Pluronic L64 Research Articles

Production of stabilized quercetin aqueous suspensions by supercritical fluid extraction of emulsions

Quercetin is a flavonoid with highly promising bioactivity against a variety of diseases, due to its strong antioxidant, antiviral and antihistaminic effect, but these applications are limited by the low solubility of quercetin in gastrointestinal fluids and the correspondingly low bioavailability. The objective of this work is to produce encapsulated quercetin particles in sub-micrometric scale, in order to increase their low bioavailability. These particles were produced by extraction of organic solvent from oil in water emulsions by supercritical fluid extraction of emulsions (SFEE). Due to the rapid extraction of organic solvent by this method, the disperse organic phase becomes rapidly supersaturated, causing the precipitation of quercetin particles in sub-micrometric scale, encapsulated by the surfactant material. Two different biopolymers (Pluronic L64® poloxamers and soy bean lecithin) were used as carriers and surfactant materials. In experiments with Pluronic, needle quercetin particles were obtained after SFEE treatment, with particle sizes around 1μm and poor encapsulation efficiency. In case of soy lecithin, quercetin-loaded multivesicular liposomes were obtained, with a mean particle size around 100nm and around 70% encapsulation efficiency of quercetin, without presence of segregated quercetin crystals.

TBP induced double cloud point in aqueous EO13PO30EO13 solutions: investigating the evolution of associated micellar characteristics as a function of temperature

TBP solubilized Pluronic L64 solutions exhibit inter-micellar attraction driven micellar cluster formation upon cooling, which is unique in non-ionic micellar systems.

PPO/PEO modified hollow fiber membranes improved sensitivity of 3D cultured hepatocytes to drug toxicity via suppressing drug adsorption on membranes

The three dimensional (3D) cell culture in polymer-based micro system has become a useful tool for in vitro drug discovery. Among those polymers, polysulfone hollow fiber membrane (PSf HFM) is commonly used to create a microenvironment for cells. However, the target drug may adsorb on the polymeric surface, and this elicits negative impacts on cell exposure due to the reduced effective drug concentration in culture medium. In order to reduce the drug adsorption, PSf membrane were modified with hydrophilic Pluronic (PEO-b-PPO-b-PEO) copolymers, L121, P123 and F127 (PEO contents increase from 10%, 30% to 70%), by physical adsorption. As a result, the hydrophilicity of HFMs increased at an order of PSf<L121<P123<F127 HFMs, while the negative surface charge decreased at the order of PSf>F127>P123>L121 HFMs. The three modified membrane all showed significant resistance to adsorption of acid/neutral drugs. More importantly, the adsorption of base drugs were largely reduced to an average value of 11% on the L121 HFM. The improved resistance to drug adsorption could be attributed to the synergy of hydrophobic/neutrally charged PPO and hydrophilic PEO. The L121 HFM was further assessed by evaluating the drug hepatotoxicity in 3D culture of hepatocytes. The base drugs, clozapine and doxorubicin, showed more sensitive hepatotoxicity on hepatocytes in L121 HFM than in PSf HFM, while the acid drug, salicylic acid, showed the similar hepatotoxicity to hepatocytes in both HFMs. Our finding suggests that PSf HFM modified by PEO-b-PPO-b-PEO copolymers can efficiently resist the drug adsorption onto polymer membrane, and consequently improve the accuracy and sensitivity of in vitro hepatotoxic drug screening.

Optimization of long circulating mixed polymeric micelles containing vinpocetine using simple lattice mixture design, in vitro and in vivo characterization

The aim of this study was to increase the in vivo mean residence time of vinpocetine after IV injection utilizing long circulating mixed micellar systems. Mixed micelles were prepared using Pluronics L121, P123 and F127. The systems were characterized by testing their entrapment efficiency, particle size, polydispersity index, zeta potential, transmission electron microscopy and in vitro drug release. Simple lattice mixture design was planned for the optimization using Design-Expert® software. The optimized formula was lyophilized, sterilized and imaged by scanning electron microscope. Moreover, the in vivo behavior of the optimized formula was evaluated after IV injection in rabbits. The optimized formula, containing 68% w/w Pluronic L121 and 32% w/w Pluronic F127, had the highest desirability value (0.621). Entrapment efficiency, particle size, polydispersity index and zeta potential of the optimized formula were 50.74±3.26%, 161.50±7.39nm, 0.21±0.03 and −22.42±1.72mV, respectively. Lyophilization and sterilization did not affect the characteristics of the optimized formula. Upon in vivo investigation in rabbits, the optimized formula showed a significantly higher elimination half-life and mean residence time than the market product. Finally, mixed micelles could be considered as a promising long circulating nanocarrier for lipophilic drugs.

