As photodynamic therapy is reliant on endogenous tumor oxygen to be successful, personalizing treatment to the patient is a necessary step to improving overall efficacy. We have demonstrated the feasibility of functionally imaging PDT in real-time, by mapping oxygen saturation as a surrogate marker for PDT progression with ultrasound-guided photoacoustic imaging in vivo. Particularly, we have deciphered the spatial map of the oxygen depletion rate during PDT as a function of fluence rate. Photoacoustic imaging importantly provides the temporal and spatial resolution, providing a novel platform for monitoring that can be used to inform personalized treatment in future work.
In this study, we study the influence of dye to antibody ratio on the imaging and therapeutic performance of an EGFR-targeted dual function conjugate. We further demonstrate the efficacy of the conjugate in identifying tumor depth to guide surgical resections and the ability to eliminate residual microscopic disease through photoimmunotherapy, in a single intra-operative setting.
Within 24 hours post treatment, photodynamic therapy causes a drastic reduction in tumor oxygenation and an increase in ultrasound scattering. While these trends can lead to determining acute tumor response, monitoring vascular and structural dynamics through 72 hours post treatment using ultrasound guided photoacoustic imaging provides a better insight on long-term PDT outcomes.
KEYWORDS: Photoacoustic spectroscopy, MATLAB, Process modeling, Tissues, Monte Carlo methods, Imaging systems, Image segmentation, Visualization, Visual process modeling, Ultrasonography
This work develops a thorough and user-friendly methodology for fluence compensation of photoacoustic (PA) data from the commercial Visualsonics (VS) VevoLAZR-X System. PHANTOM (PHotoacoustic ANnotation TOolkit) has been created to help fluence compensate large PA datasets. PHANTOM provides an easy-to-use graphical user interface to assist users in segmenting US-coupled PA images into three-dimensional labeled tissue structures with assignable optical characteristics. We also modeled an MCX-compatible light source configuration that replicates the VS system's optical fiber illumination. PHANTOM exports all Monte Carlo configuration parameters which are compatible with Monte Carlo eXtreme (MCX) allowing fluence compensation of PA data.
Liquid metal nanoparticles, particularly Eutectic Gallium Indium (EGaIn), hold great potential in medical applications such as biosensors, bioelectrodes, cancer treatment, and medical imaging. This study focuses on the synthesis of EGaIn nanoparticles using sonication without harsh chemicals. The nanoparticles are surface functionalized with hyaluronan and the photosensitizer benzoporphyrin derivative (BPD) for photodynamic therapy. EGaIn nanoparticles exhibit stability, good biocompatibility, and high optical absorption for photoacoustic imaging. The singlet oxygen generation of EGaPs is compared with free BPD under physiological conditions. Additionally, in vitro and in vivo investigations confirm the photodynamic efficacy of EGaPs, making them versatile nanoparticles for targeting, imaging, drug delivery, and photodynamic therapy
Head and neck cancer (HNC) presents challenges due to its heterogeneity, high recurrence rate, and hypoxic microenvironments. Current treatments fall short, leading to high morbidity and mortality. Photodynamic therapy (PDT) offers spatiotemporal tumor control but relies on oxygenation, posing limitations in hypoxic tumors. Combining hypoxia-activated prodrugs with PDT can selectively kill hypoxic tumor cells and reduce the tumor burden. This study proposes using evofosfamide, a hypoxia-activated prodrug, in combination with benzoporphyrin derivative (BPD)-PDT in a 3D spheroids model of HNC. The growth profile, BPD uptake profile, and hypoxia development were investigated. Combination therapy enhanced PDT efficacy in hypoxic HNC spheroids.
In this work, murine xenografts of pancreatic cancer cell lines AsPC-1 and Mia PaCa-2 were used to investigate vascularization, oxygenation, and the effect of sunitinib treatment on pancreatic cancer. Ultrasound-guided photoacoustic imaging (US-PAI) at multiple wavelengths was utilized to study the tumor vascular networks, throughout the receptor tyrosine kinase inhibitor sunitinib treatment. The 3D photoacoustic data was fluence compensated and spectrally unmixed to acquire the parameters of blood oxygen saturation (StO2) and total hemoglobin concentration (HbT). A custom regional segmentation algorithm was applied to the volumetric HbT images to segment the tumor volume into areas of high vascular density and low vascular density. Regionally evaluating the changes in StO2 and HbT reveals that sunitinib is preferentially targeting areas of low vascular density.
Obstetric risks, associated with pre-eclampsia, such as maternal age and pre-existing conditions are on the rise, along with the risk of ectopic calcification and placental vascular dysfunction. We investigated Slc20a2 mice models through non-invasive commercialized and synergistic ultrasound and photoacoustic imaging, which provide dual-wavelength images and radio frequency data. The increased ectopic placental vascular calcification model exhibits reduced fetal growth and decreased postnatal bone mineral density. Our experiments established a significant difference in both placental function and structural differences between normal and diseased placentas through the provided oxygen saturation and quantitative ultrasound spectral parameters.
KEYWORDS: Tumors, Nanodroplets, Photodynamic therapy, Oxygen, Photoacoustic spectroscopy, Hypoxia, Solids, In vivo imaging, Resistance, Real time imaging
Solid tumors face the challenge of hypoxia, resulting from an imbalance between oxygen demands and supply. Hypoxia leads to resistance to conventional cancer therapies like radiation and chemotherapy, including photodynamic therapy (PDT) that relies on oxygen radicals. To address this, we developed perfluorocarbon nanodroplets for co-delivering oxygen and a photosensitizer. In vitro and in vivo studies validated oxygen release and enhanced tumor oxygenation. Histological analysis confirmed reduced hypoxic regions in nanodroplet-treated tumors. PDT using the nanodroplets demonstrated superior efficacy compared to a liposomal formulation. Overall, oxygen-loaded nanodroplets guided by photoacoustic imaging offer a promising approach for hypoxic tumor treatment.
Amongst the several biomedical imaging modalities, Photoacoustic imaging stands out due to its advantage of providing optical contrast at ultrasound resolution from deeper tissues. The optical illumination is traditionally provided by the nanosecond-pulse width lasers, but they are costly, bulky, and non-portable. Light Emitting Diode-based systems can circumvent all these issues, but they deliver low-energy that brings forth another problem of low signal-to-noise-ratio (SNR) images. Averaging several frames at the same cross-section over time removes the noise, but real-time dynamic functionalities might not be captured. The tradeoff between SNR and real-time acquisition can be mitigated with a downstream noise removal algorithm. The traditional algorithms are not efficient and require prior knowledge about the noise type distribution for which deep learning-based architectures such as U-Net and generative adversarial network (GAN) are implemented. One of the issues of these supervised networks is the requirement of paired training input-label dataset which is highly cumbersome to capture or sometimes is unavailable. The pixel-wise correspondence will act as a pre-processing overburden for acquiring training data. To mitigate this issue, we implemented a Cycle-consistent GAN denoising (DenCyc-GAN) algorithm which works on unpaired training data. We compared our network’s outputs with other traditional non-learning and deep learning network and found that our network performed similar to the supervised networks with respect to image quality metrics such as Peak SNR and structural similarity index.
