Fuping Gao

Doxorubicin loaded silica nanorattles actively seek tumors with improved anti-tumor effects

Silica nanorattles (SNs) have proven to be promising vehicles for drug delivery. In order to further enhance efficacy and minimize adverse effects, active targeted delivery to tumors is necessary. In this work, SNs modified with a tumor specific targeting ligand, folic acid (FA), was used as carrier of doxorubicin (DOX) (DOX-FA-SNs). Drug loading, cytotoxicity and cellular uptake of DOX-FA-SNs in vitro in human cervical carcinoma cells (HeLa cells) were evaluated. DOX-FA-SNs showed a higher cytotoxicity in human cervical carcinoma cells (HeLa cells) than DOX loaded carboxyl (-COOH) and poly(ethylene glycol) (PEG) modified SNs (DOX-COOH-SNs and DOX-PEG-SNs, respectively). However, DOX-FA-SNs showed lower cytotoxicity in folate receptor negative normal mouse fibroblast cells (L929 cells) compared with free DOX. In vivo tumor-targeted fluorescence imaging indicated specific tumor targeting and uptake of FA-SNs in nude mice bearing subcutaneous HeLa cell-derived xenograft tumors. In vivo anti-tumor experiments demonstrated that DOX-FA-SNs (10 mg kg(-1) of DOX) significantly regressed the tumor growth and reduced toxicity compared with free DOX. These results have great significance in developing and optimizing SNs as effective intracellular delivery and specific tumor targeting vehicles.

Ultrasmall [64Cu]Cu Nanoclusters for Targeting Orthotopic Lung Tumors Using Accurate Positron Emission Tomography Imaging

Positron emission tomography (PET) imaging has received special attention owing to its higher sensitivity, temporal resolution, and unlimited tissue penetration. The development of tracers that target specific molecules is therefore essential for the development and utility of clinically relevant PET procedures. However, (64)Cu as a PET imaging agent generally has been introduced into biomaterials through macrocyclic chelators, which may lead to the misinterpretation of PET imaging results due to the detachment and transchelation of (64)Cu. In this study, we have developed ultrasmall chelator-free radioactive [(64)Cu]Cu nanoclusters using bovine serum albumin (BSA) as a scaffold for PET imaging in an orthotopic lung cancer model. We preconjugated the tumor target peptide luteinizing hormone releasing hormone (LHRH) to BSA molecules to prepare [(64)Cu]CuNC@BSA-LHRH. The prepared [(64)Cu]Cu nanoclusters showed high radiolabeling stability, ultrasmall size, and rapid deposition and diffusion into tumor, as well as predominantly renal clearance. [(64)Cu]CuNC@BSA-LHRH showed 4 times higher tumor uptake compared with that of [(64)Cu]CuNC@BSA by analyzing the (64)Cu radioactivity of tissues via gamma counting. The PET imaging using [(64)Cu]Cu nanoclusters as tracers showed more sensitive, accurate, and deep penetration imaging of orthotopic lung cancer in vivo compared with near-infrared fluorescence imaging. The nanoclusters provide biomedical research tools for PET molecular imaging.

Peptide-Conjugated Gold Nanoprobe: Intrinsic Nanozyme-Linked Immunsorbant Assay of Integrin Expression Level on Cell Membrane

Precisely quantifying the membrane protein expression level on cell surfaces is of vital importance for early cancer diagnosis and efficient treatment. We demonstrate that gold nanoparticle bioconjugated by a rationally designed peptide as nanoprobe possesses selective labeling and accurate quantification capacity of integrin GPIIb/IIIa on the human erythroleukemia cell line. Through selective recognition and marking of integrin, two-photon photoluminescence of the nanoprobe is exploited for direct observation of protein spatial distribution on cell membrane. More importantly, utilizing intrinsic enzyme-like catalysis property of the nanoprobe, the expression level of integrin on human erythroleukemia cells can be quantitatively counted in an amplified and reliable colorimetric assay without cell lysis and protein extraction process. In addition, the analysis of the correlation between the gold nanoparticle and the membrane protein via relevant inductively coupled plasma mass spectrometry measurement verifies the reliability of the new analytical method. It is anticipated that this facile and efficient strategy holds a great promise for a rapid, precise, and reliable quantification of interested functional membrane proteins on the cell surface.

Uniform double-shelled silica hollow spheres: acid/base selective-etching synthesis and their drug delivery application

Uniform mono-disperse double-shelled silica hollow spheres (DSH) with controllable inner structure have been successfully synthesized via an alternating acid/base selective etching strategy. The void spacing between the shells can be tuned during the synthesis. The formation mechanism was proposed by monitoring the synthesis process at different reaction time and comparative investigation of the used surfactants. Anti-cancer drug, mitoxantrone (Mito) was chosen as a probing molecule in drug delivery and release experiments, and the results showed that these double-shelled silica hollow spheres exhibit higher drug loading capacity and desired release rates compared with conventional single-shelled silica hollow spheres due to their hierarchically mesoporous structures. The in vitro cellular internalization of DSH was evaluated by designing fluorescein isothiocyanate labeled DSH with green fluorescence (DSH@FITC). The in vitro study of Human liver carcinoma cells of Hep-G2 cells also proves that the DSH–Mito has reduced toxicity and enhanced therapeutic efficacy, and the DSH is an ideal carrier for drug delivery.

