Bioinspired Platelet-like Nanovector for Enhancing Cancer Therapy via P-Selectin Targeting.

Cancer is a major threat to the health of humans. Recently, various natural products including curcumin (CCM) have attracted enormous interest for efficacious cancer therapy. However, natural therapeutic agents still encounter certain challenges such as rapid clearance, low bioavailability, and poor tumor targeting. Recently, the platelet membrane (PM) camouflaged nanoparticle has provided a promising solution for cancer targeting therapy. Nevertheless, only limited efforts have been dedicated to systematically explore the mechanism of affinity between PM bioinspired nanoparticles and various tumor cells. Herein, a CCM-encapsulated platelet membrane biomimetic lipid vesicle (CCM@PL) with a size of 163.2 nm, zeta potential of -31.8 mV and encapsulation efficiency of 93.62% was developed. The values of the area under the concentration-time curve and mean residence time for CCM@PL were 3.08 times and 3.04 times those of CCM, respectively. Furthermore, this PM biomimetic carrier showed an excellent affinity against Huh-7, SK-OV-3 and MDA-MB-231 cell lines due to the biomolecular interaction between P-selectin on the PM and tumoral CD44 receptors. In addition, CCM@PL displayed enhanced cytotoxicity compared with free CCM and the synthetic formulation. Overall, our results suggest that this developed PM biomimetic lipid nanovector has great potential for targeted cancer treatment and natural components delivery.

Watson-Crick Base Pairing-Inspired Laser/GSH Activatable miRNA-Coordination Polymer Nanoplexes for Combined Cancer Chemo-Immuno-Photothermal Therapy.

The tumor immunosuppressive microenvironment (TIM) greatly hindered the efficacy of cancer immunotherapy. Overexpressed indoleamine 2,3-dioxygenase-1 (IDO1) in tumor tissues plays a vital role in TIM generation, and downregulation of IDO1 expression may reverse TIM. Inspired by the Watson-Crick base-pairing rule, a versatile noncationic miRNA vector (miDAC@PDA) is developed for cancer immunotherapy. Doxorubicin (DOX), adenosine triphosphate (ATP), and copper ions (Cu2+) are coassembled into coordination polymer nanoparticles (DAC) and bind miRNA via the hydrogen bond interaction (miDAC) between adenine residues (ATP) and uracil residues (miRNA). Polydopamine (PDA) is deposited onto the surface of miDAC for photothermal therapy. miDAC@PDA can efficiently accumulate into tumor tissues for cellular uptake. Under laser irradiation and high intracellular GSH levels, the PDA shell of miDAC@PDA can dissociate from miDAC for miRNA release due to local hyperthermia. Cu2+-mediated GSH consumption and intracellular ATP release can amplify the DOX-based immunogenic cell death (ICD) cascade, together with miR-448-mediated IDO1 inhibition, and these versatile nanoplexes will not only restrain primary tumor growth but also display a remarkable abscopal effect on distant tumors. Collectively, our study provides a unique strategy for intracellular gene delivery and an inspirational approach for multimechanism cancer management.

Enhancing TNBC Chemo-immunotherapy via combination reprogramming tumor immune microenvironment with Immunogenic Cell Death.

Tumor-associated fibroblasts (TAFs) play an important role in tumor progression and therapeutic response, especially in the immunosuppressive tumor microenvironment (TME). To remodel immunosuppressive TME of 4T1 tumor, we developed a nano liposome to deliver silybin (SLN, an anti-liver fibrosis Chinese Traditional Medicine). Liposomal silybin (SLN/LIP) possessed a spherical shape with particle sizes of 75.2 nm, high stability, and good accumulation in the tumor site. After treated with SLN/LIP, α-SMA positive TAFs and the deposition of stroma were decreased significantly. SLN/LIP also changed the tumor immune microenvironment through the increase of IFN-γ and IL-12, as well as reduced of TGF-ß, SDF-1, IL6 and TNF-α. Importantly, SLN/LIP enhanced the infiltration of cytotoxic T cells (CTLs) and transformed a "cold" tumor into a "hot" tumor. To achieve the higher antitumor efficacy, an immunogenic cell death (ICD) inducer, liposomal doxorubicin (DOX/LIP) was combined with SLN/LIP. The combination treatment led to trigger immunogenic tumor apoptosis, and enhance antitumor immunity, therefore, improved anti-tumor efficiency, and further prolonged survival duration. The combination of liposomal silybin and liposomal doxorubicin might be a new chemo-immunotherapy approach for triple negative breast cancer (TNBC) tumor treatment.

