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Two frequent problems hindering clinical translation of nanomedicine are low drug loading and low colloidal stability. Previous efforts to achieve ultrahigh drug loading (>30 %) introduce new hurdles, including lower colloidal stability and others, for clinical translation. Herein, we report a new class of drug nano-carriers based on our recent finding in protein-nanoparticle co-assembly supraparticle (PNCAS), with both ultrahigh drug loading (58 % for doxorubicin, i.e., DOX) and ultrahigh colloidal stability (no significant change in hydrodynamic size after one year). We further show that our PNCAS-based drug nano-carrier possesses a built-in environment-responsive drug release feature: once in lysosomes, the loaded drug molecules are released instantly (<1 min) and completely (∼100 %). Our PNCAS-based drug delivery system is spontaneously formed by simple mixing of hydrophobic nanoparticles, albumin and drugs. Several issues related to industrial production are studied. The ultrahigh drug loading and stability of DOX-loaded PNCAS enabled the delivery of an exceptionally high dose of DOX into a mouse model of breast cancer, yielding high efficacy and no observed toxicity. With further developments, our PNCAS-based delivery systems could serve as a platform technology to meet the multiple requirements of clinical translation of nanomedicines., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The so-called "hard-to-transfect cells" are well-known to present great challenges to intracellular delivery, but detailed understandings of the delivery behaviors are lacking. Recently, we discovered that vesicle trapping is a likely bottleneck of delivery into a type of hard-to-transfect cells, namely, bone-marrow-derived mesenchymal stem cells (BMSCs). Driven by this insight, herein, we screened various vesicle trapping-reducing methods on BMSCs. Most of these methods failed in BMSCs, although they worked well in HeLa cells. In stark contrast, coating nanoparticles with a specific form of poly(disulfide) (called PDS1) nearly completely circumvented vesicle trapping in BMSCs, by direct cell membrane penetration mediated by thiol-disulfide exchange. Further, in BMSCs, PDS1-coated nanoparticles dramatically enhanced the transfection efficiency of plasmids of fluorescent proteins and substantially improved osteoblastic differentiation. In addition, mechanistic studies suggested that higher cholesterol content in plasma membranes of BMSCs might be a molecular-level reason for the greater difficulty of vesicle escape in BMSCs.

With the development of single cell analysis techniques, the concept of precision toxicology has been proposed in recent years. Due to the heterogeneity of cells, we need to perform toxicological assessments on individual cells. Microalgae, one kind of important primary producers, play as a major pathway by which heavy metals enter the food chain and thus accumulate/transfer to higher trophic levels. Herein, the biosorption of Cd (Ex-Cd) and bioaccumulation of Cd (In-Cd) for Synechocystis sp. PCC 6803 were investigated by online 3D droplet microfluidic device combined with inductively coupled plasma mass spectrometry detection. Meanwhile, the algal toxicological responses of the algae cell to Cd 2+ exposure under different concentration (50, 100, and 150 μg L  -   1 ) and time (15 min, 24, 48 and 96 h) were studied. Combining single-cell analysis with toxicological indicators, the toxicity mechanism of Cd 2+ to algal was discussed. The single cell analysis results revealed heterogeneity in cellular uptake of Cd 2+ . The proportion of Cd-containing cells and Cd content in single algal cells all reached the maximum at 24 h. The uptake of Cd 2+ occurred within 15 min under all tested exposure concentrations and a large part of Cd 2+ were adsorbed on the algal cells surface. The Pearson correlation analysis showed that cell density, chlorophyll a and carotenoids were significantly negatively correlated with Cd accumulation, whereas ROS level and SOD activity were significantly positively correlated with Cd accumulation. It suggested that Cd 2+ accumulated intracellular would show toxic effects on the algal cells and oxidative stress is the main mechanism of Cd toxicity to algal cells. This work promotes our understanding of the toxicological responses of microalgae under Cd stress at single cells level., Competing Interests: Declaration of Competing Interest The authors declared that they have no commercial or financial conflicts of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Biological delivery remains a major challenge in biotechnology, partly because it is often not enough to overcome a single delivery barrier. It is highly desirable, yet rarely available, to design delivery carriers with both simple structures and the ability to cross multiple delivery barriers with high efficiency. Herein, we describe a distinct design (dubbed 'SDot') of delivery carriers with a single structural feature that can enhance the crossing of multiple delivery barriers. The bio-interface (the interface with a biological environment) of an SDot nanoparticle is highly hydrophobic, thus enhancing its interactions with lipid membranes, which are the primary components of many bio-delivery barriers. We used quantum dots (QDs) as the model core material of SDots and conjugated them with a RGD peptide. Thus-formed SDots-RGD demonstrated greatly improved abilities of cellular uptake and transcytosis in a brain tumor cell line, U87MG, compared with the conventional nanoparticle counterpart with a hydrophilic bio-interface (wQDs-RGD). Further, after loading a microtubule-binding anticancer drug, paclitaxel (PTX), onto the nanoparticle surface of SDots-RGD, the resulting drug formulation PTX@SDots-RGD displayed excellent ability of intracellular targeting to microtubules in U87MG cells. In a small animal cancer model, PTX@SDots-RGD exhibited significantly higher ability to slow down brain tumor growth than that of PTX@wQDs-RGD and free PTX. Taken together, these experimental results indicated the significant potential of SDots-RGD for bio-delivery, although the possible long-term toxicity of QDs used as the core material needs to be addressed in future work by replacing QDs with clinically approved materials.

