Transport of nanoparticles across an in vitro model of the human intestinal follicle associated epithelium (original) (raw)
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Journal of Controlled Release, 2007
M cells represent a potential portal for oral delivery of peptides and proteins due to their high endocytosis abilities. An in vitro model of human FAE (co-cultures) was used to evaluate the influence of M cells on the transport of free and encapsulated helodermina model peptideacross the intestinal epithelium. M cells enhanced transport of intact helodermin (18-fold, Papp = 3 × 10 − 6 cm s − 1 ). As pegylation increased nanoparticle transport by M cells, helodermin was encapsulated in 200 nm nanoparticles containing PEG-b-PLA:PLGA 1:1. Stability of the selected formulation was demonstrated in simulated gastric and intestinal fluids. M cells increased the transport of helodermin encapsulated in these nanoparticles by a factor of 415, as compared to Caco-2 cells. Transport of free and encapsulated helodermin occurred most probably by endocytosis. In conclusion, M cells improved helodermin transport across the intestinal epithelium, confirming their high potential for oral delivery of peptides.
It takes more than a coating to get nanoparticles through the intestinal barrier in vitro
European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V, 2016
Size and shape are crucial parameters which have impact on the potential of nanoparticles to penetrate cell membranes and epithelial barriers. Current research in nanotoxicology additionally focuses on particle coating. To distinguish between core- and coating-related effects in nanoparticle uptake and translocation, two nanoparticles equal in size, coating and charge but different in core material were investigated. Silver and iron oxide nanoparticles coated with poly (acrylic acid) were chosen and extensively characterized by small-angle x-ray scattering, nanoparticle tracing analysis and transmission electron microscopy (TEM). Uptake and transport were studied in the intestinal Caco-2 model in a Transwell system with subsequent elemental analysis. TEM and ion beam microscopy were conducted for particle visualization. Although equal in size, charge and coating, the behavior of the two particles in Caco-2 cells was different: while the internalized amount was comparable, only iron ...
Journal of Pharmacology and Experimental Therapeutics, 2006
The aim of this study was to identify cell adhesion molecules that could serve as targets of the human follicle-associated epithelium (FAE) overlying Peyer's patches and to assess nanoparticle uptake levels across this epithelium. We first studied the expression of the mouse M-cell marker  1 -integrin and used a model of human FAE derived from intestinal epithelial Caco-2 cells and Raji B-cells to identify additional potential targets by cDNA array. The protein expression of potential targets in the model FAE and in human ileal FAE tissues was quantified by immunofluorescence. Integrin targeting was studied by investigating the transport of Arg-Gly-Asp (RGD)-coated (integrin-binding), Arg-Gly-Glu (RGE)coated (nonintegrin-binding), and uncoated nanoparticles across ileal specimens mounted in Ussing chambers. Both  1 -integrin and the cell adhesion molecule CD9 were more abundantly ex-
A novel In Vitro Model for Studying Nanoparticle Interactions with the Small Intestine
EURO-NanoTox-Letters, 2016
Manufactured nanomaterials provide promising features for new technologies in cosmetic, food, and pharmaceutical applications. On the other hand, orally ingested nanomaterials/nanoparticles may interact with or enter intestinal cells via different mechanisms, resulting in possible injuries of the biological system. For that reason, the current study aims to provide useful information concerning physicochemical properties of nanoparticles with regard to cytotoxic effects and uptake mechanisms in the small intestine. Differently charged polystyrene nanoparticles were used and cytotoxicity and uptake were studied with an intestinal in vitro co-culture model, mimicking the villus epithelium and a triple-culture model recapitulating the follicle-associated epithelium. Mechanisms of cellular transport were investigated at 37°C and 4°C to verify that internalization mainly occurs energy-dependently. Chemical inhibitors (i.e., chlorpromazine, genistein, dynasore) were used to block dynamin-dependent endocytic pathways without affecting cell viability and membrane integrity. Qualification and quantification were performed via confocal microscopy and flow cytometry. Furthermore, co-localization studies with commonly used markers (i.e., transferrin, lactosylceramide) were carried out and co-localization was assessed via calculation of Pearson´s correlation coefficient and Mander´s overlap coefficient. The results show that size and surface chemistry play a crucial role in cytotoxic interactions and cellular uptake of nanoparticles (NPs). Independent of the surface charge, NPs strongly interact with intestinal mucus and are immobilized. Uptake predominantly occurs via M cells and is surface-charge dependent. Whereas negatively charged particles fail to enter cells, positive and neutral particles penetrate M cells energy-dependently. More precisely, both clathrin-and caveolae-mediated endocytosis are involved. It can be concluded that the presented system serves as a valuable tool to assess safety aspects of manufactured nanomaterials and hence, substantially contributes to nanosafety efforts.