SignificanceIndia has one of the highest rates of oral squamous cell carcinoma (OSCC) in the world, with an incidence of 15 per 100,000 and more than 70,000 deaths per year. The problem is exacerbated by a lack of medical infrastructure and routine screening, especially in rural areas. New technologies for oral cancer detection and timely treatment at the point of care are urgently needed.AimOur study aimed to use a hand-held smartphone-coupled intraoral imaging device, previously investigated for autofluorescence (auto-FL) diagnostics adapted here for treatment guidance and monitoring photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) fluorescence (FL).ApproachA total of 12 patients with 14 buccal mucosal lesions having moderately/well-differentiated micro-invasive OSCC lesions (<2 cm diameter and <5 mm depth) were systemically (in oral solution) administered three doses of 20 mg / kg ALA (total 60 mg / kg). Lesion site PpIX and auto-FL were imaged using the multichannel FL and polarized white-light oral cancer imaging probe before/after ALA administration and after light delivery (fractionated, total 100 J / cm2 of 635 nm red LED light).ResultsThe handheld device was conducive for access to lesion site images in the oral cavity. Segmentation of ratiometric images in which PpIX FL is mapped relative to auto-FL enabled improved demarcation of lesion boundaries relative to PpIX alone. A relative FL (R-value) threshold of 1.4 was found to segment lesion site PpIX production among the patients with mild to severe dysplasia malignancy. The segmented lesion size is well correlated with ultrasound findings. Lesions for which R-value was >1.65 at the time of treatment were associated with successful outcomes.ConclusionThese results indicate the utility of a low-cost, handheld intraoral imaging probe for image-guided PDT and treatment monitoring while also laying the groundwork for an integrated approach, combining cancer screening and treatment with the same hardware.
In this work, we evaluated the acoustic and optical properties of silk protein-based hydrogels to investigate its potential as a phantom material. Acoustic properties include the speed of sound and acoustic attenuation of silk scaffolds at various concentrations. Optical properties include optical absorption and reduced scattering measured between 400 nm to 1200 nm to coincide with common photoacoustic imaging bandwidths. The results indicate that silk is an acceptable phantom material for ultrasound and photoacoustic systems as it inherently displays similar acoustic properties and reduced scattering as various tissue types while displaying low absorption.
As medical advances allow the maternal age to rise, the risk of ectopic calcification and placental vascular dysfunction increases, both associated with preeclampsia. We investigated Slc20a2 mice models through non-invasive commercial ultrasound and photoacoustic imaging. This model exhibits increased ectopic placental vascular calcification, reduced fetal growth, and decreased postnatal bone mineral density. Our experiments established a significant difference in both placental function and structural differences between normal and diseased placentas. Dual-wavelength images and radio frequency data provided oxygen saturation and quantitative ultrasound spectral parameters that proved the control placentas can be distinguished from knockout placentas.
KEYWORDS: Light emitting diodes, Signal to noise ratio, In vivo imaging, Photoacoustic imaging, Imaging systems, Imaging arrays, Image quality, Image resolution, Signal generators, Real time imaging
Portable LED-based systems attempt to replace the bulky laser-based photoacoustic (PA) systems. The problem with LEDs is their low energy which generates low signal-to-noise-ratio (SNR) images. To obtain a high SNR image in real-time, we built a deep learning U-net model which transforms a low no. of frame-averaged image into a high no. of frame-averaged quality image. Both laser-based Vevo LAZR-X system and immunofluorescence histology staining show similar vascular organizations with hypoxic cores. We also achieved high SNR by running the algorithm on acoustic-resolution PA microscopy captured images. This generic network can be implemented in multiple scenarios.
Precision molecular imaging finds application in the delineation of tumor margins during surgical resection of head and neck cancers (HNCs). Despite the advantages of surgery, there remain challenges in successfully locating tumor margins, resecting the entire tumor volume and treatment of microscopic tumor tissue. The presence of residual tumors, post-surgery, may require additional interventions and often lead to tumor recurrence. While Epidermal growth factor receptor (EGFR) remains a receptor of choice for targeting in HNCs, the heterogeneity in the expression of EGFR often leads to variations in response to targeted therapeutics. To improve visualization and tumor margin delineation during head and neck tumor surgeries, this study demonstrates the development of a molecular targeted theranostic probe combining the complementary features of fluorescence and photoacoustic imaging. The probe: DFAC (Dual Function Antibody Conjugate) comprises of a fluorophore/photosensitizer; Benzoporphyrin derivative (BPD) and a photoacoustic contrast agent; naphthalocyanine (NC) derivative conjugated to an EGFR antibody; Cetuximab. While BPD assists in fluorescence imaging, it can also be used for inducing cytotoxicity, through photodynamic activation in target tissues. The efficacy of DFAC in selective visualization and photodynamic therapy of tumor cells is evaluated on heterocellular 3D tumor spheroids and orthotopic mouse tongue tumors developed from human oral cancer cell lines (CAL27 and SCC4), expressing different levels of EGFR. In summary, this study demonstrates the potential of the theranostic probe (DFAC) to delineate tumor regions for guiding surgical resection and eradicate residual tumor tissue (post-surgery) by photodynamic therapy.
This Conference Presentation, “Collagen detection in silk protein-based scaffolds through ultrasound and photoacoustic imaging,” was recorded at SPIE Photonics West held in San Francisco, California, United States
This Conference Presentation, “Eutectic gallium-indium alloy based liquid metal nanoparticles for photoacoustic imaging and photodynamic therapy,” was recorded at SPIE Photonics West 2022 held in San Francisco, California, United States.
KEYWORDS: Oxygen, Tumors, Photodynamic therapy, Nanodroplets, Image enhancement, Photoacoustic spectroscopy, Hypoxia, Solids, In vivo imaging, Real time imaging
This Conference Presentation, “Photoacoustic image guided oxygen enhanced photodynamic therapy of hypoxic tumors,” was recorded at SPIE Photonics West held in San Francisco, California, United States
This Conference Presentation, “3D ultrasound guided-photoacoustic vascular imaging of pancreatic tumors to predict response to tyrosine kinase inhibitor therapy,” was recorded at SPIE Photonics West held in San Francisco, California, United States
India has one of the highest rates of oral squamous cell carcinoma (OSCC) in the world, with an incidence of 15 per 100,000 and more than 70,000 deaths per year. The problem is exacerbated by lack of medical infrastructure and routine screening, especially in rural areas. This collaboration recently developed, and clinically validated, a low-cost, portable and easy-to-use platform for intraoral photodynamic therapy (PDT) specifically engineered for use in global health settings. Here, we explore the implementation of our low-cost PDT system in conjunction with a small, handheld smartphone-coupled, multichannel fluorescence and white-light oral cancer imaging probe, which was also developed for global health settings. Our study aimed to use this mobile intraoral imaging device for treatment guidance and monitoring PDT using 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PS; PpIX) fluorescence. A total of 12 patients with 14 lesions having moderately/well-differentiated micro-invasive OSCC lesions (<2 cm diameter, depth <5 mm) were systemically administered with three doses of 20mg/kg ALA (total 60mg/kg). Lesion site PpIX and auto fluorescence was analyzed before/after ALA administration, and again after light delivery (fractionated, total 100 J/cm2 of 630nm red LED light). Quantification of relative PpIX fluorescence enables lesion area segmentation to improve guidance of light delivery and reports extent of photobleaching. These results indicate the utility of this approach for image-guided PDT and treatment monitoring while also laying groundwork for an integrated approach, combining cancer screening and treatment with the same hardware.
