Photoacoustic Chemical Imaging Sodium Nano-Sensor Utilizing a Solvatochromic Dye Transducer for In Vivo Application.

Sodium has many vital and diverse roles in the human body, including maintaining the cellular pH, generating action potential, and regulating osmotic pressure. In cancer, sodium dysregulation has been correlated with tumor growth, metastasis, and immune cell inhibition. However, most in vivo sodium measurements are performed via Na23 NMR, which is handicapped by slow acquisition times, a low spatial resolution (in mm), and low signal-to-noise ratios. We present here a plasticizer-free, ionophore-based sodium-sensing nanoparticle that utilizes a solvatochromic dye transducer to circumvent the pH cross-sensitivity of most previously reported sodium nano-sensors. We demonstrate that this nano-sensor is non-toxic, boasts a 200 µM detection limit, and is over 1000 times more selective for sodium than potassium. Further, the in vitro photoacoustic calibration curve presented demonstrates the potential of this nano-sensor for performing the in vivo chemical imaging of sodium over the entire physiologically relevant concentration range.

An Automatic 3-D Ultrasound and Photoacoustic Combined Imaging System for Human Inflammatory Arthritis.

Aiming at a point-of-care device for rheumatology clinics, we developed an automatic 3-D imaging system combining the emerging photoacoustic (PA) imaging with conventional Doppler ultrasound (US) for detecting human inflammatory arthritis. This system is based on a commercial-grade GE HealthCare (GEHC, Chicago, IL, USA) Vivid E95 US machine and a Universal Robot UR3 robotic arm. This system automatically locates the patient's finger joints from a photograph taken by an overhead camera powered by an automatic hand joint identification method, followed by the robotic arm moving the imaging probe to the targeted joint to scan and obtain 3-D PA and Doppler US images. The GEHC US machine was modified to enable high-speed, high-resolution PA imaging while maintaining the features available on the system. The commercial-grade image quality and the high sensitivity in detecting inflammation in peripheral joints via PA technology hold great potential to significantly benefit clinical care of inflammatory arthritis in a novel way.

Longitudinal volumetric assessment of inflammatory arthritis via photoacoustic imaging and Doppler ultrasound imaging.

Aiming at clinical translation, we developed an automatic 3D imaging system combining the emerging photoacoustic imaging with conventional Doppler ultrasound for detecting inflammatory arthritis. This system was built with a GE HealthCare (GEHC) Vivid™ E95 ultrasound system and a Universal Robot UR3 robotic arm. In this work, the performance of this system was examined with a longitudinal study utilizing a clinically relevant adjuvant induced arthritis (AIA) murine model. After adjuvant injection, daily imaging of the rat ankle joints was conducted until joint inflammation was obvious based on visual inspection. Processed imaging results and statistical analyses indicated that both the hyperemia (enhanced blood volume) detected by photoacoustic imaging and the enhanced blood flow detected by Doppler ultrasound reflected the progress of joint inflammation. However, photoacoustic imaging, by leveraging the highly sensitive optical contrast, detected inflammation earlier than Doppler ultrasound, and also showed changes that are more statistically significant. This side-by-side comparison between photoacoustic imaging and Doppler ultrasound using the same commercial grade GEHC ultrasound machine demonstrates the advantage and potential value of the emerging photoacoustic imaging for rheumatology clinical care of arthritis.

Photoacoustic Imaging of COVID-19 Vaccine Site Inflammation of Autoimmune Disease Patients.

Based on the observations made in rheumatology clinics, autoimmune disease (AD) patients on immunosuppressive (IS) medications have variable vaccine site inflammation responses, whose study may help predict the long-term efficacy of the vaccine in this at-risk population. However, the quantitative assessment of the inflammation of the vaccine site is technically challenging. In this study analyzing AD patients on IS medications and normal control subjects, we imaged the inflammation of the vaccine site 24 h after mRNA COVID-19 vaccinations were administered using both the emerging photoacoustic imaging (PAI) method and the established Doppler ultrasound (US) method. A total of 15 subjects were involved, including 6 AD patients on IS and 9 normal control subjects, and the results from the two groups were compared. Compared to the results obtained from the control subjects, the AD patients on IS medications showed statistically significant reductions in vaccine site inflammation, indicating that immunosuppressed AD patients also experience local inflammation after mRNA vaccination but not in as clinically apparent of a manner when compared to non-immunosuppressed non-AD individuals. Both PAI and Doppler US were able to detect mRNA COVID-19 vaccine-induced local inflammation. PAI, based on the optical absorption contrast, shows better sensitivity in assessing and quantifying the spatially distributed inflammation in soft tissues at the vaccine site.

