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  • 1.
    Fontana, Jacopo Maria
    et al.
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Yin, Huijuan
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Chen, Yun
    Florez, Ricardo
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Brismar, Hjalmar
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Fu, Ying
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Transport and release of colloidal 3-mercaptopropionic acid-coated CdSe-CdS/ZnS core-multishell quantum dots in human umbilical vein endothelial cells2017In: International Journal of Nanomedicine, ISSN 1176-9114, E-ISSN 1178-2013, Vol. 12, p. 8615-8629Article in journal (Refereed)
    Abstract [en]

    Colloidal semiconductor quantum dots (QDs) have been extensively researched and developed for biomedical applications, including drug delivery and biosensing assays. Hence, it is pivotal to understand their behavior in terms of intracellular transport and toxicological effects. In this study, we focused on 3-mercaptopropionic acid-coated CdSe-CdS/ZnS core-multishell quantum dots (3MPA-QDs) converted from the as-grown octadecylamine-coated quantum dots (ODA-QDs) and their direct and dynamic interactions with human umbilical vein endothelial cells (HUVECs). Live cell imaging using confocal fluorescence microscopy showed that 3MPAQDs first attached to and subsequently aggregated on HUVEC plasma membrane similar to 25 min after QD deposition. The aggregated QDs started being internalized at similar to 2 h and reached their highest internalization degree at similar to 24 h. They were released from HUVECs after similar to 48 h. During the 48 h period, the HUVECs responded normally to external stimulations, grew, proliferated and wound healed without any perceptible apoptosis. Furthermore, 1) 3MPA-QDs were internalized in newly formed LysoTracker-stained early endosomes; 2) adenosine 5'-triphosphateinduced [Ca2+](i) modulation caused a transient decrease in the fluorescence of 3MPA-QDs that were attached to the plasma membrane but a transient increase in the internalized 3MPA-QDs; and 3) fluorescence signal modulations of co-stained LysoTracker and QDs induced by the lysosomotropic agent Gly-Phe-beta-naphthylamide were spatially co-localized and temporally synchronized. Our findings suggest that 3MPA-QDs converted from ODA-QDs are a potential nontoxic fluorescent probe for future use in clinical applications. Moreover, the photophysical strategy and techniques reported in this work are easily applicable to study of direct interactions between other nanoparticles and live cells; contributing to awareness and implementation of the safe applications of nanoparticles.

  • 2.
    Yin, Huijuan
    et al.
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Fontana, Jacopo M.
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Solandt, Johan
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab. AstraZeneca R&D, Sweden.
    Jussi, Johnny Israelsson
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Xu, Hao
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Brismar, Hjalmar
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Fu, Ying
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Quantum dots modulate intracellular Ca2+ level in lung epithelial cells2017In: International Journal of Nanomedicine, ISSN 1176-9114, E-ISSN 1178-2013, Vol. 12, p. 2781-2792Article in journal (Refereed)
    Abstract [en]

    While adverse effects of nanoparticles on lung health have previously been proposed, few studies have addressed the direct effects of nanoparticle exposure on the airway epithelium. In this work, we examine the response of the pulmonary airway to nanoparticles by measuring intracellular Ca2+ concentration ([Ca2+](i)) in the Calu-3 epithelial layer stimulated by 3-mercaptopropionic-acid (3MPA) coated CdSe-CdS/ZnS core-multishell quantum dots (QDs). Simultaneous transient transepithelial electrical resistance (TEER) decrease and global [Ca2+](i) increase in Calu-3 epithelial layer, accompanied by cell displacements, contraction, and expansion, were observed under QD deposition. This suggests that a QD-induced global [Ca2+](i) increase in the Calu-3 epithelial layer caused the transient TEER decrease. The [Ca2+](i) increase was marked and rapid in the apical region, while [Ca2+](i) decreased in the basolateral region of the epithelial layer. TEER transient response and extracellular Ca2+ entry induced by QD deposition were completely inhibited in cells treated with stretched-activated (SA) inhibitor GdCl3 and store-operated calcium entry (SOCE) inhibitor BTP2 and in cells immersed in Ca2+-free medium. The voltage-gated calcium channel (VGCC) inhibitor nifedipine decreased, stabilized, and suppressed the TEER response, but did not affect the [Ca2+](i) increase, due to QD deposition. This demonstrates that the Ca2+ influx activated by QDs' mechanical stretch occurs through activation of both SA and SOCE channels. QD-induced [Ca2+](i) increase occurred in the Calu-3 epithelial layer after culturing for 15 days, while significant TEER drop only occurred after 23 days. This work provides a new perspective from which to study direct interactions between airway epithelium and nanoparticles and may help to reveal the pathologies of pulmonary disease.

  • 3.
    Yin, Huijuan
    et al.
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics.
    Shi, Xiafei
    Wang, Hong
    Jin, Wendong
    Li, Yingxin
    Fu, Ying
    KTH, School of Engineering Sciences (SCI), Applied Physics, Cellular Biophysics.
    Photodynamic therapy targeting VCAM-1-expressing human umbilical vein endothelial cells using a PpIX-VCAM-1 binding peptide-quantum dot conjugate2017In: RSC Advances, ISSN 2046-2069, E-ISSN 2046-2069, Vol. 7, no 80, p. 50562-50570Article in journal (Refereed)
    Abstract [en]

    With increasing knowledge of the relevance of vascular cell adhesion molecule 1 (VCAM-1) for tumor growth, metastasis, angiogenesis, and related processes, it has become an attractive anti-tumor strategy to target VCAM-1 expression on the tumor vasculature. We designed a new targeted nanodrug, denoted PVQ, based on a photosensitizer (for the photodynamic effect), VCAM-1 target and quantum dot (QD) carrier, using conjugated water-dispersible colloidal CdSe-CdS/ZnS QDs, protoporphyrin IX (PpIX) photosensitizers, and VCAM-1 binding peptides. Its targeting ability and photodynamic therapy (PDT) efficiency against VCAM-1 expression in human umbilical vein endothelial cells (HUVECs) were then investigated. Conjugates of QD-VCAM-1 binding peptide (VQ), PpIX-VCAM-1 binding peptide (PV), and PVQ prepared using amide coupling were verified by agarose gel electrophoresis, Fourier transform infrared spectroscopy, and fluorescence spectrometry. VCAM-1 expression in HUVECs was induced by TNF-alpha treatment. PVQ conjugates were co-cultured with VCAM-1 expressing (VCAM-1(+)) and non-expressing (VCAM-1(-)) HUVECs, and target imaging, ROS generation, cell death, and apoptosis were analyzed using confocal fluorescence microscopy. VCAM-1 target imaging could not distinguish between VCAM-1(+) and VCAM-1(-) HUVECs after only 6 h of incubation; however it could distinguish between the cells after incubation for 24 h. After incubation for ca. 30 min, PVQ generated a significantly higher yield of ROS (3.6 fold) in VCAM-1(+) HUVECs compared with VCAM-1(-) cells, during 10 min of irradiation at a wavelength of 405 nm, and this was followed by a second rise in ROS at 30 min after irradiation. Moreover, cell destruction was observed clearly in VCAM-1(+) cells treated with PVQ and almost all cells became round after 30 min of irradiation at 405 nm. PVQ-induced PDT effects caused a significant apoptosis (onset and late apoptosis) in VCAM-1(+) HUVECs at 6 h after PDT treatment. In conclusion, PVQ shows a great potential for targeted PDT in cancer therapy.

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