Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: Preliminary evaluation of pharmaceutical excipient based paclitaxel nanoparticles for developing breast cancer therapeutics.

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques: Binding Assay, Fluorescence

Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: Formation and optimization of TAP NPs: A) Space-filling energy-minimized (MM2) molecular modeling shows generation of thermodynamically stable TA-PTX self-assembly. Various ratios as shown above were employed to generate MM2 models of TA-PTX self-assembly. B) Hypothetical representation of 24:1 ratio of TA: PTX self- assembly, so called Tannic acid Nanoparticles (TAP NPs). For visual clarity, PTX in hypothetical structure is represented as.

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques:

Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: A-B) Dynamic light scattering analysis of TAP NPs: stability over 1-week and in different pH solutions. 50 µL of TAP NPs were added to 1 mL of ultrapure water or pH solutions and probe sonicated for 30 s and particle size was measured using Zetasizer (Nano ZS, Malvern Instruments, Malvern, UK) at 25 °C. A) TAP NPs exhibited particle size of 102.22±14.05 nm. Inset: representative particle size of TAP NPs stored at 25 °C for 1-week, demonstrating stability and no significant change in size. B) Stability of TAP NPs in HEPES buffer solutions (pH 6, 7, 7.4 and 8). Data presented as mean ± standard error of the mean (n = 3). C) A representative transmission electron microscopic (TEM) image of TAP NPs. UranyLess EM Stain solution was used to achieve a better contrast purpose for nanoparticles. Image was acquired by using JEOL 200EX TEM at a direct magnification of 100,000× (scale = 100 nm). D) Fourier-transform Infrared (FTIR) spectral analysis of TA, PTX and TAP NPs acquired on a Universal Attenuated Total Reflectance (UATR) accessory plate by a Spectrum 100 FTIR spectrophotometer (Waltham, MA), between 4000 and 650 cm−1 at a scanning speed of 4 cm−1 for 32 scans. TAP NPs show characteristic TA peaks at 1603 and 1700 cm−1 and PTX peaks at 3350 and 1650 cm–1, confirming the presence of functional moieties of TA and PTX in TAP NPs. E) Thermogravimetric analysis (TGA) was recorded for TA, PTX and TAP NPS from 50 to 500 °C by a Rigaku D/Max-B diffractometer (Rigaku Americas Corp, Woodlands, TX) with cobalt-alpha radiation (k = 1.5 Å). There is no significant change in thermograms of TAP NPs in comparison to TA and PTX. F) X-ray diffraction (XRD) was acquired at 2θ range of 25 – 70 oC suggesting TAP NPs has both TA and PTX present in amorphous or dissolution state.

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques: Sonication, Transmission Assay, Staining, Spectrophotometry, Functional Assay

Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: A) Cellular uptake of Coumarin 6 uptake in breast cancer MDA-MB-231 cells were confirmed by fluorescence microscopy an EVOS® 214 FL Imaging System, in a time dependent manner with dye loaded nanoparticles. B) Intracellular uptake of TAP NPs by LC-MS/MS in MDA-MB-231 after treatment with PTX and TAP NPs for 2, 4 and 6 hours. Cells were treated with PTX and TAP NPs (equivalent to 500 ng of PTX) resulting in 95.52% of drug internalization by TAP NPs in 6 hours in contrast to only 57.19% by native drug PTX for the same time. Data presented as mean ± standard error of the mean (n = 3). A Liquid chromatography–tandem mass spectrometry (LC-MS/MS) (Shimadzu Corporation, Kyoto, Japan) connected to Triple Quad 5500 tandem mass spectrometer (AB SCIEX, Framingham, MA, at a flow rate of 0.8 mL/min was used to determine drug concentration(s) in cells.

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques: Fluorescence, Microscopy, Imaging, Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Mass Spectrometry, Concentration Assay

Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: Cell proliferation measured by MTT assay. A) TAP NPs markedly decreased proliferation of breast cancer cells (MDA-MB-231 and MCF7) after treatment with 1–50 nmol/L PTX or TAP NPs (equivalent amount of drug) for 48 hours. Untreated cells were used as control. Effect of TAP NPs and PTX were assessed by MTT reagent and absorbance was recorded by a Microplate Reader (BioTeK Cytation 3, Winooski, VT, USA) at 570 nm. Data presented as mean ± standard error of the mean (n = 3). B) Bright field microscopy images after MTT analysis show significant changes in cell morphology after 48 hours treatment with TAP NPs and PTX. Imaging was done by an EVOS® FL Imaging System (20X magnification, scale = 200 μm).

