Green and highly effective extraction of bioactive flavonoids from Fructus aurantii employing deep eutectic solvents-based ultrasonic-assisted extraction protocol.

In China, Jiang Fructus aurantii (JFA) has attracted increasing interest as a famous traditional herbal medicine and valuable economic food for its valuable medicinal and industrial properties. In the current work, contrasted with conventional extraction techniques, natural flavonoids from JFA (naringin and neohesperidin) were extracted with remarkable effectiveness utilizing a sustainable deep eutectic solvents combined ultrasonic-assisted extraction (DESs-UAE) protocol. The optimal extraction capacity can be achieved by mixing 30 % water with a molar ratio of 1:3 for choline chloride and ethylene glycol, as opposed to the classical extraction solvents of 95 % ethanol, methanol, and water. Moreover, the DESs-UAE extraction programs were also systematically optimized employing Box-Behnken design (BBD) trials, and the eventual findings suggested that the best parameters were a 27 % water content in DES, a 16 mL/g liquid-solid ratio, a 72 min extraction time, and a 62 °C extraction temperature, along with the corresponding greatest contents of NAR (48.18 mg/g) and NEO (34.50 mg/g), respectively. Notably, by comparison with the pre-optimization data, the optimized DES extraction efficiency of flavonoids is markedly higher. Thereafter, the characterization of the solvents before and after extraction, as well as the differences between the four extraction solvent extracts, were compared using the FT-IR analyses. Furthermore, SEM results suggested that the penetration and erosion abilities of the plant cell wall of DES-1 were stronger than those of the other three traditional solvents, thus allowing more release of flavonoid compounds. In conclusion, the present research develops a straightforward, sustainable, and exceedingly efficient approach for the extraction of bioactive flavonoids from JFA, which has the potential to facilitate the efficient acquisition of active ingredients from TCM.

Mn3O4 nanoparticles decorated porous reduced graphene oxide with excellent oxidase-like activity for fast colorimetric detection of ascorbic acid.

Mn3O4 nanoparticles composed of porous reduced graphene oxide nanosheets (Mn3O4@p-rGO) with enhanced oxidase-like activity were successfully fabricated through an in-situ approach for fast colorimetric detection of ascorbic acid (AA). The residual Mn2+ in the GO suspension of Hummers method was directly reused as the manganese source, improving the atom utilization efficiency. Benefiting from the uniform distribution of Mn3O4 nanoparticles on the surface of p-rGO nanosheets, the nanocomposite exhibited larger surface area, more active sites, and accelerated electron transfer efficiency, which enhanced the oxidase-like activity. Mn3O4@p-rGO nanocomposite efficiently activate dissolved O2 to generate singlet oxygen (1O2), leading to high oxidation capacity toward the substrate 3,3',5,5'-tetramethylbenzidine (TMB) without the extra addition of H2O2. Furthermore, the prominent absorption peak of the blue ox-TMB at 652 nm gradually decreased in the presence of AA, and a facile and fast colorimetric sensor was constructed with a good linear relationship (0.5-80 µM) and low LOD (0.278 µM) toward AA. Owing to the simplicity and excellent stability of the sensing platform, its practical application for AA detection in juices has shown good feasibility and reliability compared with HPLC and the 2, 4-dinitrophenylhydrazine colorimetric method. The oxidase-like Mn3O4@p-rGO provides a versatile platform for applications in food testing and disease diagnosis.

Partially oxidized MoS2 nanosheets with high water-solubility to enhance the peroxidase-mimic activity for sensitive detection of glutathione.

Surface oxidation engineering is an effective strategy to construct nanomaterials with enhanced biocatalytic activity. In this study, a facile one-pot oxidation strategy was proposed to synthesize partially oxidized molybdenum disulfide nanosheets (ox-MoS2 NSs), which exhibit good water solubility and can be used as an excellent peroxidase substitute. Under the oxidation process, Mo-S bonds are partially broke and S atoms are replaced by excess oxygen atoms, and the released abundant heat and gases efficiently expended the interlayer distance and weaken the van der Waals forces between adjacent layers. Porous ox-MoS2 NSs can be easily exfoliated by further sonication, and the nanosheets exhibits excellent water dispersibility and no obvious sediment appear even after store for months. Benefiting from the desirable affinity property with enzyme substrates, optimized electronic structure and prominent electron transfer efficiency, the ox-MoS2 NSs exhibit enhanced peroxidase-mimic activity. Furthermore, the ox-MoS2 NSs catalyzed 3,3',5,5'-tetramethylbenzidine (TMB) oxidation reaction could be inhibited by the redox reaction that take place between glutathione (GSH) as well as the direct interaction between GSH and ox-MoS2 NSs. Thus, a colorimetric sensing platform was constructed for GSH detection with good sensitivity and stability. This work provides a facile strategy for engineering structure of nanomaterials and improving enzyme-mimic performance.

