This PDF file contains the front matter associated with SPIE Proceedings Volume 9338, including the Title Page, Copyright information, Table of Contents, Introduction (if any), and Conference Committee listing.

Current advances in nanotechnology hold the promises to greatly impact on current medical practice. Since nanometric materials interact with cells, tissue and organs at a molecular level, they may be used as probes for ultrasensitive molecular sensing and diagnostic imaging or carriers for drug and gene delivery. However, along with the excitement that has driven the development of novel nanocarriers, there have been increasing concerns regarding the risks these materials may generate. As these nanostructures are intentionally engineered to target specific cells or tissues, it is imperative to ensure their safety. The optimal design of safe and functional nanocarriers for medicine requires a better understanding of the interaction between the physical-chemistry properties of the nanoparticle surface with the complex protein machinery existing at the cell membrane. In particular the effect of the particles properties (charge, shape, protein coating) on the mechanism of cellular uptake is highly relevant both to assess the real biological risks coupled with the use of nanomaterial (nanopathology and nanotoxicology) and to engineer carriers able to improve the medical practice. The nanometric size and the surface molecular decoration may activate mechanisms of cellular uptake different from those commonly used by cells: these open the possibility to activated/modulated the membrane crossing by tuning chemical-physical properties of nanometric materials. In this work, the design and production of novel degradable polymeric nanocavities via layer-by-layer and temperature induced phase separation technology will be presented along with a detailed characterization of their in vitro performances. Furthermore, possible mechanisms of cellular uptake will be discussed and critically presented. The effect of surface bioconjugation on cell membrane crossing will be exploited and elucidated. Particular attention will be devoted to surface molecular decoration able to guide the nanoparticle throughout the cytosol.

Numerous nanomaterials have been developed for biomedical application, especially cancer therapy. Visualizing cancer therapy is highly promising now because of the potential ability to realize accurate, localized treatment. In this work, we firstly synthesized metal nanorattles (MNRs), which utilized porous gold shells capable of photothermal therapy to carry multiple superparmagnetic iron oxide nanoparticles (SPIONs) as MR imaging contrast agents inside. As shown in the infrared light, these metal rattle-typed nanostructures were able to convert to heat to kill cells, and inhibit tumor growth. As a carrier for multiple SPIONs, it also performed a good behavior for T2-weighted MR imaging in tumor site. Moreover, the rest of the inner space of the gold shell also introduced potential ability as nanocarriers for other cargos such as chemotherapeutic drugs, which is still under investigation. This metal-rattle-type nanocarriers is highly potential as a novel platforms for cancer therapy in the future.

In this paper we present current and previous methods developed in our laboratory to program the assembly of nanoparticles into oligomers or more sophisticated structures. To direct the assembly of nanoparticles we used biomolecular and chemical tools ranging from peptides to oligonucleotides and photosensitive ligands.

Photodynamic therapy (PDT) for cancer is based on the use of a light sensitive molecule to produce, under specific irradiation, toxic reactive oxygen species (ROS). A way to improve the therapy efficiency is to increase the amount of produced ROS near cancer cells. This aim can be achieved by using a metal enhanced process arising when an optically active molecule is located near a metallic nanoparticle (NP). Here, the coupling effect between silver (Ag) NPs and protoporphyrin IX (PpIX) molecules, a clinically approved photosensitizer, is studied compared first, to PpIX fluorescence yield and second, to ROS production efficiency. By applying a modified Stöber process, PpIX was encapsulated into a silica (SiO₂) shell, surrounding a 60 nm sized Ag core. We showed that, compared to SiO₂-PpIX NPs, Ag coated SiO₂-PpIX NPs dramatically decreased PpIX fluorescence together with singlet oxygen production efficiency. However, after incubation time in the dark, the amount of superoxide anions generated by the Ag doped sample was higher than the control sample one.

