Polyvinyl-Pyrrolidone-Coated Silver Nanoparticles-The Colloidal, Chemical, and Biological Consequences of Steric Stabilization under Biorelevant Conditions.

(1) Background: Several properties of silver nanoparticles (AgNPs), such as cytotoxic, anticancer, and antimicrobial activities, have been subjects of intense research; however, important aspects such as nanoparticle aggregation are generally neglected, although a decline in colloidal stability leads to a loss of the desired biological activities. Colloidal stability is affected by pH, ionic strength, or a plethora of biomolecules that interact with AgNPs under biorelevant conditions. (2) Methods: As only a few studies have focused on the relationship between aggregation behavior and the biological properties of AgNPs, here, we have systematically evaluated this issue by completing a thorough analysis of sterically (via polyvinyl-pyrrolidone (PVP)) stabilized AgNPs that were subjected to different circumstances. We assessed ultraviolet-visible light absorption, dynamic light scattering, zeta potential measurements, in vitro cell viability, and microdilution assays to screen both colloidal stability as well as bioactivity. (3) Results: The results revealed that although PVP provided outstanding biorelevant colloidal stability, the chemical stability of AgNPs could not be maintained completely with this capping material. (4) Conclusion: These unexpected findings led to the realization that stabilizing materials have more profound importance in association with biorelevant applications of nanomaterials than just being simple colloidal stabilizers.

The Synthesis Methodology of PEGylated Fe3O4@Ag Nanoparticles Supported by Their Physicochemical Evaluation.

Many investigations are currently being performed to develop the effective synthesis methodology of magnetic nanoparticles with appropriately functionalized surfaces. Here, the novelty of the presented work involves the preparation of nano-sized PEGylated Fe3O4@Ag particles, i.e., the main purpose was the synthesis of magnetic nanoparticles with a functionalized surface. Firstly, Fe3O4 particles were prepared via the Massart process. Next, Ag+ reduction was conducted in the presence of Fe3O4 particles to form a nanosilver coating. The reaction was performed with arabic gum as a stabilizing agent. Sound energy-using sonication was applied to disintegrate the particles' agglomerates. Next, the PEGylation process aimed at the formation of a coating on the particles' surface using PEG (poly(ethylene glycol)) has been performed. It was proved that the arabic gum limited the agglomeration of nanoparticles, which was probably caused by the steric effect caused by the branched compounds from the stabilizer that adsorbed on the surface of nanoparticles. This effect was also enhanced by the electrostatic repulsions. The process of sonication caused the disintegration of aggregates. Formation of iron (II, III) oxide with a cubic structure was proved by diffraction peaks. Formation of a nanosilver coating on the Fe3O4 nanoparticles was confirmed by diffraction peaks with 2θ values 38.15° and 44.35°. PEG coating on the particles' surface was proven via FT-IR (Fourier Transform Infrared Spectroscopy) analysis. Obtained PEG-nanosilver-coated Fe3O4 nanoparticles may find applications as carriers for targeted drug delivery using an external magnetic field.

PEG-OligoRNA Hybridization of mRNA for Developing Sterically Stable Lipid Nanoparticles toward In Vivo Administration.

Lipid nanoparticles (LNPs) exhibit high potential as carriers of messenger RNA (mRNA). However, the arduous preparation process of mRNA-loaded LNPs remains a huge obstacle for their widespread clinical application. Herein, we tackled this issue by mRNA PEGylation through hybridization with polyethylene glycol (PEG)-conjugated RNA oligonucleotides (PEG-OligoRNAs). Importantly, mRNA translational activity was preserved even after hybridization of 20 PEG-OligoRNAs per mRNA. The straightforward mixing of the PEGylated mRNA with lipofectamine LTX, a commercial lipid-based carrier, just by pipetting in aqueous solution, allowed the successful preparation of mRNA-loaded LNPs with a diameter below 100 nm, whereas the use of non-PEGylated mRNA provided large aggregates above 100- and 1000-nm. In vivo, LNPs prepared from PEG-OligoRNA-hybridized mRNA exhibited high structural stability in biological milieu, without forming detectable aggregates in mouse blood after intravenous injection. In contrast, LNPs from non-PEGylated mRNA formed several micrometer-sized aggregates in blood, leading to rapid clearance from blood circulation and deposition of the aggregates in lung capillaries. Our strategy of mRNA PEGylation was also versatile to prevent aggregation of another type of mRNA-loaded LNP, DOTAP/Chol liposomes. Together, our approach provides a simple and robust preparation method to LNPs for in vivo application.

