To improve drug efficacy and minimize side effects for treating human promyelocytic leukemia cells, we explored a strategy using the traditional chemotherapy drug all-trans retinoic acid (ATRA) combined with nanodiamonds (NDs) and blue light (BL) irradiation on the typical HL60 cell line. The synergistic ATRA–BL–ND could significantly induce cell inhibition ∼4 times that treated by ATRA alone. The introduction of BL plays an important role in the enhancement of cells apoptosis, improves the level of reactive oxygen species (ROS), reduces the expression of B-cell leukemia/lymphoma 2 (Bcl-2) mRNA, and increases the activation of caspase-3 of HL60 cells. The biocompatible NDs are favorable for increasing ATRA concentration and improving drug delivery. The synergistically enhanced antitumor effect was further verified by in vivo examinations. The results in this paper could be helpful to establish a high efficiency and low toxicity strategy for clinical treatment of various tumors.

Exosomes, the smallest subgroup of extracellular vesicles, have been recognized as extracellular organelles that contain genetic and proteomic information for long distance intercellular communication. Exosome-based drug delivery is currently a subject of intensive research. Here, we report a novel strategy to produce nanoscale exosome-mimics (EMs) in sufficient quantity for gene delivery in cancer both in vitro and in vivo. Size-controllable EMs were generated at a high yield by serial extrusion of non-tumorigenic epithelial MCF-10A cells through filters with different pore sizes. siRNA was then encapsulated into the EMs by electroporation. Biosafety and uptake efficiency of the EMs were evaluated both in vitro and in vivo. The mechanism underlying their cellular endocytosis was also studied.

Clinical development of siRNA has been hindered by the lack of an effective delivery system. Here, we report the construction of a novel siRNA delivery system, sTOLP, which is based on cell penetrating peptide oleoyl-octaarginine (OA-R8) modified multifunctional lipid nanoparticles. sTOLP nanoparticles are composed of a protamine complexed siRNA core, OA-R8, cationic and PEGylated lipids, and transferrin as a targeting ligand. sTOLP formulation was optimized and characterized in vitro and showed excellent gene silencing activity. In vivo, siRNA encapsulated in sTOLP exhibited potent tumor inhibition (61.7%) and was preferentially taken up by hepatocytes and tumor cells in HepG2-bearing nude mice without inducing immunogenicity or hepatic or renal toxicity. Furthermore, sTOLP-loaded siRNA had stability in circulation greater than that of free siRNA. These data demonstrated potential utility of sTOLP-mediated siRNA delivery in cancer therapy.Keywords: cancer; cell penetrating peptide; drug delivery; lipid nanoparticles; multifunctional; siRNA

Research on quantitatively controlling the ligand density on the surface of nanocarriers is in the frontier and becomes a technical difficulty for targeted delivery system designing. In this study, we developed an improved pre-conjugation (Imp) strategy, in which herceptin as a ligand was pre-conjugated with DSPE-PEG2000-Mal via chemical cross-linking, followed by conjugation onto the surface of pre-prepared paclitaxel-loaded PLGA/DODMA nanoparticles (PDNs) through hydrophobic interaction and electrostatic attraction for paclitaxel delivery. Compared with the post-conjugation (Pos) strategy, in which the ligand was conjugated onto the nanoparticle surface after the preparation of the nanoparticles, it realized a precise control targeting effect via adjustment of the herceptin density on the surface of the nanoparticles. Within the range of 0–20% of DSPE-PEG2000-herceptin in the blend, it showed a linear relation with the ligand density on the surface of the nanoparticles. The Imp strategy protected the bioactivity of the ligand during the preparation of nanoparticles. At the same time it avoided the waste of an excess amount of herceptin to drive the conjugation reaction in comparison with the post-conjugation (Pos) strategy. The nanoparticles from the Imp strategy showed much better cytotoxicity (p < 0.001), tumor targeting and cellular uptake efficiency (p < 0.001) than that of the other strategies in BT474 cells, in which BT474 cells were HER2 receptor over-expression breast cancer cell lines. A significant reduction in cellular uptake of the nanoparticles from the Imp strategy was observed in the presence of sucrose and cytochalasin D, indicating that clathrin-mediated and caveolae-dependent endocytosis was as a primary mechanism of cellular entry for these antibody-modified nanoparticles.

