Mild Hyperthermia Enhanced Liposomal Doxorubicin Delivery and CD8 + T cell Infiltration in Triple Negative Breast Cancer.

Mild hyperthermia (MHTh) is often used in combination with chemotherapy and radiotherapy for cancer treatment. In the current study, the effect of MHTh on the enhanced uptake of the FDA-approved chemotherapy drug, liposomal doxorubicin (dox) in syngeneic 4T1 tumors was investigated. Doxorubicin has inherent fluorescence properties having an emission signal at 590 nm upon excitation with a 480 nm laser. A group of mice administered with doxorubicin (dox) were exposed to MHTh (42 °C) for 30 minutes whereas control group given dox did not receive MHTh. Ex vivo optical imaging of harvested tumors confirmed higher uptake of dox in treated versus the control untreated tumors. Confocal microscopy of tumor sections indicates higher fluorescent intensity due to increased accumulation of dox in MHTh-treated compared to untreated tumors. We examined the effect of MHTh to enhance CD8 tumor infiltration, production of interferon-γ (IFN-γ) and expression of programmed death ligand-1 (PD-L1). mRNA in situ hybridization was performed to test for transcripts of CD8, IFN-γ and PD-L1. Results showed that higher expression of CD8 mRNA was observed in MHTh-administered tumors versus untreated cohorts. The signal for IFN-γ and PD-L1 in both groups were not significantly different. Taken together, our findings imply that MHTh can improve tumor uptake of dox. Importantly, our data suggests that MHTh can boost CD8 + T cell infiltration.

Breast Tomographic Ultrasound: The Spectrum from Current Dense Breast Cancer Screenings to Future Theranostic Treatments.

This review provides unique insights to the scientific scope and clinical visions of the inventors and pioneers of the SoftVue breast tomographic ultrasound (BTUS). Their >20-year collaboration produced extensive basic research and technology developments, culminating in SoftVue, which recently received the Food and Drug Administration's approval as an adjunct to breast cancer screening in women with dense breasts. SoftVue's multi-center trial confirmed the diagnostic goals of the tissue characterization and localization of quantitative acoustic tissue differences in 2D and 3D coronal image sequences. SoftVue mass characterizations are also reviewed within the standard cancer risk categories of the Breast Imaging Reporting and Data System. As a quantitative diagnostic modality, SoftVue can also function as a cost-effective platform for artificial intelligence-assisted breast cancer identification. Finally, SoftVue's quantitative acoustic maps facilitate noninvasive temperature monitoring and a unique form of time-reversed, focused US in a single theranostic device that actually focuses acoustic energy better within the highly scattering breast tissues, allowing for localized hyperthermia, drug delivery, and/or ablation. Women also prefer the comfort of SoftVue over mammograms and will continue to seek out less-invasive breast care, from diagnosis to treatment.

Niche preclinical and clinical applications of photoacoustic imaging with endogenous contrast.

In the past decade, photoacoustic (PA) imaging has attracted a great deal of popularity as an emergent diagnostic technology owing to its successful demonstration in both preclinical and clinical arenas by various academic and industrial research groups. Such steady growth of PA imaging can mainly be attributed to its salient features, including being non-ionizing, cost-effective, easily deployable, and having sufficient axial, lateral, and temporal resolutions for resolving various tissue characteristics and assessing the therapeutic efficacy. In addition, PA imaging can easily be integrated with the ultrasound imaging systems, the combination of which confers the ability to co-register and cross-reference various features in the structural, functional, and molecular imaging regimes. PA imaging relies on either an endogenous source of contrast (e.g., hemoglobin) or those of an exogenous nature such as nano-sized tunable optical absorbers or dyes that may boost imaging contrast beyond that provided by the endogenous sources. In this review, we discuss the applications of PA imaging with endogenous contrast as they pertain to clinically relevant niches, including tissue characterization, cancer diagnostics/therapies (termed as theranostics), cardiovascular applications, and surgical applications. We believe that PA imaging's role as a facile indicator of several disease-relevant states will continue to expand and evolve as it is adopted by an increasing number of research laboratories and clinics worldwide.

Versatile multimodal modality based on Brillouin light scattering and the photoacoustic effect.

