Multifunctional Nanoprobes Research Articles

A universal reagent for detection of emerging diseases using bioengineered multifunctional yeast nanofragments.

Accurate and early detection of biomarkers provides the molecular evidence for disease management, allowing prompt actions and timely treatments to save lives. Multivalent biomolecular interactions between the probe and biomarker as well as controlled probe orientation on material surfaces are keys for highly sensitive detection. Here we report the bioengineering of programmable and multifunctional nanoprobes, which can provide rapid, specific and highly sensitive detection of emerging diseases in a range of widely used diagnostic systems. These nanoprobes composed of nanosized cell wall fragments, termed as synthetic bionanofragments (SynBioNFs), are generated by the fragmentation of genetically programmed yeast cells. SynBioNFs display multiple copies of biomolecules for high-affinity target binding and molecular handles for the precisely orientated attachment on surfaces used in diagnostic platforms. SynBioNFs are demonstrated for the capture and detection of SARS-CoV-2 virions using multiple diagnostic platforms, including surface-enhanced Raman scattering, fluorescence, electrochemical and colorimetric-based lateral flow systems with sensitivity comparable with the gold-standard reverse-transcription quantitative polymerase chain reaction.

Multifunctional fluorescent mesoporous carbon nanoprobe for MMP-2-activated cancer cell imaging and targeted photothermal therapy

The integration of cancer imaging with therapy in a simple system is warranted for precise cancer therapy. In this study, carboxyl-functionalized mesoporous carbon nanospheres (MCN) which are efficient photothermal agents and excellent fluorescence quenchers, were used for cancer cell imaging and selective photothermal therapy (PTT) applications. Using MCN, a matrix metalloproteinase-2 (MMP-2)- responsive theranostic nanoprobe was generated by functionalizing an MMP-2-specific fluorescent-labeled PLGVR sequence on the surface of MCN. The nanoprobe not only can be used to detect MMP-2 with a low detection limit of 0.3 pg mL−1, but also can achieve the sensitive intracellular MMP-2 imaging in living cells, validating the differentiation of cancer cells from healthy cells based on the recovered fluorescence intensity. More importantly, selective cancer PTT was achieved using MMP-2-triggered cancer cell imaging. Our in vitro studies showed that by regulating the power density and irradiation time, the nanoprobe can effectively kill cancer cells via PTT. Our strategy opens new avenues for precision medicine, especially phototherapy.

Detection of H2S in Living Cells Using Escape Lysosome Technology Based on the Swelling Effect of Polymeric Nanomicelles

AbstractThe conventional fluorescent probe for sensitive detection of bioactive substance in living cells are still challenging due to the degradation and destruction of lysosomes to probes. Here, we develop a new method based on escape lysosome technology for detection of bioactive small molecules like hydrogen sulfide (H2S) in living cells. It relies on the swelling effect of polymeric nonomicelles. Specifically, PEG‐PEI/DP‐NBD complex probes were designed and prepared by integrating polyethyleneimine (PEI) and fluorescent probe dansylpiperazine nitrobenzo(c)‐(1,2,5) oxadiazole (DP‐NBD) into poly(ethylene glycol) (PEG) nanogel, which possessed lysosome‐escape ability based on swelling effect of PEG nanomicelles. Results indicate the probes we proposed can be used for sensitive tracking and long‐term visualization of H2S in living cells, and the detection limit of H2S is 0.1 μM. The multifunctional nanoprobes based on escape lysosome technology by help of swelling effect of polymeric micelles offers an important strategy for detecting bioactive substance in living cells.

Novel Self-Assembled Multifunctional Nanoprobes for Second-Near-Infrared-Fluorescence-Image-Guided Breast Cancer Surgery and Enhanced Radiotherapy Efficacy.

