Assessing the Effect of Photodynamic Therapy on Peripheral Nerve and Cancer Cells Using a Thin Tissue Engineered Collagen Culture Model

Abstract not available

Intracellular localisation of mTHPC and effect of photodynamic therapy in cells of the mammalian peripheral nervous system

Fewer nerve-related side effects have been noted after treating head and neck cancer with photodynamic therapy (PDT) compared to conventional cancer therapy. Our aim is to investigate the biological basis for any such nerve-sparing effect. In this study the intracellular localisation and effect on cell viability of the photosensitiser meta-tetrahydroxylphenylchlorin (mTHPC) was investigated in cell culture models using peripheral nerve cells. Primary cells from adult rat dorsal root ganglia (containing both neurons and glia) were used in these experiments. Localisation of mTHPC was detected using fluorescence and confocal microscopy. Levels of mTHPC fluorescence were quantified using digital image analysis. Immunocytochemistry with anti-?-III-tubulin and anti-S100 was used to distinguish neuronal and glial cell populations respectively. A cell-death assay using propidium iodide was used to evaluate neural cell susceptibility to PDT following incubation with mTHPC. The results showed that mTHPC was localised in cytoplasmic regions of neurons and glia, but was not detected in neuronal axons. Necrotic cell death was detected after PDT in these neural cell types. These results suggest that the cells of the peripheral nervous system are susceptible to PDT-mediated necrosis, but that the sparing of nerves observed during clinical PDT may be related to the heterogeneous distribution of mTHPC within neurons

Differences in sensitivity to mTHPC-mediated photodynamic therapy of neurons, glial cells and MCF7 cells in a 3-dimensional cell culture model

The effect of photodynamic therapy (PDT) on the cells of the nervous system is an important consideration in the treatment of tumours that are located within or adjacent to the brain, spinal cord and peripheral nerves. Previous studies have reported the sparing of nerves during PDT using meta-tetrahydroxyphenylchlorin (mTHPC, Foscan®) in patients and in animal models. The aim of this study was to investigate the effects of mTHPC on key nervous system cells using a 3-dimensional cell culture system for the accurate detection of differences in sensitivity

Conjugatable water-soluble Pt(ii) and Pd(ii) porphyrin complexes: Novel nano- and molecular probes for optical oxygen tension measurement in tissue engineering

Measurement of oxygen tension in compressed collagen sheets was performed using matrix-embedded optical oxygen sensors based on platinum(II) and palladium(II) porphyrins supported on polyacrylamide nanoparticles. Bespoke, fully water-soluble, mono-functionalised Pt(II) and Pd(II) porphyrin complexes designed for conjugation under mild conditions were obtained using microwave-assisted metallation. The new sensors display a linear response (1/τ vs. O₂) to varying oxygen tension over a biologically relevant range (7.0 × 10⁻⁴ to 2.7 × 10⁻¹ mM) in aqueous solutions; a behaviour that is maintained following conjugation to polyacrylamide nanoparticles, and following embedding of the nanosensors in compressed collagen sheets, paving the way to innovative approaches for real-time resolution of oxygen gradients throughout 3D matrices useful for tissue regeneration

The bactericidal activity of glutaraldehyde-impregnated polyurethane

Although glutaraldehyde is known to be bactericidal in solution, its potential use to create novel antibacterial polymers suitable for use in healthcare environments has not been evaluated. Here, novel materials were prepared in which glutaraldehyde was either incorporated into polyurethane using a simple “swell-encapsulation-shrink” method (hereafter referred to as “glutaraldehyde-impregnated polyurethane”), or simply applied to the polymer surface (hereafter referred to as “glutaraldehyde-coated polyurethane”). The antibacterial activity of glutaraldehyde-impregnated and glutaraldehyde-coated polyurethane samples was tested against Escherichia coli and Staphylococcus aureus. Glutaraldehyde-impregnated polyurethane resulted in a 99.9% reduction in the numbers of E. coli within 2 h and a similar reduction of S. aureus within 1 h, whereas only a minimal reduction in bacterial numbers was observed when the biocide was bound to the polymer surface. After 15 days, however, the bactericidal activity of the impregnated material was substantially reduced presumably due to polymerization of glutaraldehyde. Thus, although glutaraldehyde retains antibacterial activity when impregnated into polyurethane, activity is not maintained for extended periods of time. Future work should examine the potential of chemical modification of glutaraldehyde and/or polyurethane to improve the useful lifespan of this novel antibacterial polymer