We described a novel approach for survivin siRNA cellular delivery via a cationic liposome incorporating fatty acid-modified polyethylenimine. A linoleic acid derivative of branched polyethylenimine(PEI, Mw=25 kDa), PEI-LA, was synthesized and incorporated into the liposome. The properties of the liposome, cytotoxicity, cellular uptake of cancer cells for survivin siRNA, survivin protein downregulation levels were investigated. PEI-modified liposome showed a lower cytotoxicity and delivered survivin siRNA into HeLa cells and A549 cells efficiently compared with PEI-25kDa.

•We synthesized a novel antisense oligonucleotide carrier PEI-OA for the delivery of LOR-2501.•PEI-OA/LOR-2501 complexes were further coated with folic acid and evaluated in tumor cells.•The levels of cellular uptake of FA/PEI-OA/LOR-2501 were much higher than PEI.•The levels of the uptake were not affected by the expression level of folate receptor.•We found that the complexes were mainly taken up by cells via the clathrin-mediated endocytosis.Polyethylenimine (PEI) was conjugated to oleic acid (PEI-OA) and evaluated as a delivery agent for LOR-2501, an antisense oligonucleotide against ribonucleotide reductase R1 subunit. PEI-OA/LOR-2501 complexes were further coated with folic acid (FA/PEI-OA/LOR-2501) and evaluated in tumor cells. The level of cellular uptake of FA/PEI-OA/LOR-2501 was more than double that of PEI/LOR-2501 complexes, and was not affected by the expression level of folate receptor (FR) on the cell surface. Efficient delivery was seen in several cell lines. Furthermore, pathway specific cellular internalization inhibitors and markers were used to reveal the principal mechanism of cellular uptake. FA/PEI-OA/LOR-2501 significantly induced the downregulation of R1 mRNA and R1 protein. This novel formulation of FA/PEI-OA provides a reliable and highly efficient method for delivery of oligonucleotide and warrants further investigation.In this study, a novel antisense oligonucleotide carrier, polyethylenimine (PEI) conjugated with oleic acid (PEI-OA), was synthesized and evaluated for the delivery of LOR-2501. PEI-OA/LOR-2501 complexes were further coated with folic acid (FA/PEI-OA/LOR-2501) and evaluated in tumor cells. The levels of cellular uptake of FA/PEI-OA/LOR-2501 were increased and not affected by the expression level of folate receptor on the cell surface.

The siRNA LOR-1284 targets the R2 subunit of ribonucleotide reductase (RRM2) and has shown promise in cancer therapy. In this study, transferrin (Tf) conjugated lipid nanoparticles (Tf–NP–LOR-1284) were synthesized by microfluidic hydrodynamic focusing (MHF) and evaluated for the targeted delivery of LOR-1284 siRNA into acute myeloid leukemia (AML) cells. The in vitro study showed that Tf–NP–LOR-1284 can protect LOR-1284 from serum nuclease degradation. Selective uptake of Tf–NP–LOR-1284 was observed in MV4-11 cells. In addition, qRT-PCR and Western blot results revealed that Tf–NP–LOR-1284 was more effective than the free LOR-1284 in reducing the R2 mRNA and protein levels. The Tf–NP–LOR-1284 showed prolonged circulation time and increased AUC after i.v. administration relative to the free LOR-1284. Furthermore, Tf–NP–LOR-1284 facilitated increased accumulation at the tumor site along with the decreased R2 mRNA and protein expression in a murine xenograft model. These results suggest that Tf-conjugated NPs prepared by MHF provide a suitable platform for efficient and specific therapeutic delivery of LOR-1284 into AML cells.

•Vitamin E TPGS stabilized the nanoparticle formulation.•Efficient encapsulation of paclitaxel in PLGA by ethanol–water system.•PTX-TENPs showed the uniform particle size and high drug encapsulation.Nanoparticles are efficient delivery vehicles for cancer therapy such as paclitaxel (PTX). In this study, we formulated PTX into PLGA polymeric nanoparticles. Vitamin E TPGS was used as an emulsifier to stabilize the nanoparticle formulation. PTX was encapsulated in TPGS-emulsified polymeric nanoparticles (TENPs) by a nanoprecipitation method in ethanol–water system. The resultant PTX-TENPs showed a very uniform particle size (∼100 nm) and high drug encapsulation (>80%). The cytotoxicity of PTX-TENPs was examined in A549 lung cancer cell line. Preferential tumor accumulation of TENPs was observed in the A549 lung cancer xenograft model. Tumor growth was significantly inhibited by intravenous injection of PTX-TENPs. Our results suggested that the modified nanoprecipitation method holds great potential for the fabrication of the PTX loaded polymeric nanoparticles. TPGS can be used in the manufacture of polymeric nanoparticles for the controlled release of PTX and other anti-cancer drugs.Schematic illustration of the modified nanoprecipitation method to prepare PTX-TENPs. (A) The schematic diagram of nanoparticle forming process. (B) The hypothesis of PTX-TENP structure formation.