Multimodal optical techniques are useful for the comprehensive characterization of material properties. In this work, we developed a new, to the best of our knowledge, multimodal technology that can simultaneously measure a subset of mechanical, optical, and acoustical properties of the sample and is based on the integration of Brillouin (Br) and photoacoustic (PA) microscopy. The proposed technique can acquire co-registered Br and PA signals from the sample. Importantly, using synergistic measurements of the speed of sound and Brillouin shift, the modality offers a new approach to quantifying the optical refractive index, which is a fundamental property of a material and is not accessible by either technique individually. As a proof of concept, we demonstrated the feasibility of integrating the two modalities and acquired the colocalized Br and time-resolved PA signals in a synthetic phantom made out of kerosene and CuSO4 aqueous solution. In addition, we measured the refractive index values of saline solutions and validated the result. Comparison with previously reported data showed a relative error of 0.3%. This further allowed us to directly quantify the longitudinal modulus of the sample with the colocalized Brillouin shift. While the scope of the current work is limited to introducing the combined Br-PA setup for the first time, we envision that this multimodal modality could open a new path for the multi-parametric analysis of material properties.

Wavelength-dependent error minimization for quantitative spectroscopic photoacoustic tomography with a ring-array system.

PURPOSE: Photoacoustic tomography (PAT) is a non-invasive and high-resolution imaging technique that can provide functional and molecular information from the optical properties of pathological tissues, such as cancer. Spectroscopic PAT (sPAT) is capable of supplying information such as oxygen saturation (sO2), which is an important biological indicator for diseases such as cancer. However, the wavelength dependent nature of sPAT makes it challenging to provide accurate quantitative measurements of tissue oxygenation beyond shallow depths. We have previously reported the utility of combined ultrasound tomography and PAT to achieve optical and acoustic compensated PAT images at a single wavelength and for enhanced PAT images at larger depths. In this work we further explore the utility of the optical and acoustic compensation PAT algorithm to minimize the wavelength dependency in sPAT by showcasing improvements in spectral unmixing. MATERIALS AND METHODS: Two optically and acoustically characterized heterogenous phantoms were manufactured to test the ability of the system and developed algorithm to minimize the wavelength-dependence driven error in sPAT spectral unmixing. The PA inclusions within each phantom were composed of a mixture of two sulfate dyes, copper sulfate (CuSO4) and nickel sulfate (NiSO4), with known optical spectra. Improvements between uncompensated and optically and acoustically compensated PAT (OAcPAT) were quantified as the relative percent error between the measured results and the ground truth. RESULTS: The results of our phantom studies demonstrate that OAcPAT can significantly improve the accuracy of sPAT measurements in a heterogenous medium and especially at larger inclusions depths which can reach to up to 12% improvement in measurement errors. This significant improvement can play a vital role in reliability of future in-vivo biomarker quantifications. CONCLUSIONS: Utilizing UST for model-based optical and acoustic compensation of PAT images was proposed by our group previously. In this work, we further demonstrated the efficacy of the developed algorithm in sPAT by minimizing the error caused by the tissue's optical heterogeneity on improving spectral unmixing, which is a major limiting factor in reliability of sPAT measurements. Such synergistic combination of UST and PAT provides a window of opportunity to achieve bias-free quantitative sPAT measurements, which plays an important role in future pre-clinical and clinical utility of PAT.

A versatile multimodal optical modality based on Brillouin light scattering and photoacoustic effect.

Multimodal optical imaging techniques are useful for various applications, including imaging biological samples for providing comprehensive material properties. In this work, we developed a new modality that can measure a set of mechanical, optical, and acoustical properties of a sample at microscopic resolution, which is based on the integration of Brillouin (Br) and photoacoustic (PA) microscopy. The proposed multimodal imaging technique not only can acquire co-registered Br and PA signals but also allows us to utilize the sound speed measured by PA to quantify the sample’s refractive index, which is a fundamental property of the material and cannot be measured by either technique individually. We demonstrated the colocalization of Br and time-resolved PA signals in a synthetic phantom made of kerosene and CuSO 4 aqueous solution. In addition, we measured the refractive index of saline solutions and validated the result against published data with a relative error of 0.3 %. This multimodal Br-PA modality could open a new way for characterizing biological samples in physiological and pathological conditions.

Integrated Ultrasound and Photoacoustic-Guided Laser Ablation Theranostic Endoscopic System.

