Correct Identification of the Core-Shell Structure of Cell Membrane-Coated Polymeric Nanoparticles

Transmission electron microscopy (TEM) observations of negatively stained cell membrane (CM)-coated polymeric nanoparticles (NPs) reveal a characteristic core-shell structure. However, negative staining agents can create artifacts that complicate the determination of the actual NP structure. Herein, it is demonstrated with various bare polymeric core NPs, such as poly(lactic-co-glycolic acid) (PLGA), poly(ethylene glycol) methyl ether-block-PLGA, and poly(caprolactone), that certain observed core-shell structures are actually artifacts caused by the staining process. To address this issue, fluorescence quenching was applied to quantify the proportion of fully coated NPs and statistical TEM analysis was used to identify and differentiate whether the observed core-shell structures of CM-coated PLGA (CM−PLGA) NPs are due to artifacts or to the CM coating. Integrated shells in TEM images of negatively stained CM−PLGA NPs are identified as artifacts. The present results challenge current understanding of the structure of CM-coated polymeric NPs and encourage researchers to use the proposed characterization approach to avoid misinterpretations.     

Artificial Spores: Cytocompatible Coating of Living Cells with Plant‐Derived Pyrogallol

Cell nanoencapsulation, generating cell‐in‐shell structures (“artificial spores“), provides a chemical toolbox for controlling the cellular behaviors and functional characteristics of individual cells. Among the shell materials studied so far, naturally occurring polyphenolic compounds, including polydopamine and tannic acid, have intensively been employed in cell‐surface engineering, because their material‐independent coating property eliminates an extra priming step for inducing subsequent shell formation. Albeit successful in generating cell‐in‐shell structures, the coating of polyphenolic compounds generally requires alkaline conditions and/or high salt conditions, which are not compatible with certain cell types. In this work, we demonstrate that the nanocoating of individual cells with a plant‐derived phenolic compound, pyrogallol (1,2,3‐trihydroxybenzene), occurs at mildly alkaline pH of 7.8 in an isotonic buffer. Three different cell types (anucleate, microbial, and mammalian cells) are coated with pyrogallol without noticeable decrease in cell viability. The protocol developed in this work could be applied to other polyphenolic compounds, and, considering the many polyphenols identified as a coating material, provides an advanced chemical tool in cell‐surface engineering.     

Near‐Infrared Light Triggered‐Release in Deep Brain Regions Using Ultra‐photosensitive Nanovesicles

Remote and minimally‐invasive modulation of biological systems with light has transformed modern biology and neuroscience. However, light absorption and scattering significantly prevents penetration to deep brain regions. Herein, we describe the use of gold‐coated mechanoresponsive nanovesicles, which consist of liposomes made from the artificial phospholipid Rad‐PC‐Rad as a tool for the delivery of bioactive molecules into brain tissue. Near‐infrared picosecond laser pulses activated the gold‐coating on the surface of nanovesicles, creating nanomechanical stress and leading to near‐complete vesicle cargo release in sub‐seconds. Compared to natural phospholipid liposomes, the photo‐release was possible at 40 times lower laser energy. This high photosensitivity enables photorelease of molecules down to a depth of 4 mm in mouse brain. This promising tool provides a versatile platform to optically release functional molecules to modulate brain circuits.     

Nanoparticle-Decorated Biomimetic Extracellular Matrix for Cell Nanoencapsulation and Regulation

Cell encapsulation has been studied for various applications ranging from cell transplantation to biological production. However, current encapsulation technologies focus on cell protection rather than cell regulation that is essential to most if not all cell-based applications. Here we report a method for cell nanoencapsulation and regulation using an ultrathin biomimetic extracellular matrix as a cell nanocapsule to carry nanoparticles (CN²). This method allows high-capacity nanoparticle retention at the vicinity of cell surfaces. The encapsulated cells maintain high viability and normal metabolism. When gold nanoparticles (AuNPs) are used as a model to decorate the nanocapsule, light irradiation transiently increases the temperature, leading to the activation of the heat shock protein 70 (HSP70) promoter and the regulation of reporter gene expression. As the biomimetic nanocapsule can be decorated with any or multiple NPs, CN² is a promising platform for advancing cell-based applications.     

Interfacing carbon nanotubes with living cells

We developed a polymer coating for carbon nanotubes (CNTs) that mimics the mucin glycoprotein coating of mammalian cells. CNTs coated with these mucin mimic polymers have two novel properties: they can bind to carbohydrate receptors, providing a means for biomimetic interactions with cell surfaces, and, importantly, they are rendered nontoxic to cells.     