PEGylated cationic liposome-DNA complexation in brine is pathway-dependent.

Cationic liposome-DNA (CL-DNA) complexes, are regarded as promising materials for safe and efficient delivery of genes for therapeutical applications. In order to be used in vivo, these complexes may be coated with a hydrophilic polymer (e.g. polyethylene-glycol, PEG) that provides steric stabilization towards adhesion of proteins and removal by the immune system. In this work we study the influence of the initial salt concentration (Cs) - which modulates the electrostatic interaction between oppositely charged vesicles and DNA - on the structure and stability of PEGylated CL-DNA particles. Previous small-angle X-ray scattering has shown that if non-PEGylated or PEGylated CL-DNA lamellar complexes are prepared in water, their structure is well defined with a high number of lipid membrane-DNA layers (larger than 20). Here we show that if these complexes are transferred to saline media (150mM NaCl or DMEM, both near physiological conditions), this structure remains nearly unchanged. Conversely, if PEGylated complexes are prepared in saline media, their lamellar structure is much looser, with fewer number of layers. This pathway dependent behavior of PEGylated complex formation in brine is modulated by the liposome membrane charge density and the mole fraction of PEG 2000 in the membranes, with the average number of layers decreasing with increasing Cs and in going from 5mol% to 10mol% PEG-lipid. Each of these structures (high and low number of layers) is stable with time, suggesting that despite complex formation being thermodynamically favored, the complexation process in PEGylated membranes, which determines the number of layers per particle, is kinetically controlled. In the extreme case (when polymer repulsions from 10mol% PEG-lipid are maximized and electrostatic attraction between PEGylated CLs and DNA are minimized at low membrane charge density) complex formation is suppressed at high Cs=150mM.

Dispersible carbon nanotubes.

A method is proposed to produce nanoparticles dispersible and recyclable in any class of solvents, and the concept is illustrated with the carbon nanotubes. Classically, dispersions of CNTs can be achieved through steric stabilization induced by adsorbed or grafted polymer chains. Yet, the surface modification of CNTs surfaces is irreversible, and the chemical nature of the polymer chains imposes the range of solvents in which CNTs can be dispersed. To address this limitation, supramolecular bonds can be used to attach and to detach polymer chains from the surface of CNTs. The reversibility of supramolecular bonds offers an easy way to recycle CNTs as well as the possibility to disperse the same functional CNTs in any type of solvent, by simply adapting the chemical nature of the stabilizing chains to the dispersing medium. The concept of supramolecular functionalization can be applied to other particles, for example, silica or metal oxides, as well as to dispersing in polymer melts, films or coatings.

Preparation of liquid yogurt in the presence of pectin and its formation mechanism.

We had previously observed that adding pectin into milk before fermentation inhibited gelation of yogurt but did not affect the pH. Thus, this work aimed to prepare such liquid yogurt and clarify its formation mechanism. It was found that liquid yogurt was obtained in the presence of 0.10%-0.20% pectin. However, at lower or higher pectin concentrations, yogurt was gelled. Confocal laser scanning microscopy analysis demonstrated that 0.10%-0.20% pectin induced milk protein aggregating into separated particles rather than a continuous network, which explained why liquid yogurt was formed. Moreover, adding 0.10%-0.20% pectin into the casein micelle suspension induced aggregation of casein micelles at pH 6.8. After pH decreased to 4.3, casein micelles showed more aggregation but they were still separated particles, which was the same in the corresponding yogurt samples. These results suggested that pectin changed the aggregation mode of casein micelles and induced formation of liquid yogurt.

Development, Characterization and Pharmacological Evaluation of Cannabidiol-Loaded Long Circulating Niosomes.