Photoacoustic Spectral Analysis for Evaluating the Aggressiveness of Prostate Cancer Labeled by Methylene Blue Polyacrylamide Nanoparticles.

Evaluating the aggressiveness of prostate cancer (PCa) is crucial for PCa diagnosis and prognosis. Previously, studies have shown that photoacoustic spectral analysis (PASA) can assess prostate tissue microarchitecture for evaluating the aggressiveness of PCa. In this study, in a transgenic mouse (TRAMP) model of PCa, we utilized methylene blue polyacrylamide nanoparticles (MB PAA NPs) to label the cancer cells in prostate in vivo. MB PAA NPs can specifically target proliferating cancer cells as a contrast agent, allowing photoacoustic (PA) imaging to better detect PCa tumors, and also assessing prostate glandular architecture. With the PA signals from the prostates measured simultaneously by a needle hydrophone and a PA and ultrasound (US) dual-imaging system, we conducted PASA and correlated the quantified spectral parameter slopes with the cancer grading from histopathology. The PASA results from 18 mice showed significant differences between normal and cancer, and also between low-score cancer and high-score cancer. This study in the clinically relevant TRAMP model of PCa demonstrated that PA imaging and PASA, powered by MB PAA NPs that can label the PCa microarchitectures in vivo after systemic administration, can detect PCa and, more importantly, evaluate cancer aggressiveness.

Personalized Oncology by In Vivo Chemical Imaging: Photoacoustic Mapping of Tumor Oxygen Predicts Radiotherapy Efficacy.

We hereby apply the approach of photoacoustic chemical imaging, performing an in vivo chemical analysis that is spatially resolved (200 µm) and in real time, to predict a given tumor's response to therapy. Using triple negative breast cancer as a model, we took photoacoustic images of tumors' oxygen distributions in patient-derived xenografts (PDXs) in mice using biocompatible, oxygen-sensitive tumor-targeted chemical contrast nanoelements (nanosonophores), which function as contrast agents for photoacoustic imaging. Following radiation therapy, we established a quantitatively significant correlation between the spatial distribution of the initial oxygen levels in the tumor and its spatial distribution of the therapy's efficacy: the lower the local oxygen, the lower the local radiation therapy efficacy. We thus provide a simple, noninvasive, and inexpensive method to both predict the efficacy of radiation therapy for a given tumor and identify treatment-resistant regions within the tumor's microenvironment.

Characterizing the aggressiveness of prostate cancer using an all-optical needle photoacoustic sensing probe: feasibility study.

In our previous studies, we have developed a prototype interstitial needle sensing probe that can acquire broadband A-line photoacoustic (PA) signals encoding both tissue microarchitecture and histochemical information comparable to that accessible by histology. Paving the road toward clinical translation of this technology, we replaced the piezoelectric hydrophone in the needle PA probe with a fiber optic hydrophone that enabled both broader bandwidth and sufficient signal-to-noise ratio (SNR) for PA signal detection. Such an all-optical design also facilitated disposability and significantly reduced the footprint of the needle PA sensing probe. Experiments were performed on well-controlled phantoms and human prostate tissues. The microarchitectures in each sample were quantitatively evaluated by both the nonlinear spectral slope of the PA signal power spectrum and the generalized gamma (GG) parameter a by implementing envelope statistics to the PA signal. In the studies on phantoms containing optically absorbing microspheres with various sizes and concentrations, the nonlinear spectral slope showed a strong correlation of r=-0.80 with the microsphere dimensions, and a relatively weak correlation of r=-0.54 with the microsphere concentrations, while the GG parameter a showed a strong correlation with the microsphere dimensions (r=0.72) and a moderate correlation with the microsphere concentrations (r=0.63). In the studies on human prostate tissues containing progressive cancer stages, both the nonlinear spectral slope and the GG parameter a demonstrated a statistically significant difference between benign and nonaggressive cancer tissues (p<0.01), and between nonaggressive and aggressive cancer tissues (p<0.01). In addition, a multivariate analysis combining the two quantitative measurements demonstrated the boundaries among the different progressive stages of prostate cancer.