Recent Advances of Cell Membrane Coated Nanoparticles in Treating Cardiovascular Disorders

Cardiovascular diseases (CVDs) are the leading cause of death worldwide, causing approximately 17.9 million deaths annually, an estimated 31% of all deaths, according to the WHO. CVDs are essentially rooted in atherosclerosis and are clinically classified into coronary heart disease, stroke and peripheral vascular disorders. Current clinical interventions include early diagnosis, the insertion of stents, and long-term preventive therapy. However, clinical diagnostic and therapeutic tools are subject to a number of limitations including, but not limited to, potential toxicity induced by contrast agents and unexpected bleeding caused by anti-platelet drugs. Nanomedicine has achieved great advancements in biomedical area. Among them, cell membrane coated nanoparticles, denoted as CMCNPs, have acquired enormous expectations due to their biomimetic properties. Such membrane coating technology not only helps avoid immune clearance, but also endows nanoparticles with diverse cellular and functional mimicry. In this review, we will describe the superiorities of CMCNPs in treating cardiovascular diseases and their potentials in optimizing current clinical managements.     

具有内皮细胞选择性的细胞膜仿生支架材料的研究

利用AIBN引发自由基反应,由单体2-(甲基丙烯酰氧基)乙基-2-(三甲基氨基)乙基磷酸酯(MPC)、甲基丙烯酸十八酯(SMA)、对硝基苯氧羰基聚乙二醇甲基丙烯酸酯(MEONP)合成了一种新型类细胞膜仿生涂层材料.MPC可以阻抗非特异性吸附;MEONP可以结合抗体或多肽促进特异性识别.通过表面固定的方法引入多肽序列Arg-Glu-Asp-Val(REDV),使涂层具有内皮细胞选择性.核磁、紫外吸收、红外光谱表征证实聚合物的组成以及REDV多肽在表面的固定;并通过血浆复钙化实验表征涂层的血液相容性.细胞黏附与增殖实验反映REDV多肽构建的涂层表面具备良好的特异性识别并结合内皮细胞的能力.     

A Biomimetic Nanoparticle to “Lure and Kill” Phospholipase A2

Inhibition of phospholipase A2 (PLA2) has long been considered for treating various diseases associated with an elevated PLA2 activity. However, safe and effective PLA2 inhibitors remain unavailable. Herein, we report a biomimetic nanoparticle design that enables a “lure and kill” mechanism designed for PLA2 inhibition (denoted “L&K‐NP”). The L&K‐NPs are made of polymeric cores wrapped with modified red blood cell membrane with two inserted key components: melittin and oleyloxyethyl phosphorylcholine (OOPC). Melittin acts as a PLA2 attractant that works together with the membrane lipids to “lure” in‐coming PLA2 for attack. Meanwhile, OOPC acts as inhibitor that “kills” PLA2 upon enzymatic attack. Both compounds are integrated into the L&K‐NP structure, which voids toxicity associated with free molecules. In the study, L&K‐NPs effectively inhibit PLA2‐induced hemolysis. In mice administered with a lethal dose of venomous PLA2, L&K‐NPs also inhibit hemolysis and confer a significant survival benefit. Furthermore, L&K‐NPs show no obvious toxicity in mice. and the design provides a platform technology for a safe and effective anti‐PLA2 approach.     

Single step hybrid coating process to enhance the electrosteric stabilization of inorganic particles

We report on a single-step coating process and the resulting colloidal stability of silica-coated spindle-type hematite nanoparticles (NPs) decorated with a layer of poly(acrylic acid) (PAA) polyelectrolyte chains that are partially incorporated into the silica shell. The stability of PAA coated NPs as a function of pH and salt concentration in water was compared to bare hematite particles and simple silica-coated hematite NPs, studying their electrophoretic mobility and the hydrodynamic radius by dynamic light scattering. Particles coated with this method were found to be more stable upon the addition of salt at pH 7, and their aggregation at the pH of the isoelectric point is reversible. The hybrid coating appears to increase the colloidal stability in aqueous media due to the combination of the decrease of the isoelectric point and the electrosteric stabilization. This coating method is not limited to hematite particles but can easily be adapted to any silica-coatable particle.     

Controlled Assembly of Block Copolymer Coated Nanoparticles in 2D Arrays

The defined assembly of nanoparticles (NPs) in polymer matrices is an important prerequisite for next‐generation functional materials. A promising approach to control NP positions in polymer matrices at the nanometer scale is the use of block copolymers. It allows the selective deposition of NPs in nanodomains, but the final defined and ordered positioning of the NPs within the domains has not been possible. This can now be achieved by coating NPs with block copolymers. The self‐assembly of block copolymer‐coated NPs directly leads to ordered microdomains containing ordered NP arrays with exactly one NP per unit cell. By variation of the grafting density, the inter‐nanoparticle distance can be controlled from direct NP surface contact to surface separations of several nanometers, determined by the thickness of the polymer shell. The method can be applied to a wide variety of block copolymers and NPs and is thus suitable for a broad range of applications.     

