Non-functionalized Particles Research Articles

Angiopep-2-Functionalized Lipid Cubosomes for Blood-Brain Barrier Crossing and Glioblastoma Treatment.

Glioblastoma multiforme (GBM) is an aggressive brain cancer with high malignancy and resistance to conventional treatments, resulting in a bleak prognosis. Nanoparticles offer a way to cross the blood-brain barrier (BBB) and deliver precise therapies to tumor sites with reduced side effects. In this study, we developed angiopep-2 (Ang2)-functionalized lipid cubosomes loaded with cisplatin (CDDP) and temozolomide (TMZ) for crossing the BBB and providing targeted glioblastoma therapy. Developed lipid cubosomes showed a particle size of around 300 nm and possessed an internal ordered inverse primitive cubic phase, a high conjugation efficiency of Ang2 to the particle surface, and an encapsulation efficiency of more than 70% of CDDP and TMZ. In vitro models, including BBB hCMEC/D3 cell tight monolayer, 3D BBB cell spheroid, and microfluidic BBB/GBM-on-a-chip models with cocultured BBB and glioblastoma cells, were employed to study the efficiency of the developed cubosomes to cross the BBB and showed that Ang2-functionalized cubosomes can penetrate the BBB more effectively. Furthermore, Ang2-functionalized cubosomes showed significantly higher uptake by U87 glioblastoma cells, with a 3-fold increase observed in the BBB/GBM-on-a-chip model as compared to that of the bare cubosomes. Additionally, the in vivo biodistribution showed that Ang2 modification could significantly enhance the brain accumulation of cubosomes in comparison to that of non-functionalized particles. Moreover, CDDP-loaded Ang2-functionalized cubosomes presented an enhanced toxic effect on U87 spheroids. These findings suggest that the developed Ang2-cubosomes are prospective for improved BBB crossing and enhanced delivery of therapeutics to glioblastoma and are worth pursuing further as a potential application of nanomedicine for GBM treatment.

Composite Containing Calcium Phosphate Particles Functionalized with 10-MDP

The phosphate ester monomer 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP) is capable of bonding to hydroxyapatite and, for this reason, is a key component of several self-etch adhesives. In this study, dicalcium phosphate dihydrate particles (DCPD; CaHPO4.2H2O) were functionalized with 10-MDP and used to formulate an experimental composite with 50 vol% inorganic content (3:1 DCPD:silanated barium glass ratio) dispersed in a BisGMA/TEGDMA matrix. The tested hypothesis was that DCPD functionalization would improve the composite’s mechanical performance without compromising Ca2+ release. Composites containing nonfunctionalized DCPD or only reinforcing glass (in both cases, with or without 10-MDP mixed in the resin phase) were used as controls. Materials were tested for degree of conversion (DC; by Fourier transform infrared spectroscopy), water sorption (WS) and solubility (SL; according to ISO 4049), biaxial flexural strength (BFS)/modulus (FM) after 24 h and 5 mo in water, and 28-d Ca2+ release in water (by plasma-coupled optical emission spectroscopy). Data were analyzed using analysis of variance/Tukey test (alpha: 5%). DCPD functionalization did not interfere with DC. The composite containing functionalized DCPD showed significantly lower WS and SL in comparison with the material formulated with nonfunctionalized particles. The presence of 10-MDP (as a functionalizing agent or dispersed in the resin phase) reduced the composite’s initial BFS and FM. After 5 mo in water, the composite with functionalized DCPD and both glass-only composites were able to maintain their mechanical properties at levels statistically similar to what was observed after 24 h. Ca2+ release was significantly reduced in both formulations containing 10-MDP. In conclusion, DCPD functionalization with 10-MDP increased the composite’s resistance to hydrolytic degradation, improving its mechanical stability after prolonged water storage. However, the impaired water transit at the particle-matrix interface led to a reduction in Ca2+ release.

Effects of polystyrene micro/nanoplastics on liver cells based on particle size, surface functionalization, concentration and exposure period.

