Molecular Contrast Optical Coherence Tomography: A Review

This article reviews the current state of research on the use of molecular contrast agents in optical coherence tomography (OCT) imaging techniques. After a brief discussion of the basic principle of OCT and the importance of incorporating molecular contrast agent usage into this imaging modality, we shall present an overview of the different molecular contrast OCT (MCOCT) methods that have been developed thus far. We will then discuss several important practical issues that define the possible range of contrast agent choice, the design criteria for engineered molecular contrast agent and the implementability of a given MCOCT method for clinical or biological applications. We will conclude by outlining a few areas of pursuit that deserve a greater degree of research and development

Particulate airborne impurities

The cumulative effects of air pollutants are of principal concern in research on environmental protection in Sweden. Post-industrial society has imposed many limits on emitted air pollutants, yet the number of reports on the negative effects from them is increasing, largely due to human activity in the form of industrial emissions and increased traffic flows. Rising concerns over the health effects from airborne particulate matter (PM) stem from in vitro, in vivo, and cohort studies revealing effects of mostly negative nature. Full insight into the health effects from PM can only be achieved through practical investigation of the mode of toxicity from distinct types of particles and requires techniques for their identification, monitoring, and the production of model fractions for health studies. To this effect, comprehensive collection and chemical analysis of particulates at the origin of emission was performed in order to provide clearer insight into the nature of the particulates at exposure and add detail to aid risk assessment. Methods of capturing particles and analyzing their chemical nature were devised using scanning electron microscopy coupled with energy dispersive spectroscopy (SEM-EDS). Furthermore, taking the approach of in vitro cytotoxicity testing, nanoparticles of types typical to automotive emissions, were synthesized and extensively characterized using SEM-EDS, X-ray diffraction (XRD), transmission electron microscopy (TEM),dynamic light scattering (DLS), and nanoparticle tracking analysis (NTA). The produced model magnetite and palladium nanoparticles were found to induce toxicity in human pulmonary epithelial cells (A549 and PBEC) as well as impact severely on immunological and renal cells (221 B- and 293T-cells) in a dose-dependent manner

Chemical regulators of epithelial plasticity reveal a nuclear receptor pathway controlling myofibroblast differentiation

Plasticity in epithelial tissues relates to processes of embryonic development, tissue fibrosis and cancer progression. Pharmacological modulation of epithelial transitions during disease progression may thus be clinically useful. Using human keratinocytes and a robotic high-content imaging platform, we screened for chemical compounds that reverse transforming growth factor β (TGF-β)-induced epithelial-mesenchymal transition. In addition to TGF-β receptor kinase inhibitors, we identified small molecule epithelial plasticity modulators including a naturally occurring hydroxysterol agonist of the liver X receptors (LXRs), members of the nuclear receptor transcription factor family. Endogenous and synthetic LXR agonists tested in diverse cell models blocked α-smooth muscle actin expression, myofibroblast differentiation and function. Agonist-dependent LXR activity or LXR overexpression in the absence of ligand counteracted TGF-β-mediated myofibroblast terminal differentiation and collagen contraction. The protective effect of LXR agonists against TGF-β-induced pro-fibrotic activity raises the possibility that anti-lipidogenic therapy may be relevant in fibrotic disorders and advanced cancer

The Importance of Dosimetry and Radiobiology in Nuclear Medicine : Quantitative methods and modelling