The feasibility of ultrasound-assisted endovascular laser thrombolysis in an acute rabbit thrombosis model.

PURPOSE: This study aimed to test the feasibility of combined ultrasound and laser technique, namely, ultrasound-assisted endovascular laser thrombolysis (USELT), for thrombolysis by conducting in vivo tests in a rabbit thrombosis model. MATERIALS AND METHODS: An acute thrombus was created in the right jugular vein of rabbit and then was treated with ultrasound only, laser only, and USELT to dissolve the blood clot. A total of 20 rabbits were used. Out of which, the first three rabbits were used to titrate the laser and ultrasound parameters. Then, five rabbits were treated with ultrasound only, five rabbits were treated with laser only, and seven rabbits were treated with USELT. During USELT, 532-nm laser pulses were delivered endovascularly directly to the clot through a fiber optic, and 0.5 MHz ultrasound pulses were applied noninvasively to the same region. A laser fluence of 4 to 12 mJ/cm2 and ultrasound amplitude of 1 to 2 MPa were used. Recanalization of the jugular vein was assessed by performing ultrasound Doppler imaging immediately after the treatment. The maximum blood flow speed after the treatment as compared to its value before the treatment was used to calculate the blood flow recovery in vessel. RESULTS: The blood flow was fully recovered (100%) in three rabbits, partially recovered in two rabbits (more than 50% and less than 100%) with mean percentage recovery of 69.73% and poorly recovered in two rabbits (<50%) with mean percentage recovery of 6.2% in the USELT group. In contrast, the treatment group with ultrasound or laser alone did not show recanalization of vein in any case, all the five rabbits were poorly/not recovered with a mean percentage recovery of 0%. CONCLUSIONS: The USELT technology was shown to effectively dissolve the blood clots in an acute rabbit jugular vein thrombosis model.

Detecting joint inflammation by an LED-based photoacoustic imaging system: a feasibility study (Erratum).

An error in the first author's name is corrected.

Imaging of enthesitis by an LED-based photoacoustic system (Erratum).

An error in the first author's name is corrected.

Ultrasound-assisted laser thrombolysis with endovascular laser and high-intensity focused ultrasound.

PURPOSE: The combination of laser and ultrasound can significantly improve the efficiency of thrombolysis through an enhanced cavitation effect. We developed a fiber optics-based laser-ultrasound thrombolysis device and tested the feasibility and efficiency of this technology for restoring blood flow in an in vitro blood clot model. METHODS: An in vitro blood flow-clot model was setup, and then an endovascular laser thrombolysis system was combined with high-intensity focused ultrasound to remove the clot. The laser and ultrasound pulses were synchronized and delivered to the blood clot concurrently. The laser pulses of 532 nm were delivered to the blood clot endovascularly through an optical fiber, whereas the ultrasound pulses of 0.5 MHz were applied noninvasively to the same region. Effectiveness of thrombolysis was evaluated by the ability to restore blood flow, which was monitored by ultrasound Doppler. RESULTS: As laser powers increased, the ultrasound threshold pressures for effective thrombolysis decreased. For laser fluence levels of 0, 2, and 4 mJ/cm2 , the average negative ultrasound threshold pressures were 1.26 ± 0.114, 1.05 ± 0.181, and 0.59 ± 0.074 MPa, respectively. The periods of time needed to achieve effective thrombolysis were measured at 0.8, 2, and 4 mJ/cm2 laser fluence levels and 0.42, 0.70, and 0.98 MPa negative ultrasound pressures. In general, thrombolysis could be achieved more rapidly with higher laser powers or ultrasound pressures. CONCLUSIONS: Effective thrombolysis can be achieved by combining endovascular laser with noninvasive ultrasound at relatively low power and pressure levels, which can potentially improve both the treatment efficiency and safety.