Micro/nanoplastics (MP/NP) contaminate our food and drinking water but their impact on human health has not been well-documented. The liver is one of the first organs that ingested MP/NP encounter and it has a major role in the clearance of xenobiotics. Therefore, the effects of polystyrene MP/NP on liver HepG2 cells were studied. Cellular responses to particles of various sizes (50-5000 nm) and surface functionalization (aminated, carboxylated or non-functionalized) were determined at different concentrations (0.1-100 μg/mL) and exposure periods (1-24 h). Smaller sized particles were internalized by HepG2 cells more avidly than larger particles regardless of functionalization; the highest uptake being for 50 and 100 nm aminated particles at lower concentrations. Confocal microscopy images of cells corroborated quantitative uptake results. Aminated particles were more toxic to the cells than carboxylated or non-functionalized particles. Among aminated particles smaller particles (50 and 100 nm) were more detrimental to cell viability compared to larger particles (1000 or 5000 nm) with toxicity increasing with concentration. Treatment with the particles for 4 h increased intracellular concentrations of Caspase-3 by 1.5-2.8 fold, but 24 h exposure to the particles attenuated this increase in Caspase-3 concentrations. A slight trend of higher Caspase-3 concentration in cells treated with larger particles (500-5000 nm) compared to smaller particles (50-200 nm) was observed, indicating that larger particles are more likely to direct cells toward apoptotic cell death upon 4 h exposure. Exposure of cells to large PS particles (500-5000 nm) upregulated interleukin-8 and the effect was enhanced at 24 h. Overall, the study demonstrated that smaller aminated particles were most toxic to hepatocytes, but larger particles induced apoptotic cell death or an inflammatory response depending on the length of exposure.

Uptake and toxicity of polystyrene micro/nanoplastics in gastric cells: Effects of particle size and surface functionalization.

Toxicity of micro or nanoplastics (MP/NP) in aquatic life is well-documented, however, information about the consequences of exposure to these particles in terrestrial species is scarce. This study was used to evaluate the uptake and/or toxicity of polystyrene MP/NP in human gastric cells, comparing doses, particle sizes (50, 100, 200, 500, 1000 or 5000 nm) and surface functionalization (aminated, carboxylated or non-functionalized). In general, the uptake of 50 nm particles was significantly higher than 1000 nm particles. Among the 50 nm particles, the aminated particles were more avidly taken up by the cells and were cytotoxic at a lower concentration (≥ 7.5 μg/mL) compared to same sized carboxylated or non-functionalized particles (≥ 50 μg/mL). High toxicity of 50 nm aminated particles corresponded well with significantly high rates of apoptosis-necrosis induced by these particles in 4 h (29.2% of total cells) compared to all other particles (≤ 16.8%). The trend of apoptosis-necrosis induction by aminated particles in 4 h was 50 > 5000 > 1000 > 500 > 200 > 100 nm. The 50 nm carboxylated or non-functionalized particles also induced higher levels of apoptosis-necrosis in the cells compared to 100, 1000 and 5000 nm particles with same surface functionalization but longer exposure (24 h) to 50 nm carboxylated or non-functionalized particles significantly (p<0.0001) increased apoptosis-necrosis in the cells. The study demonstrated that the toxicity of MP/NP to gastric cells was dependent on particle size, dose surface functionalization and exposure period.

Surface functionalized GdVO4:Eu3+ nanocrystals as turn-off luminescent probe for selective sensing of Cu2+ ions: role of pH

Poly(ethylene glycol) bis(carboxymethyl) ether (PEG-diacid) functionalized GdVO4:Eu3+ (5 at%) nanocrystals have been synthesized by polyol method. X-ray diffraction and transmission electron microscopy were used for the structural and morphology characterization of GdVO4:Eu3+ samples. Fourier transform infrared spectroscopy confirms the functionalization of nanocrystal surface by PEG-diacid. The resonance energy transfer from the vanadate (VO43−) to Eu3+ results in enhanced red emission. The GdVO4:Eu3+ as luminescent probe can selectively detect the presence of Cu2+ ions out of various metal ions investigated. Upon addition of Cu2+ ions, photoluminescence intensity of Eu3+ reduces instantaneously more than 90%, which is related to energy transfer from Eu3+ to Cu2+ ions. This shows the fast response during sensing. Surface functionalized particles shows a significant sensitivity as compared to the non-functionalized particles. These probes show more favourable in the alkaline aqueous medium for sensing. The possibility of reabsorption is ruled out. The limit of detection is about 9 nM of Cu2+ with the first response time and this method will be useful in selective sensing of Cu2+ ions in aqueous medium.

