PEG-coated Nanoparticles Research Articles

Coarse-grained modeling and dynamics tracking of nanoparticles diffusion in human gut mucus

The gastrointestinal (GI) tract's mucus layer serves as a critical barrier and a mediator in drug nanoparticle delivery. The mucus layer's diverse molecular structures and spatial complexity complicates the mechanistic study of the diffusion dynamics of particulate materials. In response, we developed a bi-component coarse-grained mucus model, specifically tailored for the colorectal cancer environment, that contained the two most abundant glycoproteins in GI mucus: Muc2 and Muc5AC. This model demonstrated the effects of molecular composition and concentration on mucus pore size, a key determinant in the permeability of nanoparticles. Using this computational model, we investigated the diffusion rate of polyethylene glycol (PEG) coated nanoparticles, a widely used muco-penetrating nanoparticle. We validated our model with experimentally characterized mucus pore sizes and the diffusional coefficients of PEG-coated nanoparticles in the mucus collected from cultured human colorectal goblet cells. Machine learning fingerprints were then employed to provide a mechanistic understanding of nanoparticle diffusional behavior. We found that larger nanoparticles tended to be trapped in mucus over longer durations but exhibited more ballistic diffusion over shorter time spans. Through these discoveries, our model provides a promising platform to study pharmacokinetics in the GI mucus layer.

Evaluation of Antiproliferative Properties of CoMnZn-Fe2O4 Ferrite Nanoparticles in Colorectal Cancer Cells.

The PEG-coated ferrite nanoparticles Co0.2Mn0.6Zn0.2Fe2O4 (X1), Co0.4Mn0.4Zn0.2Fe2O4 (X2), and Co0.6Mn0.2Zn0.2Fe2O4 (X3) were synthesized by the coprecipitation method. The nanoparticles were characterized by XRD, Raman, VSM, XPS, and TEM. The magnetic hyperthermia efficiency (MH) was determined for PEG-coated nanoparticles using an alternating magnetic field (AMF). X2 nanoparticles displayed the highest saturation magnetization and specific absorption rate (SAR) value of 245.2 W/g for 2 mg/mL in a water medium. Based on these properties, X2 nanoparticles were further evaluated for antiproliferative activity against HCT116 cells at an AMF of 495.25 kHz frequency and 350 G strength, using MTT, colony formation, wound healing assays, and flow cytometry analysis for determining the cell viability, clonogenic property, cell migration ability, and cell death of HCT116 cells upon AMF treatment in HCT116 cells, respectively. We observed a significant inhibition of cell viability (2% for untreated control vs. 50% for AMF), colony-forming ability (530 cells/colony for untreated control vs. 220 cells/colony for AMF), abrogation of cell migration (100% wound closure for untreated control vs. 5% wound closure for AMF), and induction of apoptosis-mediated cell death (7.5% for untreated control vs. 24.7% for AMF) of HCT116 cells with respect to untreated control cells after AMF treatment. Collectively, these results demonstrated that the PEG-coated (CoMnZn-Fe2O4) mixed ferrite nanoparticles upon treatment with AMF induced a significant antiproliferative effect on HCT116 cells compared with the untreated cells, indicating the promising antiproliferative potential of the Co0.4Mn0.4Zn0.2Fe2O4 nanoparticles for targeting colorectal cancer cells. Additionally, these results provide appealing evidence that ferrite-based nanoparticles using MH could act as potential anticancer agents and need further evaluation in preclinical models in future studies against colorectal and other cancers.

Mucus-penetrating and permeation enhancer albumin-based nanoparticles for oral delivery of macromolecules: Application to bevacizumab