Cannabidiol (CBD) is a promising drug candidate with pleiotropic pharmacological activity, whose low aqueous solubility and unfavorable pharmacokinetics have presented obstacles to its full clinical implementation. The rational design of nanocarriers, including niosomes for CBD encapsulation, can provide a plausible approach to overcoming these limitations. The present study is focused on exploring the feasibility of copolymer-modified niosomes as platforms for systemic delivery of CBD. To confer steric stabilization, the niosomal membranes were grafted with newly synthesized amphiphilic linear or star-shaped 3- and 4-arm star-shaped copolymers based on polyglycidol (PG) and poly(ε-caprolactone) (PCL) blocks. The niosomes were prepared by film hydration method and were characterized by DLS, cryo-TEM, encapsulation efficacy, and in vitro release. Free and formulated cannabidiol were further investigated for cytotoxicity and pro-apoptotic and anti-inflammatory activities in vitro in three human tumor cell lines. The optimal formulation, based on Tween 60:Span60:Chol (3.5:3.5:3 molar ration) modified with 2.5 mol% star-shaped 3-arm copolymer, is characterized by a size of 235 nm, high encapsulation of CBD (94%), and controlled release properties. Niosomal cannabidiol retained the antineoplastic activity of the free agent, but noteworthy superior apoptogenic and inflammatory biomarker-modulating effects were established at equieffective exposure vs. the free drug. Specific alterations in key signaling molecules, implicated in programmed cell death, cancer cell biology, and inflammation, were recorded with the niosomal formulations.

Shielding behavior of electrokinetic properties of polystyrene latex particle by the adsorption of neutral poly(ethylene oxide).

To understand the shielding of electrokinetics of colloidal particles by polymer coating, we measured the electrophoretic mobility of negatively charged polystyrene sulfate latex (PSL) adsorbed with electrostatically neutral polyethylene oxide (PEO) chains with various molecular weights under different ionic strengths. We confirmed that substantial adsorbed neutral polymer on the particle surface would decrease the absolute value of electrophoretic mobility. Even though the polymer layer is sufficiently thicker compared to the thickness of electric double layer (EDL), the electrophoretic mobility (EPM) never vanishes, which indicates the incompleteness of electrokinetic shielding by an adsorbed neutral polymer. To relate such interesting phenomena, a simple mathematical model has been proposed to evaluate the electrophoretic mobility, assuming the presence of a scaling structure of adsorbed permeable polymer layer does not influence the Poisson-Boltzmann distribution of ions in the electric double layer (EDL). An analytical expression of electrophoretic mobility under Debye-Hu¨ckel approximation has been derived using the method of Ohshima-Kondo theory, which successfully justifies the experimentally obtained data.

Drug localization and its effect on the physical stability of poloxamer 188-stabilized colloidal lipid emulsions.

Colloidal lipid emulsions are a promising formulation option for poorly water-soluble drugs. Due to their complex composition, they provide different sites for the localization of drugs. Drug molecules can be situated in the lipid matrix, in the aqueous phase with its structures formed by an excess of emulsifier or at the droplet interface. The interface and the mechanism of stabilization is mainly characterized by the emulsifier. In this study, the main focus was on the influence of drug localization on the stability of emulsions sterically stabilized with poloxamer188. In addition to 5% of this non-ionic emulsifier, the emulsions contained 10% soybean oil. The localization of the drugs fenofibrate, curcumin, betamethasone valerate, cinnarizine, dibucaine and flufenamic acid within the emulsion system at a physiological pH of 7.4 as well as their influence on emulsion stability were examined. The results indicated that the stability of poloxamer 188-stabilized emulsions can be influenced in a positive or negative way by the localization of drug molecules in the interface of emulsion droplets. Applying cinnarizine as model substance at pH 5, 7.4 and 10, no pronounced change in the localization was detected as a result of alterations in the charge of the drug.

Colorimetric assay for the determination of molecular weight distribution and branching characteristics of starch hydrolysates.

