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Blood-brain barrier transport studies, aggregation, and molecular dynamics simulation of multiwalled carbon nanotube functionalized with fluorescein isothiocyanate.

Shityakov S, Salvador E, Pastorin G, Förster C - Int J Nanomedicine (2015)

Bottom Line: The results indicated that the MWCNT-FITC conjugate is able to penetrate microvascular cerebral endothelial monolayers; its concentrations in the Transwell(®) system were fully equilibrated after 48 hours.These microscopic techniques also revealed presumably the intracellular localization of fluorescent MWCNT-FITCs apart from their massive nonfluorescent accumulation on the cellular surface due to nanotube lipophilic properties.In addition, the 1,000 ps molecular dynamics simulation in vacuo discovered the phenomenon of carbon nanotube aggregation driven by van der Waals forces via MWCNT-FITC rapid dissociation as an intermediate phase.

View Article: PubMed Central - PubMed

Affiliation: Department of Anaesthesia and Critical Care, University of Würzburg, Würzburg, Germany.

ABSTRACT
In this study, the ability of a multiwalled carbon nanotube functionalized with fluorescein isothiocyanate (MWCNT-FITC) was assessed as a prospective central nervous system-targeting drug delivery system to permeate the blood-brain barrier. The results indicated that the MWCNT-FITC conjugate is able to penetrate microvascular cerebral endothelial monolayers; its concentrations in the Transwell(®) system were fully equilibrated after 48 hours. Cell viability test, together with phase-contrast and fluorescence microscopies, did not detect any signs of MWCNT-FITC toxicity on the cerebral endothelial cells. These microscopic techniques also revealed presumably the intracellular localization of fluorescent MWCNT-FITCs apart from their massive nonfluorescent accumulation on the cellular surface due to nanotube lipophilic properties. In addition, the 1,000 ps molecular dynamics simulation in vacuo discovered the phenomenon of carbon nanotube aggregation driven by van der Waals forces via MWCNT-FITC rapid dissociation as an intermediate phase.

No MeSH data available.


Related in: MedlinePlus

Western blot analysis of tight junction-related proteins expressed in cEND cells was performed using antibodies against occludin, claudin-5, and β-actin (internal/loading control).Notes: Expression profiles of tight junction-related proteins (occludin and claudin-5) in untreated (control) and MWCNT–FITC-treated cEND cells (A). Immunoblots analyzed and proteins quantified with the ImageJ software (B). The area is measured in arbitrary units, abbreviated as au. Data represent means ± SD of three independent experiments.Abbreviations: MWCNT–FITC, multiwalled carbon nanotube functionalized with fluorescein isothiocyanate; cEND, cerebral endothelial; SD, standard deviation.
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f3-ijn-10-1703: Western blot analysis of tight junction-related proteins expressed in cEND cells was performed using antibodies against occludin, claudin-5, and β-actin (internal/loading control).Notes: Expression profiles of tight junction-related proteins (occludin and claudin-5) in untreated (control) and MWCNT–FITC-treated cEND cells (A). Immunoblots analyzed and proteins quantified with the ImageJ software (B). The area is measured in arbitrary units, abbreviated as au. Data represent means ± SD of three independent experiments.Abbreviations: MWCNT–FITC, multiwalled carbon nanotube functionalized with fluorescein isothiocyanate; cEND, cerebral endothelial; SD, standard deviation.

Mentions: In the next step, the expression of tight junction proteins (occludin and claudin-5) in the cEND cells was analyzed by Western blotting technique in untreated control cEND cells and cells following 48 hours of MWCNT–FITC exposure in a concentration of 40 μg/mL. As shown in Figure 3, the occludin and claudin-5 protein expression profiles at the steady state were the same in both control (untreated cultures) and MWCNT-treated cells. Although a slight elevation of tight junction proteins (ZO-1) has been previously reported by Rotoli et al investigating the impairment of barrier function in Calu-3 monolayers, caused by MWCNT structures, the change did not reach statistical significance because of the variability of absolute protein expression levels in the different experiments.40


Blood-brain barrier transport studies, aggregation, and molecular dynamics simulation of multiwalled carbon nanotube functionalized with fluorescein isothiocyanate.