Breast-conserving surgery (BCS) is the predominant treatment approach for initial breast cancer. However, due to a lack of effective methods evaluating BCS margins, local recurrence caused by positive margins remains an issue. Accordingly, radiation therapy (RT) is a common modality in patients with advanced breast cancer. However, while RT also protects normal tissue and enhances tumor bed doses to improve therapeutic effects, current radiosensitizers cannot meet these urgent clinical needs. To address this, a novel self-assembled multifunctional nanoprobe (NP) gadolinium (Gd)-diethylenetriaminepentaacetic acid-human serum albumin (HSA)@indocyanine green-Bevacizumab (NPs-Bev) is synthesized to improve the efficacy of fluorescence-image-guided BCS and RT. Fluorescence image guidance of the second near infrared NP improves complete resection in tumor-bearing mice and accurately discriminates between benign and malignant mammary tissue in transgenic mice. Moreover, targeting tumors with NPs induces more reactive oxygen species under X-ray radiation therapy, which not only increases RT sensitivity, but also reduces tumor progression in mice. Interestingly, self-assembled NPs-Bev using HSA, the magnetic resonance contrast agent and Bevacizumab-targeting vascular growth factor A, which are clinically safe reagents, are safe in vitro and in vivo. Therefore, the novel self-assembled NPs provide a solid precision therapy platform to treat breast cancer.

Multifunctional nanoprobe for multi-mode imaging and diagnosis of metastatic prostate cancer

The high incidence and complex subtypes of prostate cancer put forward higher requirements for accurate diagnosis. Furthermore, advanced prostate cancer is prone to metastasis. Single biological imaging mode faces a challenge of sensitive and fast bioimaging of metastasic prostate cancer. Thus, exploring a nanoprobe with multi-mode imaging function has an important impact on preoperative imaging and intraoperative visualization guide of metastatic prostate cancer. Herein, based on the optical properties and X-ray attenuation capability of Au nanodots as well as the slow electronic relaxation of Gd3+, we designed and fabricated the multifunctional nanoprobe Au/Gd nanodots for multi-mode imaging and accurate diagnosis of bone metastatic prostate cancer. The results showed that multiple imaging modes complement each other to achieve high-precision of metastasic prostate cancer detection and accurately guide treatment. In addition, in vitro/vivo experiments showed that Au/Gd nanodots had good biocompatibility and biosafety. Therefore, the prepared multifunctional nanoprobe may provide new strategies and insights for precise diagnosis of metastatic prostate cancer in clinical practice.

Multifunctional DNA nanoprobe for tumor-targeted synergistic therapy by integrating chemodynamic therapy with gene silencing.

Due to the high complexity, diversity and heterogeneity of tumor occurrence and development, multi-mode synergistic therapy is more effective than single treatment modes to improve the antitumor efficacy. Also, multifunctional probes are crucial to realize synergistic therapy. Herein, a multifunctional DNA tetrahedron nanoprobe was ingeniously designed to simultaneously achieve chemodynamic therapy (CDT) and gene silencing for synergistic antitumor. The multifunctional DNA tetrahedron nanoprobe, DNA tetrahedron-silver nanocluster-antagomir-21 (D-sgc8-DTNS-AgNCs-Anta-21), integrated a CDT reagent (DNA-AgNCs) and miRNA-21 inhibitor (Anta-21) with a specific recognition probe (aptamer). After targeted entry in cancer cells, D-sgc8-DTNS-AgNCs-Anta-21 silenced endogenous miRNA-21 by Anta-21 and produced highly toxic ˙OH by reacting with H2O2, which induced apoptosis in the tumor cells. The targeted recognition of aptamers led to the concentration-dependent death of HeLa cells. On the contrary, the cell survival rate of normal cells was basically unaffected with an increase in the concentration of D-sgc8-DTNS-AgNCs-Anta-21. Therefore, the diverse functions, biocompatibility and programmability of DNA provide a useful and easy way to assemble multifunctional probes for synergistic therapy.

Self-driven immune checkpoint blockade and spatiotemporal-sensitive immune response monitoring in acute myeloid leukemia using an all-in-one turn-on bionanoprobe.