Mitochondria-Modulating Porous Se@SiO2 Nanoparticles Provide Resistance to Oxidative Injury in Airway Epithelial Cells: Implications for Acute Lung Injury.

BACKGROUND: Mitochondrial dysfunction played a vital role in the pathogenesis of various diseases, including acute lung injury (ALI). However, few strategies targeting mitochondria were developed in treating ALI. Recently, we fabricated a porous Se@SiO2 nanoparticles (NPs) with antioxidant properties. METHODS: The protective effect of Se@SiO2 NPs was assessed using confocal imaging, immunoblotting, RNA-seq, mitochondrial respiratory chain (MRC) activity assay, and transmission electron microscopy (TEM) in airway epithelial cell line (Beas-2B). The in vivo efficacy of Se@SiO2 NPs was evaluated in a lipopolysaccharide (LPS)-induced ALI mouse model. RESULTS: This study demonstrated that Se@SiO2 NPs significantly increased the resistance of airway epithelial cells under oxidative injury and shifted lipopolysaccharide-induced gene expression profile closer to the untreated controls. The cytoprotection of Se@SiO2 was found to be achieved by maintaining mitochondrial function, activity, and dynamics. In an animal model of ALI, pretreated with the NPs improved mitochondrial dysfunction, thus reducing inflammatory responses and diffuse damage in lung tissues. Additionally, RNA-seq analysis provided evidence for the broad modulatory activity of our Se@SiO2 NPs in various metabolic disorders and inflammatory diseases. CONCLUSION: This study brought new insights into mitochondria-targeting bioactive NPs, with application potential in curing ALI or other human mitochondria-related disorders.

Transferrin-Modified Nanoliposome Codelivery Strategies for Enhancing the Cancer Therapy.

Chemotherapy remains one of the most effective treatments for many cancers in a clinic. At present, various targets have been used to modify the PEGylated liposomes for doxorubicin (Dox) delivery, but the antitumor effect of Dox is not satisfactory. Therefore, combination chemotherapeutics has been considered as a promising method to improve tumor treatment. These years, RAF/MEK/ERK-mediated cell signaling pathway has been discovered to inhibit the growth of tumors. Thus, Sorafenib tosylate (Sor) was used in this study, which directly inhibited tumor cell proliferation through blocking RAF/MEK/ERK-mediated cell signaling pathway and indirectly inhibited tumor cell growth through blocking angiogenesis by VEGFR and PDGF. In this article, we develop a "combination delivery system" to deliver the hydrophobic drug (Sor) in phospholipid bilayer and hydrophilic drug (Dox) in inner cores for enhancing the antitumor effect. Moreover, in vitro experiments verified whether the physicochemical properties of carriers were stable and transferrin-modified liposomes displayed the highest uptake. The results of in vivo experiments showed that the codelivery system inhibited the tumor growth more effectively than monotherapy. Overall, this combination delivery system for delivering the hydrophobic and hydrophilic drugs simultaneously may offer a novel strategy for breast cancer treatment and provide a reference for the possibility of clinical usage.

Chitosan Oligosaccharide Ameliorates Nonalcoholic Fatty Liver Disease (NAFLD) in Diet-Induced Obese Mice.