Here, we present a gold nanoparticle-based colorimetric assay for the determination of molecular weight distribution and branching characteristics of enzymatically hydrolyzed starch. The steric stabilization effect of starch hydrolysate on the colloidal stability of AuNPs was found to be proportional to the ratio of high molecular weight amylopectins, which was clearly reflected by the intensity of the characteristic surface plasmonic resonance (SPR) absorbance peak of the AuNPs. The fractional change of high molecular weight amylopectin over the course of enzymatic hydrolysis reaction could be measured based on the intensity of SPR peak, in which the results correlated well with those obtained by conventional gel permeation chromatography. With the proper calibration of a specific set of enzyme and starch type, this method would provide a fairly simple and fast means of analyzing the molecular weight distribution of starch hydrolysate on site as well as the amylose content in native starch.

Tailor-Made Single-Core PLGA Microbubbles as Acoustic Cavitation Enhancers for Therapeutic Applications.

Microbubbles (MBs), being gas bubbles encapsulated inside a solid shell, have been investigated extensively in the field of therapeutic ultrasound as acoustic cavitation enhancers. Hard-shell MBs have an advantage over soft-shell MBs due to their improved stability. Poly(lactic-co-glycolic acid) (PLGA) is one of the most attractive polymers for hard-shell MB synthesis; however, very little is known regarding the effect of synthesis parameters on the acoustic cavitation activity of PLGA MBs and the tunability of this activity. In this study, by manipulating the synthesis parameters, we were able to control the characteristics of the MBs, such as their internal structure, gas core, size distribution, and shell thickness, which significantly affect the total acoustic cavitation activity that they exhibit (i.e., their cavitation dose). We showed that single-core MBs filled with C3F8 gas can produce cavitation effects for extended periods under continuous circulation. These MBs exhibited high stability, and their cavitation activity was not affected by prior circulation in the system. Preliminary in vivo results demonstrated that intravenously injected MBs did not cause adverse effects and produced cavitation activity that increased the permeability of the pig blood-brain barrier. Although more tests should be performed to evaluate the MB long-term safety and activity in vivo, these encouraging results suggest that our PLGA MBs have potential for future therapeutic applications as cavitation enhancers.

To what extent do polymeric stabilizers affect nanoparticles characteristics?

Colloidal synthesis of nanoparticles using polymeric stabilizers as a template of a structure directing agent provided a plethora of opportunities in fabricating nanoparticles (NPs) with controlled size, shape, composition and structural characteristics. To understand the complete potency of polymeric stabilizers during the synthesis of nanoparticles, the relationship between polymer characteristics such as structure, molecular weight and concentration and nanoparticles characteristics is discussed in depth. This review portrays the use of polymers to attain nanostructured materials via covalent and non-covalent approaches. These polymers can also serve as surfaces modifier as well as the growth regulators during the synthesis of nanomaterials. The effect provided by polymers that directs the formation of nanomaterials into desired forms is otherwise hard to achieve. We especially spotlight on the approaches for tuning the characteristic properties of nanoparticles via cautious choice of the polymer system with special focus to stimuli-responsive polymers. This review mainly focusses on answering the main challenging question; what is the ideal polymeric stabilizer system to obtain specific morphology, size and phase structure of nanoparticles? Such vital information will enable rational design of nanoparticles to meet specific needs for different applications.

Ginkgolides-loaded soybean phospholipid-stabilized nanosuspension with improved storage stability and in vivo bioavailability.

