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.

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.

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.

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

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

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.

Noninvasive reflection mode photoacoustic imaging through infant skull toward imaging of neonatal brains.

The feasibility of functional imaging of neonatal brains was studied in a noninvasive transcranial manner by using reflection mode photoacoustic technique for the first time. Experiments were conducted to examine the quality of photoacoustic signals and consequent images across a newborn infant skull. With the designed system, photoacoustic imaging of blood vessels through the infant skull has been achieved with an axial resolution up to 50mum and a lateral resolution up to 420mum. Experimental results also indicate that photoacoustic imaging of neonatal brain with a depth of 21mm or more beneath the skull is feasible when working with near-infrared light. Moreover, the performance of this technique for measuring and monitoring the changes in blood oxygenation level through the newborn infant skull has also been explored. This study suggests that reflection mode photoacoustic imaging holds promise to become a novel and powerful tool for noninvasive diagnosis, monitoring and prognosis of disorders in neonatal brains.

Photoacoustic tomography of carrageenan-induced arthritis in a rat model.

Laser-based photoacoustic tomography (PAT), a novel, nonionizing, noninvasive, laser-based technology, has been adapted to the diagnosis and imaging of inflammatory arthritis. A commonly used adjuvant induced arthritis model using carrageenan was employed to simulate acute rheumatoid arthritis in rat tail joints. Cross-sectional photoacoustic images of joints affected by acute inflammation were compared to those of the control. The diameter of the periosteum and the optical absorption of intra-articular tissue were measured on each joint image. Significant differences were found on PAT imaging between the affected joints and the control for both variables measured, including enlarged periosteum diameter and enhanced intra-articular optical absorption occurring in the joints affected with carrageenan-induced arthritis. Anatomical correlation with histological sections of imaged joints and microMRI results verified the findings of PAT. This suggests that PAT has the potential for highly sensitive diagnosis and evaluation of pathologic hallmarks of acute inflammatory arthritis.

Imaging of joints with laser-based photoacoustic tomography: an animal study.

Photoacoustic tomography (PAT), a nonionizing, noninvasive, laser-based technology was adapted to joint imaging for the first time. Pulsed laser light in the near-infrared region was directed toward a joint with resultant ultrasonic signals recorded and used to reconstruct images that present the optical properties in subsurface joint tissues. The feasibility of this joint imaging system was validated on a Sprague Dawley rat tail model and verified through comparison with histology. With sufficient penetration depth, PAT realized tomographic imaging of a joint as a whole organ noninvasively. Based on the optical contrast, various intra- and extra-articular tissues, including skin, fat, muscle, blood vessels, synovium and bone, were presented successfully in images with satisfactory spatial resolution that was primarily limited by the bandwidth of detected photoacoustic signals rather than optical diffusion as occurs in traditional optical imaging. PAT, with its intrinsic advantages, may provide a unique opportunity to enable the early diagnosis of inflammatory joint disorders, e.g., rheumatoid arthritis, and to monitor therapeutic outcomes with high sensitivity and accuracy.

Characterization and treatment monitoring of inflammatory arthritis by photoacoustic imaging: a study on adjuvant-induced arthritis rat model.

Neovascularity also known as angiogenesis is an early feature of inflammatory arthritis disease. Therefore, identifying the development of neovascularity is one way to potentially detect and characterize arthritis. Laser-based photoacoustic imaging (PAI) is an emerging biomedical imaging modality which may aid in the detection of both early and continued development of neovascularity. In this work, we investigated the feasibility of PAI to measure angiogenesis, for the purpose of evaluating and monitoring inflammatory arthritis and responses to treatment. The imaging results on an arthritis rat model demonstrate that 1) there is noticeable enhancement in image intensities in the arthritic ankle joints when compared to the normal joints, and 2) there is noticeable decrease in image intensities in the arthritic ankle joints after treatment when compared to the untreated arthritic joints. In order to validate the findings from PAI, we performed positron emission tomography (PET) and histology on the same joints. The diameters of the ankle joints, as a clinical score of the arthritis, were also measured at each time point.

Dual-mode imaging with radiolabeled gold nanorods.

Many nanoparticle contrast agents have difficulties with deep tissue and near-bone imaging due to limited penetration of visible photons in the body and mineralized tissues. We are looking into the possibility of mediating this problem while retaining the capabilities of the high spatial resolution associated with optical imaging. As such, the potential combination of emerging photoacoustic imaging and nuclear imaging in monitoring of antirheumatic drug delivery by using a newly developed dual-modality contrast agent is investigated. The contrast agent is composed of gold nanorods (GNRs) conjugated to the tumor necrosis factor (TNF-α) antibody and is subsequently radiolabeled by (125)I. ELISA experiments designed to test TNF-α binding are performed to prove the specificity and biological activity of the radiolabeled conjugated contrast agent. Photoacoustic and nuclear imaging are performed to visualize the distribution of GNRs in articular tissues of the rat tail joints in situ. Findings from the two imaging modalities correspond well with each other in all experiments. Our system can image GNRs down to a concentration of 10 pM in biological tissues and with a radioactive label of 5 µCi. This study demonstrates the potential of combining photoacoustic and nuclear imaging modalities through one targeted contrast agent for noninvasive monitoring of drug delivery as well as deep and mineralized tissue imaging.