Mask-less preparation of Janus particles through ultraviolet irradiation on hydrophobic particles assembled at the air-water interface

Janus particles are a class of materials that exhibit two contrasting properties. They possess unique properties, such as the ability to self-assemble at dissimilar liquid interfaces. Achieving high production yields remains a challenge to the practical application of Janus particles, as existing methods are generally tedious in the preparation procedures. The aim of this work is to prepare Janus particles using a process that is simple to facilitate scaling up. Semiconductor titanium dioxide (TiO2) and cuprous oxide (Cu2O) particles, pre-functionalized with hydrophobic coatings, were assembled at an air-water interface. The particles were then subjected to ultraviolet irradiation, which degraded the hydrophobic coating on the exposed side, and subsequently recovered through the removal of water. Janus particles that contained contrasting wettability over their surfaces were successfully obtained; the particle surfaces were demonstrated to be partially functionalized through comparison in the self-assembly behavior with fully-functionalized and non-functionalized particles. The obtained TiO2 Janus particles have demonstrated a Janus balance factor close to 1, and remained attached to the oil/water interface with long-term stability. The presence of a photocatalytic substrate was found to be necessary to accelerate the degradation of the coating. Different alkylsilane coatings, particle types, and irradiation durations were examined, leading to different types of Janus particles. The method can be potentially extended to other semiconductor metal oxide particles to generate Janus-like properties.

The effect of polymer and gold functionalization on the magnetic properties of magnetite nanoparticles

Silica coated magnetite particles ([email protected]) functionalized with gold [email protected] + Au), or gold plus poly (vinylpyrrolidone) (PVP) (Fe3O[email protected] + Au + PVP) were synthesized. Their structural and magnetic properties were studied using a combination of experimental techniques including electron micr oscopy (EM), superconducting quantum interference device (SQUID) magnetrometry, and electron paramagnetic resonance (EPR) spectroscopy. The saturation magnetization (MS) of particles functionalized with gold and gold plus PVP were found to be 67.24 emu/g and 65.78 emu/g respectively. Both functionalized ensembles maintained a large percentage (78–80%) of their MS values compared to pristine magnetite. The coercivity (HC) for pristine magnetite was 227.25 Gauss compared to 200.00 Gauss for [email protected] + Au and 228.57 Gauss for [email protected] + Au + PVP. Furthermore, these magnetic particles, being biologically compatible and resistant to oxidation, were functionalized with an antibody designed to target A431 oral cancer cells. The result demonstrates high specificity of binding compared to non-functionalized particles, attributable to a favorable interaction between gold and the antibody making them excellent candidates for applications like bio-separation and imaging.

Development of calcium phosphate/ethylene glycol dimethacrylate particles for dental applications

This study describes the synthesis of dicalcium phosphate dihydrate (DCPD) particles in the presence of different ethylene glycol dimethacrylates (EGDMA, ethylene glycol/EG units: 1, 2, 3 or 4) at two monomer-to-ammonium phosphate molar ratios (1:1 and 2:1), as a strategy to develop CaP-monomer particles with improved interaction with resin matrices. Particles displaying high surface areas and organic contents were added to a photocurable BisGMA-TEGDMA resin and the resulting materials were tested for degree of conversion (DC), biaxial flexural strength (BFS), flexural modulus, and ion release. Data were subjected to one-way ANOVA or Kruskal-Wallis/Dunn test (alpha: 0.05). Functionalization with EGDMA derivatives was dependent upon the length of the spacer group and monomer concentration in the synthesis. No differences in DC were observed among materials (p > 0.05). A 39% increase in BFS was obtained with the use of particles with the highest functionalization level compared to non-functionalized particles (p < 0.001). The use of functionalized DCPD reduced flexural modulus in comparison to non-functionalized particles (p < 0.001). Calcium release was similar among materials and remained constant during the experiment, while phosphate release was higher at 7 days in comparison to the remaining weeks (p < 0.001). In conclusion, diethylene glycol dimethacrylate resulted in the highest functionalization levels and the highest BFS among DCPD-containing materials. Ion release was not affected by functionalization. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2018. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 708-715, 2019.