The present study aims to identify the major active component from mutant Irpex lacteus, which protects against cationic bovine serum albumin (C-BSA)-induced membranous glomerulonephropathy (MGN). The candidate component ILN3A (MW: 2264 kDa) was purified from mutant Irpex lacteus water extract. The backbone of ILN3A comprises (1→2) and (1→4) linkages, and 1H NMR spectrum suggests the existence of α- and β-glycosidic anomeric carbon. In tissue culture study, ILN3A inhibits mesangial cell proliferation. In MGN rats, ILN3A reverses structural changes in kidney, suppresses abnormal high level of urine protein and restores concentration of serum albumin. ILN3A also reduces total cholesterol, triglycerides, and creatinine in serum, and 6-keto-PGF in kidney cortex. Further study shows ILN3A restores serum Interleukin 2, Interleukin 2 receptor, Interleukin 6, tumor necrosis factor α, and renal cortical nuclear factor kappa B. Our data shows ILN3A, the major active component of mutant Irpex lacteus, is a novel candidate anti-inflammatory medicine to treat MGN in clinics.

The aim of this review is to summarize advances that have been made in the delivery of phytochemicals for cancer therapy by the use of nanotechnology. Over recent decades, much research effort has been invested in developing phytochemicals as cancer therapeutic agents. However, several impediments to their wide spread use as drugs still have to be overcome. Among these are low solubility, poor penetration into cells, high hepatic disposition, and narrow therapeutic index. Rapid clearance or uptake by normal tissues and wide tissue distribution result in low drug accumulation in the target tumor sites can result in undesired drug exposure in normal tissues. Association with or encapsulation in nanoscale drug carriers is a potential strategy to address these problems. This review discussed lessons learned on the use of nanotechnology for delivery of phytochemicals that been tested in clinical trials or are moving towards the clinic.

In our previous work, partial least squares (PLSs) were employed to develop the near infrared spectroscopy (NIRs) models for at-line (fast off-line) monitoring key parameters of Lactococcus lactis subsp. fermentation. In this study, radial basis function neural network (RBFNN) as a non-linear modeling method was investigated to develop NIRs models instead of PLS. A method named moving window radial basis function neural network (MWRBFNN) was applied to select the characteristic wavelength variables by using the degree approximation (Da) as criterion. Next, the RBFNN models with selected wavelength variables were optimized by selecting a suitable constant spread. Finally, the effective spectra pretreatment methods were selected by comparing the robustness of the optimum RBFNN models developed with pretreated spectra. The results demonstrated that the robustness of the optimal RBFNN models were better than the PLS models for at-line monitoring of glucose and pH of L. lactis subsp. fermentation.

Microfluidic systems can accelerate clinical translation of nanoparticles due to their ability to generate nanoparticles in a well-controlled and reproducible manner. In this study, a single-step process based on microfluidic focusing (MF) was employed to synthesize transferrin-conjugated lipid nanoparticles (Tf-LNPs) and the method was compared with a multi-steps bulk mixing (BM) method. The results indicate that this single-step MF process enables rapid and efficient synthesis of Tf-LNPs, which were named Tf-LNPs-MF. Tf-LNPs-MF was shown to have a smaller size and more uniform structures compared to LNPs produced by multi-steps BM method (Tf-LNPs-BM). Furthermore, efficient cellular uptake of Tf-LNPs-MF in vitro as well as greater tumor inhibition in vivo proved that Tf-LNPs-MF had higher siRNA delivery efficiency in vitro and in vivo. Taken together, this single-step microfluidic synthesis significantly simplified the Tf-LNPs production and improved their drug delivery properties and may serve as a valuable tool for developing new cancer therapies.In this study, we developed a new strategy to synthesize the Tf-LNPs in a single step process based on microfluidic technology. To have a better understanding of Tf-LNPs synthesized by this single-step MF method, we put bulk mixing (BM) method as control and evaluated the Tf-LNPs produced by this single-step MF method and BM method in vitro and in vivo. The high siRNA delivery efficiency and great tumor inhibition of Tf-LNPs-MF revealed the possibility and benefit of this single-step MF method, which is a promising strategy for synthesis of targeted LNPs.