International Journal of Pharmaceutics, 2010
The aim of the present work was to study the influence of surface hydrophilicity of biodegradable polymeric nanoparticles on cellular uptake by Caco-2 cells. Poly(d,l-lactide-co-glycolide acid) particles loaded with a fluorescent dye, 3,3′-dioctadecyloxacarbo-cyanine perchlorate (DiO), were prepared by the emulsion–evaporation process. Three batches of particles with narrow size distribution (100, 300 and 1000 nm) were produced using selective centrifugation. One set of particles was coated by adsorption of chitosan to increase the hydrophilicity of the particles. The interaction of particles with Caco-2 cells was determined by fluorescence spectroscopy and the number of particles associated with one single cell was then calculated. Interaction with cells was clearly dependant on particle size and surface hydrophilicity. Particles in the range of 100 nm presented higher interaction when compared to larger particles. Approximately 6000 uncoated particles and more than 30,000 chitosan-coated particles were quantified per cell. Confocal microscopy confirmed the spectroscopic measurements and revealed the location of the particles in the cell monolayer. Only small particles were observed intracellularly, whereas particles larger than 300 nm were associated with the apical membranes. The location of particles <300 nm appeared to be intracellular and some particles colocalized with the nucleus.
AAPS PharmSciTech, 2020
Targeting the small intestine employing nanotechnology has proved to be a more effective way for site-specific drug delivery. The drug targeting to the small intestine can be achieved via nanoparticles for its optimum bioavailability within the systemic circulation. The small intestine is a remarkable candidate for localized drug delivery. The intestine has its unique properties. It has a less harsh environment than the stomach, provides comparatively more retention time, and possesses a greater surface area than other parts of the gastrointestinal tract. This review focuses on elaborating the intestinal barriers and approaches to overcome these barriers for internalizing nanoparticles and adopting different cellular trafficking pathways. We have discussed various factors that contribute to nanocarriers' cellular uptake, including their surface chemistry, surface morphology, and functionalization of nanoparticles. Furthermore, the fate of nanoparticles after their uptake at cellular and subcellular levels is also briefly explained. Finally, we have delineated the strategies that are adopted to determine the cytotoxicity of nanoparticles.
Fc-mediated transport of nanoparticles across airway epithelial cell layers
Journal of Controlled Release, 2012
In a study directed towards non-invasive delivery of therapeutic biomacromolecules, we examined whether surface modification of sub-200 nm model nanoparticles with the Fc portion of IgG promotes their cell uptake and transport across the airway epithelial cells. The study initially confirms the expression of the relevant receptor, namely neonatal Fc receptor (FcRn), by Calu-3 cell layers simulating the airway epithelium and demonstrates FcRn-mediated cell association, internalization and transcellular transport of molecular IgG. Surface decoration of nanoparticles with the Fc portion of IgG enhanced both cell uptake and translocation of the particulate system across the cell layers, in a manner strongly suggesting FcRn involvement in these processes. The study further demonstrates the potential of Fc-modified nanoparticles to 'shuttle' a model therapeutic antibody fragment across the epithelial cell layers. Fc-modified nanoparticles are transported in the μg/h/cm 2 range, presenting a substantial increase in transport capacity in comparison to molecular IgG (ng/h/cm 2 range), therefore warranting consideration of the FcRn transcytotic pathway for further investigation as a means to achieve transmucosal delivery of nanoparticulate systems that could act as carriers of a range of biotherapeutics.
Nanoparticles as potential oral delivery systems of proteins and vaccines: A mechanistic approach
Journal of Controlled Release, 2006
Peptides and proteins remain poorly bioavailable upon oral administration. One of the most promising strategies to improve their oral delivery relies on their association with colloidal carriers, e.g. polymeric nanoparticles, stable in gastrointestinal tract, protective for encapsulated substances and able to modulate physicochemical characteristics, drug release and biological behavior. The mechanisms of transport of these nanoparticles across intestinal mucosa are reviewed. In particular, the influence of size and surface properties on their non-specific uptake or their targeted uptake by enterocytes and/or M cells is discussed. Enhancement of their uptake by appropriate cells, i.e. M cells by (i) modeling surface properties to optimize access to and transport by M cells (ii) identifying surface markers specific to human M cell allowing targeting to M cells and nanoparticles transcytosis is illustrated. Encouraging results upon in vivo testing are reported but low bioavailability and lack of control on absorbed dose slow down products development. Vaccines are certainly the most promising applications for orally delivered nanoparticles.