Detection modalities that can evaluate the early stages of dental caries are indispensable. The purpose of this thesis is to evaluate the efficacy of photothermal radiometry and modulated luminescence (PTR-LUM) to non-destructively detect and quantify simulated enamel caries. Two experiments were performed based on the PTR-LUM detection mode: back-propagation or transmission-mode. Artificial demineralized lesions were created in human molars and a subset was further exposed to an artificial remineralizing solution. PTR-LUM frequency scans were performed periodically during de/re-mineralization treatments. PTR data was fitted to a theoretical model based on optical and thermal fluxes in enamel to extract opto-thermophysical parameters. Lesion validation was performed using transverse microradiography (TMR). Optical and thermal properties changed with the development and repair of the caries lesions while theory-derived thicknesses paralleled those determined microradiographically. These trends coupled with the uniqueness-of-fit of the generated parameters illustrate the efficacy of PTR- LUM to non-destructively detect and quantify de/re-mineralized lesions.

Detection modalities that can evaluate the early stages of dental caries are indispensable. The purpose of this thesis is to evaluate the efficacy of photothermal radiometry and modulated luminescence (PTR-LUM) to non-destructively detect and quantify simulated enamel caries. Two experiments were performed based on the PTR-LUM detection mode: back-propagation or transmission-mode. Artificial demineralized lesions were created in human molars and a subset was further exposed to an artificial remineralizing solution. PTR-LUM frequency scans were performed periodically during de/re-mineralization treatments. PTR data was fitted to a theoretical model based on optical and thermal fluxes in enamel to extract opto-thermophysical parameters. Lesion validation was performed using transverse microradiography (TMR). Optical and thermal properties changed with the development and repair of the caries lesions while theory-derived thicknesses paralleled those determined microradiographically. These trends coupled with the uniqueness-of-fit of the generated parameters illustrate the efficacy of PTR- LUM to non-destructively detect and quantify de/re-mineralized lesions.

In frequency-domain photothermal radiometry (FD-PTR) a low-power intensity-modulated optical excitation generates thermal-wave field inside the sample and the subsequent infrared radiation from the sample is analyzed to detect material’s inhomogeneities. The non-contact nature of FD-PTR makes it very suitable for non-destructive evaluation of broad range of materials. Moreover, the methodology is based on intrinsic contrast of light absorption which can be used as a diagnostic tool for inspection of malignancy in biological tissues. Nevertheless, the bottom line is that the physics of heat diffusion allows for a highly damped and dispersive propagation of thermal-waves. As a result, the current FD-PTR modalities suffer from limited inspection depth and poor axial/depth resolution. The main objective of this thesis is to show that using alternative types of modulation schemes (such as linear frequency modulation and binary phase coding) and radar matched filter signal processing, one can obtain localized responses from inherently diffuse thermal wave fields. In this thesis, the photothermal responses of turbid, transparent, and opaque media to linear frequency modulated and binary phase coded excitations are analytically derived. Theoretical simulations suggest that matched-filtering in diffusion-wave field acts as constructive interferometry, localizing the energy of the long-duty excitation under a narrow peak and allowing one to construct depth resolved images. The developed technique is the diffusion equivalent of optical coherence tomography and is named thermal coherence tomography. It was found that the narrow-band binary phase coded matched filtering yields optimal depth resolution, while the broad-band linear frequency modulation can be used to quantify material properties through the multi-parameter fitting of the experimental data to the developed theory. Thermophotonic detection of early dental caries is discussed in detail as a potential diagnostic application of the proposed methodologies. The performance of the diagnostic system is verified through a controlled demineralization protocol as well as in teeth with natural caries.

Photothermal cancer therapy is a potential alternative to surgery and involves selective tissue destruction using thermal energy. Targeted photoabsorbers, used in conjunction with matching a continuous wave laser, make photothermal therapy both noninvasive and tumor-specific. However, to become clinically relevant, there is a need to develop an imaging technique to identify tissue composition and to detect the presence of photoabsorbers in the tumor volume before therapy; to monitor the temperature rise during therapy; and to assess the tumor damage after therapy. In this study, a combined ultrasound and photoacoustic imaging system was designed to assist photothermal therapy. The imaging system was tested on tissue mimicking phantoms, ex-vivo porcine tissue samples, ex-vivo mice and in-vivo mice. First, ultrasound imaging was utilized to differentiate between water-based and lipidbearing tissue. A combined ultrasound and photoacoustic imaging system was then assembled to identify the presence and spatial location of gold nanoparticles. Multiwavelength photoacoustic imaging was used to further confirm the presence of nanoparticles. Temperature monitoring algorithms, using both temperature-dependent time shifts in ultrasound signals and amplitude changes in photoacoustic signals, were developed. Finally, photothermal therapy was carried out on tumor-bearing nude mice using in-vivo ultrasound and photoacoustic imaging to identify the tumor boundary, detect the nanoparticles and monitor the temperature elevation. The results of the studies show that ultrasound and photoacoustic imaging provide complementary and clinically relevant information. Overall, there is potential of using the ultrasound and photoacoustic imaging system to plan, guide and monitor photothermal therapy. / text