Advancements in ablation techniques have paved the way towards the development of safer and more effective clinical procedures for treating various maladies such as atrial fibrillation (AF). AF is characterized by rapid, chaotic atrial activation and is commonly treated using radiofrequency applicators or laser ablation catheters. However, the lack of thermal lesion formation and temperature monitoring capabilities in these devices prevents them from measuring the treatment outcome directly. In addition, poor differentiation between healthy and ablated tissues leads to incomplete ablation, which reduces safety and causes complications in patients. Hence, a novel photoacoustic (PA)-guided laser ablation theranostic device was developed around a traditional phased-array endoscope. The proposed technology provides lesion formation, tissue distinguishing, and temperature monitoring capabilities. Our results have validated the lesion monitoring capability of the proposed technology through PA correlation maps. The tissue distinguishing capability of the theranostic device was verified by the measurable differences in the PA signal between pre-and post-ablated mice myocardial tissue. The increase in the PA signal with temperature variations caused by the ablation laser confirmed the ability of the proposed device to provide temperature feedback.

Feasibility of ultrasound tomography-guided localized mild hyperthermia using a ring transducer: Ex vivo and in silico studies.

BACKGROUND: As of 2022, breast cancer continues to be the most diagnosed cancer worldwide. This problem persists within the United States as well, as the American Cancer Society has reported that ∼12.5% of women will be diagnosed with invasive breast cancer over the course of their lifetime. Therefore, a clinical need continues to exist to address this disease from a treatment and therapeutic perspective. Current treatments for breast cancer and cancers more broadly include surgery, radiation, and chemotherapy. Adjuncts to these methods have been developed to improve the clinical outcomes for patients. One such adjunctive treatment is mild hyperthermia therapy (MHTh), which has been shown to be successful in the treatment of cancers by increasing effectiveness and reduced dosage requirements for radiation and chemotherapies. MHTh-assisted treatments can be performed with invasive thermal devices, noninvasive microwave induction, heating and recirculation of extracted patient blood, or whole-body hyperthermia with hot blankets. PURPOSE: One common method for inducing MHTh is by using microwave for heat induction and magnetic resonance imaging for temperature monitoring. However, this leads to a complex, expensive, and inaccessible therapy platform. Therefore, in this work we aim to show the feasibility of a novel all-acoustic MHTh system that uses focused ultrasound (US) to induce heating while also using US tomography (UST) to provide temperature estimates. Changes in sound speed (SS) have been shown to be strongly correlated with temperature changes and can therefore be used to indirectly monitor heating throughout the therapy. Additionally, these SS estimates allow for heterogeneous SS-corrected phase delays when heating complex and heterogeneous tissue structures. METHODS: Feasibility to induce localized heat in tissue was investigated in silico with a simulated breast model, including an embedded tumor using continuous wave US. Here, both heterogenous acoustic and thermal properties were modeled in addition to blood perfusion. We further demonstrate, with ex vivo tissue phantoms, the feasibility of using ring-based UST to monitor temperature by tracking changes in SS. Two phantoms (lamb tissue and human abdominal fat) with latex tubes containing varied temperature flowing water were imaged. The measured SS of the water at each temperature were compared against values that are reported in literature. RESULTS: Results from ex vivo tissue studies indicate successful tracking of temperature under various phantom configurations and ranges of water temperature. The results of in silico studies show that the proposed system can heat an acoustically and thermally heterogenous breast model to the clinically relevant temperature of 42°C while accounting for a reasonable time needed to image the current cross section (200 ms). Further, we have performed an initial in silico study demonstrating the feasibility of adjusting the transmit waveform frequency to modify the effective heating height at the focused region. Lastly, we have shown in a simpler 2D breast model that MHTh level temperatures can be maintained by adjusting the transmit pressure intensity of the US ring. CONCLUSIONS: This work has demonstrated the feasibility of using a 256-element ring array transducer for temperature monitoring; however, future work will investigate minimizing the difference between measured SS and the values shown in literature. A hypothesis attributes this bias to potential volumetric average artifacts from the ray-based SS inversion algorithm that was used, and that moving to a waveform-based SS inversion algorithm will greatly improve the SS estimates. Additionally, we have shown that an all-acoustic MHTh system is feasible via in silico studies. These studies have indicated that the proposed system can heat a tumor within a heterogenous breast model to 42°C within a narrow time frame. This holds great promise for increasing the accessibility and reducing the complexity of a future all-acoustic MHTh system.

Ultrasound and Photoacoustic Imaging of Breast Cancer: Clinical Systems, Challenges, and Future Outlook.