The effect of saliva on the fate of nanoparticles

ObjectivesThe design of nanocarriers for local drug administration to the lining mucosa requires a sound knowledge of how nanoparticles (NPs) interact with saliva. This contact determines whether NPs agglomerate and become immobile due to size- and interaction-filtering effects or adsorb on the cell surface and are internalized by epithelial cells. The aim of this study was to examine the behavior of NPs in saliva considering physicochemical NP properties.Materials and methodsThe salivary pore–size distribution was determined, and the viscosity of the fluid inside of the pores was studied with optical tweezers. Distinct functionalized NPs (20 and 200 nm) were dispersed in saliva and salivary buffers and characterized, and surface-bound MUC5B and MUC7 were analyzed by 1D electrophoresis and immunoblotting. NP mobility was recorded, and cellular uptake studies were performed with TR146 cells.ResultsThe mode diameter of the salivary mesh pores is 0.7 μm with a peak width of 1.9 μm, and pores are filled with a low-viscosity fluid. The physicochemical properties of the NPs affected the colloidal stability and mobility: compared with non-functionalized particles, which did not agglomerate and showed a cellular uptake rate of 2.8%, functionalized particles were immobilized, which was correlated with agglomeration and increased binding to mucins.ConclusionThe present study showed that the salivary microstructure facilitates NP adsorption. However, NP size and surface functionalization determine the colloidal stability and cellular interactions.Clinical relevanceThe sound knowledge of NP interactions with saliva enables the improvement of current treatment strategies for inflammatory oral diseases.

A novel synthetic approach of cerium oxide nanoparticles with improved biomedical activity

Cerium oxide nanoparticles (CNPs) are novel synthetic antioxidant agents proposed for treating oxidative stress-related diseases. The synthesis of high-quality CNPs for biomedical applications remains a challenging task. A major concern for a safe use of CNPs as pharmacological agents is their tendency to agglomerate. Herein we present a simple direct precipitation approach, exploiting ethylene glycol as synthesis co-factor, to synthesize at room temperature nanocrystalline sub-10 nm CNPs, followed by a surface silanization approach to improve nanoparticle dispersibility in biological fluids. CNPs were characterized using transmission electron microscopy (TEM) observations, X-ray diffraction (XRD) analysis, thermogravimetric analysis (TGA), Fourier-transform infrared (FT-IR) spectroscopy, proton nuclear magnetic resonance (1H-NMR) spectroscopy, dynamic light scattering (DLS) and zeta potential measurements. CNP redox activity was studied in abiotic systems using electron spin resonance (ESR) measurements, and in vitro on human cell models. In-situ silanization improved CNP colloidal stability, in comparison with non-functionalized particles, and allowed at the same time improving their original biological activity, yielding thus functionalized CNPs suitable for biomedical applications.

Comparison of fluorescence-based methods to determine nanoparticle uptake by phagocytes and non-phagocytic cells in vitro.

At many portals of entry the relative uptake by phagocytes and non-phagocytic cells has a prominent effect on availability and biological action of nanoparticles (NPs). Cellular uptake can be determined for fluorescence-labeled NPs. The present study compares three methods (plate reader, flow cytometry and image analysis) in order to investigate the influence of particle size and functionalization and medium content on cellular uptake of fluorescence–labeled polystyrene particles and to study the respective method’s suitability for uptake studies. For comparison between the techniques, ratios of macrophage to alveolar epithelial cell uptakes were used. Presence of serum protein in the exposure solution decreased uptake of carboxyl-functionalized and non-functionalized particles; there was no clear effect for the amine-functionalized particles. The 200 nm non- or carboxyl-functionalized NPs were taken up preferentially by phagocytes while for amine-functionalized particles preference was lowest. The presence of the serum slightly increased the preference for these particles.In conclusion, due to the possibility of calibration, plate reader measurements might present a better option than the other techniques to (semi)quantify differences between phagocytes and non-phagocytic cells for particles with different fluorescence. In order to obtain unbiased data the fluorescent labeling has to fulfill certain requirements.