Antibacterial activity of glutathione-coated silver nanoparticles against Gram positive and Gram negative bacteria

In the present paper, we study the mechanism of antibacterial activity of glutathione (GSH) coated silver nanoparticles (Ag NPs) on model Gram negative and Gram positive bacterial strains. Interference in bacterial cell replication is observed for both cellular strains when exposed to GSH stabilized colloidal silver in solution, and microbicidal activity was studied when GSH coated Ag NPs are (i) dispersed in colloidal suspensions or (ii) grafted on thiol-functionalized glass surfaces. The obtained results confirm that the effect of dispersed GSH capped Ag NPs (GSH Ag NPs) on Escherichia coli is more intense because it can be associated with the penetration of the colloid into the cytoplasm, with the subsequent local interaction of silver with cell components causing damages to the cells. Conversely, for Staphylococcus aureus, since the thick peptidoglycan layer of the cell wall prevents the penetration of the NPs inside the cytoplasm, the antimicrobial effect is limited and seems related to the interaction with the bacterial surfaces. Experiments on GSH Ag NPs grafted on glass allowed us to elucidate more precisely the antibacterial mechanism, showing that the action is reduced because of GSH coating and the limitation of the translational freedom of NPs.     

Biocompatibility and hyperthermia cancer therapy of casein‐coated iron oxide nanoparticles in mice

Magnetic nanoparticles (MNPs) can be used as heat generation source in cancer hyperthermia therapy. While iron oxide nanoparticles (NPs) are the most popular choice for magnetic hyperthermia, adding a surface enhancement can improve its performance. Furthermore, for MNPs to be used in biomedical application their cytotoxicity needs to be evaluated. In this study biocompatibility and also in vivo performance of casein‐coated MNPs were assessed. Cell viability of normal cell lines in all of tests remained above 95% for 0.5 and 1 mg/mL concentration and even the minimum recorded cell viability for normal cell lines was 84.78% at 20 mg/mL concentration. In contrast cell viability of cancer cell lines in contact with casein coated MNPs core‐shell structure except for one sample remained below 85%. By introduction of and alternating magnetic field, cell viability of samples with lower MNP concentration dropped by 20% to 30% while this drop for samples with higher concentration was 10% to 20%. Furthermore, results of in vivo trials show that just 1 week of hyperthermia treatment with casein coated MNPs core‐shell structure can reduce the tumor size of the mice by 33%. Real‐time polymerase chain reaction results further confirmed the effectiveness of this method. Moreover, findings of this study suggest that lower injection speed can improve NPs distribution and treatment effect. Results of this study suggest that core‐shell structure can positively affect the tumor growth and the combination of good biocompatibility, innate hostility toward cancer cells and good heating power makes them a good candidate for hyperthermia cancer therapy applications.     

Paramagnetic nanoparticle T(1) and T(2) MRI contrast agents

There is no doubt that magnetic resonance imaging contrast agents (MRI CAs) can play a vital role in diagnosing diseases. Therefore, demand for new MRI CAs with an enhanced sensitivity and advanced functionalities is very high. Here, paramagnetic nanoparticles (NPs) are reviewed as new potential candidates for either T(1) or T(2) MRI CAs or both. These include surface coated lanthanide (Ln) oxide NPs (Ln = Gd, Dy, and Ho) and manganese oxide NPs. Surface coating materials should be biocompatible and hydrophilic. Compared to conventional large NPs, these surface coated paramagnetic NPs can be made ultrasmall with core particle diameter ranging from 1 to 3 nm, but their magnetic properties are still sufficient for MRI CAs. At this particle diameter, they can be easily excreted from the body through the renal system, which is prerequisite for in vivo applications. Mixed lanthanide oxide NPs into which a fluorescent Ln material is incorporated will be valuable as multiple imaging agents for both MRI-fluorescent imaging (FI) and MRI-cellular imaging (CL). These paramagnetic NPs can be further functionalized towards target-specific imaging, multiplex imaging, and drug delivery.     