Immune checkpoint (ICP) blockade (ICB) is one of the most promising immunotherapies for acute myeloid leukemia (AML). However, owing to their heterogeneity, AML cells may cause uncoordinated metabolic fluxes and heterogeneous immune responses, inducing the release of a spatiotemporally sensitive immune response marker. Timely and in situ detection of immune responses in ICB therapy is important for therapeutic strategy adjustment. Herein, we constructed an all-in-one nanoprobe for self-driving ICB and simultaneously detecting an immune response in the same AML cell in vivo, thus enabling accurate evaluation of heterogenetic immune responses in living AML mice without additional drug treatment or probe processes. The nature-inspire polydopamine (PDA) nanoparticles loaded with an ICP blocker were targeted to the leukocyte immunoglobulin like receptor B4 (a new ICP) of AML cells to induce the release of immune response marker granzyme B (GrB). The PDA nanoparticles were additionally paired with carbon-derived graphene quantum dots (GQDs) to construct a full-organic 'turn-on' bionanoprobe that can transfer fluorescence resonance energy for GrB detection. This multifunctional nanoprobe was validated for triggering ICB therapy and monitoring the changes of GrB levels in real-time both in vitro and in vivo. The organic nanoprobe showed excellent permeability and retention in tumor cells and high biocompatibility in vivo. This bionanoprobe orderly interacted with the upstream ICP molecules and downstream signal molecule GrB, thereby achieving in situ immune response signals within the therapeutic efficacy evaluation window.

A covalent organic framework-derived M1 macrophage mimic nanozyme for precise tumor-targeted imaging and NIR-II photothermal catalytic chemotherapy.

Nanoprobes for efficient tumor-targeted imaging and therapy are urgently needed for clinical tumor theranostics. Herein, inspired by the heterogeneity of the tumor microenvironment, we report a covalent organic framework (COF)-derived biomimetic nanozyme for precise tumor-targeted imaging and NIR-II photothermal-catalysis-enhanced chemotherapy (PTCEC). Using a crystalline nanoscale COF as the precursor, a peroxidase-like porous N-doped carbonous nanozyme (PNC) was obtained, which was cloaked with an M1 macrophage cell membrane (M1m) to create a multifunctional biomimetic nanoprobe for tumor-targeted imaging and therapy. The M1m coating enabled the nanoprobe to target cancer cells and tumor tissues for highly efficient tumor imaging and drug delivery. The peroxidase-like activity of the PNC allowed for the conversion of intratumoral H2O2 into toxic ˙OH that synergized with its NIR-II photothermal effect to strengthen the chemotherapy. Therefore, highly efficient tumor-targeted imaging and NIR-II PTCEC were realized with an M1 macrophage mimic nanoprobe. This work provides a feasible tactic for a biomimetic theranostic nanoprobe and will inspire the development of new bioactive nanomaterials for clinical tumor theranostic applications.

Multifunctional Nanoprobe for 3D Nanoresolution Imaging of Intact Cell HER2 Protein with Hard X-ray Tomography.

Three-dimensional nondestructive, nanoresolution, and in situ visualization of protein spatial localization in a large, thick single cell remains challenging. In this study, we designed a multifunctional iron oxide (Fe@BFK) nanoprobe that possesses fluorescence and hard X-ray imaging signals. This probe can specifically target the human epidermal growth factor receptor 2 (HER2) protein and help optimize the label condition and selection of suitable samples for X-ray imaging. Combining 30 nm resolution synchrotron radiation hard X-ray nanocomputed tomography and the X-ray-sensitive Fe@BFK nanoprobe, a 3D localization of HER2 on SK-BR-3 cells was obtained for the first time. HER2 was mainly localized and cluster-distributed on the cell membrane with a heterogeneous pattern. This study provides a novel method for the in situ and nondestructive synchrotron radiation imaging of the desired protein localization in large, thick cells and evaluation of the true cellular distribution of a nanoprobe with high resolution.

An AIE luminogen self-assembled nanoprobe for efficient monitoring of the concentration and structural transition of human serum albumin

Monitoring the content and structural transition of human serum albumin (HSA) is valuable for diagnosing hypoalbuminemia and albuminuria and elucidating its transport function, respectively. Herein, we developed a multifunctional and efficient nanoprobe based on the self-assembly of an amphiphilic aggregation-induced emission luminogen, TPNN, for HSA detection and structure monitoring. TPNN molecules formed loose self-assembly in aqueous media and emitted faint fluorescence due to vigorous intramolecular motion. In the presence of HSA, a remarkable fluorescence enhancement accompanied by dissociation of TPNN assembly was observed due to the site-specific binding of TPNN to the middle long-narrow pocket of HSA. This binding blocked the intramolecular motion while producing a sensitive, selective, fast and stable fluorescence turn-on response to HSA, making TPNN assembly suitable for the HSA assay. TPNN assemblies also showed superior selectivity to HSA than bovine serum albumin (BSA) due to the distinct binding modes, and enabled us to distinguish the two homologous proteins. Furthermore, the generated TPNN-HSA complex underwent stepwise fluorescence quenching in the presence of protein denaturants and proteases, which revealed the process and kinetics of HSA unfolding and cleavage. TPNN assembly provides a powerful tool for accurate quantification of HSA in serum and urine and real-time monitoring of HSA structural transition.