Single-cell studies can help to understand individual differences and obtain atypical cellular characteristics in view of cellular heterogeneity. Herein, the accumulation of mercury (Hg) in single algae cells was studied by droplet chip-time resolved inductively coupled plasma mass spectrometry analytical system, and the relation of Hg accumulation to the physiological responses of algae cell was explored. When low concentrations of Hg 2+ (5-20 μg/L) were used in the exposure experiment, the content of Hg in single cells increased in first 2 h, then decreased with further increase of exposure time to 96 h, probably due to the growth dilution effect of the algae. When exposed to 30 μg/L Hg 2+ , the uptake of Hg by individual cells increased over time, which was associated with increased cell membrane permeability. The exposure to Hg 2+ (5-30 μg/L) inhibited the growth of algae in a concentration-dependent manner and serious growth inhibition occurred under the exposure concentration of 30 μg/L. While the exposure concentration was lower than 20 μg/L, algal cells exhibited a recover tendency due to the self-protection mechanism of algal cells. Bivariate results showed that intracellular Hg accumulation was significantly negatively correlated with cells growth in terms of OD 680 , photosynthetic pigments, F v /F m and PI abs . On the contrast, reactive oxygen species content, superoxide dismutase activity, and cell membrane permeability were significantly positively correlated with the accumulation of intracellular Hg. These results are helpful to further understand the toxic effect of Hg on algae., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Hard-to-transfect cells are cells that are known to present special difficulties in intracellular delivery of exogenous entities. However, the special transport behaviors underlying the special delivery problem in these cells have so far not been examined carefully. Here, we combine single-particle motion analysis, cell biology studies, and mathematical modeling to investigate nanoparticle transport in bone marrow-derived mesenchymal stem cells (BMSCs), a technologically important type of hard-to-transfect cells. Tat peptide-conjugated quantum dots (QDs-Tat) were used as the model nanoparticles. Two different yet complementary single-particle methods, namely, pair-correlation function and single-particle tracking, were conducted on the same cell samples and on the same viewing stage of a confocal microscope. Our results reveal significant differences in each individual step of transport of QDs-Tat in BMSCs vs a commonly used model cell line, HeLa cells. Single-particle motion analysis demonstrates that vesicle escape and cytoplasmic diffusion are dramatically more difficult in BMSCs than in HeLa cells. Cell biology studies show that BMSCs use different biological pathways for the cellular uptake, vesicular transport, and exocytosis of QDs-Tat than HeLa cells. A reaction-diffusion-advection model is employed to mathematically integrate the individual steps of cellular transport and can be used to predict and design nanoparticle delivery in BMSCs. This work provides dissective, quantitative, and mechanistic understandings of nanoparticle transport in BMSCs. The investigative methods described in this work can help to guide the tailored design of nanoparticle-based delivery in specific types and subtypes of hard-to-transfect cells.

Understanding the underlying transport mechanism of biological delivery is important for developing delivery technologies for pharmaceuticals, imaging agents, and nanomaterials. Recently reported by our group, SDots are a novel class of nanoparticle delivery systems with distinct biointerface features and excellent fusogenic capabilities (i.e., strong ability to interact with the hydrophobic portions of biomembranes). In this study, we investigate the cellular transport mechanism of SDots conjugated with Tat peptide (SDots-Tat) by live-cell spinning-disk confocal microscopy combined with molecular biology methods. Mechanistic studies were conducted on the following stages of cellular transport of SDots-Tat in HeLa cells: cellular entry, endosomal escape, nucleus entry, and intranuclear transport. A key finding is that, after escaping endosomes, SDots-Tat enter the cell nucleus via an importin β -independent pathway, bypassing the usual nucleus entry mechanism used by Tat. This finding implies a new approach to overcome the nucleus membrane barrier for designing biological delivery technologies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Dai, Wang, Yang, Xu and Ruan.)

Most neurological diseases have no cure today; innovations in neurotechnology are in urgent need. Nanomaterial-based remote neurostimulation with physical fields (NNSPs) is an emerging class of neurotechnologies that has generated tremendous interest in recent years. This perspective focuses on the clinical translation of this new class of neurotechnologies, an issue that so far has not received enough attention. We outline the major barriers in their clinical translation. We highlight our recent efforts to tackle these translational barriers, with a focus on the biological delivery problem. In particular, for the first time, we have shown that it is feasible to use noninvasive brain delivery to generate significant physiological responses in living animals by NNSP. However, much more work is needed to overcome the translational barriers., (© 2020 Author(s).)

