Transcytosis of Lipid Microspheres by Human Endothelial Cells

Takahashi, Kimiko; Suzuki, Kimihiro; Ichiki, Yayoi; Fukushima, Tsunekazu; Nakamura, Haruo; Sawasaki, Yoshio

doi:10.1159/000139413

Cited by 8 publications

(4 citation statements)

References 22 publications

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“…The binding affinity of Hn33 and DrBoNT was measured using surface plasmon resonance (SPR) using a Biacore 100T instrument. As determined with isothermal calorimetry (ITC) [ 28 ], the dissociation constant (K D ) for Hn33-BoNT/A toxin binding has been reported as 0.4 μM. The K D was determined in this study by immobilizing ligand Hn33 on the CM 3 chip through amine coupling to different concentrations of the analyte DrBoNT.…”

Section: Resultsmentioning

confidence: 99%

“…Proteins labeled with fluorescent dyes have been previously used to study transcytosis [ 26 – 28 ] and have shown proteins labeled with fluorescent dyes provide a sensitive method for measuring transcytosis. Experiments performed in this study used proteins that were conjugated to dyes that fluoresce in the NIR range and were measured using the Odyssey imaging system as previously described [ 22 , 29 ].…”

Section: Methodsmentioning

confidence: 99%

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Role of critical elements in botulinum neurotoxin complex in toxin routing across intestinal and bronchial barriers

et al. 2018

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The highly potent botulinum neurotoxin serotype A (BoNT/A) inhibits neurotransmitter release at neuromuscular junctions resulting in flaccid muscle paralysis, respiratory arrest and death. In order to reach their neuronal cell targets, BoNT/A must cross epithelial cell barriers lining the intestines and airways. The toxin is produced as a large protein complex comprised of the neurotoxin and non-toxic neurotoxin-associated proteins (NAPs). Although NAPs are known to protect the toxin from harsh environments, their role in the movement of BoNT/A across epithelial barriers has not been fully characterized. In the current study, movement of the toxin across epithelial cells was examined macroscopically using a sensitive near infrared fluorescence transcytosis assay and microscopically using fluorescently labeled toxin and confocal microscopy. The studies show that the BoNT/A complex internalizes more rapidly than the pure toxin. The studies also show that one NAP protein, hemaglutinin 33 (Hn33), enhanced both the binding and movement of a deactivated recombinant botulinum neurotoxin A (DrBoNT) across epithelial cell monolayers and that the toxin associates with Hn33 on the cell surface. Collectively, the data demonstrate that, in addition to their protective role, NAPs and Hn33 play an important role in BoNT/A intoxication.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Role of critical elements in botulinum neurotoxin complex in toxin routing across intestinal and bronchial barriers

et al. 2018

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Section: Nanoconjugate Drug-delivery Pathwaymentioning

confidence: 99%

“…Binding of mAbs to TfRs or insulin receptors that are overexpressed at the surface of tumor cells and tumor endothelium enables the nanocarrier to pass through the BTB by transcytosis [12,31,[77][78][79]. Transcytosis does not appear to cause complex degradation inside endothelial cells [80,81], especially not to cleave disulfide (S-S) bonds [82], thus enabling disulfide conjugation of the drugs to the nanocarrier platform.Once in the interstitium, the delivery vehicle penetrates through tumor tissue, probably by diffusion, its rate depending on cell adhesion and density [83]. After binding to a surface antigen of recipient tumor cells (e.g., TfR), the nanoconjugate is internalized into early endosomes, which then, by fusion with primary lysosomes, mature into lysosomes with concomitant acidification towards pH 5.…”

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confidence: 99%

Poly(Malic Acid) Nanoconjugates Containing Various Antibodies and Oligonucleotides for Multitargeting Drug Delivery