The oral administration of therapeutic proteins copes with important challenges (mainly degradation and poor absorption) making their potential therapeutic application extremely difficult. The aim of this study was to design and evaluate the potential of the combination between mucus-permeating nanoparticles and permeation enhancers as a carrier for the oral delivery of the monoclonal antibody bevacizumab, used as a model of therapeutic protein. For this purpose, bevacizumab was encapsulated in PEG-coated albumin nanoparticles as a hydrophobic ion-pairing complex with either sodium deoxycholate (DS) or sodium docusate (DOCU). In both cases, complex formation efficiencies close to 90% were found. The incorporation of either DS or DOCU in PEG-coated nanoparticles significantly increased their mean size, particularly when DOCU was used. Moreover, the diffusion in mucus of DOCU-loaded nanoparticles was significantly reduced, compared with DS ones. In a C. elegans model, DS or DOCU (free or nanoencapsulated) disrupted the intestinal epithelial integrity, but the overall survival of the worms was not affected. In rats, the relative oral bioavailability of bevacizumab incorporated in PEG-coated nanoparticles as a complex with DS (B-DS-NP-P) was 3.7%, a 1000-fold increase compared to free bevacizumab encapsulated in nanoparticles (B-NP-P). This important effect of DS may be explained not only by its capability to transiently disrupt tight junctions but also to their ability to increase the fluidity of membranes and to inhibit cytosolic and brush border enzymes. In summary, the current strategy may be useful to allow the therapeutic use of orally administered proteins, including monoclonal antibodies.Graphical

Investigation of Cubic and Spherical IONPs’ Rheological Characteristics and Aggregation Patterns from the Perspective of Magnetic Targeting

Based on our prior research, we synthesized iron-oxide nanoparticles (IONPs) in two shapes (spherical and cubic) and sized them for the current inquiry. This research examined the magnetic, rheological, and sedimentation properties of the suspensions containing PEG-coated IONPs, considering that both produced particles are intended to be employed for magnetic targeting applications. The saturation magnetization of both IONPs was lower than the magnetite domain magnetization of 92 emu/g due to the surfactant and the dead surface magnetic layer. Under each investigated magnetic field (0, 34 and 183 mT), the shear viscosity behaviour of the MNP suspensions of both kinds was comparable. Shear thinning behaviour was seen for both particle suspensions in the low shear rate area (0.1 s−1 to 1 s−1). The rheological curves from this paper show that the suspensions present a higher viscosity at lower shear rates for spherical and cubic PEG-coated nanoparticles when a magnetic field is applied. The aggregation behaviour demonstrates that cubic-shaped IONPs are more stable throughout time, with hydrodynamic diameter measurements showing a relatively slow variation of the DLS size distribution from 250 nm to 210 nm in the first 600 s; contrarily, the hydrodynamic diameter of spherical IONPs fluctuated significantly, from 855 nm to 460 nm. Another key finding relates to the sedimentation profile, specifically that PEG-coated IONPs with spherical shapes have a stronger tendency to sediment than those with cubic forms, which are more stable.

Gold Nanoparticles as Drug Carriers: The Role of Silica and PEG as Surface Coatings in Optimizing Drug Loading.

The use of gold nanoparticles as drug delivery systems has received increasing attention due to their unique properties, such as their high stability and biocompatibility. However, gold nanoparticles have a high affinity for proteins, which can result in their rapid clearance from the body and limited drug loading capabilities. To address these limitations, we coated the gold nanoparticles with silica and PEG, which are known to improve the stability of nanoparticles. The synthesis of the nanoparticles was carried out using a reduction method. The nanoparticles' size, morphology, and drug loading capacity were also studied. The SEM images showed a spherical and homogeneous morphology; they also showed that the coatings increased the average size of the nanoparticles. The results of this study provide insight into the potential of gold nanoparticles coated with silica and PEG as drug delivery systems. We used ibuprofen as a model drug and found that the highest drug load occurred in PEG-coated nanoparticles and then in silica-coated nanoparticles, while the uncoated nanoparticles had a lower drug loading capacity. The coatings were found to significantly improve the stability and drug load properties of the nanoparticles, making them promising candidates for further development as targeted and controlled release drug delivery systems.

Treatment of pollution resulting from the spillage of crude oil slicks from ship accidents in sea water using PEG-coated nanoparticles

Treatment of pollution resulting from the spillage of crude oil slicks from ship accidents in sea water using PEG-coated nanoparticles

Changeable net charge on nanoparticles facilitates intratumor accumulation and penetration.