Recently, with increasing medical practices, including organ transplantation and tumor chemotherapy, fungal infections, particularly the occurrence of drug-resistant fungal strains, remain a severe problem for the public health, which cause worse complications in the immunocompromised patients. The search for efficacious yet safe antifungal agents is in high demand in precision medicine. However, fungicides are often poorly water soluble for oral absorption, which is difficult for pharmaceutical efficacy evaluation. In this study, lipophilic oleic acid (OA)-grafted mesoporous silica (SBA-15) was facilely modified by cetyltrimethylammonium bromide (CTAB), which acts as an efficient antifungal drug matrix of itraconazole (ITZ). Characterized by physicochemical methods, the rod-like SBA-15-OA-CTAB/ITZ composite with retained mesostructural regularity shows that the loading amount of ITZ in the mesopore is ∼18%, contributing to the enhanced antifungal activity against Aspergillus fumigatus (A. fumigatus) and Candida albicans (C. albicans). The antimicrobial mechanism study suggests that the reactive oxygen species (ROS) were formed when fungal cells were incubated with the formulated ITZ, while there was no ROS formation in the presence of pure ITZ, which may result from the quaternary ammonium moieties of CTAB in the nanocomposites. Due to the potential toxicity of CTAB on mammalian cells, the as-synthesized mesoporous SBA-15-OA-CTAB/ITZ provides an alternative molecular design for the formulation improvement of a lipophilic antifungal drug applicable for external uses such as topical therapy.

Fluorescent and magnetic materials play a significant role in biosensor technology, enabling sensitive quantification and separations with applications in diagnostics, purification, quality control, and therapeutics. Here, we present a magneto-fluorescent biosensor/separations platform consisting of quantum dots (QDs) and superparamagnetic iron oxide nanoparticles (SPIONs) that are separately encapsulated in amphiphilic block co-polymer micelles conjugated to DNA or protein (i.e., single-stranded (ss) DNA derived from the mRNA of the tumor suppressor protein p53 or avidin protein). Analytes were detected via an aggregation sandwich assay upon binding of at least 1 QD and 1 SPION-containing micelle to result in a fluorescent/magnetic composite. Multiplexed isolation of protein and DNA biomolecules was demonstrated by using QDs of varying emission wavelength; QD fluorescence intensity could be correlated with analyte concentration. Sequential or parallel biomolecule separation was achieved by adding appropriately functionalized SPION-containing micelles and applying user-controlled magnetic fields via patterned magnetic disks and wires. QD fluorescence was used to continuously visualize analyte separation during this process. This QD/SPION platform is simple to use, demonstrates ∼10-16 M sensitivity in analyte detection (comparable to competing QD biosensors based on energy transfer) with specificity against 1 and 2 basepair mismatches in DNA detection, molecular separations capability in solutions of ∼10-10 M, and permits simultaneous or parallel, multiplexed separation of protein and DNA. Thus, this versatile platform enables self-assembly-based rapid, sensitive, and specific detection and separation of biomolecules, simultaneously and with real-time visualization. This technology demonstrates potential for nanoscale assembly, biosensing, and bioseparations.

Recently, supraparticle protein-nanoparticle co-assemblies (or 'supraparticle co-assemblies' for short) have attracted considerable interest due to their fundamental and technological value. However, it remains challenging to form supraparticle co-assemblies with high stability. Here, we show that using hydrophobic interaction, instead of the previously used electrostatic and van der Waals interactions, as the primary driving force can lead to instant formation of exceptionally stable supraparticle co-assemblies with minimal external energy input. Our formation method of supraparticle co-assemblies simply involves mixing globular proteins ( e.g. , bovine serum albumin) with hydrophobic nanoparticles ( e.g. , hydrophobic magnetic nanoparticles and hydrophobic quantum dots) without significant energy input ( e.g. , sonication or stirring). Upon mixing of hydrophobic nanoparticles and proteins, the formation of supraparticle co-assemblies only takes <1 minute. Further incubation of the mixture for several hours results in a gradual increase of the size uniformity of supraparticle co-assemblies. The formed supraparticle co-assemblies have been colloidally stable for 6 months and counting, and can withstand harsh environments such as basic and acidic pH, high temperature, high dilution, and serum. Co-encapsulation of different sizes/types of nanoparticles is found to be feasible and the co-encapsulation number ratio of different nanoparticles is well-controlled by the feeding ratio. Proof-of-concept studies show the potential of the supraparticle co-assemblies for biological imaging, delivery, and modulation. The combination of very rapid formation, minimal energy consumption, highly stable products, and inexpensive raw materials of this hydrophobic interaction-driven process meets many of the main goals of 'ideal' nano-manufacturing. Thus, this process could serve as the foundation of ideal manufacturing of supraparticle co-assemblies., Competing Interests: The authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)