Nuclear medicine uses radioactive pharmaceuticals for diagnostic or therapeutic purposes. The ionizing radiation emitted from the radiopharmaceutical is partially absorbed within the patient's body and internal dosimetry is the method to estimate the absorbed dose to a tumour or risk organ. This is of special importance in radiopharmaceutical therapy (RPT), where particle-emitting radionuclides are utilized for their therapeutic effect. A better understanding of where and to what extent the radiation energy is deposited, i.e. dosimetry, in combination with a better understanding of the irradiation-induced biological processes in tissues and tumours, i.e., radiobiology, is the foundation to establish an absorbed dose-effect relationship. This thesis comprises quantitative methods and modelling within dosimetry and radiobiology, with a special focus on quantitative methods for activity concentration, absorbed dose calculation and quantification of biological effects after nuclear medicine exposures. Nonuniformity of activity distribution and the biological effect of internal irradiation is considered in Paper I and Paper II. When a radiopharmaceutical primarily localizes within specific tissue substructures of an organ, the average absorbed dose to the whole organ may become insufficient for dosimetric analysis. Hence, the nonuniformities of the distribution of activity need to be considered and absorbed dose calculations to part of an organ, cellular, or a sub-cellular structure may be a better predictor of the therapy outcome or normal tissue toxicity. In Paper I, a small-scale anatomical dosimetry model of the liver tissue structure addressed the issue of activity nonuniformity. Monte Carlo simulations were performed to simulate the particle transport from various substructure sources within the organ model for some clinically available radionuclides. The model enabled comparison between the average absorbed dose to the entire organ and the local absorbed dose close to the source region, which for particle emitting radionuclides differed significantly. To address the resulting biological effect after internal irradiation, an ex vivo method using the γH2AX surrogate marker to visualize and quantify DNA double-strand breaks in in vivo-irradiated tissues was developed. The method was demonstrated to be useful for γH2AX-foci quantification in both the fast proliferating, radiosensitive testis tissue and the slow proliferating and more radioresistant liver tissue. Image-based activity quantification and absorbed dose estimation are considered in Paper III and Paper IV, using somatostatin receptor targeting agents for both diagnostic and therapeutic applications for neuroendocrine tumours. In Paper III, the quantitative accuracy of pre-therapeutic 111In-Octreoscan® SPECT/CT and [68Ga]Ga-DOTA-TATE PET/CT images was investigated due to the change in clinical method to use PET- instead of SPECT-imaging. Further, the quantitative relationship between the theragnostic pair of DOTA-TATE was investigated in Paper IV. The relationship between activity uptakes observed at [68Ga]Ga-DOTA-TATE PET imaging and absorbed doses at subsequent [177Lu]Lu-DOTA-TATE therapy was studied. The study demonstrated that on a group level, a higher tumour uptake measured from pretherapeutic PET images is associated with higher absorbed doses in subsequent therapy with [177Lu]Lu-DOTA-TATE. However, on the individual level, there are limitations of using the 68Ga PET as a predictor for therapy absorbed dose

Investigation of the interactions between selected nanoparticles and human lung carcinoma cells at the single cell and single particle level

The recent advances in nanomaterials development and applications have sparked concerns regarding the safety of these materials in living organisms. This body of work investigates specific interactions between chosen nanoparticles and living human lung carcinoma (A549) cells --Abstract, page iv

Biocompatibility of Synthetic Poly(Ester urethane)/Polyhedral Oligomeric Silsesquioxane Matrices with Embryonic Stem Cell Proliferation and Differentiation

Incorporation of polyhedral oligomeric silsesquioxanes (POSS) into poly(ester urethanes) (PEU) as a building block results in a PEU/POSS hybrid polymer with increased mechanical strength and thermostability. An attractive feature of the new polymer is that it forms a porous matrix when cast in the form of a thin film, making it potentially useful in tissue engineering. In this study, we present detailed microscopic analysis of the PEU/POSS matrix and demonstrate its biocompatibility with cell culture. The PEU/POSS polymer forms a continuous porous matrix with open pores and interconnected grooves. From SEM image analysis, it is calculated that there are about 950 pores/mm2 of the matrix area with pore diameter size in the range 1-15 μm. The area occupied by the pores represents approximately 7.6% of the matrix area. Using mouse embryonic stem cells (ESCs), we demonstrate that the PEU/POSS matrix provides excellent support for cell proliferation and differentiation. Under the cell culture condition optimized to maintain self-renewal, ESCs grown on a PEU/POSS matrix exhibit undifferentiated morphology, express pluripotency markers and have a similar growth rate to cells grown on gelatin. When induced for differentiation, ESCs underwent dramatic morphological change, characterized by the loss of clonogenecity and increased cell size, with well-expanded cytoskeleton networks. Differentiated cells are able to form a continuous monolayer that is closely embedded in the matrix. The excellent compatibility between the PEU/POSS matrix and ESC proliferation/differentiation demonstrates the potential of using PEU/POSS polymers in future ESC-based tissue engineering. Copyright (C) 2010 John Wiley & Sons, Ltd