Atherosclerosis and specifically rupture of vulnerable plaques account for 23% of all deaths worldwide, far surpassing both infectious diseases and cancer. Plaque-based macrophages, often associated with lipid deposits, contribute to atherogenesis from initiation through progression, plaque rupture and ultimately, thrombosis. Therefore, the macrophage is an important early cellular marker related to vulnerability of atherosclerotic plaques. The objective of my research is to assess the ability of multiple optical imaging modalities to detect, and further characterize the distribution of macrophages (having taken up plasmonic gold nanoparticles as a contrast agent) and lipid deposits in atherosclerotic plaques. Tissue phantoms and macrophage cell cultures were used to investigate the capability of nanorose as an imaging contrast agent to target macrophages. Ex vivo aorta segments from a rabbit model of atherosclerosis after intravenous nanorose injection were imaged by optical coherence tomography (OCT), photothermal imaging (PTW) and two-photon luminescence microscopy (TPLM), respectively. OCT images depicted detailed surface structure of atherosclerotic plaques. PTW images identified nanorose-loaded macrophages (confirmed by co-registration of a TPLM image and corresponding RAM-11 stain on a histological section) associated with lipid deposits at multiple depths. TPLM images showed three-dimensional distribution of nanorose-loaded macrophages with a high spatial resolution. Imaging results suggest that superficial nanorose-loaded macrophages are distributed at shoulders on the upstream side of atherosclerotic plaques at the edges of lipid deposits. Combination of OCT with PTW or TPLM can simultaneously reveal plaque structure and composition, permitting assessment of plaque vulnerability during cardiovascular interventions. / text

Molecular imaging is an emerging imaging principle which can visually represent the biological processes both spatially and temporally down to the sub-cellular level in vivo. The outcome of this research is expected to have a profound impact on facilitating the early diagnosis of diseases, accelerating the development of new drugs, and improving the efficacy of therapy. In general, molecular imaging highly relies on probes to sense the occurrence of molecular biological events, and to generate signals which could be picked up by diagnostic imaging modalities. The advances in the design of molecular probes not only have equipped traditional anatomical medical imaging with new capabilities but also, in some cases, stimulated developments of new imaging modalities and renaissance of existing medical imaging modalities. One of these is photoacoustic imaging, which as an emerging medical imaging modality, unites the merits from both optical imaging and ultrasound imaging. It shares with optical imaging, that it uses non-ionizing radiation and provides higher contrast and higher sensitivity than ultrasound imaging. Unlike optical imaging, which requires ballistic photons for imaging, photoacoustic imaging requires only diffusive photons to excite the ultrasound signal from the imaging target; therefore, it is capable of imaging much deeper into the tissue. In combination with molecular probes, photoacoustic molecular imaging has been demonstrated by several research groups using various photoacoustic molecular probes. However, the molecular probes used for most of these studies were contrast agents simply adopted from other optical imaging modalities. Our research on photoacoustic contrast agents indicated that the mechanism of photoacoustic signal generation from nanometer-sized contrast agents is distinct from that of optically homogeneous materials, such as tissue. We have discovered that, the amplitude of the photoacoustic signal generated from nano-contrast agents depends not only on the optical absorption of the particles, but more importantly, on the dynamic process of the heat conduction from the nanoparticles to the ambient, and the thermal properties of the surrounding materials. Based on our finding, we explored and further improved the photoacoustic response of the nanoparticles by exploiting the heat conduction process between the nanoparticle and its surrounding materials and by manipulating the excitations. This research allows to create optimized molecular specific contrast enhanced photothermal stable probes which can aid photoacoustic imaging and image guided photothermal cancer therapy. / text