Electroneutralized amphiphilic triblock copolymer with a peptide dendron for efficient muscular gene delivery.

Hydrophilic-hydrophobic-hydrophilic triblock copolymers, such as Pluronic L64, P85, and P105, have attracted more attention due to their enhancement in muscular gene delivery. In the present study, a new kind of electroneutralized triblock copolymer, LPL, dendron G2(L-lysine-Boc)-PEG2k-dendron G2(L-lysine-Boc), was designed and investigated. This hydrophobic-hydrophilic-hydrophobic copolymer is composed of a structure reverse to that of L64, one of the most effective materials for intramuscular gene delivery so far. Our results showed that LPL exhibited good in vivo biocompatibility after intramuscular and intravenous administration. LPL mediated higher reporter gene expression than L64 in assays of β-galactosidase (LacZ), luciferase, and fluorescent protein E2-Crimson. Furthermore, LPL-mediated mouse growth hormone expression significantly accelerated mouse growth within the first 10 days. Altogether, LPL-mediated gene expression in skeletal muscle exhibits the potential of successful gene therapy. The current study also presented an innovative way to design and construct new electroneutralized triblock copolymers for safe and effective intramuscular gene delivery.

Modified release from lipid bilayer coated mesoporous silica nanoparticles using PEO–PPO–PEO triblock copolymers

Triblock copolymers comprised of poly(ethylene oxide)–poly(propylene oxide)–poly(ethylene oxide) (PEO–PPO–PEO, or trade name Pluronic) interact with lipid bilayers to increase their permeability. Here we demonstrate a novel application of Pluronic L61 and L64 as modification agents in tailoring the release rate of a molecular indicator species from 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) bilayer-coated superparamagnetic Fe3O4/mesoporous silica core–shell nanoparticles. We show there is a direct relationship between the Pluronics’ concentration and the indicator molecule release, suggesting Pluronics may be useful for the controlled release of drugs from lipid bilayer-coated carriers.

Pluronic L64-mediated stable HIF-1α expression in muscle for therapeutic angiogenesis in mouse hindlimb ischemia.

Intramuscular injection of plasmid DNA (pDNA) to express a therapeutic protein is a promising method for the treatment of many diseases. However, the therapeutic applications are usually hindered by gene delivery efficiency and expression level. In this study, critical factors in a pDNA-based gene therapy system, such as gene delivery materials, a therapeutic gene, and its regulatory elements, were optimized to establish an integrated system for the treatment of mouse hindlimb ischemia. The results showed that Pluronic® L64 (L64) was an efficient and safe material for gene delivery into mouse skeletal muscle. It also showed intrinsic ability to promote in vivo angiogenesis in a concentration-dependent manner, which might be through the activation of nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB)-regulated angiogenic factors. The combination of 0.1% L64 with a hybrid gene promoter (pSC) increased the gene expression level, elongated the gene expression duration, and enhanced the number of transfected muscle fibers. In mice ischemic limbs, a gene medicine (pSC-HIF1αtri/L64) composed of L64 and pSC-based expression plasmid encoding hypoxia-inducible factor 1-alpha triple mutant (HIF-1αtri), improved the expression of stable HIF-1α, and in turn, the expression of multiple angiogenic factors. As a result, the ischemic limbs showed accelerated function recovery, reduced foot necrosis, faster blood reperfusion, and higher capillary density. These results indicated that the pSC-HIF1αtri/L64 combination presented a potential and convenient venue for the treatment of peripheral vascular diseases, especially critical limb ischemia.

Cytotoxicity and chemosensitizing activity of amphiphilic poly(glycerol)-poly(alkylene oxide) block copolymers.

