A novel insight on the intensification mechanism of sludge dewaterability by ionic liquids.

The huge output of sewage sludge has caused a remarkable environmental burden. Sludge dewatering is considered as an important way to reduce the sludge volume. Five imidazole-based ionic liquids were used to improve the dewaterability of sewage sludge. 1-ethyl-3-methylimidazolium dihydrogen phosphate ([Emim][H2PO4]) was screened out as a potential conditioner of sludge due to its excellent dewatering performance and reusability. The solid content of sludge filter cake after treatment with [Emim][H2PO4] was about 10% higher than that of sludge treated by cationic polyacrylamides (CPAM). The intensification mechanism of ionic liquids to the improvement of sludge dewatering performance was studied. The presence of acidic ionic liquids [Emim][H2PO4] resulted the increase of zeta potential from -14.57 ± 0.81 mV to -5.60 ± 0.30 mV and led to the protonation of biopolymers. Acidic ionic liquids [Emim][H2PO4] inactivated the microorganism and led to a porous and unconsolidated structure of the solid sludge particles. All these effects were conducive to destroy the microstructure of sludge and release water. However, [Emim]Cl, [Bmim][OTf] and [Hmim][OTf] showed little effect on the protonation of ionizable functional groups at near-neutral environment. The dissolution of biopolymer decreased the zeta potential and strengthened the electrostatic repulsion. So, they showed weaker intensification effects than CPAM.

Drug release evaluation of Paclitaxel/Poly-L-Lactic acid nanoparticles based on a microfluidic chip.

Paclitaxel is a commonly used drug in the medical field because of its strong anticancer effect. However, it may produce relatively severe side effects (i.e., allergic reactions). A major characteristic of paclitaxel is low solubility in water. Special solvents are used for dissolving paclitaxel and preparing the paclitaxel drugs, while the solvents themselves will cause certain effects. Polyoxyethylene castor oil, for example, can cause severe allergic reactions in some people, and the clinical use is limited. In this study, we developed a new Paclitaxel/Poly-L-Lactic Acid (PLLA) nanoparticle drug, which is greatly soluble in water, and carried out in vitro drug sustained release research on it and the original paclitaxel drug. However, because the traditional polymer drug carrier usually uses dialysis bag and thermostatic oscillation system to measure the drug release degree in vitro, the results obtained are greatly different from the actual drug release results in human body. Therefore, this paper adopts the microfluidic chip we previously developed to mimic the human blood vessels microenvironment to study the sustained-release of Paclitaxel/PLLA nanoparticles to make the results closer to the release value in human body. The experimental results showed that compared with the original paclitaxel drug, Paclitaxel/PLLA nanoparticles have a long-sustained release time and a slow drug release, realizing the sustained low-dose release of paclitaxel, a cell cycle-specific anticancer drug, and provided certain reference significance and theoretical basis for the research and development of anticancer drugs.

Dual-Stimuli-Responsive Paclitaxel Delivery Nanosystems from Chemically Conjugate Self-Assemblies for Carcinoma Treatment.

Diselenide-bond-linked poly(N-isopropylacrylamide)-paclitaxel chemical conjugates are synthesized as a drug release carrier. The conjugates can self-assemble into "core-shell" nanoscaled micelles in aqueous solution and show thermal and redox dual-responsiveness. The conjugates can afford a high encapsulation efficiency of up to 72.3%, and deliver hydrophobic anticancer drug paclitaxel in a temperature and oxidization or reduction stress-mode. The in vitro 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and in vivo anticancer assays are performed to assess the cytotoxicity and anticancer activity of the conjugates, suggesting that the developed conjugates can be used to treat carcinoma as a novel and highly efficient drug delivery system.

Engineering Halomonas species TD01 for enhanced polyhydroxyalkanoates synthesis via CRISPRi.