Shityakov S, Salvador E, Pastorin G, Förster C - Int J Nanomedicine (2015)

Western blot analysis of tight junction-related proteins expressed in cEND cells was performed using antibodies against occludin, claudin-5, and β-actin (internal/loading control).Notes: Expression profiles of tight junction-related proteins (occludin and claudin-5) in untreated (control) and MWCNT–FITC-treated cEND cells (A). Immunoblots analyzed and proteins quantified with the ImageJ software (B). The area is measured in arbitrary units, abbreviated as au. Data represent means ± SD of three independent experiments.Abbreviations: MWCNT–FITC, multiwalled carbon nanotube functionalized with fluorescein isothiocyanate; cEND, cerebral endothelial; SD, standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356663&req=5

f3-ijn-10-1703: Western blot analysis of tight junction-related proteins expressed in cEND cells was performed using antibodies against occludin, claudin-5, and β-actin (internal/loading control).Notes: Expression profiles of tight junction-related proteins (occludin and claudin-5) in untreated (control) and MWCNT–FITC-treated cEND cells (A). Immunoblots analyzed and proteins quantified with the ImageJ software (B). The area is measured in arbitrary units, abbreviated as au. Data represent means ± SD of three independent experiments.Abbreviations: MWCNT–FITC, multiwalled carbon nanotube functionalized with fluorescein isothiocyanate; cEND, cerebral endothelial; SD, standard deviation.
Mentions: In the next step, the expression of tight junction proteins (occludin and claudin-5) in the cEND cells was analyzed by Western blotting technique in untreated control cEND cells and cells following 48 hours of MWCNT–FITC exposure in a concentration of 40 μg/mL. As shown in Figure 3, the occludin and claudin-5 protein expression profiles at the steady state were the same in both control (untreated cultures) and MWCNT-treated cells. Although a slight elevation of tight junction proteins (ZO-1) has been previously reported by Rotoli et al investigating the impairment of barrier function in Calu-3 monolayers, caused by MWCNT structures, the change did not reach statistical significance because of the variability of absolute protein expression levels in the different experiments.40

Bottom Line: The results indicated that the MWCNT-FITC conjugate is able to penetrate microvascular cerebral endothelial monolayers; its concentrations in the Transwell(®) system were fully equilibrated after 48 hours.These microscopic techniques also revealed presumably the intracellular localization of fluorescent MWCNT-FITCs apart from their massive nonfluorescent accumulation on the cellular surface due to nanotube lipophilic properties.In addition, the 1,000 ps molecular dynamics simulation in vacuo discovered the phenomenon of carbon nanotube aggregation driven by van der Waals forces via MWCNT-FITC rapid dissociation as an intermediate phase.

View Article: PubMed Central - PubMed

Affiliation: Department of Anaesthesia and Critical Care, University of Würzburg, Würzburg, Germany.

ABSTRACT
In this study, the ability of a multiwalled carbon nanotube functionalized with fluorescein isothiocyanate (MWCNT-FITC) was assessed as a prospective central nervous system-targeting drug delivery system to permeate the blood-brain barrier. The results indicated that the MWCNT-FITC conjugate is able to penetrate microvascular cerebral endothelial monolayers; its concentrations in the Transwell(®) system were fully equilibrated after 48 hours. Cell viability test, together with phase-contrast and fluorescence microscopies, did not detect any signs of MWCNT-FITC toxicity on the cerebral endothelial cells. These microscopic techniques also revealed presumably the intracellular localization of fluorescent MWCNT-FITCs apart from their massive nonfluorescent accumulation on the cellular surface due to nanotube lipophilic properties. In addition, the 1,000 ps molecular dynamics simulation in vacuo discovered the phenomenon of carbon nanotube aggregation driven by van der Waals forces via MWCNT-FITC rapid dissociation as an intermediate phase.

No MeSH data available.


Related in: MedlinePlus