All polymeric chemosensitizers proposed thus far have a linear poly(ethylene glycol) (PEG) hydrophilic block. To testify whether precisely this chemical structure and architecture of the hydrophilic block is a prerequisite for chemosensitization, we tested a series of novel block copolymers containing a hyperbranched polyglycerol segment as a hydrophilic block (PPO-NG copolymers) on multi-drug-resistant (MDR) tumor cells in culture. PPO-NG copolymers inhibited MDR of three cell lines, indicating that the linear PEG can be substituted for a hyperbranched polyglycerol block without loss of the polymers' chemosensitizing activity. The extent of MDR reversal increased with the polymers affinity toward the cells and the expression level of P-glycoprotein. In contrast with Pluronic L61, which increases viability of tumor cells in the absence of drugs, PPO-NG chemosensitizers are completely devoid of this property undesired in cancer therapy, making them promising candidates for application as novel MDR reversal agents.

Docetaxel load biodegradable porous microspheres for the treatment of colorectal peritoneal carcinomatosis

Micro- and nanoparticle formulations are widely used to improve the bioavailability of low solubility drugs. In this study, biodegradable poly(L-lactide acid)–Pluronic L121–poly(L-lactide acid) (PLLA–L121–PLLA) was developed. And then a controlled drug delivery system (CDDS), docetaxel (DOC) loaded PLLA–L121–PLLA porous microsphere (DOC MS) was prepared for colorectal peritoneal carcinomatosis (CRPC) therapy. DOC MS was prepared by DOC and PLLA–L121–PLLA using an oil-in-water emulsion solvent evaporation method. The particle size, morphological characteristics, encapsulation efficiency, in vitro drug release studies and in vitro cytotoxicity of DOC MS have been investigated. In vitro release profile demonstrated a significant difference between rapid release of free DOC and much slower and sustained release of DOC MS. Furthermore, cytotoxicity assay indicated cytotoxicity was increased after DOC was encapsulated into polymeric microspheres. In addition, intraperitoneal administration of DOC MS could effectively suppress growth and metastasis of CT26 peritoneal carcinomatosis in vivo, and prolonged the survival of tumor bearing mice. Immunohistochemistry staining of tumor tissues with Ki-67 revealed that DOC MS induced a stronger anti-tumor effect by increasing apoptosis of tumor cells in contrast to other groups (P<0.05). Thus, our results suggested that DOC MS may have great potential applications in clinic.

Safe and efficient local gene delivery into skeletal muscle via a combination of Pluronic L64 and modified electrotransfer

Efficient DNA electrotransfer into muscles can be achieved by combining two types of electronic pulses sequentially: short high-voltage (HV) pulse for the cell electropermeabilization and long low-voltage (LV) pulse for the DNA electrophoresis into cells. However, the voltages currently applied can still induce histological and functional damages to tissues. Pluronic L64 has been considered as a molecule possessing cell membrane-disturbing ability. For these reasons, we hope that L64 can be used as a substitute for the HV pulse in cell membrane permeabilization, and a safe LV pulse may still keep the ability to drive plasmid DNA across the permeabilized membrane. In this work, we optimized the electrotransfer parameters to establish a safe and efficient procedure using a clinically applied instrument, and found out that the critical condition for a successful combination of electrotransfer with L64 was that the injection of plasmid/L64 mixture should be applied 1 h before the electrotransfer. In addition, we revealed that the combined procedure could not efficiently transfer plasmid into solid tumor because the uncompressed plasmid may rapidly permeate the leaky tumor vessels and flow away. Altogether, the results demonstrate that the combined procedure has the potential for plasmid-based gene therapy through safe and efficient local gene delivery into skeletal muscles.