Imaging of enthesitis by an LED-based photoacoustic system.

SIGNIFICANCE: One key pathological characteristic of seronegative spondyloarthropathy (SpA) is inflammation at the insertion of tendons and ligaments into the bone (enthesitis). AIM: We explore the potential of the emerging photoacoustic (PA) imaging in diagnosis of SpA and review its feasibility in detecting SpA-associated Achilles tendon enthesitis. APPROACH: A light-emitting diode (LED)-based PA and ultrasound combined system was employed. The PA images, both along the long and the short axes of each Achilles tendon insertion region, were acquired at 850-nm wavelength, which is sensitive in depicting increased blood volume (i.e., hyperemia). To assess the hyperemia indicating enthesis inflammation, two parameters were quantified in the imaged tendons, including the average intensity and the density of the color pixels in the pseudo-color PA images. Ten SpA patients, all of which met Assessment of SpA International Society (ASAS) criteria for SpA and were found to have Achilles enthesitis by clinical exam according to a board-certified rheumatologist, were included in the study. RESULTS: The PA and Doppler ultrasound imaging of Achilles enthesitis resulting from these 10 SpA patients were compared to those from 10 healthy volunteers, leading to statistically significant differences (p < 0.05) in the applied t-tests. CONCLUSIONS: This preliminary clinical study suggests that the LED-based PA imaging holds a promise for sensitive and objective assessment of SpA enthesitis in an outpatient setting of the rheumatology clinic.

Photoacoustic spectral analysis at ultraviolet wavelengths for characterizing the Gleason grades of prostate cancer.

The diagnosis of aggressive prostate cancer (PCa) has relied on microscopic architectures, namely Gleason patterns, of tissues extracted through core biopsies. Technology capable of assessing the tissue architecture without tissue extraction will reduce the invasiveness of PCa diagnosis and improve diagnostic accuracy by allowing for more sampling locations. Our recently developed photoacoustic spectral analysis (PASA) has achieved quantification of tissue architectural heterogeneity interstitially. Taking advantage of the unique optical absorption of cell nuclei at ultraviolet (UV) wavelengths, this study investigated PASA at 266 nm for quantifying the tissue architecture heterogeneity in prostates. The results have shown significant differences among the normal, early cancer, and late cancer stages in mouse prostates ex vivo and in vivo (n=20, p<0.05). The study with human samples ex vivo has shown a correlation of 0.80 (n=11, p<0.05) between PASA quantification and pathologic diagnosis.

In Vivo Photoacoustic Lifetime Based Oxygen Imaging with Tumor Targeted G2 Polyacrylamide Nanosonophores.

Lifetime imaging methods using phosphorescence quenching by oxygen for molecular oxygen concentration measurement have been developed and used for noninvasive oxygen monitoring. This study reports photoacoustic (PA) oxygen imaging powered by polyacrylamide (PAAm) hydrogel nanoparticles (NP) which offer advantages including improved biocompatibility, reduced toxicity, and active tumor targeting. A known oxygen indicator, oxyphor G2, was conjugated with the matrix of the NPs, giving G2-PAA NPs, followed by PEGylation for biocompatibility and F3 surface modification for tumor targeting. Using two lasers providing pump and probe pulses, respectively, PA imaging was performed so as to quantitatively map the oxygen concentration in biological tissues in vivo, including cancer tumors and normal thigh muscles. Furthermore, via the imaging at the pump wavelength and two additional wavelengths, the accumulation of the G2-PAA NPs in the tumors were also determined. The successful imaging experiment accomplished on animal models renders a method for in vivo noninvasive imaging and assessment of hypoxic tumor microenvironments, which is critical for assessing cancer progression, metastasis, and treatment.

Chemical Imaging in Vivo: Photoacoustic-Based 4-Dimensional Chemical Analysis.

We describe how 4-dimensional in vivo biochemical analysis can be performed using photoacoustic contrast nanoagents that have been designed to probe both structural and chemical information in vivo, enabling noninvasive, real time, spatially resolved chemical imaging. Early chemical imaging of a patient's tumor can inform the decision of effective treatment, regarding choices of chemotherapy, radiation, or immunotherapy.