Multi-mode atomic force microscope as a versatile tool for bionanotechnology

The kernel of this dissertation is multi-mode atomic force microscopy (AFM) which is a useful and powerful tool for characterizing and analyzing samples of nano- or micro size. Various modes can satisfy specified requirements according to different samples, i.e., topography, surface electrostatic potential, magnetic domain visual observation, single molecular force analysis and a novel real-time monitoring cell viability system based on modification of AFM. No matter whether samples are in air or in liquid, topological image can be realized. Hence, the flexibility makes AFM a universal tool for exploring the biological nano-world. The subjects consist of different working modes towards biological applications. Firstly, topography is aimed at quantitative analysis of cellular morphology and surface changes, which are effected by uptake of nanoparticles. In the case of concentration-dependent experiments, the volume and number of filopodia is calculated by analyzing topological images of AFM. It is verified that cellular morphology plays an important role for quantitative indicating of harmful effects of NPs to cells. In addition, the roughness of the cellular surface which derives from disruption of cell membrane integrity, when the cells internalized magnetic NPs subjected to a rotating magnetic field, is evaluated for exploring magneto-cell-poration and magneto-cellanalysis. Secondly, single molecule force microscopy is aimed at quantitative analysis of elasticity of gold nanoparticles (Au NPs), which are coated with polyethylene glycol (PEG), whereby the diameter of the gold cores as well as the thickness of the shell of PEG was varied. A conical tip indent into single NP and then Sneddon’s equation is employed for calculating the elasticity, which serves as one of the basic physicochemical parameters having effect on structural and functional cell parameters. Thirdly, magnetic force microscopy is aimed at qualitative visual observation of magnetic domains of the sample, which is a multifunctional co-loading NP with anti-drug tetradine and superparamagnetic iron dioxide (Fe3O4) NPs. The magnetic domains of co-loading NPs, which is reflected in phase section, can present magnetic profile which is attributed to the Fe3O4 NPs. Thus such multifunctional co-loading NPs are further used for magnetic ablation to tumor cells, so that a dual enhanced anti-cancer NP can be successfully realized. Fourthly, electrostatic force microscopy (EFM) is aimed at qualitative visual observation of electrostatic potential on surface of the sample, which is a mutant purple membrane (PM) modified by functional NPs. A bias voltage between a conductive tip and the modified PM is applied in an oscillating mode. The tip is lifted such that it can induce a long term electrostatic force without effect of molecular repulsive force. Thus electric gradient dependent on surface of the PM makes phase shift in a given frequency and then the EFM signal is extracted. Therefore, the electric property of such a novel biomembrane is characterized. Fifthly, a generally applicable quantitative real-time cell viability monitoring system which uses cell adhesion property is successfully setup based on the oscillation system of AFM. The amplitude of an oscillating cantilever at a given frequency is highly dependent on the mass of the cantilever, in this situation, the mass of attached cells on the cantilever. In our method, the dynamic toxic process can be observed and recorded, and can be analyzed even at an early stage of intoxication. Therefore, this will be a greatly promising method for real-time exploring and quantitatively analyzing of cellular toxicity

Advanced Fluorescence Microscopy Techniques-FRAP, FLIP, FLAP, FRET and FLIM

Fluorescence microscopy provides an efficient and unique approach to study fixed and living cells because of its versatility, specificity, and high sensitivity. Fluorescence microscopes can both detect the fluorescence emitted from labeled molecules in biological samples as images or photometric data from which intensities and emission spectra can be deduced. By exploiting the characteristics of fluorescence, various techniques have been developed that enable the visualization and analysis of complex dynamic events in cells, organelles, and sub-organelle components within the biological specimen. The techniques described here are fluorescence recovery after photobleaching (FRAP), the related fluorescence loss in photobleaching (FLIP), fluorescence localization after photobleaching (FLAP), Forster or fluorescence resonance energy transfer (FRET) and the different ways how to measure FRET, such as acceptor bleaching, sensitized emission, polarization anisotropy, and fluorescence lifetime imaging microscopy (FLIM). First, a brief introduction into the mechanisms underlying fluorescence as a physical phenomenon and fluorescence, confocal, and multiphoton microscopy is given. Subsequently, these advanced microscopy techniques are introduced in more detail, with a description of how these techniques are performed, what needs to be considered, and what practical advantages they can bring to cell biological research