Sensitive detection of organophosphorus pesticides based on the localized surface plasmon resonance and fluorescence dual-signal readout

In this work, a dual-signal visual biosensor was designed for organophosphorus pesticides (OPs) detection using DNA functionalized Ag/Au bimetallic nanoparticles (Ag/Au NPs) as multifunctional nanoprobe. The dual-signal detection strategy was based on the inhibition of enzyme-induced H2O2 generation by OPs in the detection solution containing acetylcholinesterase (AChE), choline oxidase (CHO), acetylcholine (ACh) and nanoprobe. H2O2 produced by enzyme-catalyzed reaction could trigger the etching of Ag and dissociation of carboxyfluorescein (FAM)-labeled aptamer from the nanoprobe, resulting in significant localized surface plasmon resonance (LPRR) and fluorescence (FL) signal responses. In the presence of OPs, AChE activity was inhibited to disrupt the enzymatic generation of H2O2, which allowed to simultaneous quantitative measure OPs through the LSPR peak shifts and FL intensity variations of the nanoprobe. The LSPR/FL dual-signal biosensor showed great selectivity and sensitivity for OPs detection. In addition, two distinct colour changes were visually observed to match the LSPR/FL spectra signal responses, which was a feasible means for visual analysis of OPs. Consequently, the work provided a dual-signal visual biosensor via the combination of multifunctional nanoprobe, and had significant potential to monitor pesticide residue with high anti-interference capability and detection accuracy.

Multifunctional nanoprobe for real-time in vivo monitoring of T cell activation

Genetically engineered T cells are a powerful new modality for cancer immunotherapy. However, their clinical application for solid tumors is challenging, and crucial knowledge on cell functionality in vivo is lacking. Here, we fabricated a nanoprobe composed of dendrimers incorporating a calcium sensor and gold nanoparticles, for dual-modal monitoring of engineered T cells within a solid tumor. T cells engineered to express a melanoma-specific T-cell receptor and loaded with the nanoprobe were longitudinally monitored within melanoma xenografts in mice. Fluorescent imaging of the nanoprobe's calcium sensor revealed increased intra-tumoral activation of the T cells over time, up to 24 h. Computed tomography imaging of the nanoprobe's gold nanoparticles revealed the cells' intra-tumoral distribution pattern. Quantitative analysis revealed the intra-tumoral T cell quantities. Thus, this nanoprobe reveals intra-tumoral persistence, penetration and functional status of genetically engineered T cells, which can advance T cell-based immunotherapy and promote next-generation live cell imaging.

Synchronous enhancement of upconversion and NIR-IIb photoluminescence of rare-earth nanoprobes for theranostics.

This study develops a multifunctional molecular optical nanoprobe (SiO2@Gd2O3: Yb3+/Er3+/Li+@Ce6/MC540) with a unique core-satellite form. The rare-earth doped nanodots with good crystallinity are uniformly embedded on the surface of a hydrophilic silica core, and the nanoprobe can emit near-infrared-IIb (NIR-IIb) luminescence for imaging as well as visible light that perfectly matches the absorption bands of two included photosensitizers under 980 nm irradiation. The optimal NIR-IIb emission and upconversion efficiency are attainable via regulating the doping ratios of Yb3+, Er3+ and Li+ ions. The relevant energy transfer mechanism was addressed theoretically that underpins rare-earth photoluminescence where energy back-transfer and cross relaxation processes play pivotal roles. The nanoprobe can achieve an excellent dual-drive photodynamic treatment performance, verified by singlet oxygen detections and live-dead cells imaging assays, with a synergistic effect. And a brightest NIR-IIb imaging was attained in tumoral site of mouse. The nanoprobe has a high potential to serve as a new type of optical theranostic agent for tumor.

