Method for extraction of nanoscale plastic debris from soil.

Sample preparation for extraction of nanoscale plastic debris (NPD, size < 1 μm) from environmental samples is a critical step to prepare NPD for further identification and quantification. Developing a NPD extraction method from soil matrices is particularly challenging due to the complexity of solid matrices. In the present study, we built upon the lessons learned from method development for extraction of microplastics and nanomaterials from environmental samples to develop a sample preparation method for extraction of NPD from soil matrices. The evaluation criteria for the extraction method are size distribution, particle number recovery, and particle mass recovery. Since there is no validated method available to trace and quantify the mass of NPD in complex matrices, we applied polystyrene particles doped with europium (Eu-PS NPs). Standard LUFA soil and field soil were spiked and mixed for 24 h with 1 mg of Eu-PS NPs and the particles were extracted from the matrices of the soils. The extraction method did not significantly influence the size distribution of the particles and the extraction agents did not degrade the Eu-PS NPs. Mass balance calculation suggested recoveries of 82 and 77% of the added Eu-PS NPs in LUFA soil and field soil, respectively. The number recoveries of the particles were 81 and 85% for LUFA soil and field soil, respectively. This method can be further optimized and used as the first building block to develop a generic sample preparation method for the extraction of NPD from soil samples. By combining this developed and verified extraction method with identification and quantification techniques, a fit-for-purpose workflow can be developed to quantify and subsequently understand the fate of NPD in soil.Environmental Biolog

Classification of cilio-inhibiting effects of nasal drugs

Objective/Hypothesis: Nasal drug formulations are widely used for a local therapeutic effect, but are also used for systemic drug delivery. In the development of new nasal drugs, the toxic effects on the mucociliary clearance and therefore on the ciliated tissue is of importance. In this study, the effect of nasal drugs and their excipients on the ciliary beat frequency (CBF) is investigated. Study Design: Experimental, in vitro. Methods: CBF is measured by a photograph-electric registration method. Excised ciliated chicken trachea tissue is incubated for 15 minutes in the formulation, followed by a reversibility test. To estimate the ciliostatic potential, a classification is given of all tested formulations. According to the CBF, after 60 minutes every drug or excipient could be classified as follows: cilio-friendly: after 60 minutes the CBF has regained 75% or more of its initial frequency; cilio-inhibiting: after 60 minutes the CBF has regained between 25% and 75% of its initial frequency; or ciliostatic: after 60 minutes the CBF has regained 25% or less of its initial frequency. Results: Most formulations used are cilio-friendly or cilio-inhibiting. Only some are ciliostatic. Preservatives have a major role in the cilio-inhibiting effect of the drug. Also, other additives can contribute to the toxicity profile of nasal drug formulations. Conclusion: This classification of the cilio-inhibiting potential of nasal drug formulations is a valuable tool in the design of safe nasal drugs. The number of animal studies in vivo can be reduced substantially by using this in vitro screening technique. This study demonstrates that the effect on ciliary movement of most drug formulations is due to the preservatives and/or additives and mostly not to the drug itself

Uptake of melatonin into the cerebrospinal fluid after nasal and intravenous delivery: Studies in rats and comparison with a human study

Purpose. To investigate the possibility of direct transport of melatonin from the nasal cavity into the cerebrospinal fluid (CSF) after nasal administration in rats and to compare the animal results with a human study. Methods. Rats (n = 8) were given melatonin both intranasally in one nostril (40 μg/rat) and intravenously by bolus injection (40 μg/rat) into the jugular vein using a Vascular Access Port. Just before and after drug administration, blood and CSF samples were taken and analyzed by HPLC. Results. Melatonin is quickly absorbed in plasma (Tmax = 2.5 min) and shows a delayed uptake into CSF (Tmax = 15 min) after nasal administration. The melatonin concentration-time profiles in plasma and CSF are comparable to those after intravenous delivery. The AUCCSF/AUCplasma ratio after nasal delivery (32.7 ± 6.3%) does not differ from the one after intravenous injection (46.0 ± 10.4%), which indicates that melatonin enters the CSF via the blood circulation across the blood-brain barrier. This demonstrates that there is no additional transport via the nose-CSF pathway. These results resemble the outcome of a human study. Conclusions. The current results in rats show that there is no additional uptake of melatonin in the CSF after nasal delivery compared to intravenous administration. This is in accordance with the results found in humans, indicating that animal experiments could be predictive for the human situation when studying nose-CSF transport

Development of methods for extraction and analytical characterization of carbon-based nanomaterials (nanoplastics and carbon nanotubes) in biological and environmental matrices by asymmetrical flow field-flow fractionation.