We present the results of combined fluorescence and optoacoustic monitoring of tumor treatment using novel photoactivatable multi-inhibitor liposomes with BPD and Irinotecan providing a synergetic effect of PDT and chemotherapeutic impact.
KEYWORDS: Light emitting diodes, Photoacoustic imaging, 3D image processing, Imaging arrays, Deep tissue imaging, Tissue optics, Ultrasonography, Tumors, Switches, Signal to noise ratio
Low cost and high-power LED’s are replacing highly expensive and bulky Lasers in photoacoustic imaging (PAI) technology. LEDs bring the opportunity to image samples at a higher repetition rate (in kHz) compared to the conventional Q switched lasers (typically upto 20 Hz) and thus offer high frame rate and averaging power while maintain real-time imaging capability. In order to make a complete switch from Laser to LEDs, a proper understanding of this newly emerging class of LED arrays is important, for e.g., beam divergence, beam focus, coherence, time-dependent power and wavelength stability, penetration depth and sensitivity, etc. We performed characterization and optimization experiments on different tissue mimicking phantoms with various optical scattering and absorbing properties using a commercially available AcousticX PAI system with customized 3D printed holders for the LED array. The axial and lateral positions of LED arrays with respect to the acoustic probe to image deep lesions upto 1.5 cm were optimized. We also report the sensitivity of the system and Signal-to-noise-ratio (SNR) dependency of the LED arrays’ axial and lateral positions. Finally, we also demonstrate the adaptability of our custom designed, 3D printed variable angle and distance holder to perform deep tissue ultrasound and photoacoustic imaging probe on phantoms of with absorbers at different depth and in subcutaneous tumors and compare it with traditional laser based photoacoustic imaging systems.
Photodynamic therapy (PDT) is a photochemistry based cytotoxic technique that imparts cellular damage via excitation of a photosensitizer with drug-specific wavelength of light. Previously we and other groups have shown the efficacy of photoacoustic imaging in predicting treatment response of vascular targeted photodynamic therapy. In this work we evaluate the efficacy of photoacoustic imaging to monitor cellular targeted photodynamic therapy and its role in predicting recurrence. The dose at the treatment site for I PDT is determined by three factors: photosensitizer (PS) concentration, oxygenation status and delivered light irradiance. Most of the FDA approved photosensitizers in their triplet-excited state generate cytotoxic species by reacting with the ground state oxygen that is available in the surrounding environment. Given the inter- and intra-subject variability in the uptake of the photosensitizer and the distribution of oxygen in the tumor, understanding the interplay between these dose parameters could aid in determining photodynamic therapy efficacy. Using various subcutaneous and orthotopic mouse models we demonstrate both oxygenation status of the tumor prior to the treatment and the change in oxygen saturation 48 hrs post treatment can predict efficacy of Aminolevulinic acid (ALA) based cellular PDT with <92% sensitivity and specificity. We further compare the predictive capability of photoacoustic imaging with the more predominantly used fluorescence imaging and immunohistochemistry techniques.
This Conference Presentation, “Photoacoustic nanodroplets for oxygen enhanced photodynamic therapy,” was recorded for the Photonics West 2021 Digital Forum.
In this study, we evaluated the capability of an LED-based photoacoustic imaging system (AcousticX, 7MHz ultrasound probe, 850 nm LED arrays) in imaging heterogenous vasculature of subcutaneous tumors in mice. For evaluating the functional imaging capability, we used AcousticX with dual-wavelength LED arrays (750/850 nm) and monitored oxygen saturation of the tumor volume. We also compared the tumor images acquired by AcousticX and a commercial laser-based photoacoustic imaging system. Our results show that LED-based photoacoustic imaging can obtain similar imaging performance as a laser-based system and thus holds good promise in preclinical cancer research in an affordable setting.
Significance: India has one of the highest rates of oral cancer incidence in the world, accounting for 30% of reported cancers. In rural areas, a lack of adequate medical infrastructure contributes to unchecked disease progression and dismal mortality rates. Photodynamic therapy (PDT) has emerged as an effective modality with potential for treating early stage disease in resource-limited settings, while photosensitizer fluorescence can be leveraged for treatment guidance.
Aim: Our aim was to assess the capability of a simple smartphone-based device for imaging 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) fluorescence for treatment guidance and monitoring as part of an ongoing clinical study evaluating low-cost technology for ALA-based PDT treatment of early oral cancer.
Approach: A total of 29 subjects with <2 cm diameter moderately/well-differentiated microinvasive ( < 5 mm depth) oral squamous cell carcinoma lesions (33 lesions total, mean area ∼1.23 cm2) were administered 60 mg / kg ALA in oral solution and imaged before and after delivery of 100 J / cm2 total light dose to the lesion surface. Smartphone-based fluorescence and white light (WL) images were analyzed and compared with ultrasound (US) imaging of the same lesions.
Results: We present a comparative analysis of pre- and post-treatment fluorescence, WL, and US images of oral lesions. There was no significant difference in the distribution of lesion widths measured by fluorescence and US (mean widths of 14.5 and 15.3 mm, respectively) and linear regression shows good agreement (R2 = 0.91). In general, PpIX fluorescence images obtained prior to therapeutic light delivery are able to resolve lesion margins while dramatic photobleaching (∼42 % ) is visible post-treatment. Segmentation of the photobleached area confirms the boundaries of the irradiated zone.
Conclusions: A simple smartphone-based approach for imaging oral lesions is shown to agree in most cases with US, suggesting that this approach may be a useful tool to aid in PDT treatment guidance and monitoring photobleaching as part of a low-cost platform for intraoral PDT.