Multifunctional Magnetic CuS/Gd2O3 Nanoparticles for Fluorescence/Magnetic Resonance Bimodal Imaging-Guided Photothermal-Intensified Chemodynamic Synergetic Therapy of Targeted Tumors.

Chemodynamic therapy (CDT), which consumes endogenous hydrogen peroxide (H2O2) to generate reactive oxygen species (ROS) and causes oxidative damage to tumor cells, shows tremendous promise for advanced cancer treatment. However, the rate of ROS generation based on the Fenton reaction is prone to being restricted by inadequate H2O2 and unattainable acidity in the hypoxic tumor microenvironment. We herein report a multifunctional nanoprobe (BCGCR) integrating bimodal imaging and photothermal-enhanced CDT of the targeted tumor, which is produced by covalent conjugation of bovine serum albumin-stabilized CuS/Gd2O3 nanoparticles (NPs) with the Cy5.5 fluorophore and the tumor-targeting ligand RGD. BCGCR exhibits intense near-infrared (NIR) fluorescence and acceptable r1 relaxivity (∼15.3 mM-1 s-1) for both sensitive fluorescence imaging and high-spatial-resolution magnetic resonance imaging of tumors in living mice. Moreover, owing to the strong NIR absorbance from the internal CuS NPs, BCGCR can generate localized heat and displays a high photothermal conversion efficiency (30.3%) under 980 nm laser irradiation, which enables photothermal therapy and further intensifies ROS generation arising from the Cu-induced Fenton-like reaction for enhanced CDT. This synergetic effect shows such an excellent therapeutic efficacy that it can ablate xenografted tumors in vivo. We believe that this strategy will be beneficial to exploring other advanced nanomaterials for the clinical application of multimodal imaging-guided synergetic cancer therapies.

Handheld pH‐Meter‐based Electrochemical Aptasensing of Carcinoembryonic Antigen on Multifuctional Magnetic Beads

AbstractMethods derived from immunoassays and aptasensors have been established for monitoring of carcinoembryonic antigen (CEA) in biological fluids, but most involved complex procedure and various reactions. We designed an uncomplicated and convenient electrochemical aptasensing platform for the quantitative monitoring of CEA on multifunctionalized magnetic beads with a handheld pH meter as the read‐out. Initially, CEA‐specific aptamers were covalently conjugated to magnetic beads, and then hybridized with glucose oxidase‐labeled complementary strands to construct multifunctional nanoprobes. After addition of target CEA, the analyte interacts with the corresponding aptamer and then release the complementary strand. After magnetic separation, the released glucose oxidase‐labeled complementary strands hydrolysed glucose, thereby leading to a change in pH of the detection solution, which was quantitatively monitored on a hand‐held pH meter. Under the optimum conditions, the aptasensor based on pH meter exhibited well responses relative to CEA concentrations inside the dynamic range between 0.01 and 100 ng mL−1 and detection limit at least 9.3 pg mL−1. Good duplicability, high specificity and ability for long‐term stable storage are obtained for the as‐prepared aptasensors. Additionally, human serum specimens were determined through pH‐meter‐based aptasensors, and the results closely matched the commercial enzyme‐linked immunosorbent assay (ELISA) of human CEA.

A nanoplatform of hollow Ag2S/Ag nanocomposite shell for photothermal and enhanced sonodynamic therapy mediated by photoacoustic and CT imaging