125I-labeled gold nanorods for targeted imaging of inflammation.

For better examination of inflammation, we designed inflammation-targeted nuclear and optical dual-modality contrast agents prepared by I-125 radiolabeling of gold nanorods (GdNRs) conjugated with anti-intercellular adhesion molecule 1 (ICAM-1) antibody. The bioactivity and specific binding of the PEGylated (125)I-ICAM-GdNR conjugates to the ICAM-1 was validated through ELISA testing. Inflammation-targeted imaging was then conducted on an adjuvant-induced arthritic rat model which demonstrated an elevation of ICAM-1 level in the affected ankle joints. Facilitated by the I-125 radioisotope and the whole-body imaging via the Gamma camera, the time-dependent distribution of the systemically injected agent as well as the uptake of the agent in the inflammatory articular tissues could be examined conveniently and quantitatively. The success in targeted delivery of gold nanoparticles to inflammatory tissue enables both nuclear and optical imaging of inflammation at molecular or cellular level. Other than diagnosis, radiolabeled gold nanoparticles also hold promise for targeted therapy of a variety of disorders.

Optical imaging: new tools for arthritis.

Conventional radiography, ultrasound, CT, MRI, and nuclear imaging are the current imaging modalities used for clinical evaluation of arthritis which is highly prevalent and a leading cause of disability. Some of these types of imaging are also used for monitoring disease progression and treatment response of arthritis. However, their disadvantages limit their utilities, such as ionizing radiation for radiography, CT, and nuclear imaging; suboptimal tissue contrast resolution for radiography, CT, ultrasound, and nuclear imaging; high cost for CT and MRI and nuclear imaging; and long data-acquisition time with ensuing patient discomfort for MRI. Recently, there have been considerable advances in nonionizing noninvasive optical imaging which has demonstrated promise for early diagnosis, monitoring therapeutic interventions and disease progression of arthritis. Optical based molecular imaging modalities such as fluorescence imaging have shown high sensitivity in detection of optical contrast agents and can aid early diagnosis and ongoing evaluation of chronic inflammatory arthritis. Optical transillumination imaging or diffuse optical tomography may differentiate normal joint clear synovial fluid from turbid and pink medium early in the inflammatory process. Fourier transform infrared spectroscopy has been used to evaluate fluid composition from joints affected by arthritis. Hemodynamic changes such as angiogenesis, hypervascularization, and hypoxia in arthritic articular tissue can potentially be observed by diffuse optical tomography and photoacoustic tomography. Optical measurements could also facilitate quantification of hemodynamic properties such as blood volume and oxygenation levels at early stages of inflammatory arthritis. Optical imaging provides methodologies which should contribute to detection of early changes and monitoring of progression in pathological characteristics of arthritis, with relatively simple instrumentation.

Photoacoustic tomography of joints aided by an Etanercept-conjugated gold nanoparticle contrast agent-an ex vivo preliminary rat study.

Monitoring of anti-rheumatic drug delivery in experimental models and in human diseases would undoubtedly be very helpful for both basic research and clinical management of inflammatory diseases. In this study, we have investigated the potential of an emerging hybrid imaging technology-photoacoustic tomography-in noninvasive monitoring of anti-TNF drug delivery. After the contrast agent composed of gold nanorods conjugated with Etanercept molecules was produced, ELISA experiments were performed to prove the conjugation and to show that the conjugated anti-TNF-α drug was biologically active. PAT of ex vivo rat tail joints with the joint connective tissue enhanced by intra-articularly injected contrast agent was conducted to examine the performance of PAT in visualizing the distribution of the gold-nanorod-conjugated drug in articular tissues. By using the described system, gold nanorods with a concentration down to 1 pM in phantoms or 10 pM in biological tissues can be imaged with good signal-to-noise ratio and high spatial resolution. This study demonstrates the feasibility of conjugating TNF antagonist pharmaceutical preparations with gold nanorods, preservation of the mechanism of action of TNF antagonist along with preliminary evaluation of novel PAT technology in imaging optical contrast agents conjugated with anti-rheumatic drugs. Further in vivo studies on animals are warranted to test the specific binding between such conjugates and targeted antigen in joint tissues affected by inflammation.

Noninvasive photoacoustic tomography of human peripheral joints toward diagnosis of inflammatory arthritis.

The feasibility of photoacoustic tomography (PAT) in imaging human peripheral joints in a noninvasive manner was demonstrated through studies of cadaver human fingers. Based on the intrinsic optical contrast, intra- and extra-articular tissue structures in the finger at the levels of the joints were visualized successfully with satisfactory spatial resolution. The imaging depth of PAT in the near-infrared region enables the cross-sectional imaging of a human finger as a whole organ. As a novel technology with unique advantages, PAT holds promise for early diagnosis of inflammatory joint disorders and accurate monitoring of disease progression and response to therapy.