Photodynamic therapy (PDT), a spatially localized phototoxic therapy that involves irradiation of a photosensitizer (PS) with specific wavelengths of light, has shown exceptional promise in impacting cancer treatment outcomes, particularly oral cancer. To reduce PDT outcome variability, attempts toward image-guided personalized PDT are being pursued by monitoring PS uptake either via fluorescence or photoacoustic imaging (PAI), a nonionizing modality dependent on optical absorption properties of the tissue. PAI-guided PDT requires a near-infrared contrast agent for deep tissue imaging with minimal photobleaching effect. We evaluate the impact of PDT agent, benzoporphyrin derivative (BPD), on PAI agent indocyanine green (ICG) and vice versa, given that they have different optical absorption properties and singlet oxygen quantum yields for PDT. Specifically, we demonstrate in two oral squamous cell carcinoma lines (FaDu and SCC4) that ICG has minimal effect on BPD PDT efficacy when irradiated with either a continuous or pulsed laser. Furthermore, the impact of BPD on ICG photodegradation was monitored with PAI in tissue-mimicking phantoms. These studies inform us that the combination of BPD and ICG can be utilized for PAI-guided PDT. However, researchers need to consider the photodegradation effects of ICG in the presence of BPD when designing their drug delivery strategies for PAI-guided PDT.
Molecular Imaging techniques are a hotspot in oncology for their inherent ability to detect cancer early as well as characterize and stage them. The prognosis and treatment response can also be monitored with these imaging techniques. Specifically, photoacoustic imaging is a technique that recently had exponential growth in several biomedical applications, particularly for imaging tumors. Photoacoustic imaging involves excitation of the tissue with the nanosecond laser pulses and subsequent generation of acoustic waves with an ultrasound transducer. to reconstruct the image, has potential to detect and monitor cancer prognosis by using the optical contrast of hemoglobin. In this work, we evaluated the potential of LED-based photoacoustics in imaging heterogenous microvasculature in tumors.
A novel approach to monitor photosensitizer accumulation and photobleaching in the course of photodynamic therapy (PDT) with the use of nanoconstructs based on the simultaneous fluorescence (FL) and optoacoustic (OA) imaging is implemented. A liposome nanoconstructs employed in this studies contain benzoporphyrin derivatives (BPD) which serve as a photosensitizer and secondly, as a fluorophore, and the fluorescent IRDye800 dye acting as an additional contrasting agent due to its high quantum yield. FL provides visualization of BPD and IRDye800 distribution, while OA principle allows for BPD-absorption based imaging of tumor and its vascular environment. We demonstrate the results of a preliminary in vivo study with combined FL and OA custom-made setups on a NUDE mouse with human glioblastoma U- 87. The results of this studies show a hemorrhage in the tumor area on the OA images obtained @532 nm after PDT that is not visually detected, but confirmed with the following histological verification. Fast nanoconstructs accumulation (< 10 min) was observed using FL imaging with the concentration in tumor only 10% higher than in surrounding tissues. We believe that the ratio of nanoconstructs accumulation in tumor can be significantly increased using target approach.
Synovial angiogenesis and hypoxia in the joints are biomarkers of Rheumatoid Arthritis (RA). The ability to probe blood and accurately estimate the oxygen concentration make multiwavelength Photoacoustic (PA) imaging a potential tool for early detection of RA. In this work, a multiwavelength LED-based PA imaging system was characterized based on its imaging depth, resolution and accuracy of oxygen saturation estimation. A multicenter 3R (Replace, Refine and Reduce) focused small animal study was conducted. The 3R strategy was devised by reusing RA animal models, in vivo imaging of healthy animals and ex vivo studies with human blood. RA animal cadaver models with different levels of synovial angiogenesis (control, positive RA and treated) were imaged and compared against results from a previous study using the same samples. An ex vivo PA oxygen saturation imaging using human blood was validated against oximeter readings and further verified it with in vivo animal studies. An imaging depth of 8 mm with an SNR of 10 dB was achieved for RA samples. A difference in PA intensity was observed for RA models compared to control and treated group. The PA oxygen saturation estimation correlates with oximeter readings, which is confirmed with in vivo studies. The results show the efficacy of the LED-based PA imaging system in RA diagnosis based on synovial angiogenesis and hypoxia. The imaging depth, resolution and oxygen saturation estimate are sufficient to differentiate RA samples from control. Our future work will focus on validating the method using arthritis animal models and demonstrating the 3R potential.
Imaging technologies such as Ultrasound, OCT, MRI and CT are useful for diagnostics and tomographic assessment of therapy response. Fluorescence imaging with its high sensitivity is a promising approach and is extensively used for lesion localization, surgical guidance and monitoring response to therapies. While conjugating fluorophores to antibodies improves specificity, further conjugation of drugs provides a dual function, where fluorescence monitoring may simultaneously reveal drug pharmacokinetics. Fluorescence imaging, however, is limited by low penetration of light, which can be partially overcome by photoacoustic imaging (PAI). In this study, by conjugating a fluorophore and photoacoustic molecule to an antibody (Dual Function Antibody Conjugate (DFAC)), we evaluate whether PAI can significantly improve deep-tissue imaging.
To provide a systematic comparison of these imaging modalities we developed a DFAC, comprising of Cetuximab (anti-EGFR antibody) conjugated with a fluorophore (AF647) and a photoacoustic dye (IRDye800) in a 1:2:2 ratio. We hypothesize that, conjugating quantifiable probes to an antibody, would provide information about different depths within the confines of optical approaches. Such quantification is particularly important in photodynamic therapy, for determination of tissue concentration of photosensitizers and in chemotherapy for quantification of drug concentrations non-invasively. The relationship between the photoacoustic and fluorescence signals from the DFAC is demonstrated through spectroscopic techniques and their EGFR specificity along with deep tissue photoacoustic quantification is established using EGFR positive/negative cell lines and tissue mimicking phantoms.
The DFAC, presented in this study, demonstrates a combination of two complimentary imaging modalities for non-invasive determination of pharmacokinetics and in vivo drug quantification.
Oral cancer represents over 30% of cancers reported in low middle-income countries (LMIC), like India and is the leading cause of cancer death among Indian men. Surgery, radiation and chemo therapies are the mainstay of management but are either too expensive, unavailable for people or have extensive side effects. An alternate effective therapy for oral cancer is photodynamic therapy (PDT), a light based spatially targeted cytotoxic therapy that has shown excellent healing of the oral mucosa post treatment. We here combined engineering, optics and biochemistry to produce a low-cost, mobile LED-based light source with 3D printed light applicators for smart phone-based, image-guided PDT. After validating the devices in preclinical models, we performed an ergonomics study on 10 healthy volunteers at the MGH, where the comfort level of the applicators (anterior buccal cheek, posterior buccal cheek and retromolar positions) and presence of fatigue or numbness in the mouth due to the applicators was evaluated. We found that the retromolar and posterior applicators were the most comfortable and well tolerated. After these initial steps, the device was tested in clinical studies of early oral cancer in India. We observe in subjects with T1N0M0 oral lesions that our applicator and light system combination delivered light to cover the entire lesion area and yielded effective PDT response. Of the 18 treatments so far, 14 subjects have responded, with no residual/recurrent disease in follow-up biopsy. The significance of this work is that it offers an alternative treatment modality for early disease without associated morbidities.