Polymer-based material containing calcium phosphate particles functionalized with a dimethacrylate monomer for use in restorative dentistry.

Dicalcium phosphate dihydrate particles functionalized with triethyleneglycol dimethacrylate were synthesized and added to a photocurable mixture of bisphenol-A glycidyl dimethacrylate and triethyleneglycol dimethacrylate with the purpose of developing a resin composite capable of releasing calcium and phosphate ions to foster dental remineralization. Particle functionalization would minimize the deleterious effect of adding low cohesive strength nano-structured particles with no chemical interaction with the organic matrix on the material's mechanical properties. The results showed that calcium release over 28 days was not impaired by particle functionalization. A statistically significant 32% increase in strength was recorded with the use of functionalized dicalcium phosphate dihydrate in comparison to the material containing non-functionalized particles. However, the strength of the unfilled resin was not matched by the composite with functionalized particles. Elastic modulus increased with particle incorporation, regardless of functionalization. Degree of conversion and optical properties (total transmittance and color change/ΔE) of the resin-based materials were not affected by the addition of dicalcium phosphate dihydrate particles (functionalized or not).

Bioactive composites containing TEGDMA-functionalized calcium phosphate particles: Degree of conversion, fracture strength and ion release evaluation

ObjectiveTo evaluate the strength and ion release of experimental composites containing TEGDMA-functionalized calcium phosphate particles. MethodsSeven composites containing equal parts (in mols) of BisGMA and TEGDMA and 60vol% of fillers were manipulated. Filler phase was constituted by silanized barium glass and 0% (control), 10% or 20% (volume) of dicalcium phosphate dihydrate (DPCD) particles, either non-functionalized or functionalized with two different TEDGMA contents. DCPD particles were synthesized and characterized by X-ray diffraction (XRD), elemental analysis, surface area and dynamic light scattering. Composites were tested for degree of conversion (DC) by near-FTIR. Biaxial flexural strength (BFS) was determined after 24h and 28days in water. Calcium and phosphate release after 7days was assessed using inductively coupled plasma optical emission spectrometry (ICP-OES). Data were analyzed by ANOVA/Tukey test (alpha:5%). ResultsXRD confirmed the crystalline structure corresponding to DCPD. Elemental analysis revealed particles with zero, 14% or 22% TEGDMA, with similar D50 (around 19μm) and surface areas from 3.5 to 11.4m2/g. The presence of DCPD did not reduce DC. After 24h, functionalization (both 14% and 22% TEGDMA) improved composite strength in comparison to non-functionalized DCPD, both at 10% and 20% levels. After 28days, BFS of materials containing 10% functionalized DCPD were statistically similar to the control containing only barium glass. Among composites containing 10% DCPD, particle functionalization with 14% TEGDMA did not jeopardize ion release. SignificanceAt 10vol%, the use of TEGDMA-functionalized CaP particles improved composite strength in relation to non-functionalized particles, while maintaining similar ion release levels.

Functionalization of emissive conjugated polymer nanoparticles by coprecipitation: consequences for particle photophysics and colloidal properties

The functionalization of polyfluorene (PFO) nanoparticles by coprecipitation of the conjugated polymer with an amphiphilic comb polymer, consisting of a hydrophobic polystyrene backbone with hydrophilic, carboxylic acid-terminated polyethylene oxide side-chains (PS-PEG-COOH), is investigated. The comb polymer affects the properties of the formed hybrid nanoparticles. Non-functionalized particles are typically larger (28 nm) than functionalized ones (20 nm); peak molar extinction coefficients are found to differ in a similar trend. Zeta potentials are negative, consistent with negative surface charge on PFO particles due to chemical defect formation, with additional charge on functionalized particles due to the pendant carboxylic acid groups. Emission quantum yields of functionalized particles are typically larger, consistent with lower efficiency of energy transfer to quenchers in smaller particles and weaker PFO interchain interactions due to chain dilution. The trend in per-particle fluorescence brightness values, as confirmed by single particle fluorescence imaging, reflects the nanoparticle extinction coefficients. Photostability studies on aqueous dispersions of hybrid particles indicate mild photobrightening under continuous illumination while PFO particles exhibit slow exponential emission decay. Functionalized particles are also resistant to aggregation during exposure to adenocarcinoma cells. Generally, the hybrid particles exhibit more favorable time-, pH- and medium-dependent stabilities, likely due to steric and electrostatic stabilization by PEG-carboxylic acid functionalities. Overall, the functionalized particles exhibit attractive properties: Reasonably small size, tight size distribution, high absorption cross section, radiative rate and emission quantum yield, excellent brightness and photostability, and good colloidal stability.