BACKGROUND: Clustered regularly interspaced short palindromic repeats interference (CRISPRi) has provided an efficient approach for targeted gene inhibition. A non-model microorganism Halomonas species TD01 has been developed as a promising industrial producer of polyhydroxyalkanoates (PHA), a family of biodegradable polyesters accumulated by bacteria as a carbon and energy reserve compound. A controllable gene repression system, such as CRISPRi, is needed for Halomonas sp. TD01 to regulate its gene expression levels. RESULTS: For the first time CRISPRi was successfully used in Halomonas sp. TD01 to repress expression of ftsZ gene encoding bacterial fission ring formation protein, leading to an elongated cell morphology with typical filamentous shape similar to phenomenon observed with Escherichia coli. CRISPRi was employed to regulate expressions of prpC gene encoding 2-methylcitrate synthase for regulating 3-hydroxyvalerate monomer ratio in PHBV copolymers of 3-hydroxybutyrate (HB) and 3-hydroxyvalerate (HV). Percentages of HV in PHBV copolymers were controllable ranging from less than 1 to 13%. Furthermore, repressions on gltA gene encoding citrate synthase channeled more acetyl-CoA from the tricarboxylic acid (TCA) cycle to poly(3-hydroxybutyrate) (PHB) synthesis. The PHB accumulation by Halomonas sp. TD01 with its gltA gene repressed in various intensities via CRISPRi was increased by approximately 8% compared with the wild type control containing the CRISPRi vector without target. CONCLUSIONS: It has now been confirmed that the CRISPRi system can be applied to Halomonas sp. TD01, a promising industrial strain for production of various PHA and chemicals under open and continuous fermentation process conditions. In details, the CRISPRi system was successfully designed in this study to target genes of ftsZ, prpC and gltA, achieving longer cell sizes, channeling more substrates to PHBV and PHB synthesis, respectively. CRISPRi can be expected to use for more metabolic engineering applications in non-model organisms.

Soluplus/TPGS mixed micelles for dioscin delivery in cancer therapy.

BACKGROUND: Dioscin has shown cytotoxicity against cancer cells, but its poor solubility and stability have limited its clinical application. In this study, we designed mixed micelles composed of TPGS and Soluplus® copolymers entrapping the poorly soluble anticancer drug dioscin. METHOD: In order to improve the aqueous solubility and bioactivity of dioscin, TPGS/Soluplus® mixed micelles with an optimal ratio were prepared using a thin-film hydration method, and their physicochemical properties were characterized. Cellular cytotoxicity and uptake of the dioscin-loaded TPGS/Soluplus® mixed micelles were studied in MCF-7 breast cancer cells and A2780s ovarian cancer cells. The pharmacokinetics of free dioscin and dioscin-loaded TPGS/Soluplus® mixed micelles was studied in vivo in male Sprague-Dawley rats via a single intravenous injection in the tail vein. RESULTS: The average size of the optimized mixed micelle was 67.15 nm, with 92.59% drug encapsulation efficiency and 4.63% drug loading efficiency. The in vitro release profile showed that the mixed micelles presented sustained release behavior compared to the anhydrous ethanol solution of dioscin. In vitro cytotoxicity assays were conducted on human cancer cell lines including A2780s ovarian cancer cells and MCF-7 breast cancer cells. The mixed micelles exhibited better antitumor activity compared to free dioscin against all cell lines, which may benefit from the significant increase in the cellular uptake of dioscin from mixed micelles compared to free dioscin. The pharmacokinetic study showed that the mixed micelle formulation achieved a 1.3 times longer mean residual time (MRT) in circulation and a 2.16 times larger area under the plasma concentration-time curve (AUC) than the free dioscin solution. CONCLUSION: Our results suggest that the dioscin-loaded mixed micelles developed in this study might be a potential nano drug-delivery system for cancer chemotherapy.

A Promising Emodin-Loaded Poly (Lactic-Co-Glycolic Acid)-d-α-Tocopheryl Polyethylene Glycol 1000 Succinate Nanoparticles for Liver Cancer Therapy.