Oral cancers are among the most prevalent malignancies in males. In developing countries like India where chewing tobacco and other carcinogen containing mixtures is common, incidence of oral cancers is high. Photodynamic therapy is emerging as a promising method for non-disfiguring treatment for early malignant lesions of the buccal mucosa with relatively few effects and potential for implementation in settings with limited medical infrastructure
We evaluate ultrasonography as an approach for assessment of PDT response in oral malignancies.In our study we selected stage 1 malignancy of the buccal mucosa using both ultrasonography followed by histopathology.The criterion for selection on USG was lesion length of 2cm or less and depth of 5mm confirmed by biopsy and histopathology analysis. Subsequently, the patients found positive on biopsy were treated with ALA PDT with a LED light source. Following treatment these patients were reassessed with ultrasound at day 10. The images obtained were compared and evaluated for change in the size of the lesion, their echopattern, vascularity, mucosal status etc.
It was found, lesions completely hypoechoic signifying necrosis came out negative on biopsy every single time, confirming hypoechogenecity as the single best indicator for success of PDT treatment. However, those lesions which did not develop necrosis but were negative on biopsy, the indirect markers were small initial size of the lesion, a lack of vascularity within the lesion and surrounding inflammation.
To conclude, ultrasonography is a convenient, reliable and radiation free method for post PDT evaluation of lesions of buccal mucosa.
Oral cancers are the 8th most common cancer among males. In India, this corresponds to almost 80,000 new cases per year. With such a disease burden, oral cancers are a prime cause of morbidity and functional disability.
Conventional treatment for oral cancers is surgery and radiation therapy which are fraught with physical and functional side effects, and the high cost and infrastructure requirements present barriers to timely intervention for patients in rural and/or resource-limited areas. Motivated by these considerations we sought to evaluate photodynamic therapy (PDT) as an approach which is inherently conducive to adaptation for resource-limited settings and has previously shown promising clinical results for early stage oral lesions. In this study we evaluate a low-cost platform for aminolevulinic acid (ALA)-based PDT consisting of a portable 635nm fiber-coupled LED light source integrated with 3D printed applicators for stable intraoral light delivery and smartphone-based fluorescence imaging for treatment guidance.
Using this technology, we treated 18 subjects with histologically confirmed T1N0M0 lesions of the buccal mucosa with a mean diameter of 1.38 cm2 and micro-invasive (≤ 5mm depth) disease. A total light dose of 100 J/cm2 was delivered in 3 to 5 fractions to the buccal mucosa after oral administration of 60mg/kg ALA (in 3 aliquots of 20mg/kg). The post-PDT investigations showed a 72% success rate (no residual malignancy in follow-up biopsy).
The treatment was very well tolerated and has potential for broader dissemination into primary care sites and as an early intervention for pre-malignant conditions including submucous fibrosis and leucoplakia.
Standard of care in the management of cancer of the oral tongue is complete extirpation with clear margins. There is a direct correlation between the radial margin distance and local failure and death from disease.
Surgical resection of this site is balanced between resection of enough tissue to ensure clear margins and excessive resection that would result in unnecessary oral dysfunction. The optimal margin distance is 5 mm on permanent fixed histological assessment
Mucosal margins are obtained primarily using visual inspection with a reasonable degree of accuracy. More difficult is the deep margin which cannot be seen during the resection, where surgeons must rely on manual palpation to estimate the depth invasion and the needed deep margin tissue thickness.
Over the last three years we have utilized intraoperative ultrasound to improve the likelihood of a clear deep margin. Preliminary analysis has shown that using this technique we are able to reliably clear the deep margin with an improved rate of local control.
Drawing upon our preliminary findings using intraoperative ultrasound for surgical navigation, we have designed a study to employ dual functional antibody conjugates as a theranostic tool to allow molecular based optical imaging of tumor margins, improving precision of the resection and utilizing a benzoporphyrin derivative to provide for a tumor specific photodynamic therapy treatment at the time of resection.
Techniques and preliminary oncologic and margin outcomes of intraoperative ultrasound as well as the concepts of the use of dual functional antibody conjugates in surgical navigation will be discussed.
India has one of the highest rates of oral cancer incidence in the world, with an estimated 80,000 new cases per year, accounting for 30% of reported cancers. In rural areas, a lack of adequate medical infrastructure contributes to unchecked disease progression and dismal mortality rates. PDT emerges as a potential modality which can be implemented in resource limited settings, while photosensitizer fluorescence can be leveraged for treatment guidance. Here, as part of an ongoing clinical study evaluating low-cost technology for ALA PDT treatment, we evaluated the capability of a simple smartphone-based device for imaging ALA-induced PpIX fluorescence. The imaging device itself consists of an annulus of 405nm LEDs for PpIX excitation with emission filter in the center mounted over the phone camera. 18 subjects having <2 cm diameter (mean size; ~1.38 cm2) lesions with micro-invasive (≤5 mm. depth) moderately/well-differentiated squamous cell carcinoma were administered 60 mg/kg ALA in oral solution and imaged before and after delivery of 100 J/cm2 total light dose to the lesion surface. We will present comparative analysis of pre-and post-treatment fluorescence, white light, and ultrasound images. In general, PpIX fluorescence images obtained prior to therapeutic light delivery are able to resolve lesion margins while dramatic photobleaching in post-treatment images confirms the irradiated zone. Overall this approach is able to generate sufficient fluorescence contrast for treatment guidance and monitoring photobleaching while the use of a smartphone-based device provides a low-cost, widely available platform with potential for telemedicine integration.
We propose a new approach to monitoring of photodynamic therapy (PDT) of glioblastoma with the use of targeted nanoconstructs containing a photosensitizer (PS) benzoporphyrin derivative (BPD) and IRDye800 dye, antibodies for efficient accumulation of the drug in a tumor, and a chemotherapeutic agent for combined effect on tumor cells. Monitoring of PDT is based on the simultaneous fluorescent and optoacoustic (OA) imaging. Fluorescent imaging provides visualization of fluorescence agents with high molecular sensitivity, and monitoring of the effectiveness of PDT by PS photobleaching. OA allows to examine the vascular pattern of the tumor environment, as well as assess the tumor depth. IRDye800 is a better contrast agent in comparison to BPD due to red shifted spectral characteristics and higher fluorescence quantum yield. The results of numerical simulations have been verified in phantom studies using fluorescence and optoacoustic experimental setups and an agar phantom with optical characteristics similar to those of murine brain.
Standard chemoradiation often enriches drug-resistant tumor cell populations that can lead to recurrent and treatment-refractory disease. Preclinical models of glioblastoma brain tumors, for instance, suggest that the cancer stem cell subpopulation becomes enriched and re-populates the tumor milieu following conventional therapies. Here, we show evidence that photodynamic therapy (PDT) is effective against several patient-derived glioblastoma stem cell cultures. Moreover, sub-lethal PDT results in re-sensitization of cancer stem cell phenotypes with induced drug-resistance to chemotherapy.