보조계면활성제가 Pluronic L64 비이온 계면활성제에 의한 탄화수소 오일 가용화에 미치는 영향

본 연구에서는 보조계면활성제 첨가가 n-octane, n-decane, n-dodecane 등의 탄화수소 오일의 가용화에 미치는 영향에 관하여 살펴보았다. 탄화수소의 가용화 속도는 보조계면활성제로 첨가한 알코올의 사슬 길이와 첨가량이 증가함에 따라 증가하였으며, 특히 사술 길이가 비교적 긴 알코올을 첨가하는 경우, 탄화수소 오일의 가용화 속도가 크게 증가하였다. 이는 1-butanol과 같이 짧은 사슬을 가진 수용성 알코올을 첨가할 경우에는 첨가한 대부분의 알코올이 수용액상에 존재하여 탄화수소 오일의 가용화에 큰 영향을 주지 못하는 반면에, 1-hexanol이나 1-octanol과 같이 비교적 사슬길이가 긴 알코올을 보조계면활성제로 첨가한 경우에는 첨가한 대부분의 알코올이 마이셀 상에 위치하여 마이셀을 보다 flexible한 packing을 갖게 함으로써, 가용화를 용이하게 한 것으로 생각된다. 가용화 속도 실험 결과는 spinning drop tensiometer를 사용하여 탄화수소 오일과 계면활성제 수용액 사이의 동적 계면장력을 측정한 결과와 동일한 경향을 나타내었다. 즉, 첨가한 알코올의 사슬 길이가 증가할수록 평형에서의 계면장력 값은 감소하며, 또한 계면장력 값이 평형에 도달하는 시간은 감소하였다. In this study, effect of cosurfactant on the solubilization rate of n-octane, n-decane and n-dodecane oil was performed by micellar solutions of polymeric nonionic surfactant Pluronic L64(<TEX>$EO_{13}PO_{30}EO_{13}$</TEX>) at room temperature. It has been found that the solubilization rate of a hydrocarbon oil was enhanced with an increase in both chain length and amount of alcohol added. In case of addition of a short chain alcohol such as 1-butanol, the solubilization rate of a hydrocarbon oil was slightly increased since most of alcohol molecules remained in an aqueous surfactant solution. On the other hand, the addition of a relatively long chain alcohol such as 1-hexanol and 1-octanol produced a big increase in solubilization rate of a hydrocarbon oil mainly due to incorporation of alcohol molecules into micelles and thus producing more flexible micellar packing density. Dynamic interfacial tension measurements showed the same trend found in solubilization rate measurement. Both interfacial tension value at equilibrium and time required to reach equilibrium decreased with an increase in chain length of an alcohol.

Pluronics Suppress Association of Low-Density Lipoproteins Inducing Atherogenesis

We studied the effect of pluronics P85, L61, and F68 with different hydrophilic-lipophilic characteristics on association of LDL. It was found that pluronics with pronounced hydrophobic properties (P85 and L61) in concentrations close to or surpassing the critical concentration of micelle formation inhibited LDL association, while hydrophilic pluronic F68 in all concentrations had no effect on LDL association.

Mesoporous tantalum oxide as catalyst for dehydration of glucose to 5-hydroxymethylfurfural

Mesoporous tantalum oxide was prepared by acid hydrolysis of tantalum penta-ethoxide in the presence of a triblock co-polymer Pluronic L-121, a non-ionic surfactant, at room temperature and subsequent calcination at 550°C for 6h. This solid exhibits a suitable specific surface area (79m2g−1) and a high acidity (353μmolNH3g−1) with the presence of both Brönsted and Lewis acid sites, demonstrating to be active as solid acid catalyst in the dehydration of glucose to 5-hydroxymethylfurfural (HMF), in a biphasic water/methyl-iso-butyl ketone (MIBK) system. Thus, by using a glucose:catalyst weight ratio of 3:1, a glucose conversion of 69% and a HMF yield of 23% were achieved at 175°C, and after only 90min of reaction time. The catalytic process is selective toward HMF, which is preserved from ulterior hydration to levulinic acid. Fructose was also detected as by-product of glucose isomerisation with 14% of selectivity. The catalyst is very stable, since no leaching of tantalum species to the liquid phase was found; moreover, the catalytic performance of this acid solid is well recovered after calcination at 550°C for 2h.