In vivo hepatocyte MR imaging using lactose functionalized magnetoliposomes

The aim of this study was to assess a novel lactose functionalized magnetoliposomes (MLs) as an MR contrast agent to target hepatocytes as well as to evaluate the targeting ability of MLs for in vivo applications. In the present work, 17 nm sized iron oxide cores functionalized with anionic MLs bearing lactose moieties were used for targeting the asialoglycoprotein receptor (ASGP-r), which is highly expressed in hepatocytes. Non-functionalized anionic MLs were tested as negative controls. The size distribution of lactose and anionic MLs was determined by transmission electron microscopy (TEM) and dynamic light scattering (DLS). After intravenous administration of both MLs, contrast enhancement in the liver was observed by magnetic resonance imaging (MRI). Label retention was monitored non-invasively by MRI and validated with Prussian blue staining and TEM for up to eight days post MLs administration. Although the MRI signal intensity did not show significant differences between functionalized and non-functionalized particles, iron-specific Prussian blue staining and TEM analysis confirmed the uptake of lactose MLs mainly in hepatocytes. In contrast, non-functionalized anionic MLs were mainly taken up by Kupffer and sinusoidal cells. Target specificity was further confirmed by high-resolution MR imaging of phantoms containing isolated hepatocytes, Kupffer cell (KCs) and hepatic stellate cells (HSCs) fractions. Hypointense signal was observed for hepatocytes isolated from animals which received lactose MLs but not from animals which received anionic MLs. These data demonstrate that galactose-functionalized MLs can be used as a hepatocyte targeting MR contrast agent to potentially aid in the diagnosis of hepatic diseases if the non-specific uptake by KCs is taken into account.

Functionalization of polyanhydride microparticles with di-mannose influences uptake by and intracellular fate within dendritic cells

Innovative vaccine delivery platforms can facilitate the development of effective single-dose treatment regimens to control emerging and re-emerging infectious diseases. Polyanhydride microparticles are promising vaccine delivery vehicles due to their ability to stably maintain antigens, provide tailored release kinetics and function as adjuvants. A major obstacle for the use of microparticle-based vaccines, however, is their limited uptake by dendritic cells (DCs). In this study, we functionalized the microparticle surface with di-mannose in order to target C-type lectin receptors (CLRs) on DCs. Polyanhydride particles based on sebacic acid (SA), 1,6-bis(p-carboxyphenoxy)hexane (CPH) and 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) were evaluated. Co-incubation of di-mannose-functionalized microparticles up-regulated the expression of CLRs on DCs. More importantly, di-mannose functionalization increased the uptake, as measured by the percentage of cells internalizing particles. The uptake of CPH:SA microparticles increased ∼20-fold, from 0.82% (non-functionalized) to 20.2%, and internalization of CPTEG:CPH microparticles increased ∼7-fold from 1.35% (non-functionalized) to 9.3% upon di-mannose functionalization. Both di-mannose-functionalized and non-functionalized particles trafficked to lysosomes. Together, these studies demonstrate that employing rational vaccine design principles, such as the targeting of CLRs on antigen-presenting cells, can enhance delivery of encapsulated antigens and potentially induce a more robust adaptive immune response.

Nanodiamond-Based Nanolubricants

In the current paper, it is demonstrated that nanoadditives based on detonation nanodiamonds used in combination with other antifriction\\antiwear additives provide from modest to significant improvements of the tribological properties of the commercial lubricating oils depending on the initial formulated oil properties. It was also demonstrated that functionalization of DND surface with fluorine provides beneficial effect. Fluorine-containing DNDs demonstrated superior tribological properties as compared to the pristine non-functionalized particles.