The Enhanced Permeability and Retention (EPR) effect is a golden strategy for the nanoparticle (NP)-based targeting of solid tumors, and the surface property of NPs might be a determinant on their targeting efficiency. Poly(ethylene glycol) (PEG) is commonly used as a shell material; however, it has been pointed out that PEG-coated NPs may exhibit accumulation near tumor vasculature rather than having homogenous intratumor distribution. The PEG shell plays a pivotal role on prolonged blood circulation of NPs but potentially impairs the intratumor retention of NPs. In this study, we report on a shell material to enhance tumor-targeted delivery of NPs by maximizing the EPR effect: polyzwitterion based on ethylenediamine-based carboxybetaine [PGlu(DET-Car)], which shows the changeable net charge responding to surrounding pH. The net charge of PGlu(DET-Car), is neutral at physiological pH7.4, allowing it to exhibit a stealth property during the blood circulation; however, it becomes cationic for tissue-interactive performance under tumorous acidic conditions owing to the stepwise protonation behavior of ethylenediamine. Indeed, the PGlu(DET-Car)-coated NPs (i.e., gold NPs in the present study) exhibited prolonged blood circulation and remarkably enhanced tumor accumulation and retention than PEG-coated NPs, achieving 32.1% of injected dose/g of tissue, which was 4.2 times larger relative to PEG-coated NPs. Interestingly, a considerable portion of PGlu(DET-Car)-coated NPs clearly penetrated into deeper tumor sites and realized the effective accumulation in hypoxic regions, probably because the cationic net charge of PGlu(DET-Car) is augmented in more acidic hypoxic regions. This study suggests that the changeable net charge on the NP surface in response to tumorous acidic conditions is a promising strategy for tumor-targeted delivery based on the EPR effect.

Comparison of the influence of superparamagnetic nanoparticles concentration and coverage on the alternating magnetic field thermal effect

Magnetic nanoparticles (MNP)s of Fe3O4 were synthesized in the co-precipitation reaction of iron chlorides dissolved in water and ammonia water. To produce polyethylene glycol (PEG) coatings of the NMPs, we added PEG to the ammonia water during the fabrication process. Magnetic fluids, ferrofluids, for research were prepared as oleic acid suspension of bare and PEG-coated nanoparticles at four concentrations. The results of the conducted calorimetric experiments confirmed that the efficiency of heating ferrofluids strongly depends on the concentration of nanoparticles in fluids. The results also indicate that the ferrofluids containing PEG-coated NPs heat up more than ferrofluids with bare nanoparticles. They are characterized by a higher specific absorption rate (SAR) value calculated from the initial slope of temperature versus time curve during heating.

Combating Complement's Deleterious Effects on Nanomedicine by Conjugating Complement Regulatory Proteins to Nanoparticles.

Complement opsonization is among the biggest challenges facing nanomedicine. Nearly instantly after injection into blood, nanoparticles are opsonized by the complement protein C3, leading to clearance by phagocytes, fouling of targeting moieties, and release of anaphylatoxins. While surface polymers such as poly(ethylene glycol) (PEG) partially decrease complement opsonization, most nanoparticles still suffer from extensive complement opsonization, especially when linked to targeting moieties. To ameliorate the deleterious effects of complement, two of mammals' natural regulators of complement activation (RCAs), Factors H and I, are here conjugated to the surface of nanoparticles. In vitro, Factor H or I conjugation to PEG-coated nanoparticles decrease their C3 opsonization, and markedly reduce nanoparticle uptake by phagocytes. In an in vivo mouse model of sepsis-induced lung injury, Factor I conjugation abrogates nanoparticle uptake by intravascular phagocytes in the lungs, allowing the blood concentration of the nanoparticle to remain elevated much longer. For nanoparticles targeted to the lung's endothelium by conjugation to anti-ICAM antibodies, Factor I conjugation shifts the cell-type distribution away from phagocytes and toward endothelial cells. Finally, Factor I conjugation abrogates the severe anaphylactoid responses common to many nanoparticles, preventing systemic capillary leak and preserving blood flow to visceral organs and the brain. Thus, conjugation of RCAs, like Factor I, to nanoparticles is likely to help in nanomedicine's long battle against complement, improving several key parameters critical for clinical success.

Graphene Oxide Nanoparticels Interaction with Jurkat Cell Line in Cell-IQ System.

In recent years, materials based on graphene oxide (GO) have been actively studied for their use in biomedicine. The aim of our study was to investigate the increase in cell mass and viability of Jurkat tumor line T cells during 24 h of contact with GO nanoparticles in the Cell-IQ system of intravital observation. We used nanoparticles of different sizes coated with linear or branched polyethylene glycol (PEG) at concentrations of 5 and 25 μg/mL. It was shown for the first time that direct contact with GO nanoparticles reduced the growth in cell mass at the visualization points by more than twofold, regardless of nanoparticle size and concentration. Moreover, the number of live cells in the culture decreased by 5-9% after 24 h of monitoring. Thus, PEG-coated GO nanoparticles were found to suppress the proliferation and viability of Jurkat cell line T lymphocytes.