一种仿细胞膜聚合物用于碳纳米管表面改性

以新型含有磷酸胆碱基的仿细胞膜两亲聚合物——胆固醇封端的聚(2-甲基丙烯酰氧基乙基磷酸胆碱)(CPMPC)为表面稳定剂实现碳纳米管的表面改性,利用两亲聚合物中的胆固醇疏水段与碳纳米管表面进行非共价键的稳定结合,通过两亲聚合物中聚(2-甲基丙烯酰氧基乙基磷酰胆碱)(PMPC)亲水段实现其水溶性和生物相容性.并以商业可获得的典型两亲分子,末端为胆固醇的聚氧乙烯(CPEG)和卵磷脂,为对照进行研究.研究表明CPMPC和CPEG均具有比卵磷脂更高的对碳纳米管进行分散的能力.而CPMPC改性的碳纳米管比CPEG改性的碳纳米管具有更优的稳定性和生物相容性,通过新型仿细胞膜聚合物改性的碳纳米管在生物医用领域有潜在应用.     

Hydrophilization of Magnetic Nanoparticles with Modified Alternating Copolymers. Part 1: The Influence of the Grafting

Iron oxide nanoparticles (NPs) with a diameter 21.6 nm were coated with poly(maleic acid-alt-1-octadecene) (PMAcOD) modified with grafted 5,000 Da poly(ethyelene glycol) (PEG) or short ethylene glycol (EG) tails. The coating procedure utilizes hydrophobic interactions of octadecene and oleic acid tails, while the hydrolysis of maleic anhydride moieties as well as the presence of hydrophilic PEG (EG) tails allows the NP hydrophilicity. The success of the NP coating was found to be independent of the degree of grafting which was varied between 20 and 80% of the -MacOD-units, but depended on the length of the grafted tail. The NP coating and hydrophilization did not occur when the modified copolymer contained 750 Da PEG tails independently of the grafting degree. To explain this phenomenon the micellization of the modified PMAcOD copolymers in water was analyzed by small angle x-ray scattering (SAXS). The PMAcOD molecules with the grafted 750 Da PEG tails form compact non-interacting disk-like micelles, whose stability apparently allows for no interactions with the NP hydrophobic shells. The PMAcOD containing the 5,000 Da PEG and EG tails form much larger aggregates capable of an efficient coating of the NPs. The coated NPs were characterized using transmission electron microscopy, dynamic light scattering, ζ-potential measurements, and thermal gravimetry analysis. The latter method demonstrated that the presence of long PEG tails in modified PMAcOD allows the attachment of fewer macromolecules (by a factor of ~20) compared to the case of non-modified or EG modified PMAcOD, emphasizing the importance of PEG tails in NP hydrophilization. The NPs coated with PMAcOD modified with 60% (towards all -MAcOD- units) of the 5,000 PEG tails bear a significant negative charge and display good stability in buffers. Such NPs can be useful as magnetic cores for virus-like particle formation.     

Cholesterol-rich membrane coatings for interaction studies in capillary electrophoresis: application to red blood cell lipid extracts

The purpose was to develop a stable biological membrane coating for CE useful for membrane interaction studies. The effect of cholesterol (chol) on the stability of dipalmitoylphosphatidylcholine (DPPC) and sphingomyelin (SM) coatings was studied. In addition, a fused-silica capillary for CE was coated with human red blood cell (RBC) ghost lipids. Liposomes prepared of DPPC/SM with and without chol or RBC ghost lipids were flushed through the capillary and the stability of the coating was measured electrophoretically. Similar mixtures of DPPC/SM with and without chol were further studied by differential scanning calorimetry. The presence of phosphatidylcholine as a basic component in the coating solution of DPPC/SM/chol was found to be essential to achieve a good and stable coating. The results also confirmed the stability of coatings obtained with solutions of DPPC with 0-30 mol% of chol and SM in different ratios, which more closely resemble natural membranes. Finally, the electrophoretic measurements revealed that a stable coating is formed when capillaries are coated with liposomes of RBC ghost lipids.     

Colloidal stability of magnetic iron oxide nanoparticles: influence of natural organic matter and synthetic polyelectrolytes