Conductance Phases in Aharonov-Bohm Ring Quantum Dots

The regimes of growing phases (for electron numbers N~0-8) that pass into regions of self-returning phases (for N>8), found recently in quantum dot conductances by the Weizmann group are accounted for by an elementary Green function formalism, appropriate to an equi-spaced ladder structure (with at least three rungs) of electronic levels in the quantum dot. The key features of the theory are physically a dissipation rate that increases linearly with the level number (and tentatively linked to coupling to longitudinal optical phonons) and a set of Fano-like meta-stable levels, which disturb the unitarity, and mathematically the change over of the position of the complex transmission amplitude-zeros from the upper-half in the complex gap-voltage plane to the lower half of that plane. The two regimes are identified with (respectively) the Blaschke-term and the Kramers-Kronig integral term in the theory of complex variables.Comment: 20 pages, 4 figure

Renormalized Vacuum Polarization and Stress Tensor on the Horizon of a Schwarzschild Black Hole Threaded by a Cosmic String

We calculate the renormalized vacuum polarization and stress tensor for a massless, arbitrarily coupled scalar field in the Hartle-Hawking vacuum state on the horizon of a Schwarzschild black hole threaded by an infinte straight cosmic string. This calculation relies on a generalized Heine identity for non-integer Legendre functions which we derive without using specific properties of the Legendre functions themselves.Comment: This is an expanded version of a previous submission, we have added the calculation of the stress tensor. 28 pages, 7 figure

Model-Independent Sum Rule Analysis Based on Limited-Range Spectral Data

Partial sum rules are widely used in physics to separate low- and high-energy degrees of freedom of complex dynamical systems. Their application, though, is challenged in practice by the always finite spectrometer bandwidth and is often performed using risky model-dependent extrapolations. We show that, given spectra of the real and imaginary parts of any causal frequency-dependent response function (for example, optical conductivity, magnetic susceptibility, acoustical impedance etc.) in a limited range, the sum-rule integral from zero to a certain cutoff frequency inside this range can be safely derived using only the Kramers-Kronig dispersion relations without any extra model assumptions. This implies that experimental techniques providing both active and reactive response components independently, such as spectroscopic ellipsometry in optics, allow an extrapolation-independent determination of spectral weight 'hidden' below the lowest accessible frequency.Comment: 5 pages, 3 figure

Efficacy of photochemical internalisation using disulfonated chlorin and porphyrin photosensitisers: An in vitro study in 2D and 3D prostate cancer models

This study shows the therapeutic outcome of Photochemical Internalisation (PCI) in prostate cancer in vitro surpasses that of Photodynamic Therapy (PDT) and could improve prostate PDT in the clinic, whilst avoiding chemotherapeutics side effects. In addition, the study assesses the potential of PCI with two different photosensitisers (TPCS2a and TPPS2a) in prostate cancer cells (human PC3 and rat MatLyLu) using standard 2D monolayer culture and 3D biomimetic model. Photosensitisers were used alone for photodynamic therapy (PDT) or with the cytotoxin saporin (PCI). TPPS2a and TPCS2a were shown to be located in discrete cytoplasmic vesicles before light treatment and redistribute into the cytosol upon light excitation. PC3 cells exhibit a higher uptake than MatLyLu cells for both photosensitisers. In the 2D model, PCI resulted in greater cell death than PDT alone in both cell lines. In 3D model, morphological changes were also observed. Saporin-based toxicity was negligible in PC3 cells, but pronounced in MatLyLu cells (IC50 = 18 nM). In conclusion, the study showed that tumour features such as tumour cell growth rate or interaction with drugs determine therapeutic conditions for optimal photochemical treatment in metastatic prostate cancer

The hybrid spectral problem and Robin boundary conditions

The hybrid spectral problem where the field satisfies Dirichlet conditions (D) on part of the boundary of the relevant domain and Neumann (N) on the remainder is discussed in simple terms. A conjecture for the C_1 coefficient is presented and the conformal determinant on a 2-disc, where the D and N regions are semi-circles, is derived. Comments on higher coefficients are made. A hemisphere hybrid problem is introduced that involves Robin boundary conditions and leads to logarithmic terms in the heat--kernel expansion which are evaluated explicitly.Comment: 24 pages. Typos and a few factors corrected. Minor comments added. Substantial Robin additions. Substantial revisio