Nonalcoholic fatty liver disease (NAFLD) is a global epidemic, and there is no standard and efficient therapy for it. Chitosan oligosaccharide (COS) is widely known to have various biological effects, and in this study we aimed to evaluate the liver-protective effect in diet-induced obese mice for an enzymatically digested product of COS called COS23 which is mainly composed of dimers and trimers. An integrated analysis of the lipidome and gut microbiome were performed to assess the effects of COS23 on lipids in plasma and the liver as well as on intestinal microbiota. Our results revealed that COS23 obviously attenuated hepatic steatosis and ameliorated liver injury in diet-induced obese mice. The hepatic toxic lipids-especially triglycerides (TGs) and free fatty acids (FFAs)-were decreased dramatically after COS23 treatment. COS23 regulated lipid-related pathways, especially inhibiting the expressions of FFA-synthesis-related genes and inflammation-related genes. Furthermore, COS23 could alter lipid profiles in plasma. More importantly, COS23 also decreased the abundance of Mucispirillum and increased the abundance of Coprococcus in gut microbiota and protected the intestinal barrier by up-regulating the expression of tight-junction-related genes. In conclusion, COS23, an enzymatically digested product of COS, might serve as a promising candidate in the clinical treatment of NAFLD.

Binding effect of fluorescence labeled glycyrrhetinic acid with GA receptors in hepatocellular carcinoma cells.

Glycyrrhetinic acid (GA) is a natural active component from licorice, which is broadly used in traditional Chinese medicine. Lots of glycyrrhetinic acid receptors (GA-R) are proved to locate on the surface of liver cells. Many reports about the hepatocellular carcinoma (HCC) treatment were dependent on GA modified carriers. However, the reality of GA-R in HCC cells was not clear. In this paper, 18ß-glycyrrhetinic acid (18ß-GA) was labeled with fluorescence (FITC) by chemical synthesis. Together with the binding effect of fluorescence labeled glycyrrhetinic acid (FITC-GA), the competitive action of 18ß-GA with GA-R was investigated in HCC cells. The results showed that in HepG2 cells, 18ß-GA and FITC-GA presented similar cytotoxicity. The specific binding saturation of GA showed the dissociation constant (Kd) was 7.457±2.122pmol/L and the maximum binding counts (Bmax) was 2.385±0.175pmol/2.5×106 cells, respectively. FITC-GA bound to cytomembrane specifically and 18ß-GA competed to bind the sites significantly in HepG2 cells. Therefore, there is binding effect between fluorescence labeled GA and GA-R. The GA-R on HCC cells is confirmed as expected, which provides a useful reference of active target modified by GA and a novel approach for receptors and ligands study.

Gold nanorods/mesoporous silica-based nanocomposite as theranostic agents for targeting near-infrared imaging and photothermal therapy induced with laser.

Photothermal therapy (PTT) is widely regarded as a promising technology for cancer treatment. Gold nanorods (GNRs), as excellent PTT agent candidates, have shown high-performance photothermal conversion ability under laser irradiation, yet two major obstacles to their clinical application are the lack of selective accumulation in the target site following systemic administration and the greatly reduced photothermal conversion efficiency caused by self-aggregating in aqueous environment. Herein, we demonstrate that tLyp-1 peptide-functionalized, indocyanine green (ICG)-containing mesoporous silica-coated GNRs (I-TMSG) possessed dual-function as tumor cells-targeting near-infrared (NIR) fluorescent probe and PTT agents. The construction of the nanostructure began with synthesis of GNRs by seed-mediated growth method, followed by the coating of mesoporous silica, the chemical conjugation of PEG and tLyp-1 peptide, and the enclosure of ICG as an NIR imaging agent in the mesoporous. The as-prepared nanoparticles could shield the GNRs against their self-aggregation, improve the stability of ICG, and exhibit negligible dark cytotoxicity. More importantly, such a theranostic nanocomposite could realize the combination of GNRs-based photothermal ablation under NIR illumination, ICG-mediated fluorescent imaging, and tLyp-1-enabled more easy endocytosis into breast cancer cells. All in all, I-TMSG nanoparticles, in our opinion, possessed the strong potential to realize the effective diagnosis and PTT treatment of human mammary cancer.