LHRH-PE40 Fusion Protein Tethered Silica Nanorattles for Imaging-Guided Tumor-Specific Drug Delivery and Bimodal Therapy

Docyanine green (ICG) and LHRH-PE40 fusion protein are tethered onto drug carriers of silica nanorattles for imaging-guided tumor-specific drug delivery and bimodal therapy. The synergistic therapeutic effect of toxin PE40 and the chemotherapeutic drug docetaxel (Dtxl), specifically directed by LHRH to cancer, improves cancer treatment. Simultaneously, ICG enables real-time monitoring of the silica nanocomposites and therapeutic response.

Bio-inspired peptide-Au cluster applied for mercury (II) ions detection

Mercury ion (Hg2+ ) pollution exists in water, soil, and food. By interacting with the thiol groups in protein, Hg2+ ions can accumulate in ways that cause serious damage to the central nervous system and threaten human health and natural environment. Undoubtedly, Hg2+ ion detection is a significant issue in environment and health monitoring. A variety of sensor platforms for Hg2+ ion detection based on organic molecules, DNA, oligonucleotides, inorganic materials, etc, have been reported. In this paper, an artificial peptide PHg, with a cluster bio-mineralize sequence (CCY) and a multi-charge hydrophilic sequence is designed as a template for the one-step synthesis of a peptide-Au cluster probe. Briefly: the peptide PHg in situ anchors Au ions to form a peptide-Au (I) intermediate and the reaction pH with NaOH is adjusted; after 12 h incubation at room temperature, the peptide PHg-Au nanocluster probe with red fluorescence is obtained. The probe has a super-small core size of approximately 1.26 nm and a maximum emission peak at 650 nm. The presence of Hg2+ ions cause the fluorescence of the probe to greatly decrease. Based on the differences in fluorescence intensity of the PHg-Au nanocluster in the absence and presence of Hg2+ ions, Hg2+ ions could be quantitatively detected in concentrations ranging from 5 nmol/L to 1 µmol/L. The limit of detection (LOD) is 7.5 nmol/L. Compared with some interference ions such as, K+, Mg2+, Ca2+, Pb2+, Ni2+, Fe3+, and Cu2+, the selectivity was excellent. The sensing of Hg2+ ion is not affected by the chelate agents: EDTA, which imparts a significant advantage in a range of applications. As a result, a simple, sensitive and selective fluorescent assay based on peptide PHg-Au cluster is developed for the detection of Hg2+ ions.

Au Nanoclusters and Photosensitizer Dual Loaded Spatiotemporal Controllable Liposomal Nanocomposites Enhance Tumor Photodynamic Therapy Effect by Inhibiting Thioredoxin Reductase

Photodynamic therapy (PDT) is a minimally invasive therapeutic procedure of tumors with high selectivity and low side effect. However, it is usually not efficient in long-lasting tumor control. One of the main reasons is tumor cells develop some protective mechanisms that help them to deal with oxidative stress in the environment. The thioredoxin system in cancer is an important antioxidant defense system. Au nanoclusters could effectively inhibit thioredoxin reductase (TrxR) in tumor cell cytoplasm. Herein, Au nanoclusters and photosensitizer Chlorine 6 (Ce6) are co-loaded in spatiotemporal controllable liposomal nanocomposites. pH responsive molecule inserted in lipid bilayer greatly contributes to the instability of the lipid membrane in lysosomal at low pH environment. Then the payloads can rapidly release into cytoplasm. Au nanoclusters effectively inhibit TrxR in cytoplasm and enhance the photodynamic-induced intracellular reactive oxygen-free radical concentration, improving the effect of PDT. Breast cancer is chosen as a tumor model and the Au nanoclusters and photosensitizer co-loaded liposomal nanocomposites are studied to improve the effect of PDT both in vitro and in vivo, and its corresponding mechanism is investigated. This study develops a new application of gold nanoclusters and provides a new train of thoughts for enhancing the effect of PDT.

The Precise Diagnosis of Cancer Invasion/Metastasis via 2D Laser Ablation Mass Mapping of Metalloproteinase in Primary Cancer Tissue

Cancer invasion and metastasis remain the major causes of over 90% of patient deaths. Molecular imaging methods such as computed tomography (CT)/magnetic resonance imaging (MRI) can precisely assess primary regional lymph node invasion and distant organ metastasis via body scanning; however, such diagnostic methods are often utilized too late for cancer therapy. To date, pathologic methods mainly provide information on differentiation/proliferation and potential drug therapy biomarkers of primary tumors rather than precisely reveal tumor regional invasion and distant metastasis in the body. We hypothesized that quantification of membrane type-1 matrix metalloproteinase (MT1-MMP) levels in primary tumor tissue will provide a precise assessment of tumor regional lymph node invasion and remote organ metastasis. In this work, we developed peptide-coated Au clusters with intrinsic red fluorescence and a specific mass signal. When these clusters labeled MT1-MMP in tumor tissue sections derived from the xenograft lung carcinoma model, human lung carcinoma and human renal carcinoma, we could directly observe MT1-MMP via optical fluorescence microscopy and quantitatively detect the MT1-MMP expression level via laser ablation inductively coupled plasma mass spectrometry 2D mapping (2D-LA-Mass Mapping). By observing and quantifying the MT1-MMP expression level in primary human lung carcinoma and human renal carcinoma tissue sections, we precisely assessed the risk of primary tumor invasion/metastasis. Importantly, the accuracy of this pathologic method was verified by CT/MRI molecular imaging of cancer patients and traditional hematoxylin and eosin (H&E) staining/immunohistochemistry (IHC)/immunofluorescence (IF) pathologic studies of primary tumor tissues.