Microfluidic platform for controlled synthesis of polymeric nanoparticles

A central challenge in the development of drug-encapsulated polymeric nanoparticles is the inability to control the mixing processes required for their synthesis resulting in variable nanoparticle physicochemical properties. Nanoparticles may be developed by mixing and nanoprecipitation of polymers and drugs dissolved in organic solvents with nonsolvents. We used rapid and tunable mixing through hydrodynamic flow focusing in microfluidic channels to control nanoprecipitation of poly(lactic- co-glycolic acid)- b-poly(ethylene glycol) diblock copolymers as a model polymeric biomaterial for drug delivery. We demonstrate that by varying (1) flow rates, (2) polymer composition, and (3) polymer concentration we can optimize the size, improve polydispersity, and control drug loading and release of the resulting nanoparticles. This work suggests that microfluidics may find applications for the development and optimization of polymeric nanoparticles in the newly emerging field of nanomedicine.     

Bioerodible polymeric nanoparticles for targeted delivery of proteic drugs

Significant efforts are being devoted to develop nanotechnology for drug delivery, mainly because of the distinct advantages offered by nanometer-size polymeric systems. Moreover, targeted drug delivery can be obtained by polymer conjugation to biospecific ligands. The present investigation was aimed mainly at determining the targeting ability of hybrid nanoparticles based on synthetic polymer/protein hybrid matrices. These nanoparticles were designed for liver targeted release of proteic drugs with antiviral activity, such as alpha-interferon. Human serum albumin and the monoesters of alternating copolymers of maleic anhydride/alkyl vinyl ethers of oligo(ethylene glycol) were selected as proteic and synthetic components, respectively. Digalactosyl diacyl glycerol, a natural glycolipid selectively recognized by the asialofetuin receptor present on liver hepatocytes was used as active targeting agent. Nanoparticles of 100-300 nm average size were obtained by controlled coprecipitation method. Investigation of nanoparticle surface properties by spectroscopic analysis and by biological tests indicated that the synthesized nanoparticles do expose on their surface targeting moieties that selectively interact with liver hepatocytes receptors.     

Rationally engineered polymeric cisplatin nanoparticles for improved antitumor efficacy

The use of cisplatin, a first line chemotherapy for most cancers, is dose-limited due to nephrotoxicity. While this toxicity can be addressed through nanotechnology, previous attempts at engineering cisplatin nanoparticles have been limited by the impact on the potency of cisplatin. Here we report the rational engineering of a novel cisplatin nanoparticle by harnessing a novel polyethylene glycol-functionalized poly-isobutylene-maleic acid (PEG-PIMA) copolymer, which can complex with cis-platinum (II) through a monocarboxylato and a coordinate bond. We show that this complex self-assembles into a nanoparticle, and exhibits an IC(50) = 0.77 ± 0.11 μM comparable to that of free cisplatin (IC(50) = 0.44 ± 0.09 μM). The nanoparticles are internalized into the endolysosomal compartment of cancer cells, and release cisplatin in a pH-dependent manner. Furthermore, the nanoparticles exhibit significantly improved antitumor efficacy in a 4T1 breast cancer model in vivo, with limited nephrotoxicity, which can be explained by preferential biodistribution in the tumor with reduced kidney concentrations. Our results suggest that the PEG-PIMA-cisplatin nanoparticle can emerge as an attractive solution to the challenges in cisplatin chemotherapy.     