Preparation, optimization and characteristic of huperzine a loaded nanostructured lipid carriers.

The objective of the present work was to study the preparation, optimization and characteristics of Huperzine A (Hup A), an effective Traditional Chinese Medicine treatment of Alzheimer's disease (AD), loaded nanostructured lipid carriers (NLC). NLC were successfully prepared by a modified method of melt ultrasonication followed by high pressure homogenization using Cetyl Palmitate (CP) as the solid lipid, Miglyol((R))812 as the liquid lipid, Soybean phosphatidylcholine (Spc) and Solutol HS15 as the emulsifiers. The best formulation was optimized with a three-factor, three-level Box-Behnken design. Independent variables studied were the amount of the mixed lipid, the amount of the emulsifier mixture and lipid/drug ratio in the formulation. The dependent variables were the particle size, entrapment efficiency (EE) and drug loading (DL). Properties of NLC such as the morphology, particle size, zeta potential, EE, DL and drug release behavior were investigated, respectively. As a result, the designed nanoparticles showed nearly spherical particles with a mean particle size of 120 nm and -22.93 + or - 0.91 mV. The EE (%) and DL (%) could reach up to 89.18 + or - 0.28% and 1.46 + or - 0.05%, respectively. Differential scanning calorimetry (DSC) of Hup A loaded NLC indicated no tendency of recrystallisation. In vitro release studies showed a burst release at the initial stage and followed by a prolonged release of Hup A from NLC up to 96 h. The results suggest that the presented Hup A loaded NLC system is a potential delivery system for improving drug loading capacity and controlled drug release.

Preparation and characterization of nanostructured lipid carriers loaded traditional Chinese medicine, zedoary turmeric oil.

BACKGROUND: The objective of this work was to study the preparation and characteristics of zedoary turmeric oil (ZTO), a traditional Chinese oily medicine, loaded with nanostructured lipid carriers (NLCs). METHOD: Aqueous dispersions of NLC were successfully prepared by melt-emulsification technique using Crodamol SS as the solid lipid, Miglyol 812N as the liquid oil, and soybean phosphatidylcholine (SbPC) as the emulsifier. Properties of NLC such as the particle size and its distribution, the transmission electron microscope (TEM), drug entrapment efficiency (EE), and drug release behavior were investigated, respectively. The Germacrone blood concentration after intravenous administration of ZTO-NLC was determined and compared with that of ZTO-injection. RESULT: As a result, the drug EEs were improved by adding the liquid lipid into the solid lipid of nanoparticles (SLNs). In vitro drug release experiments indicated that the prepared NLC could enhance the drug release rate over the SLN, and the drug release rate could be adjusted by the liquid lipid content in lipid nanospheres. X-ray and differential scanning calorimetry (DSC) measurements revealed that imperfect crystallization occurred in the inner core of the NLC particles. CONCLUSION: The results suggest that the presented NLC system might be a promising intravenous dosage form of water-insoluble oily drugs.

[Preparation and stability of digalactosyl diglyceride as emulsifier for sub-microemulsion].

OBJECTIVE: To study the feasibility of digalactosyl diglyceride (DGDG) , which was used as a new type of emulsifier to prepare submicro-emulsion of bay oil. METHOD: Bay oil was employed as the model drug, emulsifer in oil method was used to prepare foremilk. Through single factor investigation and central composite design-response surface methodology (CCD-RSM) , we optimized the preparation technology and formula respectively. The stability of sub-microemulsion was studied. RESULT: The optima technology was following: emulsifer in oil method was used to prepare foremilk, temp was 60 degrees C, the micro emulsion was prepared by two-step high pressure homogen method with that the pressure was 80 Pa, 10 times, micropore film was used to sterilize, filling and sealing at the preservation of nitrogen. The best formula was following: soybean oil was 1.1%, DGDG was 1.6%, and sodium oleate was 0. 16%. The particle size of three batch submicro-emulsions were from 168.0 to 169.3 nm; Zeta potential were from-25.53 to 24.90 mV, pH value were from 8.48 to 8.52. The deviation between measured value and predictive value was 1.8%. It was stable in high temperature and illumination. CONCLUSION: DGDG can be used as the emulsifier of bay oil sub-microemulsion.