Presently, breast cancer diagnostic methods are dominated by mammography. Although drawbacks of mammography are present including ionizing radiation and patient discomfort, not many alternatives are available. Ultrasound (US) is another method used in the diagnosis of breast cancer, commonly performed on women with dense breasts or in differentiating cysts from solid tumors. Handheld ultrasound (HHUS) and automated breast ultrasound (ABUS) are presently used to generate reflection images which do not contain quantitative information about the tissue. This limitation leads to a subjective interpretation from the sonographer. To rectify the subjective nature of ultrasound, ultrasound tomography (UST) systems have been developed to acquire both reflection and transmission UST (TUST) images. This allows for quantitative assessment of tissue sound speed (SS) and acoustic attenuation which can be used to evaluate the stiffness of the lesions. Another imaging modality being used to detect breast cancer is photoacoustic tomography (PAT). Utilizing much of the same hardware as ultrasound tomography, PAT receives acoustic waves generated from tissue chromophores that are optically excited by a high energy pulsed laser. This allows the user to ideally produce chromophore concentration maps or extract other tissue parameters through spectroscopic PAT. Here, several systems in the area of TUST and PAT are discussed along with their advantages and disadvantages in breast cancer diagnosis. This overview of available systems can provide a landscape of possible intersections and future refinements in cancer diagnosis.

Photoacoustic-MR Image Registration Based on a Co-Sparse Analysis Model to Compensate for Brain Shift.

Brain shift is an important obstacle to the application of image guidance during neurosurgical interventions. There has been a growing interest in intra-operative imaging to update the image-guided surgery systems. However, due to the innate limitations of the current imaging modalities, accurate brain shift compensation continues to be a challenging task. In this study, the application of intra-operative photoacoustic imaging and registration of the intra-operative photoacoustic with pre-operative MR images are proposed to compensate for brain deformation. Finding a satisfactory registration method is challenging due to the unpredictable nature of brain deformation. In this study, the co-sparse analysis model is proposed for photoacoustic-MR image registration, which can capture the interdependency of the two modalities. The proposed algorithm works based on the minimization of mapping transform via a pair of analysis operators that are learned by the alternating direction method of multipliers. The method was evaluated using an experimental phantom and ex vivo data obtained from a mouse brain. The results of the phantom data show about 63% improvement in target registration error in comparison with the commonly used normalized mutual information method. The results proved that intra-operative photoacoustic images could become a promising tool when the brain shift invalidates pre-operative MRI.

Magnetic particles in motion: magneto-motive imaging and sensing.

Superparamagnetic nanoparticles have become an important tool in biomedicine. Their biocompatibility, controllable small size, and magnetic properties allow manipulation with an external magnetic field for a variety of diagnostic and therapeutic applications. Recently, the magnetically-induced motion of superparamagnetic nanoparticles has been investigated as a new source of imaging contrast. In magneto-motive imaging, an external, time-varying magnetic field is applied to move a magnetically labeled subject, such as labeled cells or tissue. Several major imaging modalities such as ultrasound, photoacoustic imaging, optical coherence tomography, and laser speckle tracking can utilize magneto-motive contrast to monitor biological events at smaller scales with enhanced contrast and sensitivity. In this review article, an overview of magneto-motive imaging techniques is presented, including synthesis of superparamagnetic nanoparticles, fundamental principles of magneto-motive force and its utility to excite labeled tissue within a viscoelastic medium, current capabilities of magneto-motive imaging modalities, and a discussion of the challenges and future outlook in the magneto-motive imaging domain.

The role of noninvasive diagnostic imaging in monitoring pregnancy and detecting patients at risk for preterm birth: a review of quantitative approaches.

Preterm birth (PTB) is the leading cause of neonatal morbidity and mortality worldwide. The ability to predict patients at risk for preterm birth remains a major health challenge. The currently available clinical diagnostics such as cervical length and fetal fibronectin may detect only up to 30% of patients who eventually experience a spontaneous preterm birth. This paper reviews ongoing efforts to improve the ability to conduct a risk assessment for preterm birth. In particular, this work focuses on quantitative methods of imaging using ultrasound-based techniques, magnetic resonance imaging, and optical imaging modalities. While ultrasound imaging is the major modality for preterm birth risk assessment, a summary of efforts to adopt other imaging modalities is also discussed to identify the technical and diagnostic limits associated with adopting them in clinical settings. We conclude the review by proposing a new approach using combined photoacoustic, ultrasound, and elastography as a potential means to better assess cervical tissue remodeling, and thus improve the detection of patients at-risk of PTB.