We report progress towards combining radiation therapy (RT) and photodynamic therapy (PDT) using scintillating nanoparticle (NP)-photosensitizer conjugates. In this approach, scintillating NPs are excited by clinically relevant ionizing radiation sources and subsequently transfer energy to conjugated photosensitizers via FRET, acting as an energy mediator between ionizing radiation and photosensitizer molecules. The excited photosensitizers generate reactive oxygen species that can induce local damage and immune response. Advantages of the scheme include: 1) Compared with traditional radiation therapy, a possible decrease of the total radiation dose needed to eliminate the lesion; 2) Compared with traditional PDT, the ability to target deeper and more highly pigmented lesions; 3) The possibility of additional photosensitizing effects due to the scintillation of the nanoparticles. In this work, the photosensitizer molecule chlorin e6 was covalently bound to the surface of LaF3:Ce NPs. After conjugation, the photoluminescence intensity of NPs decreased, and fluorescence lifetime of conjugated chlorin e6 became sensitive to excitation wavelength, suggesting rapid FRET. In addition, scintillation spectra of nanoparticles were measured. Preliminary calculations suggest that the observed scintillation efficiencies are sufficient to enhance RT. In vitro cancer cell studies suggest conjugates are taken up by cells. Survival curves with radiation exposure suggest that the particles alone cause radiosensitization comparable to that seen with gold nanoparticles.

The goals of this work were to determine whether conjugation of any of four selected peptides to Au nanoparticles improved their delivery to B16 melanoma in vitro and in vivo. In in vitro cytotoxicity assays, peptides and conjugates were endocytosed but did not escape from endosomes. None of the peptides showed any cytotoxicity, with or without conjugation to the nanoparticles. The combination of peptides and doxorubicin did not improve upon the cytotoxicity of gold-doxorubicin alone. We then tested targeting in vivo using inductively coupled plasma mass spectrometry to quantify the concentration of Au in the organs of B16 tumor-bearing mice 4, 24, and 72 h after intravenous Au nanoparticle injection. These experiments showed that in some cases, peptide conjugation improved upon the enhanced permeability and retention (EPR) effect. A peptide based upon the myxoma virus and the cyclic RGD peptide were both effective at tumor targeting; myxoma was more effective with un-PEGylated particles, and cRGD with PEGylated particles. The FREG and melanocyte stimulating hormone (MSH) peptides did not improve targeting. These results suggest that these peptides may improve delivery of Au particles to tumors, but also may prevent entry of particles into cell nuclei.

Fluorescent nanoparticles (NPs) are promising optical probes for biological and biomedical applications, thanks to their excellent photophysical properties, color tunability and facile bioconjugation. It still remains unclear, however, how fluorescent NPs behave in the complex biological environment. Our group has quantified interactions of different fluorescent NPs (i.e., semiconductor quantum dots and metal nanoclusters) with serum proteins and living cells by the combined use of different spectroscopic and microscopic techniques. Our studies show that (1) interactions with proteins may significantly alter the photophysical properties of the NPs as well as the responses of cells internalizing them; (2) protein surface charge distributions play an important role in the interactions of NPs with proteins and cells; (3) ultrasmall NPs (diameter less than 10 nm) show a cellular internalization behavior that is distinctly different from the one observed with larger particles (diameter ~100 nm).

Interactions between luminescent fluorophores and redox active molecules often involve complex charge transfer processes, and have great ramifications in biology. Dopamine is a redox active neurotransmitter involved in a range of brain activities. We used steady-state and time-resolved fluorescence along with transient absorption bleach measurements, to probe the effects of changing the QD size and valence on the rate of photoluminescence quenching in QD-dopamine conjugates, when the pH of the medium was varied. In particular, we measured substantially larger quenching efficiencies, combined with more pronounced shortening in the PL lifetime decay when smaller size QDs and/or alkaline pH were used. Moreover, we found that changes in the nanocrystal size alter both the electron and hole relaxation of photoexcited QDs but with very different extents. For instance, a more pronounced change in the hole relaxation was recorded in alkaline buffers and for green-emitting QDs compared to their red-emitting counterparts. We attributed these results to the more favorable electron transfer pathway from the reduced form of the complex to the valence band of the QD. This process benefits from the combination of lower oxidation potential and larger energy mismatch in alkaline buffers and for green-emitting QDs. In comparison, the effects on the rate of electron transfer from excited QDs to dopamine are less affected by QD size. These findings provide new insights into the mechanisms that drive charge transfer interactions and the ensuing quenching of QD emission in such assemblies.