Detecting joint inflammation by an LED-based photoacoustic imaging system: a feasibility study.

Light-emitting diode (LED) light sources have recently been introduced to photoacoustic imaging (PAI). The LEDs enable a smaller footprint for PAI systems when compared to laser sources, thereby improving system portability and allowing for improved access. An LED-based PAI system has been employed to identify inflammatory arthritis in human hand joints. B-mode ultrasound (US), Doppler, and PAIs were obtained from 12 joints with clinically active arthritis, five joints with subclinically active arthritis, and 12 normal joints. The quantitative assessment of hyperemia in joints by PAI demonstrated statistically significant differences among the three conditions. The imaging results from the subclinically active arthritis joints also suggested that the LED-based PAI has a higher sensitivity to angiogenic microvascularity compared to US Doppler imaging. This initial clinical study on arthritis patients validates that PAI can be a potential imaging modality for the diagnosis of inflammatory arthritis.

Light Emitting Diodes based Photoacoustic Imaging and Potential Clinical Applications.

Using low cost and small size light emitting diodes (LED) as the alternative illumination source for photoacoustic (PA) imaging has many advantages, and can largely benefit the clinical translation of the emerging PA imaging technology. Here, we present our development of LED-based PA imaging integrated with B-mode ultrasound. To overcome the challenge of achieving sufficient signal-to-noise ratio by the LED light that is orders of magnitude weaker than lasers, extensive signal averaging over hundreds of pulses is performed. Facilitated by the fast response of the LED and the high-speed driving as well as the high pulse repetition rate up to 16 kHz, B-mode PA images superimposed on gray-scale ultrasound of a biological sample can be achieved in real-time with frame rate up to 500 Hz. The LED-based PA imaging could be a promising tool for several clinical applications, such as assessment of peripheral microvascular function and dynamic changes, diagnosis of inflammatory arthritis, and detection of head and neck cancer.

Photoacoustic tomography for human musculoskeletal imaging and inflammatory arthritis detection.

With the capability of assessing high resolution optical contrast in soft tissues, photoacoustic imaging (PAI) can offer valuable structural and functional information of human joints, and hold potential for diagnosis and treatment monitoring of inflammatory arthritis. Recent studies have demonstrated that PAI can map 2D and 3D morphology of the cartilage, synovium, vascularity, and bone tissue in human peripheral joints. Initial trials with patients affected by inflammatory arthritis have also suggested that PAI can detect the hemodynamic properties in articular tissues as well as their changes due to active inflammation. This review focuses on the recent progress in technical development of PAI for human musculoskeletal imaging and inflammation detection. PAI can provide non-invasive and non-ionizing serial measurements for monitoring of therapeutic interventions with the potential for higher sensitivity than existing imaging modalities such as ultrasound. However, further investigation is needed to validate the value of PAI in rheumatology clinical settings.

A Functional Study of Human Inflammatory Arthritis Using Photoacoustic Imaging.

By using our dual-modality system enabling simultaneous real-time ultrasound (US) and photoacoustic (PA) imaging of human peripheral joints, we explored the potential contribution of PA imaging modality to rheumatology clinic. By performing PA imaging at a single laser wavelength, the spatially distributed hemoglobin content reflecting the hyperemia in synovial tissue in metacarpophalangeal (MCP) joints of 16 patients were imaged, and compared to the results from 16 healthy controls. In addition, by performing PA imaging at two laser wavelengths, the spatially distributed hemoglobin oxygenation reflecting the hypoxia in inflammatory joints of 10 patients were imaged, and compared to the results from 10 healthy controls. The statistical analyses of the PA imaging results demonstrated significant differences (p < 0.001) in quantified hemoglobin content and oxygenation between the unequivocally arthritic joints and the normal joints. Increased hyperemia and increased hypoxia, two important physiological biomarkers of synovitis reflecting the increased metabolic demand and the relatively inadequate oxygen delivery in affected synovium, can both be objectively and non-invasively evaluated by PA imaging. The proposed dual-modality system has the potential of providing additional diagnostic information over the traditional US imaging approaches and introducing novel imaging biomarkers for diagnosis and treatment evaluation of inflammatory arthritis.