Methods for Monitoring Risk of Hypoxic Damage in Fetal and Neonatal Brains: A Review.

Fetal, perinatal, and neonatal asphyxia are vital health issues for the most vulnerable groups in human beings, including fetuses, newborns, and infants. Severe reduction in oxygen and blood supply to the fetal brain can cause hypoxic-ischemic encephalopathy (HIE), leading to long-term neurological disorders, including mental impairment and cerebral palsy. Such neurological disorders are major healthcare concerns. Therefore, there has been a continuous effort to develop clinically useful diagnostic tools for accurately and quantitatively measuring and monitoring blood and oxygen supply to the fetal and neonatal brain to avoid severe consequences of asphyxia HIE and neonatal encephalopathy. Major diagnostic technologies used for this purpose include fetal heart rate monitoring, fetus scalp blood sampling, ultrasound imaging, magnetic resonance imaging, X-ray computed tomography, and nuclear medicine. In addition, given the limitations and shortcomings of traditional diagnostic methods, emerging technologies such as near-infrared spectroscopy and photoacoustic imaging have also been introduced as stand-alone or complementary solutions to address this critical gap in fetal and neonatal care. This review provides a thorough overview of the traditional and emerging technologies for monitoring fetal and neonatal brain oxygenation status and describes their clinical utility, performance, advantages, and disadvantages.

Endocavity ultrasound and photoacoustic system for fetal and maternal imaging: design, implementation, and ex-vivo validation.

Purpose: Transvaginal ultrasound (TVUS) is a widely used real-time and non-invasive imaging technique for fetal and maternal care. It can provide structural and functional measurements about the fetal brain, such as blood vessel diameter and blood flow. However, it lacks certain biochemical estimations, such as hemoglobin oxygen saturation ( SO 2 ), which limits its ability to indicate a fetus at risk of birth asphyxia. Photoacoustic (PA) imaging has been steadily growing in recognition as a complement to ultrasound (US). Studies have shown PA imaging is capable of providing such biochemical estimations as SO 2 at relatively high penetration depth (up to 30 mm). Approach: In this study, we have designed and developed a multi-modal (US, PA, and Doppler) endocavity imaging system (ECUSPA) around a commercialized TVUS probe (Philips ATL C9-5). Results: The integrated system was evaluated through a set of in-vitro, ex-vivo, and in-vivo studies. Imaging of excised sheep brain tissue demonstrated the system's utility and penetration depth in transfontanelle imaging conditions. The accuracy of using the spectroscopic PA imaging (sPA) method to estimate SO 2 was validated by comparing sPA oximetry results with the gold standard measurements indicated by a blood gas analyzer. The ability of US and Doppler to measure moving blood volume was evaluated in-vivo. Spectral unmixing capabilities were tested using fluorophores within sheep brains. Conclusion: The developed system is a high resolution (about 200 µ m at 30 mm depth), real-time (at 30 Hz), and quantitative ( SO 2 estimation error < 10 % ) imaging tool with a total diameter less than 30 mm, making it suitable for intrapartum applications such as fetal and maternal diagnostics.

Photoacoustic-guided endovenous laser ablation: Characterization and in vivo canine study.

Endovenous laser ablation (EVLA) is a minimally invasive surgical procedure, often guided by ultrasound (US) imaging, for treating venous insufficiencies. US imaging limitations in accurately visualizing the catheter and the lack of a temperature monitoring system can lead to sub-optimal outcomes. An integrated photoacoustic (PA)-guided EVLA system has been previously developed and reported to overcome the shortcomings of US-guided procedure. In this study, we further characterized the system and tested the in vivo utility. In addition, PA thermometry was further explored by compensating the variation of PA signal with temperature with respect to the temperature-dependent absorption of blood and water. In vivo imaging results indicated that the PA-guided EVLA system can provide high contrast and accurate images of the ablation catheter tip overlaid on US images of the background tissue. Additionally, absorption-compensated PA signal amplitudes over a relevant range of temperature were measured and demonstrated.

Model-based optical and acoustical compensation for photoacoustic tomography of heterogeneous mediums.

