A multifunctional protein-based hydrogel with Au nanozyme-mediated self generation of H2S for diabetic wound healing.

Diabetics usually suffer from chronic impaired wound healing due to facile infection, excessive inflammation, diabetic neuropathy, and peripheral vascular disease. Hence, the development of effective diabetic wound therapy remains a critical clinical challenge. Hydrogen sulfide (H2S) regulates inflammation, oxidative stress, and angiogenesis, suggesting a potential role in promoting diabetic wound healing. Herein, we propose a first example of fabricating an antibiotic-free antibacterial protein hydrogel with self-generation of H2S gas (H2S-Hydrogel) for diabetic wound healing by simply mixing bovine serum albumin­gold nanoclusters (BSA-AuNCs) with Bis[tetrakis(hydroxymethyl)phosphonium] sulfate (THPS) at room temperature within a few minutes. In this process, the amino group in BAS and the aldehyde group in THPS are crossed together by Mannich reaction. At the same time, tris(hydroxymethyl) phosphorus (trivalent phosphorus) from THPS hydrolysis could reduce disulfide bonds in BSA to sulfhydryl groups, and then the sulfhydryl group generates H2S gas under the catalysis of BSA-AuNCs. THPS in H2S-Hydrogel can destroy bacterial biofilms, while H2S can inhibit oxidative stress, promote proliferation and migration of epidermal/endothelial cells, increase angiogenesis, and thus significantly increase wound closure. It would open a new perspective on the development of effective diabetic wound dressing.

Association between oxidative balance score in adults with and without chronic kidney disease: 2011-2028 NHANES.

Introduction: Oxidative stress status is associated with CKD; however, few studies have investigated this association. The oxidative balance score (OBS) reflects systemic stress status and consists of 16 anti-and pro-oxidant dietary factors and four anti-and pro-oxidant lifestyle factors. Higher OBS implies exposure to more antioxidants. The purpose of this study was to explore the association between OBS and CKD. Methods: We enrolled 8,134 study participants from the 2011-2018 National Health and Nutrition Examination Survey and obtained OBS by adding the 20 dietary and lifestyle factors. Based on OBS, the participants were divided into three groups. We performed logistic regression, subgroup analyzes, and restricted cubic spline regression to explore the association between OBS and CKD. In addition, we tested the adjusted model. Results: OBS was negatively associated with CKD (OR: 0.54; 0.66, 0.82). After adjusting for all confounders, when dietary OBS was >20, the prevalence of CKD was reduced by 42% for each unit increase in OBS (p < 0.05). The negative associations of total OBS, dietary OBS, and lifestyle OBS with CKD were more significant in the female group. When the total OBS was ~20, the trend of decreasing prevalence in the female group was more significant. Conclusion: OBS is negatively associated with chronic kidney disease.

Preparation of thiol-free amino acid plasma matrix for quantification of thiol amino acids and their oxidized forms in human plasma by UPLC-MS/MS.

Thiol amino acids, with great physiological significance, are unstable, and have small molecular weights, as well as very low endogenous concentrations. Therefore, to quantitatively and directly analyze them using liquid chromatography-tandem mass spectrometry is difficult. To overcome these problems, we aimed to prepare a thiol-free amino acid plasm matrix as blank sample to reduce the background for the first time. Using compounds with maleimide group that react with classical thiols to generate water-insoluble products. Reducing agents Tris(2-carboxyethyl)phosphine (TCEP) was applied to cooperate with bismaleimide (DM) for elimination of thiol amino acids from plasma 10 min at room temperature and pH 7. Further, the residual TCEP from plasma were removed using an anion exchange resin within 10 min. Methodological validation analysis revealed good performance in linearity, precision, extraction recovery (≥ 82 %), and stability (except oxidized glutathione). This quantitative analysis was successfully applied to blood samples of 9 people in good health. This study provides a foundation for the development of accurate and rigorous quantitative analysis methods targeting thiol amino acids in different body fluids or tissues. Moreover, it paves the way toward realizing several clinical applications.

Au nanozyme-based multifunctional hydrogel for inflammation visible monitoring and treatment.

Chronic inflammation can delay wound healing, eventually leading to tissue necrosis and even cancer. Developing real-time intelligent inflammation monitoring and treatment to achieve effective wound management is important to promote wound healing. In this study, a smart multifunctional hydrogel (Hydrogel@Au NCs&DG) was proposed to monitor and treat the wound inflammation. It was prepared by mixing 3-carboxy-phenylboronic acid modified chitosan (CS-cPBA), ß-glycerophosphate (ß-GP), albumin-protected gold nanoclusters (BSA-Au NCs), and dipotassium glycyrrhizinate (DG) about 10 s. In this hydrogel, CS-cPBA and ß-GP are crosslinked together by boric acid ester bond and hydrogen bond to form the main hydrogel network, endowing the hydrogel with self-healing and injectable properties to adapt irregular wounds. Importantly, the as-prepared hydrogel with good biocompatibility and excellent adhesion property could directly determine the H2O2 to monitor the wound microenvironment by visible fluorescence change of BSA-Au NCs and then guide the frequency of dressing change to eliminate inflammation. The results demonstrated that the as-prepared smart hydrogel could be expected to serve as an intelligent wound dressing to promote inflammation-infected wound healing.