The purpose of this study was to investigate the effects of soybean phospholipid, as a steric stabilizer, on improving dissolution rate, storage stability and bioavailability of ginkgolides. The ginkgolides coarse powder, hydroxypropyl methylcellulose (HPMC), soybean phospholipid and sodium dodecyl sulfate (SDS) were mixed and wet-milled to prepare nanosuspension S1. Nanosuspension S2 was obtained by the same technique except adding the soybean phospholipid. Results of particle size showed that particle size (D50) of S1 significantly decreased from 44.25 µm to 0.373 µm. Results of differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD) and transmission electron microscope (TEM) showed that ginkgolides in nanosuspension still maintained its crystallinity, and the nanoparticles were all nearly circular and uniformly dispersed. Then, pellets F1 and F2 were prepared by layering S1 and S2 onto the microcrystalline cellulose (MCC) spheres, respectively. The dissolution rate of ginkgolide A (GA) and ginkgolide B (GB) in F1 was 98.3% and 97.7% in 30 min, respectively. It was much higher than F2 (89.0% and 86.5%) and coarse powder of ginkgolides (22.3% and 24.6%). According to the results of stability test, the storage stability of F1 was improved compared with F2. In addition, compared with coarse powder of ginkgolides, the relative bioavailability of GA and GB in F1 were up to (221.84 ±â€¯106.67) % and (437.45 ±â€¯336.43) %, respectively. The above results demonstrated that soybean phospholipid added to the nanosuspension played an important role in improving drug dissolution rate, storage stability and in vivo bioavailability: (1) The amphiphilic soybean phospholipid interacted with the drug, with the hydrophobic part adsorbed on the surface of the poorly soluble drug and the hydrophilic part exposed to the aqueous medium. This increases the wettability of the nanoparticles, which ensure a good redispersibility of the drug particles. (2) It could self-assemble to form an interfacial phospholipid film by surrounding the individual nanoparticles, which can produce enough steric hindrance to prevent nanoparticles from aggregation and ensure a rapid dissolution rate. (3) Soybean phospholipid and its hydrolysate formed strong micellar solubilizing vehicles with bile salts in vivo, stimulated the absorption process of ginkgolides. Thus, soybean phospholipid was a promising steric stabilizer in nanosuspension drug delivery system.

Synthesis and application of PEGylated tracer particles for measuring protein solution viscosities using Dynamic Light Scattering-based microrheology.

The measurement of flow properties, such as the zero shear viscosity, of protein solutions is of paramount importance for many applications such as pharmaceutical formulations, where the syringeability of physiologically effective doses is a key property. However, the determination of these properties with classical rheological methods is often challenging due to e.g. detrimental surface effects or simply the lack of sufficient material. A possible alternative is Dynamic Light Scattering-based microrheology, where the Brownian motion of tracer particles embedded in the protein solution is monitored to access the zero shear viscosity of the sample. The prime advantages of this method compared to classical rheology are the absence of disturbing surface effects and the up to two orders of magnitude smaller protein quantities needed for an entire concentration series. This Protocol provides a detailed description of the synthesis of sterically stabilized tracer particles with surface and overall particle properties specifically designed to investigate the viscosity of protein solutions up to concentrations close to the arrest transition. These particles are tailored to avoid protein-particle as well as particle-particle aggregation at various sample conditions and thus allow for an artifact-free application of Dynamic Light Scattering-based tracer microrheology to determine the flow behaviour of biological samples. The Protocol concludes with step by step instructions for the characterization of protein solutions using a combination of the tracer particles and an advanced dynamic light scattering technique yielding the concentration-dependent zero shear viscosity.

Site-specific pharmaco-laser therapy: A novel treatment modality for refractory port wine stains.

Despite extensive efforts to optimize laser therapy, i.e., the current gold standard treatment, a majority of port wine stain (PWS) patients responds suboptimally to laser therapy. This paper describes the niceties of a novel PWS treatment modality termed site-specific pharmaco-laser therapy (SSPLT). In contrast to the classic approach of enhancing the extent of intravascular photocoagulation (the photothermal response), SSPLT focuses on optimization of post-irradiation thrombus formation (i.e., the hemodynamic response) by combining conventional laser therapy with the administration of thermosensitive drug delivery systems that encapsulate prothrombotic and antifibrinolytic drugs. The aim of SSPLT is to instill complete lumenal occlusion in target vessels, which has been linked to optimal PWS blanching. RELEVANCE FOR PATIENTS: The current treatment options for PWS patients are limited in efficacy. Novel therapeutic modalities are needed to more effectively treat patients with recalcitrant PWSs. SSPLT is an experimental-stage treatment modality that could serve as an adjuvant to pulsed dye laser therapy for a selected group of patients whose PWS is ill-responsive to standard treatment. The expected clinical result of SSPLT is improved lesional blanching.