Photoacoustic tomography (PAT) is a non-invasive, high-resolution imaging modality, capable of providing functional and molecular information of various pathologies, such as cancer. One limitation of PAT is the depth and wavelength dependent optical fluence, which results in reduced PA signal amplitude from deeper tissue regions. These factors can therefore introduce errors into quantitative measurements such as oxygen saturation (sO2) or the localization and concentration of various chromophores. The variation in the speed-of-sound between different tissues can also lead to distortions in object location and shape. Compensating for these effects allows PAT to be used more quantitatively. We have developed a proof-of-concept algorithm capable of compensating for the heterogeneity in speed-of-sound and depth dependent optical fluence. Speed-of-sound correction was done by using a straight ray-based algorithm for calculating the family of iso-time-of-flight contours between the transducers and every pixel in the imaging grid, while fluence compensation was done by utilizing the graphics processing unit (GPU) accelerated software MCXCL for Monte Carlo modeling of optical fluence variation. This algorithm was tested on a polyvinyl chloride plastisol (PVCP) phantom, which contained cyst mimics and blood inclusions to test the algorithm under relatively heterogeneous conditions. Our results indicate that our PAT algorithm can compensate for the speed-of-sound variation and depth dependent fluence effects within a heterogeneous phantom. The results of this study will pave the way for further development and evaluation of the proposed method in more complex in-vitro and ex-vivo phantoms, as well as compensating for the wavelength-dependent optical fluence in spectroscopic PAT.

Efficacy of High Temporal Frequency Photoacoustic Guidance of Laser Ablation Procedures.

Inaccurate placement of the ablation catheter and the inability to monitor the real-time temperature within the tissue of interest such as veins curbs the treatment efficacy of laser ablation procedures during thermal therapies. Our previous studies have validated the efficacy of photoacoustic (PA) imaging during endovenous laser ablation (EVLA) procedures. However, the PA-guided therapies suffer from low temporal resolution, due to the low pulse repetition rates of pulsed lasers, which could cause a problem during fast catheter motion and rapid temperature changes. Herein, to enhance the accuracy and sensitivity for tracking the ablation catheter tip and temperature monitoring, we proposed to develop a high frame rate (500 Hz), combined ultrasound (US), and PA-guided ablation system. The proposed PA-guided ablation system was evaluated in a set of ex vivo tissue studies. The developed system provides a 2 ms temporal resolution for tracking and monitoring the ablation catheter tip's location and temperature, which is 50 times higher temporal resolution compared to the previously proposed 10 Hz system. The proposed system also provided more accurate feedback about the temperature variations during rapid temperature increments of 10°C per 250 ms. The co-registered US and PA images have an imaging resolution of about 200 µm and a field of view of 45 × 40 mm2. Tracking the ablation catheter tip in an excised tissue layer shows higher accuracy during a relatively fast catheter motion (0.5-3 mm/s). The fast US/PA-guided ablation system will potentially enhance the outcome of ablation procedures by providing location and temperature feedback.

Dual-Illumination Ultrasound/ Photoacoustic System for Cervical Cancer imaging.

Early stage cancer detection technologies can provide functional information and potentially decrease the mortality rate caused by cervical cancer. In our previous work, a miniaturized ultrasound and photoacoustic endoscopic system has been developed to image the cervical tissue through the cervical canal to fulfills the need for a safe, low-cost, and high-resolution functional diagnostic system. However, the miniaturized size of endoscope and American National Standards Institute safety limits cause constraints of using high-intensity illumination during imaging. In addition, the strong light scattering of tissues limits the light penetration depth. Fortunately, the cervix anatomy allows for the delivery of additional light from the ectocervix by using an external illumination system. Here we propose a dual, co-planar illumination system, which can provide adequate illumination to the cervical tissue via combined internal and external light delivery strategies. Therefore, an increase in the area of light-tissue interaction allows us to raise the laser light energy while keeping fluence under safety limits. Thus, a reliable PA imaging can be obtained for the whole cervical tissue thickness. The system performance was tested using a Monte Carlo simulation, and laser-light fluence was calculated and compared at different depths within a simulated cervical-tissue model. The results indicated a higher and more uniform fluence in the Monte Carlo simulations. In addition, the photoacoustic imaging of the proposed system was evaluated by two cervical tissue-mimicking phantoms with human blood and graphite rods as inclusions inside it. In accordance with the simulations, the phantom study revealed a more reliable photoacoustic signal for the entire depth of the phantoms with an improved contrast to noise ratio and signal to noise ratio, and a higher coverage ratio of the imaging field of view. In summary, the dual-mode illumination system can provide more realistic information of inclusions within the tissue while considering safety limits, which can lead to more accuracy in biomarker detection for cervical cancer diagnostics.