Glioblastoma (GBM) has less than one-year survival rate due to local recurrence post treatment or growth of “left-over” residual disease post surgical resection. To personalize treatment strategies and reduce the rate of recurrence, it is important to develop imaging based therapeutic strategies and prediction markers to identify GBM recurrence. Given the role of the vasculature in tumor growth and survival, here we utilize multi-parametric ultrasound and photoacoustic imaging to validate if changes in vascular structure and function could be predictive of treatment response in patient- derived orthotopic Glioblastoma xenograft models.
Patient-derived GBM cell lines were implanted in the brain of Swiss nu/nu mice. After the tumors reached 2-4 mm in diameter, the mice were divided into 4 groups namely – no treatment, surgical resection, therapy and surgical resection with therapy. Photodynamic therapy (PDT), a light based cytotoxic therapy, with photosensitizer Benzoporphyrin derivative (BPD) or FDA approved chemotherapy temozolomide were administered in the mice. Fujifilm VisualSonics LAZR system with a 20 MHz transducer was used to obtain power Doppler (%vascularity in tumors) and photoacoustic oxygen saturation (StO2) maps of the brain tumors at different time points pre and post treatment. The mice were either euthanized for immunofluorescence to validate the imaging markers or longitudinally monitored for tumor volume until moribund.The no-treatment group or the surgery only group did not have significant changes in vascular density or StO2. We observed a statistically significant decrease in StO2 immediately post BPD-PDT (primarily a vascular therapy) but not with temozolomide (a cellular therapy). Furthermore, we also observed that the sustenance of hypoxia or low StO2 in tumors for 24-72 hours can be a predictive biomarker for tumor recurrence. Overall, these results suggest the utility of ultrasound and photoacoustic imaging in monitoring treatment response and developing treatment prediction strategies for glioblastoma.
It is increasingly evident that the most effective cancer treatments will involve interactive regimens that target multiple non-overlapping pathways, preferably such that each component enhances the others to improve outcomes while minimizing systemic toxicities. Toward this goal, we developed a combination of photodynamic therapy and irinotecan, which mechanistically cooperate with each other, beyond their individual tumor destruction pathways, to cause synergistic reduction in orthotopic pancreatic tumor burden. A three-way mechanistic basis of the observed the synergism will be discussed: (i) PDT downregulates drug efflux transporters to increase intracellular irinotecan levels. (ii) Irinotecan reduces the expression of hypoxia-induced marker, which is upregulated by PDT. (iii) PDT downregulates irinotecan-induced survivin expression to amplify the apoptotic and anti-proliferative effects. The clinical translation potential of the combination will also be highlighted.
Photodynamic therapy (PDT) is a photochemistry based cytotoxic technique that imparts cellular damage via excitation of a photosensitizer with drug-specific wavelength of light. The dose at the treatment site for type II PDT is determined by three factors: photosensitizer (PS) concentration, oxygenation status and delivered light irradiance. Most of the FDA approved photosensitizers in their triplet-excited state generate cytotoxic species by reacting with the ground state oxygen that is available in the surrounding environment. Given the inter- and intra-subject variability in the uptake of the photosensitizer and the distribution of oxygen in the tumor, understanding the interplay between these dose parameters could aid in determining photodynamic therapy efficacy. Previously several studies have discussed the interplay between the dose parameters using shown point measurements and 2D imaging systems. Using various subcutaneous and orthotopic mouse models we will demonstrate the utility of a non-invasive non-ionizing photoacoustic imaging modality to determine efficacy and predict treatment response in Benzoporphyrin derivative (BPD) or Aminolevulinic acid (ALA) based PDT. We further compare the predictive capability of photoacoustic imaging with the more predominantly used fluorescence imaging and immunohistochemistry techniques.
Photodynamic therapy (PDT) is a light-based modality that shows promise for adaptation and implementation as a cancer treatment technology in resource-limited settings. In this context PDT is particularly well suited for treatment of pre-cancer and early stage malignancy of the oral cavity, that present a major global health challenge, but for which light delivery can be achieved without major infrastructure requirements. In recent reports we demonstrated that a prototype low-cost batterypowered 635nm LED light source for ALA-PpIX PDT achieves tumoricidal efficacy in vitro and vivo, comparable to a commercial turn-key laser source. Here, building on these reports, we describe the further development of a prototype PDT device to enable intraoral light delivery, designed for ALA- PDT treatment of precancerous and cancerous lesions of the oral cavity. We evaluate light delivery via fiber bundles and customized 3D printed light applicators for flexible delivery to lesions of varying size and position within the oral cavity. We also briefly address performance requirements (output power, stability, and light delivery) and present validation of the device for ALA-PDT treatment in monolayer squamous carcinoma cell cultures.
In view of the increase in cancer-related mortality rates in low- to middle-income countries (LMIC), there is an urgent need to develop economical therapies that can be utilized at minimal infrastructure institutions. Photodynamic therapy (PDT), a photochemistry-based treatment modality, offers such a possibility provided that low-cost light sources and photosensitizers are available. In this proof-of-principle study, we focus on adapting the PDT light source to a low-resource setting and compare an inexpensive, portable, battery-powered light-emitting diode (LED) light source with a standard, high-cost laser source. The comparison studies were performed in vivo in a xenograft murine model of human squamous cell carcinoma subjected to 5-aminolevulinic acid-induced protoporphyrin IX PDT. We observed virtually identical control of the tumor burden by both the LED source and the standard laser source. Further insights into the biological response were evaluated by biomarker analysis of necrosis, microvessel density, and hypoxia [carbonic anhydrase IX (CAIX) expression] among groups of control, LED-PDT, and laser-PDT treated mice. There is no significant difference in the percent necrotic volume and CAIX expression in tumors that were treated with the two different light sources. These encouraging preliminary results merit further investigations in orthotopic animal models of cancers prevalent in LMICs.
The need for patient-specific photodynamic therapy (PDT) in dermatologic and oncologic applications has triggered several studies that explore the utility of surrogate parameters as predictive reporters of treatment outcome. Although photosensitizer (PS) fluorescence, a widely used parameter, can be viewed as emission from several fluorescent states of the PS (e.g., minimally aggregated and monomeric), we suggest that singlet oxygen luminescence (SOL) indicates only the active PS component responsible for the PDT. Here, the ability of discrete PS fluorescence-based metrics (absolute and percent PS photobleaching and PS re-accumulation post-PDT) to predict the clinical phototoxic response (erythema) resulting from 5-aminolevulinic acid PDT was compared with discrete SOL (DSOL)-based metrics (DSOL counts pre-PDT and change in DSOL counts pre/post-PDT) in healthy human skin. Receiver operating characteristic curve (ROC) analyses demonstrated that absolute fluorescence photobleaching metric (AFPM) exhibited the highest area under the curve (AUC) of all tested parameters, including DSOL based metrics. The combination of dose-metrics did not yield better AUC than AFPM alone. Although sophisticated real-time SOL measurements may improve the clinical utility of SOL-based dosimetry, discrete PS fluorescence-based metrics are easy to implement, and our results suggest that AFPM may sufficiently predict the PDT outcomes and identify treatment nonresponders with high specificity in clinical contexts.