AS1411-conjugated doxorubicin-loaded silver nanotriangles for targeted chemo-photothermal therapy of breast cancer.

Background: Combination therapy has attracted tremendous interest for its great potential in treating cancers. Materials & methods: Based on chitosan-coated silver nanotriangles, polyethylene glycol, AS1411 aptamer and doxorubicin, a multifunctional nanocomposite (AS1411-DOX-AgNTs) was constructed and characterized. Then the photothermal properties, ability to target breast cancer cells and anti-breast cancer effect of AS1411-DOX-AgNTs were evaluated. Results: AS1411-DOX-AgNTs were successfully fabricated and showed excellent photothermal conversion efficiency, breast cancer cell and tumor targeting ability. Compared with single treatments, the combination of AS1411-DOX-AgNTs with near-infrared irradiation possessed the strongest anti-breast cancer effect in vitro and in vivo. Conclusion: AS1411-DOX-AgNTs hold great potential in targeted DOX delivery and combined chemo-photothermal therapy for breast cancer.

This article focuses on nanomaterials, nanomedicine and photothermal therapy (PTT) to treat breast cancer. Nanomaterials refer to materials with at least one dimension in nanometer size (1­100 nm) or materials composed as basic units in a 3D space. Nanomedicine is the application of nanomaterials in medicine. Nanoparticles can deliver drugs to areas that are difficult for the drugs themselves to reach. PTT is a noninvasive tumor therapy that uses photothermal conversion agents to convert light energy into heat energy to kill tumor cells under the irradiation of external near-infrared (NIR) light. In recent years, combination therapy for cancers has drawn more and more attention. In the current study, we investigated the in vitro and in vivo anticancer effects of silver nanocomposites combined with chemotherapy and PTT. The prepared silver nanocomposites showed excellent physicochemical properties and possessed good anti-breast cancer efficacy combined with PTT and chemotherapy drug in vitro and in vivo. The results of this study demonstrated that these prepared silver nanocomposites had exceptional anti-breast cancer effects in combination with PTT and could be promising drug-loaded photothermal conversion agents.

GMT8 aptamer conjugated PEGylated Ag@Au core-shell nanoparticles as a novel radiosensitizer for targeted radiotherapy of glioma.

Radiotherapy is one of the main treatment modalities for glioma, but the therapeutic efficacy is often limited by the radioresistance of tumor cells. The radiosensitization effects of silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) on tumors have been confirmed by previous studies. To enhance the specific killing effect of irradiation on tumor cells, targeted modification of radiosensitizers is urgently needed. Herein, we developed polyethylene glycol (PEG)-coated Ag@Au core-shell nanoparticles (PSGNPs) modified with GMT8 aptamer (GSGNPs) and evaluated their radiosensitization effects on glioma cells through in vivo and in vitro experiments. Transmission electron microscope image showed that the prepared PSGNPs had a spherical core-shell structure with an average size of 11 nm. The ultraviolet-visible absorption spectra and Fourier transform infrared spectra displayed that GMT8 was successfully conjugated to PSGNPs. The results of dark-field imaging revealed that the targeting ability of GSGNPs to U87 glioma cells was much better than that to normal human microvascular endothelial cells. Additionally, it was also found that the endocytic pathways of GSGNPs mainly involved clathrin-mediated endocytosis and macropinocytosis. The sensitization enhancement ratio of GSGNPs was calculated to be 1.62, which was higher than that of PSGNPs. In vivo imaging results showed that GSGNPs exhibited good tumor targeting and retention capabilities, and the fluorescence intensity ratio of Cy5-GSGNPs to Cy5-PSGNPs reached a peak at 4 h after injection. More importantly, the median survival time of mice bearing U87 glioma was significantly prolonged after intravenous administration of GSGNPs combined with radiotherapy. This work demonstrated that GSGNPs could be used as an effective nano-radiosensitizer for targeted radiotherapy of glioma.

The effect of AS1411 surface density on the tumor targeting properties of PEGylated silver nanotriangles.