Spectroscopic photoacoustic imaging of cervical tissue composition in excised human samples.

OBJECTIVE: Cervical remodeling is an important component in determining the pathway of parturition; therefore, assessing changes in cervical tissue composition may provide information about the cervix's status beyond the measurement of cervical length. Photoacoustic imaging is a non-invasive ultrasound-based technology that captures acoustic signals emitted by tissue components in response to laser pulses. This optical information allows for the determination of the collagen-to-water ratio (CWR). The purpose of this study was to compare the CWR evaluated by using spectroscopic photoacoustic (sPA) imaging in cervical samples obtained from pregnant and non-pregnant women. METHODS: This cross-sectional study comprised cervical biopsies obtained at the time of hysterectomy (n = 8) and at the scheduled cesarean delivery in pregnant women at term who were not in labor (n = 8). The cervical CWR was analyzed using a fiber-optic light-delivery system integrated to an ultrasound probe. The photoacoustic signals were acquired within the range of wavelengths that cover the peak absorption of collagen and water. Differences in the CWR between cervical samples from pregnant and non-pregnant women were analyzed. Hematoxylin and eosin and Sirius Red stains were used to compare the collagen content of cervical samples in these two groups. RESULTS: Eight cervix samples were obtained after hysterectomy, four from women ≤41 years of age and four from women ≥43 years of age; all cervical samples (n = 8) from pregnant women were obtained after 37 weeks of gestation at the time of cesarean section. The average CWR in cervical tissue samples from pregnant women was 18.7% (SD 7.5%), while in samples from non-pregnant women, it was 55.0% (SD 20.3%). There was a significantly higher CWR in the non-pregnant group compared to the pregnant group with a p-value <0.001. A subgroup analysis that compared the CWR in cervical samples from pregnant women and non-pregnant women ≤41 years of age (mean 46.3%, SD 23.1%) also showed a significantly higher CWR (p <0.01). Lower collagen content in the pregnancy group was confirmed by histological analysis, which revealed the loss of tissue composition, increased water content, and collagen degradation. CONCLUSION: The proposed bimodal ultrasound and sPA imaging system can provide information on the biochemical composition of cervical tissue in pregnant and non-pregnant women. Photoacoustic imaging showed a higher collagen content in cervical samples from non-pregnant women as compared to those from pregnant women, which matched with the histological analysis. This novel imaging method envisions a new potential for a sensitive diagnostic tool in the evaluation of cervical tissue composition.

Synthesis of Self-Targeted Carbon Dot with Ultrahigh Quantum Yield for Detection and Therapy of Cancer.

This study aims to engineer a new type of ultrahigh quantum yield carbon dots (CDs) from methotrexate (MTX-CDs) with self-targeting, imaging, and therapeutic effects on MDA-MB 231 breast cancer cells. CDs were synthesized via a straightforward thermal method using a methotrexate (MTX) drug source. The physicochemical characteristics of the prepared MTX-CDs were studied using Fourier transform infrared (FT-IR) spectroscopy, transmission electron microscopy (TEM), dynamic light scattering (DLS), X-ray powder diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). TEM and DLS revealed which MTX-CDs have homogeneous spherical morphology with a smaller average size of 5.4 ± 2.2 nm, polydispersity index (PDI) of 0.533, and positive surface charge of around +3.93 mV. Results of FT-IR spectroscopy and high-resolution XPS indicated the presence of residues of MTX on CDs. Therefore, the synthesized MTX-CDs could be targeted and be taken up by FR-positive cell lines without the aid of additional targeting molecules. In vitro epifluorescence images demonstrated high-contrast cytoplasm biodistribution of MTX-CDs after 2 h of treatment. A much stronger fluorescent signal was detected in MDA-MB 231 compared to MCF 7, indicating their ability to precisely target FR. The highest cytotoxic and apoptotic effects were observed in MTX-CDs compared to free MTX obtained by the MTT assay, cell cycle arrest, and annexin V-FITC apoptosis techniques. Results revealed that the novel engineered MTX-CDs were capable of inducing apoptosis (70.2% apoptosis) at a lower concentration (3.2 µM) compared to free MTX, which was proved by annexin V and cell cycle. This work highlights the potential application of CDs for constructing an intelligent nanomedicine with integration of diagnostic, targeting, and therapeutic functions.