Originally discovered in fundamental research of nanomaterial-biomolecule interactions, protein-nanoparticle co-assembly supraparticles (PNCAS) have become an emerging class of nanomaterials with various biological applications. We apply the interfacial instability process, which was originally reported for forming nanoparticles-encapsulated polymeric micelles, to produce PNCAS. By doing so hydrophobic nanoparticles, which are often the product formed from the upstream nanoparticle synthesis step, can be directly used as the raw materials of the production process of PNCAS. On the other hand, we take advantage of the structural features of protein molecules, in comparison with amphiphilic block copolymers, to mitigate two common problems encountered in the original interfacial instability-mediated nanoparticle encapsulation process, namely (1) poor encapsulation number control and (2) inconvenience and high cost to vary the assembly size. Additionally, we achieve semi-continuous and scalable production of PNCAS by combining the electrospray process and the interfacial instability process. We also conduct proof-of-concept studies of biological applications of the PNCAS products.

Transcranial magnetic stimulation (TMS) is a non-invasive and clinically approved method for treating neurological disorders. However, the relatively weak intracranial electric current induced by TMS is an obvious inferiority which can only produce limited treatment effects in clinical application. The present study aimed to investigate the possibility of enhancing the effects of TMS with intravenously administrated magnetic nanoparticles. To facilitate crossing of the blood-brain barrier (BBB), the superparamagnetic iron oxide nanoparticles (SPIONs) were coated with carboxylated chitosan and poly(ethylene glycol). To aid the nanoparticles in crossing the BBB and targeting the predesigned brain regions, an external permanent magnet was attached to the foreheads of the rats before the intravenous administration of SPIONs. The electrophysiological tests showed that the maximum MEP amplitude recorded in an individual rat was significantly higher in the SPIONs + magnet group than in the saline group (5.78 ± 2.54 vs. 1.80 ± 1.55 mV, P = 0.015). In the M1 region, biochemical tests detected that the number density of c-fos positive cells in the SPIONs + magnet group was 3.44 fold that of the saline group. These results suggest that intravenously injected SPIONs can enhance the effects of TMS in treating neurological disorders.

Semiconductor quantum dots (QDs) possess outstanding optical properties as fluorescent probes, but their applications in live cell intracellular imaging are hindered by various cellular transport barriers. Inspired by membrane proteins inserting their nanometer-scale hydrophobic surface into biomembranes, the present work aims to investigate the possibility that bare hydrophobic QDs could penetrate through live cell membranes without disrupting the membrane integrity. We utilize live cell spinning disk confocal microscopy to image and track the cellular transport process of bare hydrophobic QDs in the presence of a small percentage of three different organic cosolvents, namely, tetrahydrofuran (THF), chloroform, and hexane. A major finding is that, under certain cosolvent conditions, bare hydrophobic QDs can indeed penetrate through biomembranes in a noninvasive manner. Results of this work offer us guidance to design a new class of nanobioprobes based on combining hydrophobic nanoscale surface and cosolvent, and they provide key new pieces to the emerging complex and sophisticated picture of nanostructure-biosystem interactions.

Quantum dots (QDs) have emerged as a major class of fluorescent probes with unique optical properties, but applying QDs for imaging specific intracellular entities in live cells has been hindered by the poor performance of targeted intracellular delivery of QDs due to various cellular transport barriers. We describe a novel QD nanoprobe design, which is termed a cosolvent-bare hydrophobic QD-biomolecule (cS-bQD-BM, or 'SDot' for short), combining a cosolvent, a bare hydrophobic nanoparticle surface, ultrasmall size and biomolecular function. SDots show extraordinary intracellular targeting performance with the nucleus as the model target, including near-perfect specificity, excellent efficiency and reproducibility, high-throughput ability, minimal toxicity, and ease of operation, as well as superb optical properties and colloidal stability. We introduce integrated single-particle tracking and pair-correlation function analysis of a spinning-disk confocal microscope platform (iSPT-pCF-SDCM) to study SDot's cellular transport. Endocytosed SDots can undergo a highly potent and noninvasive process of vesicle escape, yielding complete vesicle escape with no serious vesicle disruption. We exploit SDots' unprecedented ability to overcome cellular transport barriers to enhance drug and macromolecule delivery.