Gold nanoparticles, a class of plasmonic nanoparticle, have increasingly been explored as an imaging and therapeutic agent to treat cancer due to their characteristic surface plasmon resonance phenomenon and penchant for tumor accumulation. Photothermal therapy has been shown as a promising cancer treatment by delivering heat specifically to the tumor site via gold nanoparticles. In this study, we demonstrate that gold nanorod (GNR)-mediated photothermal therapy can be more effective through the understanding of cell death mechanisms. By targeting GNRs to various cellular localizations, we explored the association of GNR localization with cell death pathway response to photothermal therapy. Furthermore, we compared the 2D monolayer experiments with 3D in vitro tumor models, multicellular tumor spheroids (MCTS), to mimic the structure of in vivo tumors. With MCTS, we evaluated the cell death response with GNRs distributed only on the periphery, as seen in typical in vivo studies, and distributed evenly throughout the tumor. We demonstrated that GNR localization influences the cell death response to photothermal therapy by showing the power threshold necessary to induce significant apoptotic and necrotic increases was lower for internalized GNRs than membrane-bound GNRs. Furthermore, apoptosis was found to increase with increasing laser power until the necrotic threshold and decreased above it, as necrosis became the dominant cell death pathway response. A similar trend was revealed with the 3D MCTS; however, the overall cell death percentages were lower, most likely due to the upregulated cell repair response and varied GNR distributions due to the presence of cell-cell and cell-matrix interactions. Furthermore, the uniformly distributed GNRs induced more apoptosis and necrosis than GNRs located in the MCTS periphery. In conclusion, we quantitatively analyzed the cell death pathway response to GNR-mediated photothermal therapy to establish that it has some dependence on GNR localization and distribution to gain a more thorough understanding of this response for photothermal therapy optimization. / text

The persistence of high global mortality rates directly attributable to cardiovascular disease drives ongoing research into novel approaches for improved diagnosis and treatment of its primary underlying cause, atherosclerosis. Combined intravascular ultrasound and photoacoustic (IVUS/IVPA) imaging is one such modality, actively being developed as a tool for improved characterization of high-risk atherosclerotic plaques. The pathophysiology associated with progression and destabilization of atherosclerotic plaques leads to characteristic changes in arterial morphology and composition. IVUS/IVPA imaging seeks to expand upon the ability of clinically utilized intravascular ultrasound (IVUS) imaging to assess vessel anatomy by adding improved sensitivity to image the underlying cellular and molecular composition through intravascular photoacoustic (IVPA) imaging of either endogenous chromophores (e.g. lipid) or exogenously delivered contrast agents. This dissertation focuses on the expansion of IVUS/IVPA imaging using exogenous contrast agents to enable the detection and subsequent optically-triggered therapy of atherosclerotic plaques. The passive extravasation and aggregation of systemically injected plasmonic gold nanorods absorbing within the near infrared tissue optical window within plaques of atherosclerotic rabbit models is first demonstrated, along with the ability to localize the contrast agents using ex vivo IVUS/IVPA imaging. The motivation for nanoparticle labeling of atherosclerosis is then expanded from that of purely image contrast agents to vehicles for image-guided, dual-modality phototherapy. The integrated IVUS/IVPA imaging catheter is utilized for photothermal delivery with simultaneous IVPA temperature monitoring using the high optical absorption of gold nanorod contrast agents to enable localized heating. Subsequently, the potential role for IVUS/IVPA-guided phototherapy is further expanded through the characterization and in vitro assessment of novel multifunctional theranostic nanoparticles comprised of a gold nanorod core with a degradable, photosensitizer-doped silica shell. Together, the results presented within this dissertation provide a framework for ongoing research into the expansion of IVUS/IVPA imaging as a platform for complimentary diagnosis and local treatment of atherosclerotic plaques using multifunctional theranostic nanoparticle contrast agents. / text

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<p>The advancement in nanotechnology creates a new perspective on future medicine. With more and more understanding on controlling the functional behavior of the nanoplatform, scientists nowadays are aiming to improve the health care system by offering personalized medicine through nanotechnology. Lots of emphasis have been placed on a promising field called theranostics, which integrate imaging and therapeutic functions into one, that not only offers monitoring and imaging of the biological process, but also provides diagnosis and drug delivery simultaneously. Plasmonic gold nanostars, because of its anisotropic geometry and unique plasmonic property, have become one of the most anticipated nanoplatform in the field of nanotheranostics, aiming to achieve superior plasmonic properties for biomedical applications. The work herein will provide an introduction to the related field on plasmonics, nanobiophotonics and nanotheranostics. A facile plasmon-tunable surfactant-free nanostars synthesis method is described followed by an extensive characterization both computationally and experimentally. Its superior plasmon behavior on two-photon photoluminescence imaging and surface-enhanced Raman scattering detection are demonstrated both in cells and in animals. Therapeutic function assessment is carried out both as drug carriers (photodynamic therapy) and as endogenous stimulus responsive agents (photothermal therapy). Finally, the nanostars' cellular uptake mechanism is investigated based on nanostars' endogenous contrast; an enhanced photothermal therapy is achieved using an ultralow irradiance that has ever published. With nanostars being a novel and powerful theranostic agent, the potentials implication lies in the study of their pharmacokinetics, targeted delivery, diagnostic imaging, and toxicity.</p> / Dissertation