Cytocompatible chitosan-graft-mPEG-based 5-fluorouracil-loaded polymeric nanoparticles for tumor-targeted drug delivery

Biodegradable materials like chitosan (CH) and methoxy polyethylene glycol (mPEG) are widely being used as drug delivery carriers for various therapeutic applications. In this study, copolymer (CH-g-mPEG) of CH and carboxylic acid terminated mPEG was synthesized by carbodiimide-mediated acid amine reaction. The resultant hydrophilic copolymer was characterized by Fourier transform infrared spectroscopy and ¹H NMR studies, revealing its relevant functional bands and proton peaks, respectively. Blank polymeric nanoparticles (B-PNPs) and 5-fluorouracil loaded polymeric nanoparticles (5-FU-PNPs) were formulated by ionic gelation method. Furthermore, folic acid functionalized FA-PNPs and FA-5-FU-PNPs were prepared for folate receptor-targeted drug delivery. FA-5-FU-PNPs were characterized by particle size, zeta potential, and in vitro drug release studies, resulting in 197.7?nm,?+29.9?mv, and sustained drug release of 88% in 24?h, respectively. Cytotoxicity studies were performed for FA-PNPs and FA-5-FU-PNPs in MCF-7 cell line, which exhibited a cell viability of 80 and 41%, respectively. In vitro internalization studies were carried out for 5-FU-PNPs and FA-5-FU-PNPs which demonstrated increased cellular uptake of FA-5-FU-PNPs by receptor-mediated transport. Significant (p?相似文献   

Controlled Fab installation onto polymeric micelle nanoparticles for tuned bioactivity

Abstract

Antibodies and antigen-binding fragments (Fabs) can be used to modify the surface of nanoparticles for enhanced target binding. In our previous work, site-specific conjugation of Fabs to polymeric micelles using conventional methods was limited to approximately 30% efficiency, possibly due to steric hindrance related to macromolecular reactants. Here, we report a new method that enables conjugation of Fabs onto a micelle surface in a controlled manner with up to quantitative conversion of nanoparticle reactive groups. Variation of (i) PEG spacer length in a heterofunctionalized cross-linker and (ii) Fab/polymer feed ratios resulted in production of nanoparticles with a range of Fab densities on the surface up to the theoretical maximum value. The biological impact of variable Fab density was evaluated in vitro with respect to cell uptake and cytotoxicity of a drug-loaded (SN38) targeted polymeric micelle bearing anti-EphA2 Fabs. Fab conjugation increased cell uptake and potency compared with non-targeted micelles, although a Fab density of 60% resulted in decreased uptake and potency of the targeted micelles. Altogether, our findings demonstrate that conjugation strategies can be optimized to allow control of Fab density on the surface of nanoparticles and also that Fab density may need to be optimized for a given cell-surface target to achieve the highest bioactivity.     

SiRNA drug delivery by biodegradable polymeric nanoparticles

RNA interference (RNAi) is an emerging technology in which the introduction of double-stranded RNA (dsRNA) into a diverse range of organisms and cell types causes degradation of the complementary mRNA. It offers a broad spectrum of applications in both biological and medical research. Small interference RNA (siRNA) was recently explored for its therapeutical potential. However, the drug delivery of siRNA oligos is very novel and is in great need of future research. To this end, a biodegradable poly(D,L-lactide-co-glycolide) (PLGA) nanoparticle drug carrier system was prepared to load siRNA oligos with desired physicochemical properties. The nanoparticles were characterized by scanning electron microscopy and laser diffraction particle sizer. The delivery of siRNA into the targeted 293T cells was observed using fluorescent-labeled double-stranded Cy3-oligos. The model siRNA oligos, si-GFP-RNA, were also successfully loaded into PLGA nanoparticles and delivered in 293T cells. The gene silencing effect and the inhibition of GFP expression were investigated using fluorescent microscopy. Both positive and negative controls were used to compare with the new siRNA nanoparticle delivery system. It was found that nanoparticles offered both effective delivery of siRNA and prominent GFP gene silencing effect. Compared to conventional carrier systems, the new biodegradable polymeric nanoparticle system may also offer improved formulation stability, which is practically beneficial and may be used in the future clinical studies of siRNA therapeutics.     