Targeted delivery of nanomaterials to specific intracellular locations is essential for the development of many nanomaterials-based biological applications. Thus far the targeting performance has been limited due to various intracellular transport barriers, especially intracellular vesicle trapping. Here we report the application of permeation enhancers based on organic solvents in small percentage to enhance the intracellular targeted delivery of nanomaterials. Previously permeation enhancers based on organic solvents and ionic liquids have been used in overcoming biological transport barriers at tissue, organ, and cellular levels, but this strategy has so far rarely been examined for its potential in facilitating transport of nanometer-scale entities across intracellular barriers, particularly intracellular vesicle trapping. Using the cell nucleus as a model intracellular target and Tat peptide-conjugated quantum dots (QDs-Tat) as a model nanomaterial-based probe, we demonstrate that a small percentage (e.g. 1%) of organic solvent greatly enhances nucleus targeting specificity as well as increasing endocytosis-based cellular uptake of QDs. We combine vesicle colocalization (DiO dye staining), vesicle integrity (calcein dye release), and single-particle studies (pair-correlation function microscopy) to investigate the process of organic solvent-enhanced vesicle escape of QDs-Tat. The organic solvent based vesicle escape-enhancing approach is found to be not only very effective but minimally invasive, resulting in high vesicle escape efficiency with no significant disruption to the membrane integrity of either intracellular vesicles or cells. This approach drastically outperforms the commonly used vesicle escape-enhancing agent (i.e., chloroquine, whose enhancement effect is based on disrupting vesicle integrity) in both potency and minimal invasiveness. Finally, we apply organic solvent-based targeting enhancement to improve the intracellular delivery of the anticancer drug doxorubicin (DOX).

Micellar nanocrystals (micelles with encapsulated nanocrystals) have become an emerging major class of nanobiomaterials. We describe a method of fabricating micellar nanocrystals based on combining top-down electrospray, bottom-up self-assembly, and solvent-based structure control. This method involves first using electrospray to generate uniform ultrafine liquid droplets, each of which functions as a micro-reactor in which self-assembly reaction occurs forming micellar nanocrystals, with the structures (micelle shape and nanocrystal encapsulation) controlled by the organic solvent used. This method is largely continuous and produces high quality micellar nanocrystal products with an inexpensive structure control approach. By using a water-miscible organic solvent tetrahydrofuran (THF), worm-shaped micellar nanocrystals can be produced due to solvent-induced/facilitated micelle fusion. Compared with the common spherical micellar nanocrystals, worm-shaped micellar nanocrystals can offer minimized non-specific cellular uptake, thus enhancing biological targeting. By co-encapsulating multiple nanocrystals into each micelle, multifunctional or synergistic effects can be achieved. Current limitations of this fabrication method, which will be part of the future work, primarily include imperfect encapsulation in the micellar nanocrystal product and the incompletely continuous nature of the process.

The interfacial instability process is an emerging general method to fabricate nanocrystal-encapsulated micelles (also called micellar nanocrystals) for biological detection, imaging, and therapy. The present work utilized fluorescent semiconductor nanocrystals (quantum dots or QDs) as the model nanocrystals to investigate the interfacial instability-based fabrication process of nanocrystal-encapsulated micelles. Our experimental results suggest intricate and intertwined roles of the emulsion droplet size and the surfactant poly (vinyl alcohol) (PVA) used in the fabrication process of QD-encapsulated poly (styrene-b-ethylene glycol) (PS-PEG) micelles. When no PVA is used, no emulsion droplet and thus no micelle is successfully formed; Emulsion droplets with large sizes (~25 μm) result in two types of QD-encapsulated micelles, one of which is colloidally stable QD-encapsulated PS-PEG micelles while the other of which is colloidally unstable QD-encapsulated PVA micelles; In contrast, emulsion droplets with small sizes (~3 μm or smaller) result in only colloidally stable QD-encapsulated PS-PEG micelles. The results obtained in this work not only help to optimize the quality of nanocrystal-encapsulated micelles prepared by the interfacial instability method for biological applications but also offer helpful new knowledge on the interfacial instability process in particular and self-assembly in general.

Quantum dots (QDs) have tremendous potential for biomedical imaging, including super-resolution techniques that permit imaging below the diffraction limit. However, most QDs are produced via organic methods, and hence require surface treatment to render them water-soluble for biological applications. Previously, we reported a micelle-templating method that yields nanocomposites containing multiple core/shell ZnS-CdSe QDs within the same nanocarrier, increasing overall particle brightness and virtually eliminating QD blinking. Here, this technique is extended to the encapsulation of Mn-doped ZnSe QDs (Mn-ZnSe QDs), which have potential applications in super-resolution imaging as a result of the introduction of Mn(2+) dopant energy levels. The size, shape and fluorescence characteristics of these doped QD-micelles were compared to those of micelles created using core/shell ZnS-CdSe QDs (ZnS-CdSe QD-micelles). Additionally, the stability of both types of particles to photo-oxidation was investigated. Compared to commercial QDs, micelle-templated QDs demonstrated superior fluorescence intensity, higher signal-to-noise ratios, and greater stability against photo-oxidization,while reducing blinking. Additionally, the fluorescence of doped QD-micelles could be modulated from a bright 'on' state to a dark 'off' state, with a modulation depth of up to 76%, suggesting the potential of doped QD-micelles for applications in super-resolution imaging.