Mechanistic study of cationic dye interactions with clay‐polymer dispersions via metachromatic effect, aggregation, and surface charge

ABSTRACTInteractions for a series of aqueous dispersions of a clay, Laponite, and nonionic difunctional triblock copolymer, Pluronic L62, with two cationic dyes, crystal violet and brilliant green, are studied using UV–vis spectroscopy, dynamic light scattering (DLS), and electrophoretic mobility (EPM) to better understand the adsorption mechanisms. Different concentrations for clay, 0.1–2.0 g/L, mixed with polymer, 0.125–0.5 g/L to produce an organoclay, were tested with a fixed dye concentration, 2.0 × 10−5 mol/L. The aggregation states of the cationic dyes interacting with the clay–polymer dispersions were characterized by UV–vis absorbance. The dyes changed aggregation states from monomer to dimer and higher aggregate states upon addition to varying Laponite–polymer dispersion concentrations. DLS was used to understand the adsorbed polymer and dye interactions on the clay particle showing longer relaxation times for the largest aggregates observed from UV–vis absorbance. The EPM was used to characterize the surface charge of the clay relative to the polymer–dye interactions, which showed that the surface charge approached zero as the dyes interacted with clay–polymer dispersions. The most significant changes occurred with a reduction in light scattering intensity and longer relaxation decay at 0.1 g/L Laponite and 0.5 g/L Pluronic L62 concentrations in the presence of cationic dyes. The study of the clay–polymer–dye interactions help to better understand and design chemical templates for removal of contaminates in waste‐water treatment. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 40141.

Lipid-like trifunctional block copolymers of ethylene oxide and propylene oxide: Effective and cytocompatible modulators of intracellular drug delivery

A new glycerol-based trifunctional block copolymer (TBC) of propylene oxide and ethylene oxide and its conjugate with succinic acid (TBC-SA) were studied as a drug delivery system and compared with Pluronic L61. TBCs have multiple effects on the plasma membrane of human cells, e.g. increasing its fluidity and ion permeability, inhibiting ATPase activity of efflux transporter P-glycoprotein through reversible membrane destabilization. Such membrane-modulating properties attributed to the unimer form of copolymers increase in the order Pluronic L61≪TBC<TBC-SA and correlate with an ability of TBCs to promote the accumulation of P-glycoprotein substrates in lung cancer A549 cells. Furthermore, TBC, and especially TBC-SA, exhibit substantially lower hemolytic, cytotoxic and proapoptotic activities in comparison with Pluronic L61. Our results demonstrate that TBCs are promising analogs of bifunctional Pluronics in anticancer drug delivery.

Transferrin-Conjugated Pluronic Niosomes as a New Drug Delivery System for Anticancer Therapy

An efficient tumor-targeted niosomal delivery system for the vehiculation of doxorubicin hydrochloride as an anticancer agent was designed. Niosomes were prepared from a mixture of an opportunely modified Pluronic L64 surfactant and cholesterol as a membrane additive and characterized in terms of size and related distribution function and drug entrapment efficiency. After the preparation, transferrin was conjugated to niosomes to produce transferrin (Tf) niosomes, and the cytotoxicity of the final formulation was studied. The specific uptake of Tf niosomes into cells was evaluated via incubation of MCF-7 and MDA-MB-231 cells with fluorescently rhodamine-loaded Tf niosomes for various times and concentration intervals and further investigated by fluorescence microscopy. Results showed that doxorubicin can be easily encapsulated into niosomes, which are regular and spherical in shape. Moreover, transferrin conjugate niosomes demonstrated far greater extents of cellular uptake by MCF-7 and MDA-MB-231 cells, suggesting that they were mainly taken up by transferrin receptor-mediated endocytosis. Doxorubicin-loaded niosome anticancer activity was also achieved against MCF-7 and MDA-MB-231 tumor cell lines, and a significant reduction in viability in a dose- and time-related manner was observed. Finally, our formulation could be potentially useful as a target doxorubicin delivery system in anticancer therapy.

Equilibrium structure of a triblock copolymer system revealed by mesoscale simulation and neutron scattering

We have performed both mesoscale simulations and neutron scattering experiments on Pluronic L62, a poly(ethylene oxide)–poly(propylene oxide)–poly(ethylene oxide) (PEO–PPO–PEO) triblock copolymer system in aqueous solution. The influence of simulation variables such PEO/PPO block ratio, interaction parameters, and coarse-graining methods is extensively investigated by covering all permutations of parameters found in the literatures. Upon increasing the polymer weight fraction from 50wt% to 90wt%, the equilibrium structure of the isotropic, reverse micellar, bicontinuous, worm-like micelle network, and lamellar phases are respectively predicted from the simulation depending on the choices of simulation parameters. Small angle neutron scattering (SANS) measurements show that the same polymer systems exhibit the spherical micellar, lamellar, and reverse micellar phases with the increase of the copolymer concentration at room temperature. Detailed structural analysis and comparison with simulations suggest that one of the simulation parameter sets can provide reasonable agreement with the experimentally observed structures.