The colloidal behavior of natural organic matter (NOM) and synthetic poly(acrylic acid) (PAA)-coated ferrimagnetic (γFe(2)O(3)) nanoparticles (NPs) was investigated. Humic acid (HA), an important component of NOM, was extracted from a peat soil. Two different molecular weight PAAs were also used for coating. The colloidal stability of the coated magnetic NPs was evaluated as a resultant of the attractive magnetic dipolar and van der Waals forces and the repulsive electrostatic and steric-electrosteric interactions. The conformational alterations of the polyelectrolytes adsorbed on magnetic γFe(2)O(3) NPs and their role in colloidal stability were determined. Pure γFe(2)O(3) NPs were extremely unstable because of aggregation in aqueous solution, but a significant stability enhancement was observed after coating with polyelectrolytes. The steric stabilization factor induced by the polyelectrolyte coating strongly dictated the colloidal stability. The pH-induced conformational change of the adsorbed, weakly charged polyelectrolytes had a significant effect on the colloidal stability. Atomic force microscopy (AFM) revealed the stretched conformation of the HA molecular chains adsorbed on the γFe(2)O(3) NP surface at pH 9, which enhanced the colloidal stability through long-range electrosteric stabilization. The depletion of the polyelectrolyte during the dilution of the NP suspension decreased the colloidal stability under acidic solution conditions. The conformation of the polyelectrolytes adsorbed on the NP surface was altered as a function of the substrate surface charge as viewed from AFM imaging. The polyelectrolyte coating also led to a reduction in magnetic moments and decreased the coercivity of the coated γFe(2)O(3) NPs. Thus, the enhanced stabilization of the coated maghematite NPs may facilitate their delivery in the groundwater for the effective removal of contaminants.     

Multilayers and poly(allylamine hydrochloride)-graft-poly(ethylene glycol) modified bovine serum albumin nanoparticles: Improved stability and pH-responsive drug delivery

To improve the colloidal stability of bovine serum albumin(BSA) nanoparticles(NPs) in diverse mediums, poly(allylamine hydrochloride)(PAH)/sodium poly(4-styrene sulfonate)(PSS) multilayers and poly(allylamine hydrochloride)-graft-poly(ethylene glycol)(PAH-g-PEG) coating were coated on the surface of BSA NPs.Stabilities of the BSA NPs in diverse mediums with different surfaces were detected by dynamic light scattering(DLS).Multilayers and PAH-g-PEG coated BSA NPs can be well dispersed in various mediums with a narrow polydispersity index(PDI).The BSA NPs with the highest surface density of PEG show the best stability.The multilayers and PAH-g-PEG coating do not deter the pH-dependent loading and release property of BSA NPs.At pH 9,the encapsulation efficiency of doxorubicin reaches almost 99%,and the release rate at pH 5.5 is significantly higher than that at pH 7.4.     

Boehmite-coated microporous membrane for enhanced electrochemical performance and dimensional stability of lithium-ion batteries

A heat-resistant boehmite-coated polypropylene (PP) membrane has been successfully fabricated and its potential application as a promising separator in the lithium-ion battery was explored. The boehmite powders with average sizes of 0.78, 1.03, and 1.72 μm, respectively, were used to fabricate the coated membrane. It was demonstrated that the coated membrane prepared by boehmite with a 0.78-μm size showed superior heat tolerance and proper air permeability. As compared to the commercialized PP membrane, such coated membrane presented improved electrolyte uptake, better interface stability, and enhanced ionic conductivity. In addition, the lithium iron phosphate (LiFePO₄)/Li cell using this composite membrane exhibited better rate capability and cycling retention than that using PP membrane owing to its facile ion transport and excellent interfacial compatibility. The coating layer showed an advantage on solid electrolyte interface film formation and greatly reduced charge transfer resistance. All these fascinating characteristics would boost the application of this composite membrane for high-performance lithium-ion battery.     

Fertility and Iron Bioaccumulation in Drosophila melanogaster Fed with Magnetite Nanoparticles Using a Validated Method

Research on nanomaterial exposure-related health risks is still quite limited; this includes standardizing methods for measuring metals in living organisms. Thus, this study validated an atomic absorption spectrophotometry method to determine fertility and bioaccumulated iron content in Drosophila melanogaster flies after feeding them magnetite nanoparticles (Fe₃O₄NPs) dosed in a culture medium (100, 250, 500, and 1000 mg kg⁻¹). Some NPs were also coated with chitosan to compare iron assimilation. Considering both accuracy and precision, results showed the method was optimal for concentrations greater than 20 mg L⁻¹. Recovery values were considered optimum within the 95–105% range. Regarding fertility, offspring for each coated and non-coated NPs concentration decreased in relation to the control group. Flies exposed to 100 mg L⁻¹ of coated NPs presented the lowest fertility level and highest bioaccumulation factor. Despite an association between iron bioaccumulation and NPs concentration, the 500 mg L⁻¹ dose of coated and non-coated NPs showed similar iron concentrations to those of the control group. Thus, Drosophila flies’ fertility decreased after NPs exposure, while iron bioaccumulation was related to NPs concentration and coating. We determined this method can overcome sample limitations and biological matrix-associated heterogeneity, thus allowing for bioaccumulated iron detection regardless of exposure to coated or non-coated magnetite NPs, meaning this protocol could be applicable with any type of iron NPs.