PURPOSE: Emodin (EMO) has multi-targets and multi-way antitumor effect, which was limited by the instability and poor solubility of EMO. The aim of this study was to formulate EMO-loaded poly (lactide-co-glycolide)-d-α-tocopheryl polyethylene glycol 1000 succinate (PLGA-TPGS) nanoparticles (EPTN) to increase the liver targeting of EMO for cancer therapy. METHODS: EMO/coumarin-6-loaded PLGA-TPGS nanoparticles (ECPTN) and EMO-loaded PLGA nanoparticles (EPN) were also prepared as comparison. The cellular uptake of ECPTN by HepG2 and HCa-F cells was investigated using Confocal laser scanning microscopy. The apoptosis of HepG2 cells handled with EPTN was assayed by flow cytometry. The liver targeting property of ECPTN in mice was evaluated using the drug concentration determined by RP-HPLC and the freezing slices were investigated via fluorescence inversion microscopy. The blood samples were obtained from vein intubation to illustrate the pharmacokinetics process of EPTN. The tumor-bearing mice model was established to elucidate the in vivo therapeutic effect of EPTN. RESULTS: The results demonstrated that ECPTN could be internalized by HepG2 and HCa-F cells respectively. The ratio of apoptosis cells was increased after dealing with EPTN. The detection indexes of drug concentration and fluorescence inversion microscopy images indicated ECPTN had an excellent effect on liver targeting property than EMO solutions (EMS). The pharmacokinetics process of EPTN showed obvious sustained-release effect than EMS. Compared with EPN, the in vivo antitumor activity of EPTN against tumor cells were better. CONCLUSIONS: In conclusion, EPTN could be used in the treatment of liver cancer acted as a kind of promising intravenous dosage forms.

Application of CRISPRi for prokaryotic metabolic engineering involving multiple genes, a case study: Controllable P(3HB-co-4HB) biosynthesis.

Clustered regularly interspaced short palindromic repeats interference (CRISPRi) is used to edit eukaryotic genomes. Here, we show that CRISPRi can also be used for fine-tuning prokaryotic gene expression while simultaneously regulating multiple essential gene expression with less labor and time consumption. As a case study, CRISPRi was used to control polyhydroxyalkanoate (PHA) biosynthesis pathway flux and to adjust PHA composition. A pathway was constructed in Escherichia coli for the production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] from glucose. The native gene sad encoding E. coli succinate semi-aldehyde dehydrogenase was expressed under the control of CRISPRi using five specially designed single guide RNAs (sgRNAs) for regulating carbon flux to 4-hydroxybutyrate (4HB) biosynthesis. The system allowed formation of P(3HB-co-4HB) consisting of 1-9mol% 4HB. Additionally, succinate, generated by succinyl-coA synthetase and succinate dehydrogenase (respectively encoded by genes sucC, sucD and sdhA, sdhB) was channeled preferentially to the 4HB precursor by using selected sgRNAs such as sucC2, sucD2, sdhB2 and sdhA1 via CRISPRi. The resulting 4HB content in P(3HB-co-4HB) was found to range from 1.4 to 18.4mol% depending on the expression levels of down-regulated genes. The results show that CRISPRi is a feasible method to simultaneously manipulate multiple genes in E. coli.

Microplastic release and sulfate reduction response in the early stage of a simulated landfill.

Landfills are essential facilities for treating and disposing municipal solid waste. They emit sulfur-containing odors and serve as an important sink for a new type of pollutant called microplastics (MPs). This study focused on the initial stage of anaerobic degradation to establish the relationship between the release of MPs and odor generation. Our findings show the rapid release of MPs into the leachate in the early stage of landfill and their predominant accumulation in the leachate sediment. The circulating leachate contained 1.45 times higher concentrations of MPs than the noncirculating leachate, with a peak concentration of 39 items·L-1. In addition, fragmentation of MPs occurred. The percentage of MPs with particle sizes of 2.5-5 mm decreased from 66.70 % to 22.32 %, while those measuring 0.1-0.5 mm increased by 33.12 %. A positive correlation was observed between MP release and sulfate reduction. Although leachate circulation increased the release of MPs, it also reduced the overall release time and total amount of MPs exported from the landfill. Compared with the initial landfill waste, the leachate operation mode, regardless of circulation, resulted in a 6.15-8.93-fold increase in MP release. These findings provide a valuable foundation for the simultaneous regulation of traditional pollutant odor and new pollutants (MPs) in landfills.

Effect of central screw taper angles on the loosening performance and fatigue characteristics of dental implants.

Biomechanical performance plays an important role in the long-term service of dental implants. Loosening and fatigue damage of the central screw are the most common problems. This research investigated the effect of the central screw taper angle on the loosening performance and fatigue characteristics of dental implants. Central screws with four taper angles, 30°, 60°, 90° and 180°, were processed and tested. The loosening performance of the screws under initial and postload conditions was compared. Then, the fatigue characteristics of dental implants was measured. Finally, the wear and fracture modes of the screws were observed. The damage locations were verified by finite element analysis (FEA). The results showed that the central screws with 30° taper had substantially better anti-loosening performance and less fretting wear. The central screws with 180° taper had a higher preload, resulting in a longer fatigue life. Furthermore, the fatigue fracture of the central screw occurred at the level of the first thread position, consistent with the FEA results. In the future clinical applications, central screws with a 30° taper angle may improve anti-loosening performance and prolong fatigue life by increasing the tightening torque.