A complete understanding of the biological mechanisms regulating devastating disease such as cancer remains
elusive. Pancreatic and brain cancers are primary among the cancer types with poor prognosis. Molecular biomarkers
have emerged as group of proteins that are preferentially overexpressed in cancers and with a key role in driving disease
progression and resistance to chemotherapy. The epidermal growth factor receptor (EGFR), a cell proliferative
biomarker is particularly highly expressed in most cancers including brain and pancreatic cancers. The ability of EGFR
to sustain prolong cell proliferation is augmented by biomarkers such as Bax, Bcl-XL and Bcl-2, proteins regulating the
apoptotic process. To better understand the role and effect of the microenvironment on these biomarkers in pancreatic
cancer (PaCa); we analysed two pancreatic tumor lines (AsPc-1 and MiaPaCa-2) in 2D, 3D in-vitro cultures and in
orthotopic tumors at different growth stages. We also investigated in patient derived glioblastoma (GBM) tumor cultures,
the ability to utilize the EGFR expression to specifically deliver photosensitizer to the cells for photodynamic therapy.
Overall, our results suggest that (1) microenvironment changes affect biomarker expression; thereby it is critical to
understand these effects prior to designing combination therapies and (2) EGFR expression in tumor cells indeed could
serve as a reliable and a robust biomarker that could be used to design targeted and image-guided photodynamic therapy.
Glioblastoma (GBM) is an aggressive cancer with dismal survival rates and few new treatment options. Fluorescence
guided resection of GBM followed by photodynamic therapy (PDT) has shown promise in several chemo- or
radiotherapy non-responsive GBM treatments clinically. PDT is an emerging light and photosensitizer (PS) mediated
cytotoxic method. However, as with other therapeutic modalities, the outcomes are variable largely due to the nonpersonalization
of dose parameters. The variability can be attributed to the differences in heterogeneous photosensitizer
accumulation in tumors. Building upon our previous findings on utilizing PS fluorescence for designing tumor-specific
PDT dose, we explore the use of photoacoustic imaging, a technique that provides contrast based on the tissue optical
absorption properties, to obtain 3D information on the tumoral photosensitizer accumulation. The findings of this study
will form the basis for customized photodynamic therapy for glioblastoma and have the potential to serve as a platform
for treatment of other cancers.
Ultrasound imaging can provide excellent resolution at reasonable depths while retaining the advantages of being nonionizing,
cost-effective and portable. However, the contrast in ultrasound imaging is limited, and various ultrasoundbased
techniques such as photoacoustic (PA) and magneto-motive ultrasound (MMUS) imaging have been developed to
augment ultrasound imaging. Photoacoustic imaging enhances imaging contrast by visualizing the optical absorption of
either tissue or injected contrast agents (e.g., gold or silver nanoparticles). MMUS imaging enhances the sensitivity and
specificity of ultrasound based on the detection of magnetic nanoparticles perturbed by an external magnetic field. This
paper presents integrated magneto-photo-acoustic (MPA) imaging - a fusion of complementary ultrasound-based
imaging techniques. To demonstrate the feasibility of MPA imaging, porcine ex-vivo tissue experiments were performed
using a dual contrast (magnetic/plasmonic) agent. Spatially
co-registered and temporally consecutive ultrasound,
photoacoustic, and magneto-motive ultrasound images of the same
cross-section of tissue were obtained. Our ex-vivo
results indicate that magneto-photo-acoustic imaging can be used to detect magnetic/plasmonic nanoparticles with high
resolution, sensitivity and contrast. Therefore, our study suggests that magneto-photo-acoustic images can identify the
morphological properties, molecular information and complementary functional information of the tissue.
Metallic nanoparticles have been widely used in a variety of imaging and therapeutic applications due to their unique
optical properties in the visible and near-infrared (NIR) regions - for example, various plasmonic nanoparticles are used
for molecular photoacoustic imaging and photothermal therapy. However, there are concerns that these agents may not
be safe under physiological conditions, because these nanoparticles are not biodegradable, could accumulate and,
therefore, could be toxic long-term. We investigate the feasibility of using biodegradable gold nanoclusters as a contrast
agent for highly sensitive photoacoustic imaging. The size of these biodegradable nanoclusters, consisting of sub-5 nm
primary gold particles and a biodegradable polymer binder, is less than 100 nm. Due to plasmon coupling, these
nanoclusters are characterized by a broad extinction spectrum that extends to the near infrared (NIR) spectral range.
Photoacoustic imaging of tissue models containing inclusions with different concentrations of nanoparticles was
performed using a tunable pulsed laser system. The results indicate that the biodegradable nanoclusters, comprised of
small gold nanoparticles, can be used as contrast agents in photoacoustic imaging.
Quantitative and qualitative monitoring of neovascular growth is required in many vascular tissue engineering
applications. For example, the contribution of progenitor cells in growing microvasculature has been demonstrated;
however, the process of vascularization from progenitor cells is not well understood. Therefore, there is a need for an
imaging technique that is consistent, easy to use, and can quantitatively assess the dynamics of vascular growth or
regression in a three-dimensional environment. In this study, we evaluate the ability of combined ultrasound and
photoacoustic imaging to assess the dynamics of vascular growth. The experiments were performed using hydrogels that
spontaneously promote tube formation from implanted mesenchymal stem cells (MSCs). Specifically, PEGylated fibrin
gels, supporting the development of capillary growth were implanted in a Lewis rat. After one week, the rat was
euthanized and the gel implants were excised and positioned in water cuvettes for imaging. Simultaneous ultrasound and
photoacoustic images were obtained using single-element, focused ultrasound transducers interfaced with a nanosecond
pulsed laser source. To image samples, ultrasound transducers operating at either 25 MHz or 48 MHz and interfaced
with laser sources operating at either 532 nm or within 680-800 nm wavelengths were used. The 3-D ultrasound and
photoacoustic images were acquired by mechanically scanning the transducer over the region of interest and capturing
spatially co-registered and temporally consecutive photoacoustic transients and ultrasound pulse-echo signals. The
ultrasound and photoacoustic images agree well with the overall anatomy and vascular structure in the gel samples. The
results suggest that the photoacoustic and ultrasound imaging could be used to sequentially monitor the growth of
neovasculature in-vivo.
