bragg Gratings (FBGs), Chirped Fibre Bragg Gratings (CFBGs) and Tilted Fibre Gratings (TFGs) with small, large and 45º tilted structures inscribed in normal silica fibre. Three fabrication techniques including holographic, phase-mask and blank beam exposure scanning, which were employed to fabricate a range of gratings in standard single mode fibre, are fully discussed. The thesis reports the creation of smart structures with self-sensing capability by embedding FBG-array sensors in Al matrix composite. In another part of this study, we have demonstrated the particular significant improvements made in sensitising standard FBGs to the chemical surrounding medium by inducing microstructure to the grating by femtosecond (fs) patterning assisted chemical etching technique. Also, a major work is presented for the investigation on the structures, inscription methods and spectral Polarisation Dependent Loss (PDL) and thermal characteristics of different angle TFGs. Finally, a very novel application in realising stable single polarisation and multiwavelength switchable Erbium Doped Fibre Lasers (EDFLs) using intracavity polarisation selective filters based on TFG devices with tilted structures at small, large and exact 45° angles forms another important contribution of this thesis.

Applications of ultralong Raman fibre lasers in photonics

This thesis presents a numerical and experimental investigation on applications of ultralong Raman fibre lasers in optical communications, supercontinuum generation and soliton transmission. The research work is divided in four main sections. The first involves the numerical investigation of URFL intra-cavity power and the relative intensity noise transfer evolution along the transmission span. The performance of the URFL is compared with amplification systems of similar complexity. In the case of intracavity power evolution, URFL is compared with a first order Raman amplification system. For the RIN transfer investigation, URFL is compared with a bi-directional dual wavelength pumping system. The RIN transfer function is investigated for several cavity design parameters such as span length, pump distribution and FBG reflectivity. The following section deals with experimental results of URFL cavities. The enhancement of the available spectral bandwidth in the C-band and its spectral flatness are investigated for single and multi-FBGs cavity system. Further work regarding extended URFL cavity in combination with Rayleigh scattering as random distributed feedback produced a laser cavity with dual wavelength outputs independent to each other. The last two sections relate to URFL application in supercontinuum (SC) generation and soliton transmission. URFL becomes an enhancement structure for SC generation. This thesis shows successful experimental results of SC generation using conventional single mode optical fibre and pumped with a continuous wave source. The last section is dedicated to soliton transmission and the study of soliton propagation dynamics. The experimental results of exact soliton transmission over multiple soliton periods using conventional single mode fibre are shown in this thesis. The effect of the input signal, pump distribution, span length and FBGs reflectivity on the soliton propagation dynamics is investigated experimentally and numerically.

Tunable compensation of second and third order dispersion by non-uniformly strained chirped fibre Bragg grating

A novel fibre grating device is demonstrated with tuneable chromatic dispersion slope. The tuning range is 70 to 190 ps/nm and 0 to 25 ps/nm2 for the second and third order dispersion, respectively.

Extended range interrogation of wavelength division multiplexed fibre Bragg grating sensors using an arrayed waveguide grating

A technique for interrogating multiplexed fibre Bragg grating (FBG) sensors using an arrayed waveguide grating (AWG) is described. The approach considerably extends the sensing range from that achieved previously, while providing a strain resolution of 17nevHz at 30 Hz.

Bend and index insensitive long period grating in progressive three layered optical fibre

A long period fibre grating written in a progressive three-layered optical fibre is shown to exhibit an attenuation band that has a very low bending sensitivity (5.1×10 –2 nm m) compared to normal step-index fibre, and is also insensitive to changes in the refractive index of the surrounding medium. Applications to sensing and telecommunications are discussed.

All-Raman amplified transmission at 40 Gbit/s in standard single mode fibre

We investigate a 40 Gbit/s all-Raman amplified standard single mode fibre (SMF) transmission system with the mid-range amplifier spacing of 80-90 km. The impact of span configuration on double Rayleigh back scattering (DRBS) was studied. Four different span configurations were compared experimentally. A transmission distance of 1666 km in SMF has been achieved without forward error correcting (FEC) for the first time. The results demonstrate that the detrimental effects associated with high pump power Raman amplification in standard fibre can be minimised by dispersion map optimisation. © 2003 IEEE.

Extended-range, low coherence dual wavelength interferometry using a superfluorescent fibre source and chirped fibre Bragg gratings

We describe an all-fibre, passive scheme for making extended range interferometric measurements based on the dual wavelength technique. The coherence tuned interferometer network is illuminated with a single superfluorescent fibre source at 1.55 µm and the two wavelengths are synthesised at the output by means of chirped fibre Bragg gratings. We demonstrate an unambiguous sensing range of 270 µm, with a dynamic range of 2.7 × 10 5.