et al. 2008

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Nanoconjugates are emerging as promising drug-delivery vehicles because of their multimodular structure enabling them to actively target discrete cells, pass through biological barriers and simultaneously carry multiple drugs of various chemical nature. Nanoconjugates have matured from simple devices to multifunctional, biodegradable, nontoxic and nonimmunogenic constructs, capable of delivering synergistically functioning drugs in vivo. This review mainly concerns the Polycefin family of natural-derived polymeric drug-delivery devices as an example. This type of vehicle is built by hierarchic conjugation of functional groups onto the backbone of poly(malic acid), an aliphatic polyester obtained from the microorganism Physarum polycephalum. Particular Polycefin variants target human brain and breast tumors implanted into animals specifically and actively and could be detected easily by noninvasive imaging analysis. Delivery of antisense oligonucleotides to a tumor-specific angiogenic marker using Polycefin resulted in significant inhibition of tumor angiogenesis and increase of animal survival. Keywordsbiodegradable; brain cancer; breast cancer; imaging analysis; multiple antibodies; multiple drug delivery; multitargeting; Polycefin; poly(malic acid); tumor angiogenesis Engineering of polymeric biodegradable nanoconjugates represents one of the rapidly emerging and important areas of nanotechnology and nanomedicine. In the context of this review, these nanoconjugates will be discussed mostly in relation to anticancer drug delivery. Polymeric nanoconjugates are characterized by multiple covalently attached NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript functional moieties that can target cellular and molecular markers of diseased tissues specifically to achieve maximum treatment efficacy. An outstanding feature of polymeric nanoconjugates is that they can carry a variety of covalently bound drugs that may act simultaneously on several targets (e.g., mRNA and/or protein). The benefit of such drug carriers is that they deliver more than one prodrug by single conjugate molecule, increasing the probability of coordinate and synergistic action and thus providing an efficient inhibition of multiple aberrant tumor pathways. Owing to the specific targeting of tumor tissue/cells, nanoconjugates provide therapeutically efficacious drug concentrations at the site of treatment and minimal side effects on healthy tissue. Nanoconjugate delivery systems are significantly different from nonconjugated nanodelivery vehicles, for example, micelles and liposomes, which also combine drugs, targeting and/or other functional moieties but do not form a covalently linked chemical entity.Devices lacking this entity are prone to continuously lose their constituents by spontaneous or damage-induced leakage and this could be a source of toxicity to healthy tissue or disparities in initially balanced drug compositions.Biodegradability and lack of immunogenicity are other criteria important for treatment tha...

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Preservation of the Characteristics of the Cultured Human Type II Alveolar Epithelial Cells

Takahashi

Mitsui

Takeuchi

et al. 2004

Lung

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The human type II alveolar epithelial cells lost their specific characteristics during cultivation. We examined the ultrastructural and biochemical nature of the human type II cells cultured by two culture systems. To make a physiological alveoli model, the epithelial cells were seeded onto the cell culture insert and allowed contact with the air directly. The cells exposed to the air expressed polarity and immature lamellar bodies in their cytoplasm. Separately, the alveolar epithelial cells were cultured as spheroids to construct the three-dimensional condition. These cells expressed mature morphological characteristics as epithelial cells and lamellar bodies. The expression of the surfactant apoprotein-A (SP-A) and -C (SP-C) mRNA was compared in the cells cultured as a monolayer, the air exposed and the spheroids. SP-A mRNA was detected in all the cultured epithelial cells, but SP-C mRNA, a specific protein for the type II cells, was expressed only in the cells forming spheroids. The expression of uPA, one of the fibrinolytic enzymes, its receptor (uPAR) and its inhibitor-1 (PAI-1) were also examined. The epithelial cells exposed to the air and formed spheroids expressed a larger amount of uPA mRNA than the monolayer, although the amount of uPAR mRNA were comparable in these cells. The amount of PAI-1 mRNA significantly increased when the epithelial cells were exposed to the air. These results indicate that the type II alveolar epithelial cells induced and preserved their specific characteristics by taking the physiological three-dimensional structure, and these characteristics were partially restored by exposure to the air. Those findings suggest that the alveolar epithelial cells should be cultivated in three-dimensional form with contact to the air to regenerate an appropriate alveolar tissue.

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Transcytosis of Lipid Microspheres by Human Endothelial Cells

Cited by 8 publications

References 22 publications

Role of critical elements in botulinum neurotoxin complex in toxin routing across intestinal and bronchial barriers

Role of critical elements in botulinum neurotoxin complex in toxin routing across intestinal and bronchial barriers

Poly(Malic Acid) Nanoconjugates Containing Various Antibodies and Oligonucleotides for Multitargeting Drug Delivery

Preservation of the Characteristics of the Cultured Human Type II Alveolar Epithelial Cells

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