Effect of Loading Angles and Implant Lengths on the Static and Fatigue Fractures of Dental Implants.

Mechanical properties play a key role in the failure of dental implants. Dental implants require fatigue life testing before clinical application, but this process takes a lot of time. This study investigated the effect of various loading angles and implant lengths on the static fracture and fatigue life of dental implants. Implants with lengths of 9 mm and 11 mm were prepared. Static fracture tests and dynamic fatigue life tests were performed under three loading angles (30°, 40°, and 50°), and the level arm and bending moment were measured. After that, the fracture morphology and fracture mode of the implant were observed. The results showed that 9 mm length implants have a higher static failure load and can withstand greater bending moments, while 11 mm length implants have a longer fatigue life. In addition, as the loading angle increases, the static strength and bending moment decrease linearly, and the fatigue life shows an exponential decrease at a rate of three times. Increasing the loading angle reduces the time of the implant fatigue test, which may be an effective method to improve the efficiency of the experiment.

Highly soluble PEGylated pyrene-gold nanoparticles dyads for sensitive turn-on fluorescent detection of biothiols.

Highly soluble fluorescent pyrene derivative with substantially improved fluorescence intensity in aqueous buffer was obtained via PEGylation strategy. The highly soluble PEGylated pyrene (PEO-Py) non-covalently adsorbed onto the surface of gold nanoparticles (Au NPs) to form dyads with quenched fluorescence due to highly efficient energy transfer between PEO-Py and Au NPs. The PEO-Py/Au NPs dyads were used for the sensitive turn-on fluorescent detection of biothiols. The fluorescence of PEO-Py was restored by the addition of cysteine (Cys), indicating that Cys can modulate the energy transfer between PEO-Py and Au NPs. This phenomenon then allowed for the sensitive detection of Cys with a limit of detection (LOD) of 11.4 nM. The linear range of determination of Cys was from 1.25 x 10(-8) to 2.25 x 10(-7) M. None of the other amino acids found in proteins showed obvious interference with the determination. It was important to note that the detection sensitivity of the PEO-Py/Au NPs system was more than 5-fold improved compared with the Py/Au NPs system. In addition, other biothiol molecules, such as glutathione, could also be detected by this sensor system. The method was also successfully applied to the determination of the total content of aminothiols in human plasma. Therefore an easily prepared, inexpensive, high solubility fluorescent probe has been realized and is also expected to detect other biological analytes of interest.

Determination of freely dissolved polycyclic aromatic hydrocarbons in human serum using core-shell Fe3O4@polyacrylate magnetic microspheres by exclusive volume effect.

Freely dissolved concentration is an important parameter for evaluating the bioavailability of compounds. Negligible-depletion solid-phase microextraction (nd-SPME) has been widely used for the determination of freely dissolved compounds but suffered from long equilibrium time. Multifunctional mesoporous composite microspheres have large specific surface area and therefore extraction equilibrium could be reached in a short time. In this study, a novel method was developed for quick determination of freely dissolved polycyclic aromatic hydrocarbons (PAHs) in human serum using core-shell polyacrylate-ferriferous oxide magnetic microspheres (Fe3O4@PA). Mass transfer of PAHs from sample solution to Fe3O4@PA was greatly increased owing to unique properties including large surface area (58.5 m2 g-1), high pore volume (0.10 cm3 g-1) and thin coating layer (50 nm). Freely dissolved PAHs can be selectively extracted because of the mesoporous structure of PA coating layer with uniform pore size of 7.08 nm. However, bound forms of PAHs would not be able to access into pore channels due to size exclusion. In comparison with long equilibration time (139 h) by nd-SPME, equilibrium can be reached within 29 min (t90%) using Fe3O4@PA as novel sorbents. The sorption coefficients (log KBSA) of PAHs to bull serum albumin (BSA) ranged from 3.36 to 4.87, which are consistent with the values measured by nd-SPME (log KBSA = 3.64-5.12). Finally, the freely dissolved PAHs (Cfree) measured by the proposed method (0.69-1.92 µg L-1) have a good agreement with that by nd-SPME (0.56-2.11 µg L-1), indicating that it is feasible for rapid determination of free forms of compounds in real samples by Fe3O4@PA.