多响应性聚合物微球的制备与光学性质研究

通过分散聚合法合成了形态规则、平均粒径在370nm左右、粒径分布均匀的聚（N-异丙基丙烯酰胺甜丙烯酸（P（NIAM-co-AAc））交联型聚合物微球．在，此基础上选择不同浓度的Tb（Ⅲ）离子与P（NIPAM-co-AAc）微球进行复配，制备了P（NIPAM-co-AAc）-Yb（Ⅲ）的复配微球．通过紫外光谱和荧光光谱对复配微球的表征发现：该复配微球具有温敏性和pH响应性：P（NIPAM-CO-AAc）微球和Tb（Ⅲ）之间发生了有效的能量传递，明显增强了Yb（Ⅲ）的特征荧光发射．     

Spontaneous formation of silver nanoparticles on polymeric supports

Silver nanoparticles with a size range of 2-4 nm were prepared on polyethylene glycol-coupled 2-chlorotrityl resins without using any reducing agents. In contrast to the polyol process, silver nanoparticles were simply reduced in the tetrahydrofuran/chloroform system on the resin without using any alcohols, and the resulting silver nanoparticles exhibited a uniform size distribution. The polyethylene glycol spacer on the resin played an important role in obtaining silver nanoparticles, probably acting as a polydentate-chelating agent for the silver ions.     

Fluorescence near gold nanoparticles for DNA sensing

We investigated fluorescence quenching and enhancement near gold nanoparticles (GNP) of various sizes using fluorescently labeled hairpin DNA probes of different lengths. A closed hairpin caused intimate contact between the fluorophore and the gold, resulting in an efficient energy transfer (quenching). Upon hybridization with complementary DNA, the DNA probes were stretched yielding a strong increase in fluorescence signal. By carefully quantifying the amount of bound fluorescent probes and the GNP concentrations, we were able to determine the quenching and enhancement efficiencies. We also studied the size and distance dependence theoretically, using both FDTD simulations and the Gersten-Nitzan model and obtained a good agreement between experiments and theory. On the basis of experimental and theoretical studies, we report over 96.8% quenching efficiency for all particle sizes tested and a maximal signal increase of 1.23 after DNA hybridization. The described results also demonstrate the potential of gold nanoparticles for label free DNA sensing.     

New polyamide-containing sorbents for one-step isolation of DNA

A composite sorbent modified with fluorinated aromatic polyamide, for single-step isolation of DNA from biological mixtures is developed. The sorbent is prepared by the polymer deposition from solution onto the surface of porous silica. It is shown that the polymer coating with a thickness of 4 nm is formed not only on the outer surface of the carrier particles but also on the inner pore surfaces. It is found that the material produced largely retained porosity of the pristine support and the material is selective in a one-step separation of nucleic acids and proteins by passing the bacterial lysate through a sorbent. The degree of purification of the isolated preparations of DNA is sufficient to be used directly in PCR analysis.     

Formulation of insulin-loaded polymeric nanoparticles using response surface methodology

The objective of this work was to formulate new oral insulin-loaded nanoparticules using the response surface methodology. The insulin nanoparticles were prepared by a water-in-oil-in-water emulsification and evaporation method. The polymers used for the encapsulation were blends of biodegradable poly-epsilon-caprolactone (PCL) and of positively-charged, nonbiodegradable polymer (Eudragis RS®). A central composite design has been built to investigate the effects of three controlled variables: ratio of polymers (PCL/RS ratio), volume, and pH of the aqueous solution of polyvinyl alcohol. The nanoparticles were characterized by measuring the amount of entrapped insulin, the particle size, the polydispersity of the obtained particles, the zeta potential, and the amount of insulin released after 7 hours. A second-order model was evaluated by multiple regression and was statistically tested for each of the studied controlled variable. The obtained polynomials proved efficient to localize an optimal operating area highlighted by the use of three-dimensional response surfaces and their corresponding isoresponse curves. An interesting formulation given by the models was selected, prepared, and evaluated. The corresponding quantity of entrapped insulin was 25 IU per 100 mg of polymer, and the particle size was 350 nm with a polydispersity of 0.21. The quantity of released insulin was 4.8 IU per 100 mg of polymer after 7 hours and the zeta potential was + 44 mV. All these collected values were in perfect accordance with values estimated by the models. Finally, the results suggested that PCL/RS 50/50 nanoparticles might represent a promising formulation for oral delivery of insulin.     

