Cell-Membrane Coated Nanoparticles for Tumor Delineation and Qualitative Estimation of Cancer Biomarkers at Single Wavelength Excitation in Murine and Phantom Models

Abstract

Real-time guidance through fluorescence imaging improves the surgical outcomes of tumor resections, reducing the chances of leaving positive margins behind. As tumors are heterogeneous, it is imperative to interrogate multiple overexpressed cancer biomarkers with high sensitivity and specificity to improve surgical outcomes. However, for accurate tumor delineation and ratiometric detection of tumor biomarkers, current methods require multiple excitation wavelengths to image multiple biomarkers, which is impractical in a clinical setting. Here, we have developed a biomimetic platform comprising near-infrared fluorescent semiconducting polymer nanoparticles (SPNs) with red blood cell membrane (RBC) coating, capable of targeting two representative cell-surface biomarkers (folate, αυβ3 integrins) using a single excitation wavelength for tumor delineation during surgical interventions. We evaluate our single excitation ratiometric nanoparticles in in vitro tumor cells, ex vivo tumor-mimicking phantoms, and in vivo mouse xenograft tumor models. Favorable biological properties (improved biocompatibility, prolonged blood circulation, reduced liver uptake) are complemented by superior spectral features: (i) specific fluorescence enhancement in tumor regions with high tumor-to-normal tissue (T/NT) ratios in ex vivo samples and (ii) estimation of cell-surface tumor biomarkers with single wavelength excitation providing insights about cancer progression (metastases). Our single excitation, dual output approach has the potential to differentiate between the tumor and healthy regions and simultaneously provide a qualitative indicator of cancer progression, thereby guiding surgeons in the operating room with the resection process.

Near-Infrared Afterglow Luminescence Amplification via Albumin Complexation of Semiconducting Polymer Nanoparticles for Surgical Navigation in Ex Vivo Porcine Models

“…The absorption spectra revealed a prominent absorbance peak at 535nm for both nanoparticles, consistent with previous reports and our bandgap measurements. 29 Upon excited at 605nm, both nanoparticles exhibited an emission peak at 820nm. However, BSA@SP1 displayed a significantly higher emission intensity compared to DSPE-PEG@SP1.…”

“…This understanding is crucial for guiding their future clinical applications. To investigate this behavior of BSA@SP1 across various tissue types, we utilized 3D tumor-mimicking phantom models using a methodology previously developed by us 29,30 comprising intralipids to simulate fat content (scattering component), hemoglobin to mimic blood vessels (absorption component), and tumor cells labeled with nanoparticles or solely nanoparticles at in vivo concentration levels (fluorescence component). 36 It offers distinct advantages over commonly adopted methods, such as utilizing intralipid solutions for simulation 31 because tumor-mimicking phantoms can more accurately replicate the optical properties of real tumors, including their scattering and absorption coefficients.…”

“…Our preliminary molecular docking studies reveal that PFODBT preferentially inserts into the hydrophobic pocket of BSA, imparting BSA@SP1 nanoparticles with improved colloidal stability and biocompatibility. Moreover, we showcase enhanced tissue permeation profiling by leveraging the afterglow capabilities of BSA@SP1 through experiments conducted on ex vivo tumor cell-mimicking phantoms 29,30 and various porcine tissue types (including skin, muscle, and fat), revealing their superior signal-to- background ratio compared to fluorescence imaging. Expanding on this, to assess its effectiveness in image-guided cancer surgeries, we implanted tumor-mimicking phantoms at various locations within the porcine lungs: (i) 0.5mm below the right inferior lobe, (ii) 5mm below the left inferior lobe, and (iii) within the thoracic cavity.…”

Recent Advances in Fluorescent Probes for Cancer Biomarker Detection

“…Compared with small molecule fluorescent probes, nanomaterials have a large specific surface area with a variety of loaded signaling sensors, thereby reducing the LOD of the sensors. Second, nanomaterials can be chemically modified easily to enable multi-channel simultaneous detection of a single analyte or simultaneous detection of multiple analytes [ 45 , 46 , 47 ]. In this section, we present recent advances and applications of fluorescent nano probes such as metal nanoclusters (MNCs), fluorescent nanoparticles (FNPs), and quantum dots (QDs), for cancer biomarkers.…”

Biomimetic-Membrane-Protected Plasmonic Nanostructures as Dual-Modality Contrast Agents for Correlated Surface-Enhanced Raman Scattering and Photoacoustic Detection of Hidden Tumor Lesions

“…Image-guided surgery (IGS) has emerged as an innovative approach for cancer treatment that combines advanced imaging technologies with surgical techniques to improve the precision and accuracy of cancer removal in an intraoperative setting. − IGS takes advantage of different modalities including optical imaging and spectroscopic modalities that can create detailed images of the cancer tumor and surrounding tissues, helping the surgeon navigate during the resection process to locate and remove cancerous tissues with greater accuracy and efficiency. − Surface-enhanced Raman scattering (SERS) is one such spectroscopic modality that arises due to the plasmonic effect where molecular dye reporters are in very close contact with a nanoroughened metal surface (gold and silver), which results in the enhancement of Raman signals. , The proximity of such reporters to gold nanoparticles (AuNPs) culminates in a strong SERS response that could be leveraged for IGS by providing real-time, quantitative, and spectroscopic information on the reporter molecules in the samples (cells and tissues). − Furthermore, compared to other optical imaging approaches like fluorescent imaging, SERS possesses distinct advantages of higher sensitivity, low photobleaching, and the potential for multiplexing. − Recently, near-infrared (NIR) light-excited SERS garnered significant attention for in vivo studies such as visualizing inflammatory lesions in animal models with high sensitivity and noninvasively imaging mouse liver with five SERS nanoprobes to obtain unmixed, fingerprint-like Raman spectra …”