Peptide-Mediated Nanoengineering of Inorganic Particle Surfaces: A General Route toward Surface Functionalization via Peptide Adhesion Domains

The peptide-mediated functionalization of inorganic particle surfaces is demonstrated on gadolinium oxide (GdO) particles, revealing specific means to functionalize nano- or microparticles. Phage display screening is exploited to select 12mer peptides, which exhibit sequence-specific adhesion onto surfaces of GdO particles. These peptide adhesion domains are exploited to effectively decorate GdO particles with fluorescently labeled poly(ethylene oxide) (PEO), proving to result in a stable surface modification as shown by significant reduction of protein adsorption by 80%, compared to nonfunctionalized particles. Peptide adhesion and stability of the noncovalent coating are investigated by adsorption/elution experiments and Langmuir isotherms. Fluorescence microscopy, contact angle, and energy dispersive X-ray (EDX) measurements confirmed the sequence specificity of the interactions by comparing adhesion sequences with scrambled peptide sequences. Noncovalent, but specific modification of inorganic particle surfaces represents a generic strategy to modulate functionality and function of nano- or microparticle surfaces.

Amino-Functionalized Polystyrene Nanoparticles Activate the NLRP3 Inflammasome in Human Macrophages

Specifically designed and functionalized nanoparticles hold great promise for biomedical applications. Yet, the applicability of nanoparticles is critically predetermined by their surface functionalization. Here we demonstrate that amino-functionalized polystyrene nanoparticles (PS-NH(2)) of ∼100 nm in diameter, but not carboxyl- or nonfunctionalized particles, trigger NLRP3 inflammasome activation and subsequent release of proinflammatory interleukin 1β (IL-1β) by human macrophages. PS-NH(2) induced time-dependent proton accumulation in lysosomes associated with lysosomal destabilization, release of cathepsin B, and damage of the mitochondrial membrane. Accumulation of mitochondrial reactive oxygen species was accompanied by oxidation of thioredoxin, a protein playing a central role in maintaining the cellular redox balance. Upon oxidation, thioredoxin dissociated from the thioredoxin-interacting protein (TXNIP). Liberated TXNIP, in turn, interacted with the NLRP3 protein, resulting in a conformational change of the pyrin domain of the NLRP3 protein, as was predicted by molecular modeling. Consequently, this prompted assembly of the NLRP3 inflammasome complex with recruitment and activation of caspase-1, inducing IL-1β release by cleavage of pro-IL-1β. The central role of the NLRP3 inflammasome for cytokine production was confirmed by in vitro knockdown of NLRP3 and of the adaptor protein ASC, confirming that other inflammasomes were not activated by PS-NH(2). The PS-NH(2)-mediated proinflammatory macrophage activation could be antagonized by the radical scavenger N-acetyl-L-cysteine, which prevented mitochondrial damage, caspase-1 activation, and the subsequent release of IL-1β. Our study reveals the molecular mechanism of NLRP3 inflammasome activation by amino-functionalized nanoparticles and suggests a strategy as to how such adverse effects could be antagonized.

Identification of Peptide Ligands Facilitating Nanoparticle Attachment to Erythrocytes

Peptide ligands capable of mediating nanoparticle adhesion to human red blood cells (RBCs) were identified from a large bacterial display peptide library. Peptides were displayed on the surface of fluorescent Escherichia coli, enabling quantitative measurement of RBC binding and high-throughput screening using fluorescence-activated cell sorting. One of the isolated clones remained attached to RBCs under high-shear stresses equivalent to those encountered in vivo. Furthermore, nanoparticles functionalized with the identified RBC-binding peptides exhibited nearly 100-fold increased RBC binding relative to nonfunctionalized particles in the presence of physiologically relevant concentrations of human serum albumin, indicating that peptides remained functional in the absence of the protein scaffold used for display. The RBC-binding peptides identified here provide new opportunities for sustained therapeutic delivery applications whereby nanoparticulate drug carriers can be attached to RBCs to achieve long-circulating carrier systems.