Successive grafting of poly(hydroxyethyl methacrylate) brushes and melamine onto chitosan microspheres for effective Cu(II) uptake.

Cross-linked chitosan (CCS) microspheres tethered with melamine-conjugated poly(hydroxyethyl methacrylate) (PHEMA) brushes were synthesized by combination of surface-initiated atom transfer radical polymerization (ATRP) of HEMA and subsequent covalent immobilization of melamine onto the chain ends of PHEMA brushes. The as-synthesized CCS microsphere was used as a novel adsorbent for effective uptake of Cu(II) ions from aqueous solution. Success in each functionalization step was ascertained by SEM, ATR-FTIR and XPS characterization. Batch adsorption experimental results demonstrated that the adsorption equilibrium of Cu(II) ions on the melamine-grafted CCS microsphere was rapidly established within 20 min, and the adsorption process was found to be governed by intra-particle diffusion and chemisorption processes. The Langmuir-fitted maximum adsorption capacity of Cu(II) ions on the as-synthesized CCS microspheres was as high as circa 4.67 mmol L-1 (299 mg g-1). The calculated thermodynamic parameters revealed an endothermic and spontaneous adsorption process of Cu(II) ions on the melamine-grafted CCS microspheres. XPS spectra revealed that the adsorption mechanism was attributed to coordination (or chelation) interactions between amino (or hydroxyl) groups with cationic Cu(II) ions.

Self-Nanoemulsifying Drug Delivery System of Tetrandrine for Improved Bioavailability: Physicochemical Characterization and Pharmacokinetic Study.

The main purpose of this study was to investigate the potential of self-nanoemulsified drug delivery system (SNEDDS) to improve the oral bioavailability of tetrandrine (Tet). SNEDDS was developed by using rational blends of excipients with good solubilizing ability for Tet which was selected based on solubility studies. Further ternary phase diagram was constructed to determine the self-emulsifying region. The optimal formulation with the best self-nanoemulsified and solubilization ability consisted of 40% (w/w) oleic acid as oil, 15% (w/w) SPC and 30% (w/w) Cremophor RH-40 as surfactant, and 15% (w/w) PEG400 as cosurfactant. The average droplet size and zeta-potential of the optimal Tet SNEDDS were 19.75±0.37 nm and 1.87±0.26 mv, respectively. The dissolute rate of Tet SNEDDS in various dissolution media was remarkably faster than Tet commercial tablet. Moreover, in vivo pharmacokinetic study results show that significant increase (p≤ 0.05) in the peak concentration (Cmax) and the area under the curve (AUC) of Tet was observed after the oral administration of Tet SNEDDS and the absorption of Tet from SNEDDS resulted in approximately 2.33-fold increase in oral bioavailability compared with the commercial tablet. Our research suggests that the prepared Tet SNEDDS could be a good candidate for improved the dissolution and oral bioavailability of Tet.

Pharmacokinetics of Quercetin-Loaded Methoxy Poly(ethylene glycol)-b-poly(L-lactic acid) Micelle after Oral Administration in Rats.

The purpose of this study was to evaluate the potential of micelle to change the pharmacokinetics of quercetin (QUT), with a primary goal of enhancing its oral bioavailability. QUT-loaded methoxy poly(ethylene glycol)-b-poly(L-lactic acid) micelle (QUT-loaded MPEG-b-PLLA micelle) was prepared by a thin-film hydration method, resulting in a particle size of 88.5 nm. A liquid chromatography tandem-mass spectrometry (LC-MS/MS) method was developed and validated for determination of QUT in rat plasma. The chromatographic separation was performed on an Agilent Eclipse-C18 (4.6 mm × 50 mm, 3.5 µm) with an isocratic mobile phase system consisting of water and methanol (30 : 70, v/v) at a flow rate of 0.4 mL/min. Calibration curves were linear over the concentration ranges of 2.5-2000 ng/mL for QUT. The micelle was orally administered at a single does in rats, and the pharmacokinetic parameters were evaluated and compared with that administered with the QUT aqueous suspension. The results show that the micelle was able to increase the QUT's oral bioavailability 9-fold compared to the QUT aqueous suspension. These results suggest that methoxy poly(ethylene glycol)-b-poly(L-lactic acid) is a potential carrier for the oral delivery of QUT.