Pluronic-poly (acrylic acid)-cysteine/Pluronic L121 mixed micelles improve the oral bioavailability of paclitaxel

The aim of the study is to synthesize a thiolated Pluronic copolymer, Pluronic-poly (acrylic acid)-cysteine copolymer, to construct a mixed micelle system with the Pluronic-poly (acrylic acid)-cysteine copolymer and Pluronic L121 (PL121) and to evaluate the potential of these mixed micelles as an oral drug delivery system for paclitaxel. Compared with Pluronic-poly (acrylic acid)-cysteine micelles, drug-loading capacity of Pluronic-poly (acrylic acid)-cysteine/PL121 mixed micelles was increased from 0.4 to 2.87%. In vitro release test indicated that Pluronic-poly (acrylic acid)-cysteine/PL121 mixed micelles exhibited a pH sensitivity. The permeability of drug-loaded micelles in the intestinal tract was studied with an in situ perfusion method in rats. The presence of verapamil and Pluronic both improved the intestinal permeability of paclitaxel, which further certified the inhibition effect of thiolated Pluronic on P-gp. In pharmacokinetic study, the area under the plasma concentration-time curve (AUC0→∞) of paclitaxel-loaded mixed micelles was four times greater than that of the paclitaxel solution (p < 0.05). In general, Pluronic-poly (acrylic acid)-cysteine/PL121 micelles were proven to be a potential oral drug delivery system for paclitaxel.

Purification of a novel protease enzyme from kesinai plant (Streblus asper) leaves using a surfactant–salt aqueous micellar two-phase system: a potential low cost source of enzyme and purification method

Serine protease from kesinai leaves was purified for the first time by a surfactant–polymer aqueous micellar two-phase system. The effectiveness of different types and concentrations of non-ionic surfactants (Pluronic series and X-114) on the partitioning behaviour of the protease was evaluated. The results showed that the enzyme preferentially partitioned into the bottom surfactant-rich phase, while the hydrophilic amino acid preferred the top aqueous phase. This distribution of the enzyme is due to the hydrophobic interaction of the serine protease with the hydrophobic lid of the micelle core in the bottom phase. The influence of different types of salts (K2SO4, KH2PO4, KCl and KNO3) on the purification and selectivity of the enzyme was determined. The protease partitioning in the bottom phase increased in the presence of KNO3, which confirmed that the salt was able to improve the protein solubility in bottom phase and increase the hydrophobic interaction between the two phases. In addition, the protease from the bottom phase was re-extracted to a new aqueous phase solution to remove and recycle the surfactant. Addition of potassium thiocyanate led to the partitioning of the enzyme in top aqueous phase due to high ionic strength of SCN−, which forced the lighter micellar phase toward the upper position of the system. A high purification factor (10.3) and yield of 92 % of the enzyme were achieved in a solution of 31 % of Pluronic L61 using 0.3 % KNO3 and 50 % crude feedstock at pH 7.0.

A Hybrid Particle‐Field Coarse‐Grained Molecular Model for Pluronics Water Mixtures

AbstractIn the framework of a hybrid particle‐field simulation technique where self‐consistent field (SCF) theory and molecular dynamics (MD) are combined, specific coarse‐grained (CG) models for Pluronic block copolymers are developed. In particular, the behavior of the model in the correct reproduction of micellar and non‐micellar phases has been tested for Pluronic L62 and L64. At different temperature and polymer contents of the water–polymer mixture, the proposed model is able to correctly describe the different morphologies that are experimentally found. The proposed CG models, still very close to atomistic ones, allow the reconstruction of full atomistic structures by applying suitable reverse mapping techniques. This opens the way to all atom description of these systems.magnified image