Effective extraction of bioactive alkaloids from the roots of Stephania tetrandra by deep eutectic solvents-based ultrasound-assisted extraction.

The efficient and green extraction of bioactive ingredients from natural plants play a vital role in their corresponding drug effects and subsequent studies. Recently, deep eutectic solvents (DESs) have been considered promising new green solvents for efficiently and selectively extracting substances from varied plants. In this work, an environment-friendly DESs-based ultrasonic-assisted extraction (DESs-UAE) procedure was developed for highly efficient and non-polluting extraction of alkaloids from the roots of Stephania tetrandra (ST). A total of fifteen different combinations of DESs, compared with traditional organic solvents (methanol and 95% ethanol) and water, were evaluated for extraction of bioactive alkaloids (FAN and TET) from ST, and the results revealed that DESs system made up of choline chloride and ethylene glycol with mole ratio of 1:2 exhibited the optimal extraction efficiency for alkaloids. Additionally, a four-factor and three-level Box-Behnken design (BBD), a particular pattern of response surface methodology (RSM), was used to optimize extraction conditions. RSM results indicated that the maximum extraction yields of FAN, TET, and TA were attained 7.23, 13.36, 20.59 mg/g, respectively, within extraction temperature of 52 °C, extraction time of 82 min, DES water content of 23% (v/v), and liquid-solid ratio of 23 mL/g. The measured results were consistent with the predicted values. Notably, the optimized DES extraction efficiency of TA, according to the experimental data analysis, is 2.2, 3.3 and 4.1 times higher than methanol, 95% ethanol and water, respectively. Meanwhile, based on 3D response surface plots, interactive effects plots and contour maps, the effects of the aforementioned four essential factors on the extraction yield and their interactions on the response were visualized. The results revealed that the mutual interactions between extraction temperature and liquid-solid ratio exhibited positive effects on all responses, while extraction time and water content in DES posed a negative effect. Therefore, these results suggest that DESs, as a class of novel green solvents, with the potential to substitute organic solvent and water, can be widely and effectively applied to extract bioactive compounds from natural plants.

A novel metal-organic framework of Ba-hemin with enhanced cascade activity for sensitive glucose detection.

Early glucose detection is important in both healthy people and diabetic patients. Glucose biosensing based on glucose oxidase (GOX) is a common method. However, native proteins are mostly membrane impermeable and are prone to degradation in complex sample environments. Herein, we report a facile one-step biomineralization method by simply mixing aqueous solutions of hemin and barium nitrate with glucose oxidase (GOX) to form Ba-hemin@GOX composites. Glucose (Glu) is introduced through self-driven sampling to trigger the GOX-catalysed production of hydrogen peroxide, which could help the subsequent 3,3',5,5'-tetramethylbenzidine (TMB) oxidation reaction catalysed by Ba-hemin to yield the blue-coloured product. The sensor exhibited a detection limit as low as 3.08 µM. The operability and accuracy of the Ba-hemin@GOX biosensor were confirmed by the quantitative determination of glucose in real samples, such as tap water, serum and drinks. Moreover, the Ba-hemin@GOX-based colorimetric biosensor showed good selectivity, storage stability and recoverability. The experimental results reveal that a GOX activity of more than 90% was still maintained even after being incubated at 60 °C for 30 minutes, and Ba-hemin@GOX could be reused for glucose detection at least six times. Even after 30 days of storage, the relative activity was still more than 90%. Overall, the developed Ba-hemin@GOX biosensor provides a valuable and general platform for applications in colorimetric biosensing and medical diagnostics.

Kidney-targeted triptolide-encapsulated mesoscale nanoparticles for high-efficiency treatment of kidney injury.