Treatment of deep venous thrombosis (DVT)—a primary cause of potentially fatal pulmonary embolism (PE)—depends on the age of the thrombus. The existing clinical imaging methods are capable of visualizing a thrombus but cannot determine the age of the blood clot. Therefore, there is a need for an imaging technique to reliably diagnose and adequately stage DVT. To stage DVT (i.e., to determine the age of the thrombus, and therefore, to differentiate acute from chronic DVT), we explored photoacoustic imaging, a technique capable of noninvasive measurements of the optical absorption in tissue. Indeed, optical absorption of the blood clot changes with age, since maturation of DVT is associated with significant cellular and molecular reorganization. The ultrasound and photoacoustic imaging studies were performed using DVT-mimicking phantoms and phantoms with embedded acute and chronic thrombi obtained from an animal model of DVT. The location and structure of the clots were visualized using ultrasound imaging, while the composition, and therefore age, of thrombi were related to the magnitude and spatiotemporal characteristics of the photoacoustic signal. Overall, the results of our study suggest that combined ultrasound and photoacoustic imaging of thrombi may be capable of simultaneous detection and staging of DVT.
Gold nanoparticles functionalized with anti-EGFR antibodies undergo molecular specific aggregation on
the cellular membrane and later within the cell that leads to a red shift in the plasmon resonance frequency
of the gold nanoparticles. Capitalizing on this effect, we previously demonstrated on tissue phantoms that
highly sensitive and selective detection of cancer cells can be achieved using the combination of
photoacoustic imaging and molecular specific gold nanoparticles. To further evaluate the efficacy of
molecular specific photoacoustic imaging technique in detecting deeply situated tumors, small animal
experiments were performed. In this study, two gelatin solutions mixed with cells labeled with gold
nanoparticles and cells mixed with polyethylene glycol-thiol
(mPEG-SH) coated gold nanoparticles were
injected in a mouse abdomen ex-vivo. The photoacoustic and ultrasound images from the same crosssection
of the region before and after the injections were obtained using a 25 MHz single element
ultrasound transducer interfaced with pulsed laser system. The results of our study suggest that the
molecular specific photoacoustic imaging with plasmonic nanosensors could be used to detect deeply
embedded tumors.
Diagnosis and treatment of atherosclerosis necessitates the detection and differentiation of rupture prone plaques. In
principle, intravascular photoacoustic (IVPA) imaging has the ability to simultaneously visualize the structure and
composition of atherosclerotic plaques by utilizing the difference in optical absorption. Extensive studies are required to
validate the utility of IVPA imaging in detecting vulnerable plaques and address issues associated with the clinical
implementation of the technique. In this work, we performed ex vivo imaging studies using a rabbit model of
atherosclerosis. The intravascular photoacoustic (IVPA) and ultrasound (IVUS) images of the normal aorta and aorta
with plaque were obtained and compared with histological slices of the tissue. The results indicate that IVPA imaging is
capable of detecting plaques and showed potential in determining the composition. Furthermore, we initially addressed
several aspects of clinical implementation of the IVPA imaging. Specifically, the configuration of combined IVPA and
IVUS catheter was investigated and the effect of the optical absorption of the luminal blood on the IVPA image quality
was evaluated. Overall, this study suggests that IVPA imaging can become a unique and important clinical tool.
In many clinical and research applications including cancer diagnosis, tumor response to therapy, reconstructive
surgery, monitoring of transplanted tissues and organs, and quantitative evaluation of angiogenesis, sequential and
quantitative assessment of microcirculation in tissue is required. In this paper we present an imaging technique capable
of spatial and temporal measurements of blood perfusion through microcirculation. To demonstrate the developed
imaging technique, studies were conducted using phantoms with modeled small blood vessels of various diameters
positioned at different depths. A change in the magnitude of the photoacoustic signal was observed during vessel
constriction and subsequent displacement of optically absorbing liquid present in the vessels. The results of the study
suggest that photoacoustic, ultrasound and strain imaging could be used to sequentially monitor and qualitatively assess
blood perfusion through microcirculation.
To perform ultrasound imaging using an array transducer, a focused ultrasound beam is transmitted in a particular direction within the tissue and the received backscattered ultrasound wave is then dynamically focused at every position along the beam. The ultrasound beam is scanned over the desired region to form an image. The photoacoustic imaging, however, is distinct from conventional ultrasound imaging. In photoacoustic imaging the acoustic transients are generated simultaneously in the entire volume of the irradiated tissue - no transmit focusing is possible due to light scattering in the tissue. The photoacoustic waves are then recorded on every element of the ultrasound transducer array at once and processed to form an image. Therefore, compared to ultrasound imaging, photoacoustic imaging can utilize dynamic receive focusing only. In this paper, we describe the image formation algorithms of the array-based photoacoustic and ultrasound imaging system and present methods to improve the quality of photoacoustic images.
To evaluate the performance of photoacoustic imaging using an array transducer, numerical simulations and phantom experiments were performed. First, to evaluate spatial resolution, a point source was imaged using a combined ultrasound and photoacoustic imaging system. Next, image quality was assessed by imaging tissue imaging phantoms containing a circular inclusion. Finally, the photoacoustic and ultrasound images from the combined imaging system were analyzed.
Tissue engineering is an interdisciplinary field that combines various aspects of engineering and life sciences and
aims to develop biological substitutes to restore, repair or maintain tissue function. Currently, the ability to have
quantitative functional assays of engineered tissues is limited to existing invasive methods like biopsy. Hence, an
imaging tool for non-invasive and simultaneous evaluation of the anatomical and functional properties of the engineered
tissue is needed. In this paper we present an advanced in-vivo imaging technology - ultrasound biomicroscopy combined
with complementary photoacoustic and elasticity imaging techniques, capable of accurate visualization of both structural
and functional changes in engineered tissues, sequential monitoring of tissue adaptation and/or regeneration, and possible
assistance of drug delivery and treatment planning. The combined imaging at microscopic resolution was evaluated on
tissue mimicking phantoms imaged with 25 MHz single element focused transducer. The results of our study
demonstrate that the ultrasonic, photoacoustic and elasticity images synergistically complement each other in detecting
features otherwise imperceptible using the individual techniques. Finally, we illustrate the feasibility of the combined
ultrasound, photoacoustic and elasticity imaging techniques in accurately assessing the morphological and functional
changes occurring in engineered tissue.
A hybrid imaging system is proposed for cancer detection, diagnosis and therapy monitoring by integrating
three complementary imaging techniques - ultrasound, photoacoustic and elasticity imaging. Indeed, simultaneous
imaging of the anatomy (ultrasound imaging), cancer-induced angiogenesis (photoacoustic imaging) and changes in
biomechanical properties (elasticity imaging) of tissue is based on many synergistic features of these modalities and
may result in a unique and important imaging tool. To facilitate the design and development of a real-time imaging
system for clinical applications, we have investigated the core components of the imaging system using numerical
simulations. Differences and similarities between each imaging technique were considered and contrasted. The results
of our study suggest that the integration of ultrasound, photoacoustic and elasticity imaging is possible using a custom
designed imaging system.
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