Annals of Biomedical Engineering

Label-Free Monitoring of Endometrial Cancer Progression Using Multiphoton Microscopy

AbstractEndometrial cancer is the most common gynecological cancer in the developed world. However, the accuracy of current diagnostic methods is still unsatisfactory and time-consuming. Here, we presented an alternate approach to monitoring the progression of endometrial cancer via multiphoton microscopy imaging and analysis of collagen, which is often overlooked in current endometrial cancer diagnosis protocols but can offer a crucial signature in cancer biology. Multiphoton microscopy (MPM) based on the second-harmonic generation and two-photon excited fluorescence was introduced to visualize the microenvironment of endometrium in normal, hyperplasia without atypia, atypical hyperplasia, and endometrial cancer specimens. Furthermore, automatic image analysis based on the MPM image processing algorithm was used to quantify the differences in the collagen morphological features among them. MPM enables the visualization of the morphological details and alterations of the glands in the development process of endometrial cancer, including irregular changes in the structure of the gland, increased ratio of the gland to the interstitium, and atypical changes in the glandular epithelial cells. Moreover, the destructed basement membrane caused by gland proliferation and fusion is clearly shown in SHG images, which is a key feature for identifying endometrial cancer progression. Quantitative analysis reveals that the formation of endometrial cancer is accompanied by an increase in collagen fiber length and width, a progressive linearization and loosening of interstitial collagen, and a more random arrangement of interstitial collagen. Observation and quantitative analysis of interstitial collagen provide invaluable information in monitoring the progression of endometrial cancer. Label-free multiphoton imaging reported here has the potential to become an in situ histological tool for effective and accurate early diagnosis and detection of malignant lesions in endometrial cancer.

Usability of Speculum-Compatible Injection Devices for Administering Ethyl Cellulose-Ethanol Ablation to Treat Cervical Neoplasia in Low- and Middle-Income Countries

Abstract Purpose Current treatments for cervical neoplasia are often inaccessible in low- and middle-income countries (LMICs), which contributes to high cervical cancer mortality. We previously developed a low-cost ablative therapy using ethanol mixed with ethyl cellulose (EC) to a form an ethanol-retaining gel that reduces injection leakage. To optimize delivery of EC-ethanol into the cervix, we developed and compared three speculum-compatible injectors that each address clinical challenges: 1) a single needle injector, which contained an adjustable depth stop to control the depth of injection, 2) a multi needle injector, which injected three locations in the cervix simultaneously, and 3) an extender injector, which included a needle extender. Methods The variability in EC-ethanol injections was evaluated through bench top and ex vivo swine testing. Usability testing was performed by gynecology (GYN) providers who used each device in a custom pelvic model. Results Both the extender and single needle devices led to consistent ejection volumes in benchtop tests with no variability between injections. All devices achieved spherical depots with minimal leakage in ex vivo tests. In usability testing, 65% of GYN providers preferred the extender device, which achieved significantly shorter injection times in the custom pelvic model compared to other injectors. Conclusion While all devices met clinical constraints, the extender device was preferred by clinicians and achieved repeatable injection distributions. This work presents a clinically informed low-cost intracervical delivery method for LMICs. Future work will include validating performance in clinical trials and assessing feasibility in clinical settings to advance global cervical neoplasia treatment.

Virus-Mimicking Nanoparticles for Targeted Near Infrared Fluorescence Imaging of Intraperitoneal Ovarian Tumors in Mice

Ovarian cancer is the most lethal malignancy affecting the female reproductive system. Identification and removal of all ovarian intraperitoneal tumor deposits during the intraoperative surgery is important towards preventing cancer recurrence and ultimately improving patient survival. Herein, we investigate the effectiveness of virus mimicking nanoparticles, derived from genome-depleted plant-infecting brome mosaic virus, and doped with near infrared (NIR) brominated cyanine dye BrCy106-NHS, for targeted NIR fluorescence imaging of intraperitoneal ovarian tumors. We refer to these nanoparticles as optical viral ghosts (OVGs). We functionalized the OVGs with antibodies against HER2 receptor, a biomarker over-expressed in ovarian cancers. We injected functionalized OVGs, non-functionalized OVGs, and non-encapsulated BrCy106-NHS intravenously in mice implanted with ovarian intraperitoneal tumors. Tumors were extracted at 2, 6, and 24 h post-injection, and quantitatively analyzed using NIR fluorescence imaging. Fluorescence emission from tumors associated with the injection of the functionalized OVGs continued to increase between 2 and 24 h post-injection. At 24 h timepoint, the average spectrally-integrated fluorescence emission from homogenized tumors containing functionalized-OVGs was about 3.5 and 19.5 times higher than those containing non-functionalized OVGs or non-encapsulated BrCy106-NHS, respectively. Similarly, by using the functionalized-OVGs, the imaging signal-to-noise ratio at 24 h timepoint was enhanced by approximately threefold and sevenfold as compared to non-functionalized OVGs and the non-encapsulated dye, respectively. These functionalized virus-mimicking NIR nano-constructs could potentially be used for intraoperative visualization of ovarian tumors implants.