Bad water solubility and undesired toxicity of triptolide (TP) still restrict its clinical applications in renal diseases. In this work, well-defined, monodispersed, and uniform-sized TP-encapsulated mesoscale nanoparticles (TP-MNPs) were fabricated through a nanoprecipitation method, which possesses special kidney-targeting capacity, slow-release property, and high-efficiency treatment for renal ischaemia-reperfusion injury (IRI). The TP-MNPs had good cytocompatibility in wide TP concentration (0-500 ng ml-1) and time ranges (6-24 h). Ex vivo organ fluorescence imaging and pharmacokinetic analysis suggested that TP-MNPs possessed excellent kidney-targeting capability with long retention time (7 days). The TP-MNPs with a very low dose of TP (0.01 mg kg-1) could effectively protect the kidney against IRI, while 0.01 mg kg-1 TP was completely ineffective. After treatment with TP-MNPs, the serum creatine, blood urea nitrogen, expression of C3 complement, and phospho-extracellular signal-regulated kinase of renal IRI mice were estimated to be 5.9-, 2.0-, 5.4-, and 2.8-fold lower than those of the mice treated with TP, respectively. Compared with TP, the TP-MNPs exhibited ignorable hepatotoxicity, reproductive toxicity, and immunotoxicity, such as lower alanine aminotransferase (0.5 fold) and aspartate aminotransferase (0.2 fold), and a higher ratio of CD4+/CD8+ (2.2 fold). Thus, the monodispersed and uniform-sized TP-MNPs with special kidney-targeting and slow-release property may pave an avenue for designing a new therapeutic strategy for renal diseases.

A stimuli-responsive drug release nanoplatform for kidney-specific anti-fibrosis treatment.

The renoprotective effects of hypoxia inducible-factor (HIF) activators have been demonstrated by improving renal hypoxia in chronic kidney disease. Cobalt chloride is one of the most widely used HIF activators in biomedicine; however, poor kidney targeting and undesirable side effects greatly limit its clinical applications. Here, we report a novel stimuli-responsive drug release nanoplatform in which glutathione (GSH)-modified Au nanoparticles (GLAuNPs) and Co2+ self-assemble into nanoassemblies (GLAuNPs-Co) through coordination interactions between empty orbitals of Co2+ and lone pairs of GSH. The GLAuNPs, when used as a drug carrier, demonstrated high drug loading capacity and pH-triggered drug release after assembling with Co2+. The acidic environment of lysosomes in renal fibrosis tissues could disassemble GLAuNPs-Co and release Co2+. Moreover, encapsulation of the Co2+ ions in the GLAuNPs greatly lowered the cytotoxicity of Co2+ in kidney tubule cells. Tissue fluorescence imaging showed that GLAuNPs-Co specifically accumulated in the kidneys, especially in the renal proximal tubules. After GLAuNPs-Co was intraperitoneally injected into ureter-obstructed mice, significant attenuation of interstitial fibrosis was exhibited. The beneficial effects can be mainly ascribed to miR-29c expression restored by HIF-α activation. These findings revealed that GLAuNPs-Co have pH-responsive drug release and renal targeting capabilities; thus, they are a promising drug delivery platform for treating kidney disease.

Hydrophilic Porous Poly(l-Lactic Acid) Nanomembranes: Self-Assembly, and Anti-Biofouling and Anti-Thrombosis Effects.

Hydrophilic PEGylated porous self-assembled nanomembranes (PSANMs) with average thickness and pore diameter of ca. 10 and 20-24 nm were successfully prepared by an emulsification-induced programmable self-assembly strategy. The hydrophilicity, anti-biofouling, and anti-thrombosis properties of PEGylated PSANMs were largely improved in comparison with the nonfunctionalized PSANMs, which could transform into hydrophilic (PEGylated PSANMs, minimum water contact angle: 38.8°) from hydrophobic units (PSANMs, maximum water contact angle: 137.5°) with increasing PEG density and length. The total protein adsorption of PEGylated PSANMs was about six times lower than that of the PSANMs, while the thrombosis of the PEGylated PSANMs (maximum R-time: 5.37 min) was also greatly relieved in comparison with the PSANMs (minimum R-time: 2.93 min). Such PEG-modified PSANMs may have applications in drug delivery and tissue and organ repair in vivo.

Folate Receptor-Mediated Renal-Targeting Nanoplatform for the Specific Delivery of Triptolide to Treat Renal Ischemia/Reperfusion Injury.

Triptolide (TP) has been widely used in clinical medicine; however, it has created a dilemma due to its toxicity and nonspecificity. Here, we reported a biocompatible and high-efficiency renal-targeting nanoplatform for renal ischemia/reperfusion injury (IRI) therapy, in which the toxic drug of TP was encapsulated into folate (FA)-modified Pluronic F127/P123 nanoparticles (FPNPs). The TP-loaded FPNPs (TP-FPNPs) had good stability and could effectively reduce the cytotoxicity of TP. Compared with the Pluronic nanoparticles (PNPs) group, cellular uptake ability of FPNPs significantly improved because of folate receptor-mediated endocytosis effect. Ex vivo organ imaging and pharmacokinetic results indicated that FPNPs possessed high kidney selectivity and long retention time. The therapeutic effect of TP-FPNPs on renal IRI was more superior to that of free TP, such as lower acute tubular injury index (2.9-fold), renal function indexes of serum creatinine (4.3-fold), urea nitrogen (2.0-fold), and Western blotting (2.4-fold). Systemic toxicity assay suggested that TP-FPNPs had much lower nephrotoxicity, hepatotoxicity, and genital system toxicity than free TP. Thus, renal-targeting FPNPs will be a potential delivery platform of hydrophobic drugs for treatment of renal diseases.