Nanoparticle encapsulation within micelles has been demonstrated as a versatile platform for creating water-soluble nanocomposites. However, in contrast to typical micelle encapsulants, such as small molecule drugs and proteins, nanoparticles are substantially larger, which creates significant challenges in micelle synthesis, especially at large scale. Here, we describe a new nanocomposite synthesis method that combines electrospray, a top-down, continuous manufacturing technology currently used for polymer microparticle fabrication, with bottom-up micellar self-assembly to yield a scalable, semicontinuous micelle synthesis method: i.e., micellar electrospray. Empty micelles and micellar nanocomposites containing quantum dots (QDs), superparamagnetic iron oxide nanoparticles (SPIONs), and their combination were produced using micellar electrospray with a 30-fold increase in yield by weight over batch methods. Particles were characterized using dynamic light scattering, transmission electron microscopy, and scanning mobility particle sizing, with remarkable agreement between methods, which indicated size distributions with variations of as little as ~5%. In addition, new methodologies for qualitatively evaluating nanoparticle loading in the resultant micelles are presented. Micellar electrospray is a broad, scalable nanomanufacturing approach that should be easily adapted to virtually any hydrophobic molecule or nanoparticle with a diameter smaller than the micelle core, potentially enabling synthesis of a vast array of nanocomposites and self-assembled nanostructures.

Quantum dots (QDs) have great promise in biological imaging, and as this promise is realized, there has been increasing interest in combining the benefits of QDs with those of other materials to yield composites with multifunctional properties. One of the most common materials combined with QDs is magnetic materials, either as ions (e.g. gadolinium) or as nanoparticles (e.g. superparamagnetic iron oxide nanoparticles, SPIONs). The fluorescent property of the QDs permits visualization, whereas the magnetic property of the composite enables imaging, magnetic separation, and may even have therapeutic benefit. In this review, the synthesis of fluorescent-magnetic nanoparticles, including magnetic QDs is explored; and the applications of these materials in imaging, separations, and theranostics are discussed. As the properties of these materials continue to improve, QDs have the potential to greatly impact biological imaging, diagnostics, and treatment., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The ability to quickly analyze, separate, and manipulate multiple types of biomarkers from small sample volumes is a significant step toward personalized medicine.

Because of their extraordinary brightness and photostability, quantum dots (QDs) have tremendous potential for long-term, particle tracking in heterogeneous systems (e.g., living cells, microfluidic flow). However, one of their major limitations is blinking, an intermittent loss of fluorescence, characteristic of individual and small clusters of QDs, that interrupts particle tracking. Recently, several research groups have reported "nonblinking QDs". However, blinking is the primary method used to confirm nanoparticle aggregation status in situ, and single or small clusters of nanoparticles with continuous fluorescence emission are difficult to discern from large aggregates. Here, we describe a new class of quantum dot-based composite nanoparticles that solve these two seemingly irreconcilable problems by exhibiting near-continuous, alternating-color fluorescence, which permits aggregation status discrimination by observable color changes even during motion across the focal plane. These materials will greatly enhance particle tracking in cell biology, biophysics, and fluid mechanics.

Controlled transport of multiple individual nanostructures is crucial for nanoassembly and nanodelivery but is challenging because of small particle size. Although atomic force microscopy and optical and magnetic tweezers can control single particles, it is extremely difficult to scale these technologies for multiple structures. Here, we demonstrate a "nano-conveyer-belt" technology that permits simultaneous transport and tracking of multiple individual nanospecies in a selected direction. The technology consists of two components: nanocontainers, which encapsulate the nanomaterials transported, and nanoconveyer arrays, which use magnetic force to manipulate individual or aggregate nanocontainers. This technology is extremely versatile. For example, nanocontainers encapsulate quantum dots or rods and superparamagnetic iron oxide nanoparticles in <100 nm nanocontainers, the smallest magnetic composites to have been simultaneously moved and optically tracked. Similarly, the nanoconveyers consist of patterned microdisks or zigzag nanowires, whose dimensions can be controlled through micropatterning. The nanoconveyer belt technology could impact multiple fields, including nanoassembly, biomechanics, nanomedicine, and nanofluidics.

A new bifunctional nanocomposite, consisting of CdTe quantum dots complexed with FeP nanoparticles, was synthesized using a one-pot, high temperature precursor decomposition method. Nanocomposites formed consisted of heterofunctional, dumbbell shaped particles, which exhibited supplementary and altered properties from those of constituent particles. For example, an additional peak was observed in the PL spectra, whose intensity was directly related to the ratio of FeP:CdTe. Also, the magnetic behavior of the particles altered from ferromagnetic to paramagnetic upon conjugation to CdTe. These results indicate that interactions between monomers in multifunctional particles can occur, which might result from the formation of a composite 'doped' phase at the interface of the individual particles. These interactions may lead to additional properties that could be exploited to increase the utility of the resultant particles.