Aim: To determine the optimal AS1411 density on polyethylene glycol (PEG)ylated silver nanotriangles (PNTs) for targeting breast cancer cells. Methods: PNTs modified with different AS1411 densities (ANTs) were constructed, characterized and evaluated for their targeting properties in breast cancer cells and a mouse model of breast cancer. Results: AS1411 was successfully conjugated to PNTs. The accumulation and cellular uptake of 10-ANTs were the highest. 10-ANTs plus near-IR laser irradiation displayed the greatest inhibitory effect on cell viability. However, 5-ANTs had the highest accumulation in tumor tissues. When combined with NIR laser, 5-ANTs exhibited the best in vivo photothermal therapy effect. Conclusion: The optimal AS1411 densities at the cellular and animal levels were 10% and 5%, respectively.

DOX-loaded silver nanotriangles and photothermal therapy exert a synergistic antibreast cancer effect via ROS/ERK1/2 signaling pathway.

The combination of multiple therapies has been proved to be more effective than a single therapy for many cancers. This study aimed to investigate the synergistic antibreast cancer effect of doxorubicin-loaded silver nanotriangles (DOX-AgNTs) combined with near-infrared (NIR) irradiation and explore the underlying mechanism. AgNTs were prepared by a chemical method and DOX was loaded via electrostatic adsorption. Characterization was performed by transmission electron microscopy, ultraviolet-visible spectroscopy and dynamic light scattering. The viability of MDA-MB-231 cells was detected by using MTT assay to evaluate the synergistic anticancer effect of DOX-AgNTs combined with NIR irradiation. The intracellular reactive oxygen species (ROS) level and cell apoptosis were analyzed by flow cytometry. Mitochondrial membrane potential (MMP) was measured with fluorescence microscopy. The mechanism was further investigated with ROS scavenger N-acetylcysteine and specific inhibitors of extracellular signal-regulated kinase 1/2 (ERK1/2), C-jun N-terminal kinase and p38 pathways. Characterization results revealed that the prepared AgNTs were mostly triangular and the mean edge length was about 126 nm. The combination of DOX-AgNTs and NIR exhibited a superior synergistic anticancer effect over single DOX-AgNTs or photothermal therapy (PTT). N-acetylcysteine and ERK1/2 inhibitor U0126 were found to significantly rescue the decreased cell viability, declined MMP and increased apoptosis induced by the combined treatment. Our results suggested that DOX-AgNTs combined with PTT performed a synergistic antibreast cancer effect. The synergy might be closely associated with the excessive production of ROS, changed MMP and the activation of ERK1/2 signaling pathway. These findings might provide a new perspective for the development of breast cancer treatments with excellent efficacy.

AS1411 and EpDT3-conjugated silver nanotriangle-mediated photothermal therapy for breast cancer and cancer stem cells.

Aim: This study aimed to construct AS1411 and EpDT3-conjugated PEGylated silver nanotriangles (AENTs) and assess their ability to target breast cancer and cancer stem cells, as well as the antitumor and antimetastatic effects of AENT-mediated photothermal therapy. Materials & methods: AENTs were constructed and characterized. The targeting properties, as well as antitumor and antimetastatic activities, were evaluated in MDA-MB-231 breast cancer cells, cancer stem cells and breast cancer-bearing mice. Results: AENTs displayed excellent targeting property to breast cancer cells and cancer stem cells. AENT-mediated photothermal therapy greatly inhibited (>45%) the migration and invasion of breast cancer cells, as well as tumor growth and lung metastasis in the mice. Conclusion: AENT-mediated photothermal therapy might be an effective strategy for the treatment of breast cancer.

Application of silver nanotriangles as a novel contrast agent in tumor computed tomography imaging.

This study aimed to prepare chitosan-coated silver nanotriangles (AgNTs) and assess their computed tomography (CT) contrast property byin vitroandin vivoexperiments. AgNTs with a range of sizes were synthesized by a seed-based growth method, and subsequently characterized by transmission electron microscopy (TEM), ultraviolet-visible absorption spectroscopy and dynamic light scattering. The x-ray attenuation capability of all prepared AgNTs was evaluated using micro CT. The CT contrast effect of AgNTs with the highest x-ray attenuation coefficient was investigated in MDA-MB-231 breast cancer cells and a mouse model of breast cancer. The TEM results displayed that all synthesized AgNTs were triangular in shape and their mean edge lengths ranged from 60 to 149 nm. All AgNTs tested exhibited stronger x-ray attenuation capability than iohexol at the same mass concentration of the active elements, and the larger the AgNTs size, the higher the x-ray attenuation coefficient. AgNTs with the largest size were selected for further research, due to their strongest x-ray attenuation capability and best biocompatibility. The attenuation coefficient of breast cancer cells treated with AgNTs increased in a particle concentration-dependent manner.In vivoCT imaging showed that the contrast of the tumor injected with AgNTs was significantly enhanced. These findings indicated that AgNTs could be a promising candidate for highly efficient tumor CT contrast agents.