• A hollow Ag 2 S/Ag nanoshell composed of elemental Ag and compound Ag 2 S is synthesized. • The probe avoids the major defect of pure Ag shell gradual oxidiation and dissolution. • The probe solves the problem of hypoxia while taking both PTT and PAI into account. • The elemental Ag can assist H 2 O 2 in tumor cells to produce O 2 to improve SDT. In this study, a hollow Ag 2 S/Ag nanocomposite shell comprised of elemental Ag and compound Ag 2 S was synthesized, after loading the sonosensitizer and CT contrast agent, a multifunctional HASAIC nanoprobe was obtained for the first time by wrapping it with a thermosensitively supported lipid bilayer. The elemental Ag in the probe can catalyze H 2 O 2 to O 2 to relieve the hypoxic environment at the tumor site and improve the effect of sonodynamic therapy. Moreover, the hollow Ag 2 S/Ag nanocomposite shell avoids the key limitation of pure Ag shell during the process of catalyzing H 2 O 2 to O 2 , the pure Ag shell will dissolve and could not be used for photothermal therapy (PTT) and photoacoustic imaging (PAI), and enables the probe to use stable Ag 2 S to employ PTT and PAI though solving the problem of hypoxia. The synergy combination of the nano size (70 nm) HASAIC probe, low cytotoxicity, good safety, and high biocompatibility boost up the tumor killing capability. Because of the high efficiency, the HASAIC nanoprobe has been employed for real-time ( in vitro and in vivo) CT and PAI imaging in a 4T1 tumor-bearing mouse. Laser and ultrasound irradiation demonstrated a higher survival rate of HASAIC-injected mice compared to a probe without elemental silver, thereby indicating the high efficiency of our diagnosis and treatment probe.

Dye Sensitization Offers a Brighter Afterglow Nanoparticle Future for in vivo Recharged Luminescent Imaging.

While concerns about improving recharged afterglow intensity in vivo still motivate further exploration, afterglow nanoparticles (AGNP) offer unique optical merit for autofluorescence-free biological imaging. Apart from efforts enhancing the afterglow emission properties of AGNP, improving afterglow excitation response to visible or near infrared light is important but has lacked success. Dye sensitization has been used to improve the optical response of photovoltaic nanomaterials and to enhance upconversion luminescence efficiency. This concept has recently been expanded and applied to AGNPs. As a new multifunctional nanoprobe, such dye-sensitized AGNP takes advantage of both high spatial resolution fluorescence imaging and sensitive afterglow imaging. This Concept introduces the background, the concept, mechanism, and related imaging application, as well as reviewing existing challenges and proposing future developmental directions for the dye-sensitized AGNPs.

Biomimetic multifunctional persistent luminescence nanoprobes for long-term near-infrared imaging and therapy of cerebral and cerebellar gliomas.

Glioma is the most common malignant primary brain tumor, and the accurate diagnosis of glioma has always been a challenge. Moreover, cerebellar glioma, which is difficult to be detected by magnetic resonance imaging, is not usually diagnosed until after the appearance of clinical symptoms. In this study, TRZD, a near-infrared (NIR) persistent luminescence (PL) nanoparticle with a dual function of imaging and therapy, was synthesized based on ZnGa2O4:Cr3+,Sn4+. TRZD showed excellent rechargeable NIR PL for more than 30 hours in vivo with good tissue penetration for long-term autofluorescence-free imaging. The tumor growth of both the subcutaneous and orthotropic glioma models was significantly inhibited by TRZD. This is the first-time approach using NIR PL nanoprobes for both diagnosis and therapy of glioma. This is also the first-time report of nanotechnology-based diagnosis and therapy of cerebellar gliomas. This study offers a highly promising multifunctional nanoparticle for theranostics of a wide range of brain diseases.

Abstract P3-02-08: Diagnosis, navigation, treatment: A versatile biocompatible and biodegradable mesoporous silica platform of NIR II fluorescence imaging for early diagnosis, breast cancer precisely surgery, and radiosensitization