There is currently interest in the development of nanoemulsions as imaging and therapeutic agents, particularly perfluorohexane (PFH) droplets, whose amphiphilic shell protects drugs against physico-chemical and enzymatic degradation. When delivered to their target sites, these perfluorocarbon (PFC) droplets can vaporize upon laser excitation, efficiently releasing their drug payload and/or imaging tracers. Due to the optical properties of gold, coupling PFC droplets with gold nanoparticles significantly reduces the energy required for vaporization. In this work, nanoemulsions with a PFC core and Zonyl FSP surfactant shell were produced using sonication. Droplets were characterized in terms of size and morphology using high resolution fluorescence microscopy (i.e. total internal reflection fluorescence microscopy, TIRFM), fluorescence correlation spectroscopy (FCS), transmission electron microscopy (TEM), and light scattering techniques (i.e. dynamic light scattering, DLS). The ability of PFC droplets to vaporize was demonstrated using optical light microscopy.

It is now well accepted that upon their entrance into the biological environments, the surface of nanomaterials would be covered by various biomacromolecules (e.g., proteins and lipids). The absorption of these biomolecules, so called ‘protein corona’, onto the surface of (nano)biomaterials confers them a new ‘biological identity’. Although the formation of protein coronas on the surface of nanoparticles has been widely investigated, there are few reports on the effect of various diseases on the biological identity of nanoparticles. As the type of diseases may tremendously changes the composition of the protein source (e.g., human plasma/serum), one can expect that amount and composition of associated proteins in the corona composition may be varied, in disease type manner. Here, we show that corona coated silica and polystyrene nanoparticles (after interaction with in the plasma of the healthy individuals) could induce unfolding of fibrinogen, which promotes release of the inflammatory cytokines. However, no considerable releases of inflammatory cytokines were observed for corona coated graphene sheets. In contrast, the obtained corona coated silica and polystyrene nanoparticles from the hypofibrinogenemia patients could not induce inflammatory cytokine release where graphene sheets do. Therefore, one can expect that disease-specific protein coronas can provide a novel approach for applying nanomedicine to personalized medicine, improving diagnosis and treatment of different diseases tailored to the specific conditions and circumstances.

Various methods have been used in medicine for more than one century to explore the lymphatic system. Radioactive colloids (RuS labelled with ^99mTc) or/and Vital Blue dye are injected around the primary tumour and detected by means of nuclear probe or visual colour inspection respectively. The simultaneous clinical use of both markers (dye and radionuclide) improves the sensitivity of detection close to 100%. Superparamagnetic iron oxides (SPIOs) are currently receiving much attention as strong T₂ weighted magnetic resonance imaging contrast agents that can be potentially used for preoperative localization of sentinel nodes, but also for peroperative detection of sentinel node using hand-held probes. In that context, we present the elaboration of dendronized iron oxide nanoparticles elaborated at the Institute of Physics and Chemistry of Materials of Strasbourg.

There is considerable interest in using Quantum Dots (QDs) as fluorescent probes such for cellular imaging due to unique advantages in comparison with conventional molecular dyes. However, cytosolic delivery of QDs into live cells remains a major challenge. Here we demonstrate highly efficient delivery of PEG-coated QDs into live cells by means of laser-induced vapour nanobubbles. Using this procedure we succeeded in high-throughput loading of ~80% of cells while maintaining a cell viability of ~85%.