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques: MTT Assay, Microscopy, Imaging

Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: Inhibition of clonogenicity was measured by colony formation assay. A) Effect of TAP NPs and PTX (equivalent to 0.25, 0.5 and 1 nmol/L PTX) on MDA-MB-231 and MCF7 cells for 15 days were observed for the colony forming ability of breast cancer cells. Colonies were stained with hematoxylin and imaged in Multimage™ light cabinet. Data presented as mean ± standard error of the mean (n = 3). B) Number of colonies formed by each treatment groups; untreated cells (control) were considered as 100 % clonogenicity. Data presented as mean ± standard error of the mean (n = 3).

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques: Inhibition, Colony Assay, Staining

Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: A) Migration assay to determine inhibition potential of breast cancer (MDA-MB-231) cells after exposure to 5 and 10 nmol/L of PTX or TAP NPs (PTX equivalent) for 24 hours. Migratory cells were fixed with 4% formaldehyde and stained with crystal violet and imaged using EVOS® FL Imaging System (scale = 400 µm). Representative bar graphs of the number of migratory cells with each treatment group, where untreated (control) cells served as an experimental control and was considered as 100% and other treatment groups expressed as percentage with respect to the migrated control cells. B) Matrigel Invasion assay showing similar effects, upon treatment with TAP NPs and PTX (5 and 10 nmol/L equivalent to PTX) for 24 hours. Invasive cells following incubation with PTX and TAP NPs were fixed with methanol and stained using crystal violet. Imaged using EVOS® FL Imaging System (scale = 200 μm). Percentage inhibition of invasive breast cancer cells upon treatment and data represented as a bar graph with respect to untreated (control) cells which also served as our experimental control. Data represented as mean ± standard error of the mean (n = 3).

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques: Migration, Inhibition, Staining, Imaging, Invasion Assay, Incubation

Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: A) P-gp-Glo assay showing ATP consumption by different treatment groups having variation in average light units (RLU). Na3VO4 and Verapamil were negative and positive controls respectively, where higher ATP consumption signifies lower RLU. TAP NPs shows 1.42-fold higher RLU in contrast to native drug PTX, which is more effluxed out by the P-gp membranes. B) RH123 (2.62 µmol/L) dye shows higher accumulation in MDA-MB-231 cells after incubation for 30 min, when cells were treated for 48 hours with TAP NPs in contrast to PTX. TAP NPs exhibited significantly increased accumulation, suggesting TAP NPs prevent drug efflux. C) Mean fluorescence intensity in FL1 channel (488 excitation, Blue laser, 530 + 15 nm, FITC/GFP) was measured for different treatment groups. PTX 10 nmol/L treatment showed 3-fold lower intensity over TAP NPs (equivalent to 10 nmol/L PTX) which is probably because of higher efflux of PTX resulting in lower accumulation of RH123 (scale = 200 μm). Data presented as mean ± standard error of the mean (n = 3).

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques: Glo Assay, Incubation, Fluorescence

Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: A) β-tubulin stabilization assay representing confocal images after treatment with 5 and 10 nmol/L PTX or PTX equivalent TAP NPs for 8 hours on MDA-MB-231. Distinct bundles of microtubules were seen in control cells suggesting their proliferating nature, which changed to thicker clusters of microtubules after treatment with TAP NPs and PTX, suggesting stabilization of microtubules accounting for the innate nature of the taxane drug (scale = 20 μm). B) Quantitative analysis of the florescent intensity by Image J software shows significant lower Mean Intensity (arbitrary units) in contrast to TAP NPs. Data represented as mean ± standard error of the mean (n = 3).

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques: Software

Journal: Journal of colloid and interface science

Article Title: Tannic acid-inspired paclitaxel nanoparticles for enhanced anticancer effects in breast cancer cells

doi: 10.1016/j.jcis.2018.09.072

Figure Lengend Snippet: A) Western blot analysis of whole cell lysates of MDA-MB-231 that was treated with 10 nmol/L PTX or 10 nmol/L PTX equivalent TAP NPs and their respective controls for 48 hours and immunoblotted for Cleaved PARP, MDR1/ABCB1, Cleaved Caspase-7, p53, Bad, Bcl-xL and β-actin. The results were consistent in two independent sets of experiments. B) Heat map of differentially regulated gene expression of pro-apoptotic and anti-apoptotic signaling leading to apoptosis in MDA-MB-231 breast cancer cells after exposure to TAP NPs. C) Heat map representing gene expression of ABC family.

Article Snippet: Tannic acid-paclitaxel nanoparticles (TAP NPs) formation by the self-assembly process was confirmed by space-filling energy-minimized (MM2) molecular modeling using ChemBio3D Ultra 14 Suite (CambridgeSoft Corporation, A subsidiary of PerkinElmer, Inc.).

Techniques: Western Blot, Expressing