Nanoparticle shell structural cues drive in vitro transport properties, tissue distribution and brain accessibility in zebrafish

Zwitterion polymers with strong antifouling properties have been suggested as the prime alternative to polyethylene glycol (PEG) for drug nanocarriers surface coating. It is believed that PEG coating shortcomings, such as immune responses and incomplete protein repellency, could be overcome by zwitterionic polymers. However, no systematic study has been conducted so far to complete a comparative appraisal of PEG and zwitterionic-coating effects on nanoparticles (NPs) stealthness, cell uptake, cell barrier translocation and biodistribution in the context of nanocarriers brain targeting.Core-shell polymeric particles with identical cores and a shell of either PEG or poly(2-methacryloyloxyethyl phosphorylcholine (PMPC) were prepared by impinging jet mixer nanoprecipitation. NPs with similar size and surface potential were systematically compared using in vitro and in vivo assays. NPs behavior differences were rationalized based on their protein-particles interactions.PMPC-coated NPs were significantly more endocytosed by mouse macrophages or brain resident macrophages compared to PEGylated NPs but exhibited the remarkable ability to cross the blood-brain barrier in in vitro models. Nanoscale flow cytometry assays showed significantly more adsorbed proteins on PMPC-coated NPs than PEG-coated NPs. In vivo, distribution in zebrafish larvae, showed a strong propensity for PMPC-coated NPs to adhere to the vascular endothelium, while PEG-coated NPs were able to circulate for a longer time and escape the bloodstream to penetrate deep into the cerebral tissue.The stark differences between these two types of particles, besides their similarities in size and surface potential, points towards the paramount role of surface chemistry in controlling NPs fate likely via the formation of distinct protein corona for each coating.

Comparative Study of Specific Loss Power and Transverse Relaxivity of Spinel Ferrite Nanoensembles Coated With Chitosan and Polyethylene Glycol

We synthesized spinel ferrite nanoensembles (MnFe2O4, CoFe2O4, and Fe3O4) using the chemical co-precipitation method and characterized their physical, chemical, and magnetic properties by X-ray diffraction (XRD), transmission electron microscopy (TEM), physical properties measurement system (PPMS), Mössbauer spectroscopy, Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) and Raman spectroscopy. Their relaxation properties and potential for hyperthermia therapy were determined using nuclear magnetic resonance (NMR) and cell viability assay, respectively. XRD and TEM data confirmed that the particle core sizes were 6–9 nm before coating while their sizes increased to 10–14 nm and 14–20 nm after coating with chitosan and polyethylene glycol (PEG), respectively. Mössbauer spectroscopy showed superparamagnetic behavior for MnFe2O4 nanoparticles and ferrimagnetic behavior for the CoFe2O4 and Fe3O4 nanoparticles. A detailed studies of MH loops of all three ferrites before and after coating showed surface functionalization by a large reduction of coercivity and anisotropy. The successful coating was further confirmed by the peak shifts in the FTIR spectra of the particles whereas Raman spectra of coated ferrites also displayed the characteristic absorption patterns and suppression of the ferrite peaks suggesting successful coating. The induced heating profile of the nanoparticles in stable suspension was tested with a radio frequency magnetic field of 76 mT and a frequency of 400 kHz. High mortality (>98%) of 9 L gliosarcoma cancer cells by hyperthermia suggested that these nanoparticles could be used for cancer therapy. Transverse relaxivities (r2) determined by NMR for chitosan-coated MnFe2O4, CoFe2O4, and Fe3O4 nanoparticles were 297 (±22), 353 (±26), and 345 (±13), mM−1S−1, while for PEG-coated nanoparticles are 165 (±22), 146 (±14), and 159 (±07) mM−1S−1, respectively. Overall these spinel ferrite nanoensembles show great promise for cancer theranostics research applications.