Quantum dots are semiconductor nanocrystals that absorb and emit light at wavelengths tunable by the size of the crystal. Size-tuning provides access to a broad range of optical spectra, however it is fundamentally problematic for many applications because it leads to a large mismatch in absorption cross-section and fluorescence brightness across a series of colors. We have recently demonstrated engineering strategies to generate multicolor, extinction-matched, and brightness-matched quantum dots based on colloidal multi-domain core/shell structures. We use alloyed cores with composition-tunable bandgaps and finely adjust the domain size and electronic properties of the shell to precisely match both absorption cross-section and quantum yield. Using this strategy, it is possible to tune fluorescence wavelength, extinction, and quantum yield independently, vastly expanding the photophysical landscape of these materials. Moreover compared with conventional size-tuning strategies, this enables access to a wider spectral range with compact dimensions. The equalized optical properties translate from the ensemble level down to the single-molecule level, setting the stage for new possibilities in highly quantitative, multiplexed imaging in cells and tissue. However selection of appropriate structural parameters to generate specific optical properties is challenging without insight into the photophysics of these materials. Here we describe the evolution of the optical properties of alloyed cores during the shell growth process that provide new insights into general engineering strategies.

Single particle tracking has provided a wealth of information about biophysical processes such as motor protein transport and diffusion in cell membranes. However, motion out of the plane of the microscope or blinking of the fluorescent probe used as a label generally limits observation times to several seconds. Here, we overcome these limitations by using novel non-blinking quantum dots as probes and employing a custom 3D tracking microscope to actively follow motion in three dimensions (3D) in live cells. Signal-to-noise is improved in the cellular milieu through the use of pulsed excitation and time-gated detection.

Folic acid (FA) has been used as a molecular targeting strategy to improve the specificity of a CQD-protoporphyrin IX (CQD-PPIX) conjugate to folate receptor positive (FR+) HeLa cells for use in two-photon excited Photodynamic Therapy (TPE-PDT). FA was covalently attached to the CQD-PPIX conjugate to form a FA-CQD-PPIX conjugate. The uptake of the FA-CQD-PPIX conjugate in FR+ HeLa cells was shown to be 7 times greater than the CQD-PPIX conjugate, while both conjugates showed a similar uptake in FR negative (FR-) HT-47 cells. TPE-PDT experiments, using HeLa cells as a target, revealed a 30% improved cytotoxicity for cells treated with the FA-CQD-PPIX conjugate and TPE compared to controls treated with the CQD-PPIX conjugate and TPE. Collectively, these results suggest the presence of FA can facilitate targeting of CQD-sensitiser conjugates to FR+ cells resulting in an improved PDT effect.

Nanoparticles can be synthesized in a wide array of shapes and sizes to suit specific biomedical applications in therapy and imaging. Prerequisite to such applications are particle stability in biological environments, non-toxicity, and facile conjugation of the particle surface with targeting biological moieties (such as antibodies). Here we report significant flaws in the common methods used to functionalize the surface of gold nanorods (GNRs) of larger-than-usual sizes. We find that while GNRs of sizes smaller than 50 x 15 nm can be effectively stabilized by polyethylene glycol (PEG)-based methods, larger GNRs form major aggregates and crash under similar functionalization conditions. Large GNRs may provide enhanced imaging sensitivity in biological applications due to greater optical extinction cross sections, provided that the GNRs can be made biostable. In this study, GNRs of sizes up to 90 x 30 nm were synthesized using two different published methods. Particle morphology and size distributions were characterized using Transmission Electron Microscopy (TEM), and optical spectra were measured by Vis-NIR Spectrometry. The colloidal stability of different-sized GNRs was assayed at various stages of functionalization using zeta potential and Vis-NIR measurements. The results of these experiments indicate that large GNRs functionalized with PEG undergo irreversible aggregation after minimal washing. We find that coating large GNRs with polystyrene sulfonate (PSS) instead of PEG vastly improves GNR stability in water and serum. Moreover, we provide a novel platform for conjugating biomolecules of interest to PSS-coated large GNRs. We show that larger GNRs produce stronger photoacoustic signal than commonly used smaller GNRs, indicating an advantage of using large GNRs for biomedical imaging. Our observations underscore that the biomedical advantages of novel nanoparticle synthesis methods may not be realized without tailored surface functionalization methods. More generally, our results suggest that materially-identical nanoparticles (i.e. GNRs) exhibit varying stability as a function of particle size.