Increasing the accumulation of aptamer AS1411 and verapamil conjugated silver nanoparticles in tumor cells to enhance the radiosensitivity of glioma.

Radioresistance significantly decreases the efficacy of radiotherapy, which can ultimately lead to tumor recurrence and metastasis. As a novel type of nano-radiosensitizer, silver nanoparticles (AgNPs) have shown promising radiosensitizing properties in the radiotherapy of glioma, but their ability to efficiently enter and accumulate in tumor cells needs to be improved. In the current study, AS1411 and verapamil (VRP) conjugated bovine serum albumin (BSA) coated AgNPs (AgNPs@BSA-AS-VRP) were synthesized and characterized. Dark-field imaging and inductively coupled plasma mass spectrometry were applied to investigate the accumulation of AgNPs@BSA-AS and AgNPs@BSA-AS-VRP mixed in different ratios in U251 glioma cells. To assess the influences of 19:1 mixed AgNPs@BSA-AS and AgNPs@BSA-AS-VRP on the P-glycoprotein (P-gp) efflux activity, rhodamine 123 accumulation assay was carried out. Colony formation assay and tumor-bearing nude mice model were employed to examine the radiosensitizing potential of 19:1 mixed AgNPs@BSA-AS and AgNPs@BSA-AS-VRP. Thioredoxin Reductase (TrxR) Assay Kit was used to detect the TrxR activity in cells treated with different functionally modified AgNPs. Characterization results revealed that AgNPs@BSA-AS-VRP were successfully constructed. When AgNPs@BSA-AS and AgNPs@BSA-AS-VRP were mixed in a ratio of 19:1, the amount of intracellular nanoparticles increased greatly through AS1411-mediated active targeting and inhibition of P-gp activity. In vitro and in vivo experiments clearly showed that the radiosensitization efficacy of 19:1 mixed AgNPs@BSA-AS and AgNPs@BSA-AS-VRP was much stronger than that of AgNPs@BSA and AgNPs@BSA-AS. It was also found that 19:1 mixed AgNPs@BSA-AS and AgNPs@BSA-AS-VRP significantly inhibited intracellular TrxR activity. These results indicate that 19:1 mixed AgNPs@BSA-AS and AgNPs@BSA-AS-VRP can effectively accumulate in tumor cells and have great potential as high-efficiency nano-radiosensitizers in the radiotherapy of glioma.

Silver Nanotriangles and Chemotherapeutics Synergistically Induce Apoptosis in Glioma Cells via a ROS-Dependent Mitochondrial Pathway.

BACKGROUND: The synergistic effect of nanomaterials and chemotherapeutics provides a novel strategy for the treatment of tumors. Silver nanotriangles (AgNTs) exhibited some unique properties in nanomedicine. Studies on the synergy of silver-based nanomaterials and anti-tumor drugs against gliomas are rare. MATERIALS AND METHODS: Chitosan-coated AgNTs were prepared, followed by characterization using transmission electron microscopy, ultraviolet-visible spectroscopy and X-ray diffraction. The anti-glioma effect of cyclophosphamide (CTX), 5-fluorouracil (5-FU), oxaliplatin (OXA), doxorubicin (DOX) or gemcitabine (GEM) combined with AgNTs in different glioma cell lines (U87, U251 and C6) was assessed by the MTT assay to screen out a drug with the most broad-spectrum and strongest synergistic anti-glioma activity. The intracellular reactive oxygen species (ROS) level, mitochondrial membrane potential (MMP) and cell apoptosis were detected by flow cytometry. The possible underlying mechanisms of the synergy were further investigated with ROS scavenger and specific inhibitors of C-jun N-terminal kinase (JNK), p38 and extracellular signal-regulated kinase 1/2 pathways. RESULTS: The synthesized AgNTs were mainly triangular and truncated triangular with an average edge length of 125 nm. A synergistic anti-glioma effect of AgNTs combined with CTX was not observed, and the synergism between AgNTs and 5-FU was cell type-specific. AgNTs combined with OXA, DOX or GEM displayed synergistic effects in various glioma cell lines, and the combination of AgNTs and GEM showed the strongest synergistic activity. A decrease in cell viability, loss of the MMP and an increase in apoptosis rate induced by this synergy could be significantly attenuated by the ROS scavenger N-acetylcysteine and JNK inhibitor SP600125. CONCLUSION: Our results suggested that the combination of AgNTs and GEM possessed broad-spectrum and potent synergistic anti-glioma activity, resulting from cell apoptosis mediated by a ROS-dependent mitochondrial pathway in which JNK might be involved.