We report the use of Tat peptide-conjugated quantum dots (Tat-QDs) to examine the complex behavior of nanoparticle probes in live cells, a topic that is of considerable current interest in developing advanced nanoparticle agents for molecular and cellular imaging. Dynamic confocal imaging studies indicate that the peptide-conjugated QDs are internalized by macropinocytosis, a fluid-phase endocytosis process triggered by Tat-QD binding to negatively charged cell membranes. The internalized Tat-QDs are tethered to the inner vesicle surfaces and are trapped in cytoplasmic organelles. The QD loaded vesicles are found to be actively transported by molecular machines (such as dyneins) along microtubule tracks. The destination of this active transport is an asymmetric perinuclear region (outside the cell nucleus) known as the microtubule organizing center (MTOC). We also find that Tat-QDs strongly bind to cellular membrane structures such as filopodia and that large QD-containing vesicles are released from the tips of filopodia by vesicle shedding. These results provide new insights into the mechanisms of Tat peptide-mediated delivery as well as toward the design of functionalized nanoparticles for molecular imaging and targeted therapy.

A number of procedures are currently available to encapsulate and solubilize hydrophobic semiconductor Quantum Dots (QDs) for biological applications. Most of these procedures are based on the use of small-molecule coordinating ligands, amphiphilic polymers, or amphiphilic lipids. However, it is still not clear how these different surface coating molecules affect the optical, colloidal, and chemical properties of the solubilized QDs. Here we report a systematic study to examine the effects of surface coating chemistry on the hydrodynamic size, fluorescence quantum yield, photostability, chemical stability, and biocompatibility of water-soluble QDs. The results indicate that quantum dots with the smallest hydrodynamic sizes are best prepared by direct ligand exchange with hydrophilic molecules, but the resulting particles are less stable than those encapsulated in amphiphilic polymers. For stability against chemical oxidation, QDs should be protected with a hydrophobic bilayer. For high stability under acidic conditions, the best QDs are prepared by using hyperbranched polyethylenimine. For stability in high salt buffers, it is preferable to have uncharged, sterically-stabilized QDs, like those coated with polyethylene glycol (PEG). These insights are expected to benefit the development of quantum dots and related nanoparticle probes for molecular and cellular imaging applications.

Semiconductor quantum dots (QDs) are a new class of fluorescent labels with broad applications in biomedical imaging, disease diagnostics, and molecular and cell biology. In comparison with organic dyes and fluorescent proteins, quantum dots have unique optical and electronic properties such as size-tunable light emission, improved signal brightness, resistance against photobleaching, and simultaneous excitation of multiple fluorescence colors. Recent advances have led to multifunctional nanoparticle probes that are highly bright and stable under complex in vitro and in vivo conditions. New designs involve encapsulating luminescent QDs with amphiphilic block copolymers, and linking the polymer coating to tumor-targeting ligands and drug-delivery functionalities. These improved QDs have opened new possibilities for real-time imaging and tracking of molecular targets in living cells, for multiplexed analysis of biomolecular markers in clinical tissue specimens, and for ultrasensitive imaging of malignant tumors in living animal models. In this article, we briefly discuss recent developments in bioaffinity QD probes and their applications in molecular profiling of individual cancer cells and clinical tissue specimens.

Semiconductor quantum dots are luminescent nanoparticles that are under intensive development for use as a new class of optical imaging contrast agents. Their novel properties such as optical tunability, improved photostability, and multicolor light emission have opened new opportunities for imaging living cells and in vivo animal models at unprecedented sensitivity and spatial resolution. Combined with biomolecular engineering strategies for tailoring the particle surfaces at the molecular level, bio-conjugated quantum dot probes are well suited for imaging single-molecule dynamics in living cells, for monitoring protein-protein interactions within specific intracellular locations, and for detecting diseased sites and organs in deep tissue. In this article, we describe the engineering principles for preparing high-quality quantum dots and for conjugating the dots to biomolecular ligands. We also discuss recent advances in using quantum dots for in vivo molecular and cellular imaging.

Perturbative contributions to single-beam two-photon transition rates may be divided into two types. The first, involving low-energy intermediate states, require a high-order perturbation treatment, or an exact diagonalization. The other, involving high-energy intermediate states, only require a low-order perturbation treatment. We show how to partition the effective transition operator into two terms, corresponding to these two types, in such a way that a many-body perturbation expansion may be generated that obeys the linked cluster theorem and has a simple diagrammatic representation.