Multifunctional nanoparticles have attracted a lot of attention since they can combine interesting properties like magnetism, fluorescence or plasmonic effects. As a core material, iron oxide nanoparticles have been the subject of intensive research. These cost-effective and non-toxic particles are used nowadays in many applications. We developed a heterobifunctional PEG ligand that can be used to introduce functional groups (carboxylic acids) onto the surface of the NP. Via click chemistry, a siloxane functionality was added to this ligand, for a subsequent covalent ligand exchange reaction. The functionalized nanoparticles have an excellent colloidal stability in complex environments like buffers and serum or plasma. Antibodies were coupled to the introduced carboxylic acids and these NP-antibody bioconjugates were brought into contact with Legionella bacteria for magnetic separation experiments.

The self-assembly phenomena on a special class of poly-hydroxy sugar surfactant have been studied extensively. This class of material is classified as amphitropic liquid crystals since they exhibit both thermotropic and lyotropic liquid crystalline properties. Hence the potential applications of these non-ionic surfactants are more versatile than those from the conventional lyotropic liquid crystals including those from thermotropic phases, but the latters are yet to be realized. Unfortunately, due to the lack of interest (or even awareness), fundamental studies in thermotropic glycolipids are scanty to support application development, and any tangible progress is often mired by the complexity of the sugar stereochemistry. However, some applications may be pursued from these materials by taking the advantage of the sugar chirality and the tilted structure of the lipid organization which implies ferroelectric behavior. Here, we present our studies on the stereochemical diversity of the sugar units in glycosides that form the thermotropic/lyotropic phases. The structure to property relationship compares different chain designs and other popular polyhydroxy compounds, such as monooleins and alkylpolyglucosides. Different structural properties of these glycosides are discussed with respect to their self-assembly organization and potential applications, such as delivery systems and membrane mimetic study.

We have designed a new set of coordinating ligands made of a lipoic acid (LA) anchor and poly(ethylene glycol) (PEG) hydrophilic moiety appended with a terminal aldehyde for the surface functionalization of QDs. This ligand design was combined with a recently developed photoligation strategy to prepare hydrophilic CdSe-ZnS QDs with good control over the fraction of intact aldehyde (-CHO) groups per nanocrystal. We further applied the efficient hydrazone ligation to react aldehyde-QDs with 2-hydrazinopyridine (2-HP). This covalent modification produces QD-conjugates with a well-defined absorption feature at 350 nm ascribed to the hydrazone chromophore. We exploited this unique optical signature to accurately measure the number of aldehyde groups per QD when the fraction of LA-PEG-CHO per nanocrystal was varied. This allowed us to extract an estimate for the number of LA-PEG ligands per QD. These results suggest that hydrazone ligation has the potential to provide a simple and general analytical method to estimate the number of surface ligands for a variety of nanocrystals such as metal, metal oxide and semiconductor nanocrystals.