Insights into the EDC-mediated PEGylation of cellulose nanofibrils and their colloidal stability.

EDC-mediated coupling has frequently been utilized to poly(ethylene glycol) functionalize (PEGylate) cellulose-based materials, but no work has previously been reported on the direct N-(3-dimethylaminopropyl)-N-ethylcarbodiimide (EDC)-mediated PEGylation of cellulose nanofibrils (CNF). Herein, we report the first study where CNF has been directly sterically stabilized with amine-terminated PEG employing N-hydroxysuccinimide (NHS)-assisted EDC-coupling. This work has shown that this coupling reaction is highly sensitive to the reaction conditions and purification procedures, and hence an optimized coupling protocol was developed in order to achieve a reaction yield. Elemental analysis of the nitrogen content also showed the successful PEGylation. It was also shown that a surprisingly low PEGylation (1%) is sufficient to significantly improve the colloidal stability of the PEGylated samples, which reached dispersion-arrested-state-transitions at higher concentrations than neat CNF. The colloidal stability was preserved with increasing ionic strength, when comparably long polymer chains were grafted, targeting only 1% PEGylation.

Shear Thickening Electrolyte Built from Sterically Stabilized Colloidal Particles.

We present a method to prepare shear thickening electrolytes consisting of silica nanoparticles in conventional liquid electrolytes with limited flocculation. These electrolytes rapidly and reversibly stiffen to solidlike behaviors in the presence of external shear or high impact, which is promising for improved lithium ion battery safety, especially in electric vehicles. However, in initial chemistries the silica nanoparticles aggregate and/or sediment in solution over time. Here, we demonstrate steric stabilization of silica colloids in conventional liquid electrolyte via surface-tethered PMMA brushes, synthesized via surface-initiated atom transfer radical polymerization. The PMMA increases the magnitude of the shear thickening response, compared to the uncoated particles, from 0.311 to 2.25 Pa s. Ultrasmall-angle neutron scattering revealed a reduction in aggregation of PMMA-coated silica nanoparticles compared to bare silica nanoparticles in solution under shear and at rest, suggesting good stabilization. Conductivity tests of shear thickening electrolytes (30 wt % solids in electrolyte) at rest were performed with interdigitated electrodes positioned near the meniscus of electrolytes over the course of 24 h to track supernatant formation. Conductivity of electrolytes with bare silica increased from 10.1 to 11.6 mS cm-1 over 24 h due to flocculation. In contrast, conductivity of electrolytes with PMMA-coated silica remained stable at 6.1 mS cm-1 over the same time period, suggesting good colloid stability.

pH-sensitive PEGylation of RIPL peptide-conjugated nanostructured lipid carriers: design and in vitro evaluation.

BACKGROUND: RIPL peptide (IPLVVPLRRRRRRRRC)-conjugated nanostructured lipid carriers (RIPL-NLCs) can facilitate selective drug delivery to hepsin (Hpn)-expressing cancer cells, but they exhibit low stability in the blood. Generally, biocompatible and nontoxic poly(ethylene glycol) surface modification (PEGylation) can enhance NLC stability, although this may impair drug delivery and NLC clearance. To attain RIPL-NLC steric stabilization without impairing function, pH-sensitive cleavable PEG (cPEG) was grafted onto RIPL-NLCs (cPEG-RIPL-NLCs). METHODS: Various types of NLC formulations including RIPL-NLCs, PEG-RIPL-NLCs, and cPEG-RIPL-NLCs were prepared using the solvent emulsification-evaporation method and characterized for particle size, zeta potential (ZP), and cytotoxicity. The steric stabilization effect was evaluated by plasma protein adsorption and phagocytosis inhibition studies. pH-sensitive cleavage was investigated using the dialysis method under different pH conditions. Employing a fluorescent probe (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate [DiI]), in vitro drug delivery capacity of the cPEG-RIPL-NLCs under different pH conditions was also performed on Hpn-expressing SKOV3 cells and 3D-tumor spheroids. RESULTS: All prepared NLCs showed homogenous dispersion (<220 nm in size) with a negative ZP (-18 to -22 mV), except for positively charged RIPL-NLCs (~10 mV), revealing no significant cytotoxicity in either SKOV3 or RAW 264.7 cell lines. cPEG-RIPL-NLC protein adsorption was 1.75-fold less than that of RIPL-NLCs, and PEGylation significantly reduced the macrophage uptake. PEG detachment from the cPEG-RIPL-NLCs was pH-sensitive and time dependent. At 2 hours incubation, cPEG-RIPL-NLCs and PEG-RIPL-NLCs exhibited comparable cellular uptake at pH 7.4, whereas cPEG-RIPL-NLC uptake was increased over 2-fold at pH 6.5. 3D-spheroid penetration also demonstrated pH-sensitivity: at pH 7.4, cPEG-RIPL-NLCs could not penetrate deep into the spheroid core region during 2 hours, whereas at pH 6.5, high fluorescence intensity in the core region was observed for both cPEG-RIPL-NLC-and RIPL-NLC-treated groups. CONCLUSION: cPEG-RIPL-NLCs are good candidates for Hpn-selective drug targeting in conjunction with pH-responsive PEG cleavage.