Promising galactose-decorated biodegradable poloxamer 188-PLGA diblock copolymer nanoparticles of resibufogenin for enhancing liver cancer therapy.

Liver cancer is one of the major diseases affecting human health. Modified drug delivery systems through the asialoglycoprotein receptor, which is highly expressed on the surface of hepatocytes, have become a research focus for the treatment of liver cancer. Resibufogenin (RBG) is a popular traditional Chinese medicine and natural anti-cancer drug that was isolated from Chansu, but its cardiotoxicity and hydrophobicity have limited its clinical applications. Galactosyl-succinyl-poloxamer 188 and galactosyl-succinyl-poloxamer 188-polylactide-co-glycolide (Gal-SP188-PLGA) were synthesized using galactose, P188, and PLGA to achieve active liver-targeting properties. RBG-loaded Gal-SP188-PLGA nanoparticles (RGPPNs) and coumarin-6-loaded Gal-SP188-PLGA nanoparticles (CGPPNs) were prepared. The in vitro cellular uptake, cytotoxicity, and apoptosis of nanoparticles in HepG2 cells were analyzed. The in vivo therapeutic effects of nanoparticles were assessed in a hepatocarcinogenic mouse model. The results showed that Gal-SP188-PLGA was successfully synthesized. The cellular uptake assay demonstrated that CGPPNs had superior active liver-targeting properties. The ratio of apoptotic cells was increased in the RGPPN group. In comparison to the other groups, RGPPNs showed superior in vivo therapeutic effects and anticancer efficacy. Thus, the active liver-targeting RGPPNs, which can enhance the pharmacological effects and decrease the toxicity of RBG, are expected to become a promising and effective treatment for liver cancer.

A nanoparticulate pre-chemosensitizer for efficacious chemotherapy of multidrug resistant breast cancer.

Small-molecule chemosensitizers can reverse cancer multidrug resistance (MDR), thus significantly improving the in vitro effect of chemotherapy drugs for MDR cancer cells, however, their in vivo effects are not always very good, because they are difficult to effectively accumulate in tumor and enter the same cancer with chemotherapy drugs after systemic administration due to individual biopharmaceutical properties. To overcome these limitations, here we study a novel nanoparticular pre-chemosensitizer which can be also used as nanocarrier of chemotherapy drugs. We take an 'all in one' approach to develop a self-assembled nanoparticle formula of amphiphilic poly(curcumin-dithiodipropionic acid)-b-poly(ethylene glycol)-biotin. The nanoparticle is capable of tumor-targeted delivery, responsive degradation at the intracellular level of glutathione and subsequent intracellular co-release of the chemosensitizer curcumin and the encapsulated chemotherapeutic drug doxorubicin to maximize a synergistic effect of chemosensitization and chemotherapy. We demonstrate that the antitumor efficacy of nanoparticle is much superior to that of doxorubicin in the multidrug resistant MCF-7/ADR xenografted nude mice.

Oleanolic acid-loaded PLGA-TPGS nanoparticles combined with heparin sodium-loaded PLGA-TPGS nanoparticles for enhancing chemotherapy to liver cancer.

AIM: Heparin sodium (HS)-loaded polylactic-co-glycolic acid-D-α-tocopheryl polyethylene glycol 1000 succinate (PLGA-TPGS) nanoparticles (HPTNs) were prepared as sustained and targeted delivery carriers and combined with oleanolic acid (OA)-loaded PLGA-TPGS nanoparticles (OPTNs) that had been investigated in our previous work to form a combination therapy system for the treatment of liver cancer. MAIN METHODS: To inspect cellular uptake and evaluate liver-targeting performance by analysing drug concentrations and cryosections, fluorescent probe coumarin-6 and eosin was used in preparations of HS/eosin-loaded, HS/coumarin-6-loaded, and OA/coumarin-6-loaded PLGA-TPGS nanoparticles. All of these NPs were characterized in terms of size, size distribution, surface charge, drug loading, encapsulation efficiency, and in vitro release profile. The apoptosis of HepG2 cells induced by OPTNs combined with HPTNs was determined by Annexin V-FITC staining and PI labelling. KEY FINDINGS: Transmission electron microscopy indicated that all of the nanoparticles were stably dispersed spheres with diameters ranging from 100 to 200nm. The results demonstrated that fluorescent nanoparticles were efficiently internalized into HepG2 and HCa-F cells, and that they exhibited enhanced liver targeting. The combination of HPTNs and OPTNs resulted in effective cell inhibition in vitro and a remarkable synergistic anticancer effect in vivo. The cell apoptosis results indicated that OPTNs combined with HPTNs could induce HepG2 cell apoptosis and exert synergistic effects. In vivo pharmacodynamics analysis using a solid tumour-bearing mouse model indicated that OPTNs combined with HPTNs could suppress tumour growth by 67.61%. SIGNIFICANCE: This research suggests that the combined therapy system of OPTNs and HPTNs could be a new means of hepatoma therapy.