Functionalized colloidal nanoparticles for SERS serve as a promising multifunctional assay component for blood biomarker detection. Proper design of these nanoprobes through conjugation to spectral tags, protective polymers, and sensing ligands can provide experimental control over the sensitivity, range, reproducibility, particle stability, and integration with biorecognition assays. Additionally, the optical properties and degree of electromagnetic SERS signal enhancement can be altered and monitored through tuning the nanoparticle shape, size, material and the colloid’s local surface plasmon resonance (LSPR). Aptamers, synthetic affinity ligands derived from nucleic acids, provide a number of advantages for biorecognition of small molecules and toxins with low immunogenicity. DNA aptamers are simpler and more economical to produce at large scale, are capable of greater specificity and affinity than antibodies, are easily tailored to specific functional groups, can be used to tune inter-particle distance and shift the LSPR, and their intrinsic negative charge can be utilized for additional particle stability.1,2 Herein, a “turn-off” competitive binding assay platform involving two different plasmonic nanoparticles for the detection of the toxin bisphenol A (BPA) using SERS is presented. A derivative of the toxin is immobilized onto a silver coated magnetic nanoparticle (Ag@MNP), and a second solid silver nanoparticle (AgNP) is functionalized with the BPA aptamer and a Raman reporter molecule (RRM). The capture (Ag@MNP) and probe (AgNP) particles are mixed and the aptamer binding interaction draws the nanoparticles closer together, forming an assembly that results in an increased SERS signal intensity. This aptamer mediated assembly of the two nanoparticles results in a 100x enhancement of the SERS signal intensity from the RRM. These pre-bound aptamer/nanoparticle conjugates were then exposed to BPA in free solution and the competitive binding event was monitored by the decrease in SERS intensity.

Nanotechnology use in drug delivery promotes a reduction in systemic toxicity, improved pharmacokinetics, and better drug bioavailability. Liposomes continue to be extensively researched as drug delivery systems (DDS) with formulations such as Doxil^® and Ambisome^® approved by FDA and successfully marketed in the United States. However, the limited ability to precisely control release of active ingredients from these vesicles continues to challenge the broad implementation of this technology. Moreover, the full potential of the carrier to sequester drugs until it can reach its intended target has yet to be realized. Here, we describe a liposomal DDS that releases therapeutic doses of an anticancer drug in response to external stimulus. Earlier, we introduced degradable plasmon resonant liposomes. These constructs, obtained by reducing gold on the liposome surface, facilitate spatial and temporal release of drugs upon laser light illumination that ultimately induces an increase in temperature. In this work, plasmon resonant liposomes have been developed to stably encapsulate and retain doxorubicin at physiological conditions represented by isotonic saline at 37^o C and pH 7.4. Subsequently, they are stimulated to release contents either by a 5^o C increase in temperature or by laser illumination (760 nm and 88 mW/cm² power density). Successful development of degradable plasmon resonant liposomes responsive to near-infrared light or moderate hyperthermia can provide a new delivery method for multiple lipophilic and hydrophilic drugs with pharmacokinetic profiles that limit clinical utility.

Nanodrugs selectively delivered to a tumor site can be activated by radiation for drug release, or nanoparticles (NPs) can be used as a drug themselves by producing biological damage in cancer cells through thermal, mechanical ablations or charged particle emission. Radio-frequency (RF) waves have an excellent ability to penetrate into the human body without causing healthy tissue damage, which provides a great opportunity to activate/heat NPs delivered inside the body as a contrast agent for diagnosis and treatment purposes. However the heating of NPs in the RF range of the spectrum is controversial in the research community because of the low power load of RF waves and low absorption of NPs in the RF range. To resolve these weaknesses in the RF activation of NPs and dramatically increase absorption of contrast agents in tumor, we suggest aggregating the nanoclusters inside or on the surface of the cancer cells. We simulate space distribution of temperature changes inside and outside metal and dielectric nanopraticles/nanoclusters, determine the number of nanoparticles needed to form a cluster, and estimate the thermal damage area produced in surrounding medium by nanopraticles/nanoclusters heated in the RF field.

Macromolecular agents such as nucleic acids and proteins need to be delivered into living cells for therapeutic purposes. Among physical methods to deliver macromolecules across the cell membrane, laser-induced photoporation using plasmonic nanoparticles is a method that is receiving increasing attention in recent years. By irradiating gold nanoparticles bound to the cell membrane with laser light, nanosized membrane pores can be created. Pores are formed by localized heating or by vapour nanobubbles (VNBs) depending on the incident laser energy. Macromolecules in the surrounding cell medium can then diffuse through the transiently formed pores into the cytoplasm. While both heating and VNBs have been reported before for permeabilization of the cell membrane, it remains unclear which of both methods is more efficient in terms of cell loading with minimal cytotoxicity. In this study we report that under condition of a single 7 ns laser pulse VNBs are substantially more efficient for the cytosolic delivery of macromolecules. We conclude that VNB formation is an interesting photoporation mechanism for fast and efficient macromolecular delivery in live cells.