Sterically Stabilized RIPL Peptide-Conjugated Nanostructured Lipid Carriers: Characterization, Cellular Uptake, Cytotoxicity, and Biodistribution.

As a platform for hepsin-specific drug delivery, we previously prepared IPLVVPLRRRRRRRRC peptide (RIPL)-conjugated nanostructured lipid carriers (RIPL-NLCs) composed of Labrafil® M 1944 CS (liquid oil) and Precirol® ATO 5 (solid lipid). In this study, to prevent the recognition by the mononuclear phagocyte system, polyethylene glycol (PEG)-modified RIPL-NLCs (PEG-RIPL-NLCs) were prepared using PEG3000 at different grafting ratios (1, 5, and 10 mole %). All prepared NLCs showed a homogeneous dispersion (130⁻280 nm), with zeta potentials varying from -18 to 10 mV. Docetaxel (DTX) was successfully encapsulated in NLCs: encapsulation efficiency (93⁻95%); drug-loading capacity (102⁻109 µg/mg). PEG-RIPL-NLCs with a grafting ratio of 5% PEG or higher showed significantly reduced protein adsorption and macrophage phagocytosis. The uptake of PEG(5%)-RIPL-NLCs by cancer cell lines was somewhat lower than that of RIPL-NLCs because of the PEG-induced steric hindrance; however, the uptake level of PEG-RIPL-NLCs was still greater than that of plain NLCs. In vivo biodistribution was evaluated after tail vein injection of NLCs to normal mice. Compared to RIPL-NLCs, PEG(5%)-RIPL-NLCs showed lower accumulation in the liver, spleen, and lung. In conclusion, we found that PEG(5%)-RIPL-NLCs could be a promising nanocarrier for selective drug targeting with a high payload of poorly water-soluble drugs.

Simple and High Yield Synthesis of Metal-Polymer Nanocomposites: The Role of Theta-Centrifugation as an Essential Purification Step.

Nanocomposites are an important materials class, which strives to foster synergistic effects from the intimate mixture of two vastly different materials. Inorganic nanoparticles decorated with polymer ligands, for instance, aim to combine the processing flexibility of polymers with the mechanical robustness of solid state materials. The fabrication and purification of such composite nanoparticles, however, still presents a synthetic challenge. Here, we present a simple synthesis of silver polystyrene nanocomposites with a controllable interparticle distance. The interparticle distance can be well-controlled with a few nanometer precision using polystyrene ligands with various molecular weights. The nanoparticle and polymer ligand synthesis yield both materials on gram scales. Consequently, the polymer nanocomposites can also be fabricated in such large amounts. Most importantly, we introduce Θ-centrifugation as a purification method, which is capable of purifying large nanocomposite batches in a reproducible manner. We employ a range of characterization methods to prove the successful purification procedure, such as transmission electron microscopy, thermogravimetric analysis, and dynamic light scattering. Our contribution will be of high interest for many groups working on nanocomposite materials, where the sample purification has been a challenge up to now.