Abstract Background: Triple-negative breast cancer (TNBC) has the highest recurrence and distant metastasis rates among breast cancer molecular types. In breast-conserving surgery, the insensitivity discrimination of negative margins correlating with locoregional recurrence and poor patient outcomes. As for immunotherapy, only 20 %-40 % of the patients respond to the atezolizumab treatment due to the change of PDL1 receptor. Meanwhile, TNBC has radiotherapy resistance. Novel radiosensitization targets need to be found to enhance both the local and systematic benefits of radiotherapy. Objectives and rationale: We aim to construct an “all in one” multi-functional nanoprobe (MSN-Gd@Aptamer-ICG, NPs) using mesoporous silica as the framework. The mesoporous silica nanospheres are co-doped with gadolinium (Gd3+), loads indocyanine green (ICG) in its mesoporous channel, and conjugates with a specific PDL1 targeting aptamer on the surface. The biocompatible and biodegradable NPs are capable of release ICG by the reaction of intracellular high GSH and tetrasulfide bonds in silica frame. In the diagnosis, NIR II fluorescent imaging of ICG with the high resolution and deep tissue penetration depth screens out PDL1 highly expressed tumors. In surgical resection, it can show a real-time cancerous tissues margin in intraoperative scenarios and the tumors are successfully remove under the navigation of NIR II fluorescent imaging. During metastasis therapy of post-operation, the released Gd3+ effectively deposits X-rays and produces abundant hydroxyl radicals (•OH) and oxidative stress to induce radiosensitizing effects. More importantly, the therapy efficiency is monitored by NIR II fluorescent imaging. Therefore, our multifunctional NPs have roles in the accurate diagnosis of PDL1 expression of tumor in early stage, intraoperative navigation for radical excision, and metastasis eradication with radiosensitization efficiency enhanced, which lays a solid foundation for clinical TNBC therapy. Methods: The probe was 50 nm and it was successfully synthesized and certified. MTT assay suggested that it was nontoxic to normal breast cells and TNBC cells. Transmission Electron Microscope images showed that the probe was stable at the extracellular fluid and was degradable at tumor intracellular conditions. The leaching of ICG and Gd3+ was negligible at the extracellular fluid. The tumor targeting capability of the probe in vitro was evaluated on 4T1 (TNBC, PDL1 low expressed) and MDA-MB-231 (TNBC, PDL1 high expressed) cells by flow cytometry and laser confocal microscope. In vivo, the NIR II fluorescence imaging with a high resolution and tissue penetration depth successfully distinguished 4T1 and MDA-MB-231 bearing mice indicated the superior diagnostic capability of PDL1 via aptamer. In the surgical resection, the NIR II fluorescence imaging of NPs guided to remove the tumor with precisely discriminate the boundary of the tumor. When combined with RT, IHC staining of Ki67 showed that the NPs exhibited superior radiosensitizing effects under NIR II fluorescent imaging guidance in comparison with undoped Gd mesoporous silica nanoprobes. TUNEL fluorescent staining and H&E staining further confirmed that NPs radiation therapy induced more •OH and extensive DNA double-strand breaks. Results and Conclusion: Our biocompatible and biodegradable platform is nontoxic, can screen out PDL1 high expression tumors, and navigate the precise resection in the operation. Besides, it enhances the radioimmunotherapy efficacy of breast metastasis. Our platform is potential to provide new hope for breast cancer diagnostic, guiding resection, and radiosensitization under NIR II fluorescent imaging. Citation Format: Min Wei, Peiyuan Wang, Guojun Zhang. Diagnosis, navigation, treatment: A versatile biocompatible and biodegradable mesoporous silica platform of NIR II fluorescence imaging for early diagnosis, breast cancer precisely surgery, and radiosensitization [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-02-08.

Mesoporous carbon nanospheres featured multifunctional fluorescent nanoprobe: Simultaneous activation and tracing of caspase-3 involved cell apoptosis

Caspase-3 is considered to be a key regulator of cell apoptosis. Simultaneous activation and detection of caspase-3 in living cells is therefore highly important for precise cancer therapy and therapeutic efficacy evaluation but it is still a key challenge. Herein, a novel and simple multifunctional peptide-tailored mesoporous carbon nanospheres (MCN) has been developed for simultaneous cancer photothermal therapy (PTT) and real-time imaging of caspase-3 induced cell apoptosis. The nanoprobe was facilely prepared by modifying fluorescein isothiocyanate (FITC)-tagged peptide specific to caspase-3 on the surface of MCN. The signal switch relies on the unique quenching effect of MCN on the fluorescently labeled peptide, cleavage of the peptide by activated caspase-3 will result in a significant fluorescence lighting-up for sensitive caspase-3 analysis. The detection limit of 0.4 pg·mL−1 with a linear range from 10 pg·mL−1 to 1 ng·mL−1 was achieved. Owing to the intrinsic properties of MCN, the nanoprobe not only can be served as photothermal transducer to activate caspase-3, but also can simultaneously real-time monitoring caspase-3 activity in living cells with high specificity. This work demonstrates a smart molecular imaging strategy for cell apoptosis, which also provides a potential new platform for apoptosis-related diagnosis and precise cancer therapeutic applications.