Preparation and evaluation of PEG-coated zein nanoparticles for oral drug delivery purposes

The aim was to produce PEG-coated nanoparticles (NP-PEG), with mucus-permeating properties, for oral drug delivery purposes by using simple procedures and regulatory-approved compounds in order to facilitate a potential clinical development. For this purpose, zein nanoparticles were prepared by desolvation and, then, coated by incubation with PEG 35,000. The resulting nanocarriers displayed a mean size of about 200 nm and a negative zeta potential. The presence of PEG on the surface of nanoparticles was evidenced by electron microscopy and confirmed by FTIR analysis. Likely, the hydrophobic surface of zein nanoparticles (NP) was significantly reduce by their coating with PEG. This increase of the hydrophilicity of PEG-coated nanoparticles was associated with an important increase of their mobility in pig intestinal mucus. In laboratory animals, NP-PEG (fluorescently labelled with Lumogen® Red 305) displayed a different behavior when compared with bare nanoparticles. After oral administration, NP appeared to be trapped in the mucus mesh, whereas NP-PEG were capable of crossing the protective mucus layer and reach the epithelium. Finally, PEG-coated zein nanoparticles, prepared by a simple and reproducible method without employing reactive reagents, may be adequate carriers for promoting the oral bioavailability of biomacromolecules and other biologically active compounds with low permeability properties.

Laser-synthesized TiN nanoparticles for biomedical applications: Evaluation of safety, biodistribution and pharmacokinetics

Having plasmonic absorption within the biological transparency window, titanium nitride (TiN) nanoparticles (NPs) can potentially outperform gold counterparts in phototheranostic applications, but characteristics of available TiN NPs are still far from required parameters. Recently emerged laser-ablative synthesis opens up opportunities to match these parameters as it makes possible the production of ultrapure low size-dispersed spherical TiN NPs, capable of generating a strong phototherapy effect under 750–800 nm excitation. This study presents the first assessment of toxicity, biodistribution and pharmacokinetics of laser-synthesized TiN NPs. Tests in vitro using 8 cell lines from different tissues evidenced safety of both as-synthesized and PEG-coated NPs (TiN-PEG NPs). After systemic administration in mice, they mainly accumulated in liver and spleen, but did not cause any sign of toxicity or organ damage up to concentration of 6 mg kg−1, which was confirmed by the invariability of blood biochemical parameters, weight and hemotoxicity examination. The NPs demonstrated efficient passive accumulation in EMT6/P mammary tumor, while concentration of TiN-PEG NPs was 2.2-fold higher due to “stealth” effect yielding 7-times longer circulation in blood. The obtained results evidence high safety of laser-synthesized TiN NPs for biological systems, which promises a major advancement of phototheranostic modalities on their basis.

Fabrication of poly(sarcosine), poly (ethylene glycol), and poly (lactic-co-glycolic acid) polymeric nanoparticles for cancer drug delivery

The most significant advantage of polymeric nanoparticles is their systematic and target specific drug delivery properties. These polymeric nanoparticles have the potential to sustain itself within the bloodstream for a longer period so that drugs can be infused in blood steadily. To understand such a phenomenon, in this experiment, an attempt was made to prepared Docetaxel loaded poly(sarcosine)(PSar) and poly (ethylene glycol) (PEG) coated poly (lactic-co-glycolic acid) [PLGA] nanoparticles; which could efficiently encapsulate any hydrophobic drugs. These types of PEG-coated nanoparticles have a marked tendency to avoid reticuloendothelial opsonization process by restricting macrophage uptake, which ultimately leads to enhance bioavailability and tissue distribution of drugs. In this research work categorically, the concentration of PSar and PEG was optimized. The cellular uptake efficiency percentage and IC50 value of Docetaxel and Docetaxel loaded different polymeric nanoparticles evaluated in various human cancer cell lines (U-87 MG, HeLa. C2BBe1, HCT-116, NCI–N87, NCI–H929-Luc-mCh-Puro). The PSar-PLGA-PEG-NPs have shown sustainable retention in blood with minimum macrophage uptake as compared to PLGA-NPs and PSar-PLGA-NPs. Enhanced anti-tumor proliferative effects were shown in all the Docetaxel loaded nanoparticles as compared to native Docetaxel drug, which may be because of enhanced antiproliferative activities of nanoparticles. Thus, from the research outcomes, one thing is inevitable, i.e., the presence of PSar and PEG would increase the blood circulation time, and it can be used as a suitable carrier for any hydrophobic drug.

Intravenously administered d-mannitol-coated maghemite nanoparticles cause elemental anomalies in selected rat organs.