Suitable methods and fit-for-purpose techniques are required to allow characterization of carbon-based nanomaterials (CB-NMs) in complex matrices. In this study, two methods were developed; a method for extraction and characterization of CB-NMs in biological media and a method for fractionation of natural organic matter (NOM) coated CB-NMs in environmental matrices. The former method was developed by extracting carbon nanotubes (CNTs: sized 0.75 × 3000 nm) and nanoplastics (sized 60, 200 and 600 nm) from eggshells and characterizing the extracted CB-NMs in terms of particle size distribution using asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS). The latter method was developed using AF4-MALS to fraction NOM-coated CNT (sized 0.75 × 3000 nm) and nanoplastics (sized 60, 200 and 300 nm) in a simulated natural surface water and provide information about the size distribution of the CB-NM-NOM complexes. The developed AF4-MALS method successfully fractioned the CB-NM-NOM complexes based on hydrodynamic size and provided the size distribution of the complexes. The NOM corona did not shift significantly the median size of the CB-NMs. It influenced however the size distribution of the nanoplastics and CNTs. The sample preparation method failed to extract the CNTs (recovery  60%). The AF4-MALS fractogram showed that the extraction method did not significantly influence the size distribution of the nanoplastics of 60 and 200 nm size, whereas the peak of 600 nm nanoplastics shifted towards a smaller hydrodynamic size. In conclusion, the developed sample preparation method followed by the developed AF4-MALS method can be applied for extraction, separation and characterization of CB-NMs in biological and environmental matrices. Thus, the methods have a high potential to be methods of choice to investigate CB-NMs in future studies.Environmental Biolog

N-Trimethyl chitosan (TMC) nanoparticles loaded with influenza subunit antigen for intranasal vaccination: Biological properties and immunogenicity in a mouse model

In this study, the potential of N-trimethyl chitosan (TMC) nanoparticles as a carrier system for the nasal delivery of a monovalent influenza subunit vaccine was investigated. The antigen-loaded nanoparticles were prepared by mixing a solution containing TMC and monovalent influenza A subunit H3N2 with a tripolyphosphate (TPP) solution, at ambient temperature and pH 7.4 while stirring. The nanoparticles had an average size of about 800 nm with a narrow size distribution and a positive surface charge. The nanoparticles showed a loading efficiency of 78% and a loading capacity of 13% (w/w). It was shown that more than 75% of the protein remained associated with the TMC nanoparticles upon incubation of the particles in PBS for 3 h. The molecular weight and antigenicity of the entrapped hemagglutinin was maintained as shown by polyacrylamide gel electrophoresis and Western blotting, respectively. Single i.n. or i.m. immunization with antigen-loaded TMC nanoparticles resulted in strong hemagglutination inhibition and total IgG responses. These responses were significantly higher than those achieved after i.m. administration of the subunit antigen, whereas the IgG1/IgG2a profile did not change substantially. The i.n. administered antigen-TMC nanoparticles induced higher immune responses compared to the other i.n. antigen formulations, and these responses were enhanced by i.n. booster vaccinations. Moreover, among the tested formulations only i.n. administered antigen-containing TMC nanoparticles induced significant IgA levels in nasal washes of all mice. In conclusion, these findings demonstrate that TMC nanoparticles are a potent new delivery system for i.n. administered influenza antigens. (c) 2006 Elsevier Ltd. All rights reserved

Uptake studies in rat Peyer's patches, cytotoxicity and release studies of alginate coated chitosan nanoparticles for mucosal vaccination

The design of particulate vaccine delivery systems, particularly for mucosal surfaces, has been a focus of interest in recent years. In this context, we have previously described the development and the characterization of a new nanosized delivery system, consisting of a model antigen adsorbed to chitosan particles and coated with sodium alginate. In the present work the ovalbumin release profiles from these coated nanoparticles in different pH buffers were investigated and compared to those of the uncoated particles. Cytotoxicity of the polymers and nanoparticles was assessed using the MTT assay. Finally, particle uptake studies in rat Peyer's patches were performed. It was demonstrated that the coating of the nanoparticles with sodium alginate not only avoided a burst release observed with uncoated particles but also increased the stability of the particles at pH 6.8 and 7.4 at 37 °C. At neutral pH, the release was lower than 5% after 3.5 h incubation in a low ionic strength buffer. For both, chitosan and alginate polymers, and for the nanoparticles, comparable cell viability data close to 100%, were obtained. Additionally, based on confocal laser scanning microscopy observations, it was shown that alginate coated nanoparticles were able to be taken up by rat Peyer's patches, rendering them suitable carriers for intestinal mucosal vaccination.http://www.sciencedirect.com/science/article/B6T3D-4K6CHRP-1/1/a257a285900ecde6d4747e1df67e1a8