[Preparation and study in vitro of long-circulating nanoliposomes of curcumin].

OBJECTIVE: To prepare the long-circulating nanoliposomes of curcumin. METHOD: The long-circulating nanoliposomes were prepared by ethanol infusion and the encapsulation efficiency was determindated by the mini-column centrifugation. The effect of some factors on the encapsulation efficiency, such as the buffer solutions, the weight ratio of curcumin to SPC, the weight ratio of SPC to Chol, the pH of buffer solution and the iron strength of water phase, was investigated respectively. Then the formulation was optimized by orthogonal design. RESULT: The encapsulation efficiency of the curcumin liposomes was (88.27 +/- 2.16)%, and the average diameter of the liposomes was (136 +/- 18) nm. There was no change on encapsulation efficency within 30 d. CONCLUSION: The preparation of curcumin liposomes was easy and practicable and the pharmaceutical characterization showed that the curcumin liposomes are stable.

Effect of polypeptides in bee venom on growth inhibition and apoptosis induction of the human hepatoma cell line SMMC-7721 in-vitro and Balb/c nude mice in-vivo.

Polypeptides in bee venom (PBV) produced a significant growth inhibition against SMMC-7721 human hepatoma cell line. Analysis of the mechanisms of cell death indicated that PBV induced an apoptotic cell death. SMMC-7721 cells exposed to PBV (10.0 microg mL(-1)) produced an insignificant morphological change. Analysis of the cytotoxicity with the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) assay confirmed that the cytotoxic effects of PBV were dose- and timedependent. The result of Ki67 immunohistochemistry demonstrated that the proliferation of SMMC-7721 cells treated with PBV (10.0 mug mL(-1)) was inhibited. The apoptotic cell death was then confirmed by annexin V, propidium iodide staining and DNA fragmentation analysis. In in-vivo experiments, treatment with PBV (1.5 or 3 mg kg(-1)) resulted in a significant retardation of SMMC-7721 cell growth in Balb/c nude mice. These findings suggested that PBV could be used as a chemotherapeutic agent against tumours.

[Study on influence of buffer system on separate capacity of macroporous resin on ferulic liposome].

OBJECTIVE: To study the effect of buffer on separate capacity of macroporous resin. To evaluate the quality of ferulic acid liposome and determine its entrapment efficiency. METHOD: Different type of macroporous resin counterpoised by buffer system of Na2 HPO3-NaH2, PO3 was used to separate the free ferulic acid from the preparation and HPLC was used to determine the concentration of the ferulic acid to calculate the entrapment efficiency. RESULT: This method had good linearity in the range of 0.56 - 2.8 g x mL(-1) (r = 0.999 6). The precision RSD was less than 1.1%. The adsorption effect of macroporous resin on liposome was reduced while it had no effect on the absorption ability of macroporous resin on the ferulic acid by the usage of buffer. The recovery of HPD450 resin on blank liposome was between 97.2% - 100.8%, while the average recovery is 98.1%. CONCLUSION: Buffer system can enhance the separate ability of macroporous resin on liposome and free drug.

[Study on the preparation of rapid-dissoluted EGb droppills].

OBJECTIVE: To study the formulation and technique of preparation of rapid-dissoluted EGb (Extract of Ginkgo biloba) droppills. METHOD: Taking the dissolution percentage of total flavonoids in EGb and weight variation as index, the formulation and technique of EGb droppills were optimized by the orthogonal experiment. RESULT: T50 was 3.62 min and mean weight variation was 2.80%. CONCLUSION: Rapid-dissoluted EGb droppills can increase the dissoluting rate distinctly and reach the purpose of preparation.