In this study novel d-mannitol coated maghemite nanoparticles (MIONPs) are presented in terms of their influence on elemental homeostasis of living organisms and for this purpose highly sensitive total reflection X-ray fluorescence was used. Because of the biological indifference of d-mannitol and presumed lower toxicity of maghemite, compared to the most commonly used magnetite in nanomedicine, such nanoparticles seem to be promising candidates for biomedical applications. The examined dose of MIONPs was comparable with one of the lowest doses used in medical diagnostics. However, it should be emphasized that the amount of iron injected in this form is still significant compared to its total content in organs, especially in kidneys or the heart, and may easily disrupt their elemental homeostasis. The aim of the present study was to evaluate the elemental changes occurring in selected rat organs after injecting a low dose of MIONPs. The results were compared with those obtained for previously examined PEG-coated nanoparticles with magnetite cores. In the light of our findings the elemental changes observed after exposure to MIONPs were less extensive than those following PEG-coated magnetite nanoparticle administration.

PEG-coated nanoparticles detachable in acidic microenvironments for the tumor-directed delivery of chemo- and gene therapies for head and neck cancer.

Background: Head and neck cancer (HNC) is a major cause of morbidity and mortality and has a poor treatment outcome. Irinotecan, a topoisomerase-I inhibitor, induces cell death by decreasing the religation of double-strand DNA. However, epithelial-mesenchymal transition (EMT), therapy resistance, and systemic toxicity caused by available antineoplastic agents hinder the efficacy and safety of HNC treatment. Chemotherapy combined with gene therapy shows potential application in circumventing therapy resistance and EMT. miR-200 exerts a remarkable suppressing effect on EMT-associated genes. Herein, liposomes and solid lipid nanoparticles (SLNs) modified with a pH-sensitive, self-destructive polyethylene glycol (PEG) shell and different peptides were designed as irinotecan and miR-200 nanovectors to enhance tumor-specific accumulation. These peptides included one ligand targeting the angiogenic tumor neovasculature, one mitochondrion-directed apoptosis-inducing peptide, and one cell-penetrating peptide (CPP) with high potency and selectivity toward cancer cells.Methods: Physicochemical characterization, cytotoxicity analysis, cellular uptake, regulation mechanisms, and in vivo studies on miR-200- and irinotecan-incorporated nanoparticles were performed to identify the potential antitumor efficacy and biosafety issues involved in HNC treatment and to elucidate the underlying signaling pathways.Results: We found that the cleavable PEG layer responded to low extracellular pH, and that the CPP and targeting peptides were exposed to improve the uptake and release of miR-200 and irinotecan into HNC human tongue squamous carcinoma (SAS) cells. The apoptosis of SAS cells treated with the combinatorial therapy was significantly induced by regulating various pathways, such as the Wnt/β-catenin, MDR, and EMT pathways. The therapeutic efficacy and safety of the proposed co-treatment outperformed the commercially available Onivyde and other formulations used in a SAS tumor-bearing mouse model in this study.Conclusion: Chemotherapy and gene therapy co-treatment involving pH-sensitive and targeting peptide-modified nanoparticles may be an innovative strategy for HNC treatment.

Stabilizing gold nanoparticles for use in X-ray computed tomography imaging of soil systems.

This investigation establishes a system of gold nanoparticles that show good colloidal stability as an X-ray computed tomography (XCT) contrast agent under soil conditions. Gold nanoparticles offer numerous beneficial traits for experiments in biology including: comparatively minimal phytotoxicity, X-ray attenuation of the material and the capacity for functionalization. However, soil salinity, acidity and surface charges can induce aggregation and destabilize gold nanoparticles, hence in biomedical applications polymer coatings are commonly applied to gold nanoparticles to enhance stability in the in vivo environment. Here we first demonstrate non-coated nanoparticles aggregate in soil-water solutions. We then show coating with a polyethylene glycol (PEG) layer prevents this aggregation. To demonstrate this, PEG-coated nanoparticles were drawn through flow columns containing soil and were shown to be stable; this is in contrast with control experiments using silica and alumina-packed columns. We further determined that a suspension of coated gold nanoparticles which fully saturated soil maintained stability over at least 5 days. Finally, we used time resolved XCT imaging and image based models to approximate nanoparticle diffusion as similar to that of other typical plant nutrients diffusing in water. Together, these results establish the PEGylated gold nanoparticles as potential contrast agents for XCT imaging in soil.