Functionalized magnetic nanoparticles for small-molecule isolation, identification, and quantification

Functionalized magnetic nanoparticles (MNPs) were synthesized to serve as laser desorption/ionization elements as well as solid-phase extraction probes for simultaneous enrichment and detection of small molecules in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis. Two laser-absorbing matrices were each conjugated onto MNP to give MNP@matrix which provided high ionization efficiency and background-free detection in MS leading to unambiguous identification of target small molecules in a complex mixture. MNP@matrix was demonstrated to serve as a general matrix-free additive in MALDI-TOF MS analysis of structurally distinct small molecules. Also, MNP@matrix provides a simple, rapid, and reliable quantitative assay for small molecules by mass without either the use of an internal standard or an isotopic labeling tag. Furthermore, the affinity extraction of small molecules from complex biofluid was achieved by probe protein-conjugated MNP@matrix without laborious purification. We demonstrated that a nanoprobe-based assay is a cost-effective, rapid, and accurate platform for robotic screening of small molecules.     

Hemolytic effect of polymeric nanoparticles: role of albumin

Nanoparticles (NP) have drawn increasing interest from many fields in medicine and are a relatively new class of biomedical products. Because there are concerns about the health effects of nanoparticles, it is important to understand how nanoparticles interact specifically with red blood cells (RBC), a central object in the blood circulation. As numerous studies that have examined NP/RBC interaction concentrated on the hemolytic potential of nanoparticles, we describe an investigation of hemolytic activity of polystyrene nanoparticles (PS-NP) in protein free medium and its modulation by albumin. We found that treatment of RBCs with PS-NP induces hemolysis (dose and particle size dependent) in plasma free medium but not in full plasma or in buffer, which contain albumin. Critical albumin concentration is 0.05% wt. According to our results hemolytic effect of nanoparticles is strongly modulated by protein concentration in the medium.     

Laser-induced modification of polymeric beads coated with gold nanoparticles

A direct laser writing method for modifying colloidal crystals and single colloids is presented. This method takes advantage of the highly efficient conversion of photons into heat exhibited by gold nanoparticles. The easy control of experimental parameters allowed control of the spatial resolution of the patterns. This may open the way to practical applications for the technology.     

Metal oxide nanoparticles as cross-linkers in polymeric hybrid hydrogels

A novel strategy is described for the preparation of polymeric hybrid hydrogels containing metal oxide nanoparticles as cross-linkers. TiO₂ nanoparticles were functionalized by introducing amine groups onto their surfaces. The functionalized metal oxide nanoparticles were covalently bound to the polymer chains of carboxymethylcellulose and appeared to be organized as clusters with dimensions of 30 nm to 250 nm within the hydrogel. This synthesis method, based on the use of functionalized nanoparticles as cross-linkers, is of general application and it allows for the preparation of other kinds of nanoparticle/polymer hybrid hydrogels. These hybrid hydrogels may have potential applications as novel in vitro scaffolds for tissue engineering, in which the inorganic nanoparticles can simulate the nanostructured architecture of the extra-cellular matrix.     

Photoresponsive coumarin-stabilized polymeric nanoparticles as a detectable drug carrier

The ability to create aqueous suspended stable nanoparticles of the hydrophobic homopolymer poly(?‐caprolactone) end‐functionalized with coumarin moieties (CPCL) is demonstrated. Nanoparticles of CPCL are prepared in a continuous manner using nanoprecipitation. The resulting nanoparticles are spherical in morphology, about 40 nm in diameter, and possess a narrow size distribution and excellent stability over 4 months by repulsive surface charge. Nanoparticle size can be easily controlled by manipulating the concentration of CPCL in the solution. The interparticle assembly between the nanoparticles can be reversibly adjusted with photoirradiation due to photoinduced [2 + 2] cyclodimerization and cleavage between the coumarin molecules. In addition, the CPCL nanoparticles show significant cellular uptake without cytotoxicity, and the intrinsic fluorescence of the coumarin functional group permits the direct detection of cellular internalization.     