Small-Sized Cationic miRi-PCNPs Selectively Target the Kidneys for High-Efficiency Antifibrosis Treatment.

Small-sized cationic miRi (microRNA-21 inhibitor)-PCNPs (low molecular weight chitosan (LMWC)-modified polylactide-co-glycoside (PLGA) nanoparticles (PLNPs)) with special kidney-targeting and high-efficiency antifibrosis treatment are fabricated through coupling miRi, PLGA, and LMWC. In the miRi-PCNPs, easily degraded miRi is encapsulated in PCNPs and thus prevented from degradation by nuclease. Cytotoxicity, immunotoxicity, and systemic toxicity assays and in vitro and ex vivo fluorescence imaging suggest that PCNPs possess excellent biocompatibility, higher cellular uptake efficiency, and selective kidney-targeting capacity. Western blotting, pathological staining, and real-time polymerase chain reaction analyses show that the therapeutic effect of miRi-PCNPs on kidney fibrosis is much higher than that of miRi, which is mainly through suppressing transforming growth factor beta-1/drosophila mothers against decapentaplegic protein 3 (TGF-ß1/Smad3) and extracellular signal-regulated kinases/mitogen-activated protein kinase signaling pathway by inhibiting the expression of microRNA-21. For example, the tubule damage index and tubulointerstitial fibrosis area in the miRi-PCNPs group are ≈2.5-fold lower than those in the saline and bare miRi groups. The miRi-PCNPs with special kidney-targeting and high-efficiency antifibrosis treatment may represent a promising strategy for designing and developing a therapeutic treatment for kidney fibrosis.

Highly Stabilized Core-Satellite Gold Nanoassemblies in Vivo: DNA-Directed Self-Assembly, PEG Modification and Cell Imaging.

Au nanoparticles (NPs) have important applications in bioimaging, clinical diagnosis and even therapy due to its water-solubility, easy modification and drug-loaded capability, however, easy aggregation of Au NPs in normal saline and serum greatly limits its applications. In this work, highly stabilized core-satellite Au nanoassemblies (CSAuNAs) were constructed by a hierarchical DNA-directed self-assembly strategy, in which satellite Au NPs number could be effectively tuned through varying the ratios of core-AuNPs-ssDNA and satellite-AuNPs-ssDNAc. It was especially interesting that PEG-functionalized CSAuNAs (PEG-CSAuNAs) could not only bear saline solution but also resist the enzymatic degradation in fetal calf serum. Moreover, cell targeting and imaging indicated that the PEG-CSAuNAs had promising biotargeting and bioimaging capability. Finally, fluorescence imaging in vivo revealed that PEG-CSAuNAs modified with N-acetylation chitosan (CSNA) could be selectively accumulate in the kidneys with satisfactory renal retention capability. Therefore, the highly stabilized PEG-CSAuNAs open a new avenue for its applications in vivo.

Coordinatively Self-Assembled Luminescent Gold Nanoparticles: Fluorescence Turn-On System for High-Efficiency Passive Tumor Imaging.

A fluorescence turn-on system for highly efficient and prolonged tumor imaging has been established by a Co2+-induced coordination self-assembly strategy, in which luminescent glutathione (GSH)-modified gold nanoparticles (LGAuNPs) are assembled into LGAuNPs assemblies (LGAuNPs-Co) through a coordination bond between an unoccupied orbit of Co2+ and lone pair electrons of GSH on the surface of LGAuNPs. The LGAuNPs-Co is sensitive to microenvironment pH, and its quenched luminescence will be turned on in tumor tissues (acidic microenvironment), which behaves as a fluorescence turn-on system for passive tumor imaging. The fluorescence turn-on system combines advantages of the enhanced permeability and retention (EPR) effect of NPs and pH-induced fluorescence turn-on property at the tumor site, which results in a larger fluorescence intensity (FI) difference between normal and tumor tissues as compared with that of luminescent Au NPs (LAuNPs, only with the EPR effect) (∼12-fold). Such a large FI difference results in that LGAuNPs-Co has rapid (∼1.6 h), persistent (∼24 h p.i.), and highly efficient tumor targeting capability in comparison with LGAuNPs. Moreover, the LGAuNPs-Co also has much longer tumor retention, faster renal clearance, and lower reticuloendothelial system (RES) uptake than LGAuNPs. Therefore, the fluorescence turn-on system is very promising for cancer diagnosis and therapy.