Accurate characterization of submicron particles within biological fluids presents a major challenge for a wide range of biomedical research. Detection, characterization and classification are difficult due to the presence of particles and debris ranging from single molecules up to particles slightly smaller than cells. Especial interest arises from extracellular vesicles (EVs) which are known to play a pivotal role in cell-signaling in multicellular organisms. Tunable resistive pulse sensing (TRPS) is increasingly proving to be a useful tool for high throughput particle-by-particle analysis of EVs and other submicron particles. This study examines the capability of TRPS for characterization of EVs derived from bacteria, also called outer membrane vesicles (OMVs). Measurement of a size distribution (124 ± 3 nm modal diameter) and concentration (lower bound 7.4 x 10⁹ mL^-1) are demonstrated using OMVs derived from uropathogenic Escherichia coli. Important aspects of measurement are discussed, including sample preparation and size selection. Application of TRPS to study EVs could assist the development of these particles in clinical diagnostics and therapeutics.

The penetration of spherical and rod-like gold nanoparticles into human skin is reported. Several skin preparation techniques are applied, including cryo techniques, such as plunge freezing and freeze drying, and the use of wet cells. Their advantages and drawbacks for observing nanoparticle uptake are discussed. Independent of the particle shape no uptake into intact skin is observed by a combination of imaging approaches, including scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), and scanning X-ray microscopy (STXM). These results are discussed along with suitable skin preparation approaches. Experiments on barrier-disrupted skin, i.e. mechanical lesions made by pricking, indicate, however, that gold particles can be identified deep in the dermis, as follows from STXM studies on wet skin samples.

The controlled delivery of nanomaterials to the plasma membrane is critical for the development of nanoscale probes that can eventually enable cellular imaging and analysis of membrane processes. Chief among the requisite criteria are delivery/targeting modalities that result in the long-term residence (e.g., days) of the nanoparticles on the plasma membrane while simultaneously not interfering with regular cellular physiology and homeostasis. Our laboratory has developed a suite of peptidyl motifs that target semiconductor nanocrystals (quantum dots (QDs) to the plasma membrane where they remain resident for up to three days. Notably, only small a percentage of the QDs are endocytosed over this time course and cellular viability is maintained. This talk will highlight the utility of these peptide-QD constructs for cellular imaging and analysis.

Pseudomonas aeruginosa is among the top three leading causative opportunistic human pathogens, possessing one of the largest bacterial genomes and an exceptionally large proportion of regulatory genes therein. It has been known for more than a decade that the size and complexity of the P. aeruginosa genome is responsible for the adaptability and resilience of the bacteria to include its ability to resist many disinfectants and antibiotics. We have investigated the susceptibility of P. aeruginosa bacterial biofilms to iron oxide (magnetite) nanoparticles (NPs) with and without attached drug (tobramycin). We also characterized the susceptibility of zero-valent iron NPs, which are known to inactivate microbes. The particles, having an average diameter of 16 nm were capped with natural alginate, thus doubling the hydrodynamic size. Nanoparticle-drug conjugates were produced via cross-linking drug and alginate functional groups. Drug conjugates were investigated in the interest of determining dosage, during these dosage-curve experiments, NPs unbound to drug were tested in cultures as a negative control. Surprisingly, we found that the iron oxide NPs inhibited bacterial growth, and thus, biofilm formation without the addition of antibiotic drug. The inhibitory dosages of iron oxide NPs were investigated and the minimum inhibitory concentrations are presented. These findings suggest that NP-drug conjugates may overcome the antibiotic drug resistance common in P. aeruginosa infections.