Novel biodegradable in situ forming organogel, obtained via the self-assembly of long chain fatty acid in pharmaceutical oil, was prepared and characterized. Different from traditional organogels, the use of organic solvent was avoided in this gel system, in consideration of its tissue irritation. Four kinds of fatty acids were employed as organogelators, which could successfully gel with injectable soybean oil. The gelation procedure was thermo-reversible. Phase transition temperature and time were depended on carbon chain length and concentration of gelators. Optimized formulations containing drug were then injected subcutaneously in rats for pharmacokinetic study. Results showed the steady drug release for one week with the well-controlled burst, which fitted well with the drug release mechanism of both drug diffusion and frame erosion. In vivo imaging of the organogel with fluorescence in live animals suggested that the organogel matrix was gradually absorbed and completely up-taken in nine days. Histopathological analysis of the surrounding tissues was carried out and revealed an overall good biocompatibility property of the implants over drug release period. This research demonstrates that this thermo-sensitive in situ forming organogel system represents a potentially promising platform for sustained drug delivery.Download high-res image (106KB)Download full-size image

Oral vorinostat has the remarkable curative effect on aggravated and recurrent cutaneous T-cell lymphoma (CTCL), but is accompanied by serious adverse effects. Therefore, oral vorinostat is not applicable to the treatment of early stage CTCL. The aim of this study is to develop a novel vorinostat formulation which is effective for early stage CTCL and free of the serious adverse effects. A mixed-matrix hydrogel of vorinostat was prepared and characterized as a potential topical skin delivery system. Moisture retention, swelling behavior, viscosity, real-time morphology and differential scanning calorimeter analysis (DSC) of hydrogel were evaluated to select the solvent, matrix and humectant. The optimal HPMC/HPC ratio, pH, additive, dose and drug loading of vorinostat hydrogel were determined by evaluating the cumulative vorinostat amount of skin retention and transdermal amount of vorinostat through the skin in vitro. The optimal hydrogel presented a low transdermal amount of vorinostat through the skin, suggesting that the hydrogel reduced the amount of vorinostat that was absorbed in the systemic circulation. More importantly, in vivo percutaneous permeation experiments were also performed to evaluate the permeation behavior of vorinostat into the skin. The topical application with a much lower dose showed higher AUC (the cumulative vorinostat amount of skin retention) than oral application and the hydrogel achieved a sustained permeation of vorinostat in the skin for 24 h in vivo. It indicated that a higher relative bioavailability for hydrogel was achieved compared with oral vorinostat. Moreover, there was no damage, inflammation or cell swelling of the skin after administration. Thus, the mixed-matrix vorinostat hydrogel prepared in this study could deliver vorinostat into local skin more efficiently than oral administration.Download high-res image (109KB)Download full-size image

Research on quantitatively controlling the ligand density on the surface of nanocarriers is in the frontier and becomes a technical difficulty for targeted delivery system designing. In this study, we developed an improved pre-conjugation (Imp) strategy, in which herceptin as a ligand was pre-conjugated with DSPE-PEG2000-Mal via chemical cross-linking, followed by conjugation onto the surface of pre-prepared paclitaxel-loaded PLGA/DODMA nanoparticles (PDNs) through hydrophobic interaction and electrostatic attraction for paclitaxel delivery. Compared with the post-conjugation (Pos) strategy, in which the ligand was conjugated onto the nanoparticle surface after the preparation of the nanoparticles, it realized a precise control targeting effect via adjustment of the herceptin density on the surface of the nanoparticles. Within the range of 0–20% of DSPE-PEG2000-herceptin in the blend, it showed a linear relation with the ligand density on the surface of the nanoparticles. The Imp strategy protected the bioactivity of the ligand during the preparation of nanoparticles. At the same time it avoided the waste of an excess amount of herceptin to drive the conjugation reaction in comparison with the post-conjugation (Pos) strategy. The nanoparticles from the Imp strategy showed much better cytotoxicity (p < 0.001), tumor targeting and cellular uptake efficiency (p < 0.001) than that of the other strategies in BT474 cells, in which BT474 cells were HER2 receptor over-expression breast cancer cell lines. A significant reduction in cellular uptake of the nanoparticles from the Imp strategy was observed in the presence of sucrose and cytochalasin D, indicating that clathrin-mediated and caveolae-dependent endocytosis was as a primary mechanism of cellular entry for these antibody-modified nanoparticles.