Investigation of polymeric amphiphilic nanoparticles as antitumor drug carriers

In this paper, polymeric amphiphilic nanoparticles based on oleoyl–chitosan (OCH) with different degrees of substitution (DS, 5%, 11% and 27%) were prepared by Oil/Water emulsification method. Mean diameters of the nanoparticles were 327.4 nm, 255.3 nm and 192.6 nm, respectively. Doxorubicin (DOX) was efficiently loaded into OCH nanoparticles and provided a sustained released after a burst release in PBS. These nanoparticles showed no cytotoxicity to mouse embryo fibroblasts (MEF) and low hemolysis rates (<5%). The results of SDS-PAGE indicated that bovine calf serum (BCS) adsorption on OCH nanoparticles was inhibited by smaller particle size. Cellular uptake was evaluated by incubating fluorescence labeled OCH nanoparticles with human lung carcinoma cells (A549) and mouse macrophages (RAW264.7). Cellular uptake of OCH nanoparticles was time––and concentration––dependent. Finding the appropriate incubation time and concentration of OCH nanoparticles used as drug carriers might decrease phagocytic uptake, increase cancer cell uptake and ultimately improve therapeutic efficiency of antitumor therapeutic agents.     

Electroporation of polymeric nanoparticles: an alternative technique for transdermal delivery of insulin

Purpose: The aim was to study transdermal electroporation of insulin-loaded nanocarriers as a methodology for delivering macromolecules. Methods: The efficacy of electroporation of insulin as solution and nanoparticles was compared in vitro and in vivo. Histology and confocal laser scanning microscopy were used to assess the effects of electroporation on skin structure, whereas the latter also demonstrated the depth of permeation of the nanoparticles. In vivo studies were performed on streptozotocin-diabetic male Wistar rats and compared with subcutaneous administration. Results: A linear increase in insulin flux was noted on increasing the applied voltage (R² = 0.9514), the number of pulses (R² = 0.8515), and the pulse length (R² = 0.9937). Electroporation of nanoparticles resulted in fourfold enhancement in insulin deposition in rat skin in contrast to solution. In vivo studies showed maximum reduction of 77 ± 5% (87.2 ± 6.4 mIU/mL, t = 2 hours) and 85 ± 8% (37.8 ± 10.2 mIU/mL, t = 4 hours) in blood glucose levels for solution and nanoparticles, respectively, with therapeutic levels maintained for 24 and 36 hours. Conclusion: Overall, electroporation of polymeric nanosystems proved to be an ideal alternative to injectable administration.     

Preparation of magnetic polymeric composite nanoparticles by seeded emulsion polymerization

Seeded emulsion polymerization was used to prepare magnetic polymeric composite nanoparticles (MPCNPs) with the aim to successfully encapsulate magnetite particles and to improve particle size distribution (PSD). Microscopical morphology and number-average diameter of hydrophilic magnetite particles (HMPs), magnetic seed latex nanoparticles (MSLNPs) and MPCNPs were observed and analyzed by transmission electron microscope (TEM). Weight-average diameter and PSD of MSLNPs and MPCNPs were also analyzed by TEM. Magnetic properties of MPCNPs were investigated by Vibrating Sample Magnetometry (VSM). The results showed that the encapsulation of magnetite particles was not complete by conventional emulsion polymerization but very successful by seeded emulsion polymerization, the resulted MPCNPs were nanoparticles with much narrower PSD than that of MSLNPs, and exhibited superparamagnetism and possessed a certain level of magnetic response.