Quercetin-loaded poly (lactic-co-glycolic acid)-d-α-tocopheryl polyethylene glycol 1000 succinate nanoparticles for the targeted treatment of liver cancer.

Utilization of quercetin (QT) in clinics is limited by its instability and poor solubility. To overcome these disadvantages, we prepared QT as QT-loaded PLGA-TPGS nanoparticles (QPTN) and examined its properties and therapeutic efficacy for liver cancer. QT-loaded PLGA nanoparticles (QPN) and QT/coumarin-6-loaded PLGA-TPGS nanoparticles (QCPTN) with coumarin-6 as a fluorescent marker were also prepared to investigate the cellular uptake by HepG2 and HCa-F cells using a confocal laser scanning microscope (CLSM), and their effects on apoptosis of HepG2 cells were assessed with flow cytometry. The results measured using transmission electron microscopy, scanning electron microscopy and size analyses indicated that QPTN were stably dispersed sphere with diameter in the range of 100-200 nm. It indicated that the QT loading and encapsulation efficiency in QPTN reached 21.63% and 93.74%, respectively, and the accumulative drug release of QPTN was 85.8%, the QCPTN uptake in HCa-F and HepG2 cells were 50.87% and 61.09% using HPLC analysis, respectively. The results determined using an Annexin-PI flow cytometry indicated that QPTN could induce HepG2 cell apoptosis in a dose dependent manner. The results of histological examination and HPLC analysis confirmed that QPTN was targeted to liver cells. In vivo analysis using solid tumor-bearing mouse model indicated that QPTN could suppress tumor growth by 59.07%. Moreover, all the studied properties of QPTN were more desirable than those of QT-loaded PLGA nanoparticles (QPN). In conclusion, QPTN could be used as a potential intravenous dosage form for the treatment of liver cancer owing to the enhanced pharmacological effects of QT with increased liver targeting.

Amphiphilic Copolymeric Micelles for Doxorubicin and Curcumin Co-Delivery to Reverse Multidrug Resistance in Breast Cancer.

Development of multidrug resistance against chemotherapeutic drugs is one of the major obstacles to successful cancer therapy in the clinic. Thus far, amphiphilic polymeric micelles and chemosensitizers have been used to overcome multidrug resistance in cancer. The goals of this study were to prepare poly(ethylene glycol)-bock-poly(lactide) (PEG(2k)-PLA(5k)) micelles for co-delivery of the chemotherapeutic drug doxorubicin (DOX) with a chemosensitizer curcumin (CUR), investigate the potential of the dual drug-loaded micelles ((DOX+CUR)-Micelles) to reverse multidrug resistance, and explore the underlying mechanisms. (DOX + CUR)-Micelles were prepared using an emulsion solvent evaporation method. The cellular uptake, drug efflux, down-regulation of P-glycoprotein expression and inhibition of ATP activity of (DOX+ CUR)-Micelles were studied in drug-resistant MCF-7/ADR cells. In vitro analyses demonstrated that (DOX + CUR)-Micelles were superior to free DOX, free drug combination (DOX + CUR), and DOX-loaded micelles in inhibiting proliferation of MCF-7/ADR cells. This effect of (DOX + CUR)-Micelles was partially attributable to their highest cellular uptake, lowest efflux rate of DOX, and strongest effects on down-regulation of P-glycoprotein and inhibition of ATP activity. Additionally, (DOX+CUR)-Micelles showed increased tumor accumulation and strong inhibitory effect on tumor growth in the xenograft model of drug-resistant MCF-7/ADR cells compared to that of other drug formulations. These results indicate that (DOX + CUR)-Micelles display potential for application in the therapy of drug-resistant breast carcinoma.