In the present work, we developed a novel drug delivery system, liposomes-in-microsphere (LIM) of biodegradable polymers, which is conceived from a combination of the polymer- and the lipid-based delivery systems and can thus integrate the advantages and avoid the drawbacks of the two systems. Liposomes were encapsulated into microspheres of biodegradable polymers by the solvent extraction/evaporation process to form LIMs. The integrity of the liposomes was preserved by modifying the microencapsulation process and coating the liposomes with chitosan. We demonstrated by scanning electron microscopy, laser light scattering and fluorescence spectroscopy that the particle size and surface morphology of the polymeric microspheres did not change significantly with the liposomes encapsulated, the liposomes remained intact within the polymeric matrix of the microspheres, and the encapsulated liposomes could be released from the microspheres in a controlled manner at a nearly constant release rate after an initial off-release period. Decreasing the particle size of liposomes and increasing the pore size of the polymeric matrix shortened the initial off-release period and increased the liposome release rate. In conclusion, a novel drug delivery system, liposomes-in-microsphere, was successfully developed and characterized. The liposome release kinetics could be controlled by the composition and fabrication parameters of the liposomes and polymeric microspheres. Such a novel controlled release system may have potential to be applied for drug delivery and gene therapy.

Microspheres of a new kind of copolymer, poly(lactic acid)-poly(ethylene glycol)-poly(lactic acid) (PLA-PEG-PLA), are proposed in the present work for clinical administration of an antineoplastic drug paclitaxel with hypothesis that incorporation of a hydrophilic PEG segment within the hydrophobic PLA might facilitate the paclitaxel release. Paclitaxel-loaded PLA-PEG-PLA microspheres of various compositions were prepared by the solvent extraction/evaporation method. Characterization of the microspheres was then followed to examine the particle size and size distribution, the drug encapsulation efficiency, the colloidal stability, the surface chemistry, the surface and internal morphology, the drug physical state and its in vitro release behavior. The effects of polymer types, solvents and drug loading were investigated. It was found that in the microspheres the PEG segment was homogeneously distributed and caused porosity. Significantly faster release from PLA-PEG-PLA microspheres resulted in comparison with the PLGA counterpart. Incorporation of water-soluble solvent acetone in the organic solvent phase further increased the porosity of the PLA-PEG-PLA microspheres and facilitated the drug release. A total of 49.6% sustained release of paclitaxel within 1 month was achieved. Potentially, the presence of PEG on the surface of PLA-PEG-PLA microspheres could improve their biocompatibility. PLA-PEG-PLA microspheres could thus be promising for the clinical administration of highly hydrophobic antineoplastic drugs such as paclitaxel.

Human serum albumin (HSA) was encapsulated as a model protein in microspheres of biodegradable and biocompatible polymers by the water-in-oil-in-water (w/o/w) emulsion solvent extraction/evaporation (double emulsion) technique for purpose of controlled release. To improve the properties and control the rate of drug release of the delivery vehicle, materials with different hydrophobicity from that of their conventional counterparts, such as poly(lactide-co-ethylene glycol) (PELA) in place of poly(lactide-co-glycolide) (PLGA) as the polymer matrix, ethyl acetate/acetone in place of dichloride methane (DCM) as the (co)solvent and d-alpha tocopheryl polyethylene glycol 1000 succinate (vitamin E TPGS) as the additive, were used to prepare the microspheres. It has been found that PELA microspheres, compared with PLGA ones, were slightly smaller in size if prepared at identical emulsification strength. They had more porous surface and internal structure, higher encapsulation efficiency (EE) and more rapid in vitro release rate. Furthermore, the physical properties of the microspheres were also affected by the presence of solvents and additives and their properties. Our results suggest that these materials could have interesting potential applications in preparation of polymeric microspheres for controlled protein release., (Copyright 2002 Elsevier Science B.V.)

OBJECTIVE To investigate the relationship between the Nco I, Ava II polymorphism of low density lipoprotein receptor (LDL-R) gene in patients with the occurrence of atherosclerotic cerebral infarction (ACI) among the Han nationality in Liaoning province. METHODS The polymerase chain reaction technique was used to study the polymorphisms of LDL-R gene and allele frequencies in 77 patients with ACI and in 113 age-matched Chinese healthy controls. The levels of the lipid and lipoproteins were also compared among the cases with ACI and the controls. RESULTS A(+) frequencies of LDL-R gene in healthy controls and ACI group were 0.230 and 0.125 respectively, while the N(+) frequencies of healthy control and ACI group was 0.667 and 0.662 respectively. In case of the coexistence of A(-) A(-) and N(+) N(+), the relative risk (RR) of ACI was 5.56(P<0.001), while the RR of the increase of serum levels TG, TC, LDL-C, LP(a) were 4.29, 7.67, 9.33 and 3.09(P<0.05), respectively. CONCLUSION The coexistence of A(-) A(-) and N(+) N(+) can affect the concentration of lipid and lipoprotein and is in close relationship with the occurrence of ACI.