Abstract 2583: DNA-linked gold nanoprobe assay: An efficient technology for direct and high-sensitive quantification of serum microRNAs in prostate cancer

Abstract Background: Serum-based biomarkers in conjunction with advanced biosensing techniques for early detection or screening of indolent vs. aggressive prostate cancer (PCa) can improve patient care and outcmomes. Recent discoveries have identified miRNAs (miRs) as promising biomarkers and opened potential avenues for earliest diagnosis and examining aggressiveness of PCa. However, quantitation of ultra-low levels of miRs in sera or tissue of patient remains challenging with traditional techniques. The present study describes the development and application of the DNA-linked gold nanoprobe (DNA-AuNPr)-based fluorescence assay for quantification of target miRs in serum with high sensitivity. Methodology: Thorough meta-analysis followed by global in-silico analyses were performed to evaluate the differential expression of miRs among PCa studies. Further, the identified miRs were validated using TaqMan assay in human and mouse PCa serum. Thereafter, we synthesized DNA-AuNPr specific to target miRs, by conjugating FAM-tagged DNA-probes to the PEG-coated gold nanoparticle (AuNP) via cross-linker chemistry and fully characterized with UV, DLS and TEM. Sensitivity and target specificity of nanoprobe-based assay were determined using the Synergy-H1 multimode reader. As a proof of principle, a real-time evolution of fluorescence signal upon enzymatic digestion of DNA-probe in DNA-RNA heteroduplexes was tested. Finally, we utilized human (PCa n=44, control n=43) and Pten conditional knockout mouse (PCa n=8, control n=8) sera for our training and blank validation sets to determine the diagnostic significance of miRs in PCa. Results: TaqMan assays detected a significantly higher level of both miR-21 and miR-375 in the PCa compared to normal control. Analyses of DNA-AuNPr shown uniform-sized, highly stable AuNP core tagged with multiple target-specific DNA-probes (~150±21 per NP). Additional assay validation indicated an excellent sequence specificity of DNA-AuNPr in discriminating the single-base mutation (mismatch) in miRs sequence. The uniqueness of proposed assay was its capability to quantitate miR targets in the range 50 pM to 10 nM with a lower detection limit of ~5 pM (0.3 fmol) in just ~4 h of total assay time and using ~10 μL of total RNA isolated from serum. Finally, the assay performance was validated in serum for both miR-21 and miR-375. The two miRs efficiently differentiated PCa from the healthy control in training sets (AUC=0.94) as well as in independent validation sets (AUC=0.90), which reconfirmed their diagnostic potential. Conclusion: We developed a direct (without converting target miR into cDNA), highly sensitive and technologically efficient miR-quantitation strategy for diagnosis of PCa. Citation Format: Prakash Kshirsagar, Satyanarayana Rachagani, Parthasarathy Seshacharyulu, Sakthivel Muniyan, Sunandini Sharma, Ram Mahato, Surinder K. Batra, Maneesh Jain. DNA-linked gold nanoprobe assay: An efficient technology for direct and high-sensitive quantification of serum microRNAs in prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2583.

Toxicity of surface functionalized iron oxide nanoparticles toward pure suspension culture and soil microcosm

Surface functionalized iron oxide nanoparticles are increasingly being used in environmental and medical applications, raising concerns on their fate and environmental impact. In this work, Fe3O4 nanoparticles prepared by a chemical co-precipitation method were functionalized with citric acid, polyethylene glycol of molecular weights 6000 and 8000, and methacrylic acid. Resazurin reduction-based nanoparticles toxicity assessment was performed with suspended microbial culture (50–250 mg L−1 NPs) as well as in soil microcosm (250–1250 mg kg−1). Resazurin reduction by freely suspended E. coli cultures in presence of uncoated, citric acid- and PEG-coated nanoparticles was either comparable or higher than that of the nanoparticle free control. However, the toxicity of methacrylic acid coated nanoparticles was evident as resazurin reduction was largely inhibited. Residual resazurin in test sets containing nanoparticle spiked seven days aged sterile soil were 20% to 82% that of freshly spiked control, suggesting significant attenuation of toxicity in soil media. The results obtained from this work may provide a better understanding of the impact of engineered iron-oxide nanoparticles in the soil ecosystem.