ker nps ptx  (Worthington Biochemical)

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Antiproliferative activity of PTX in a free form, HSA-NPs-PTX, and KER-NPs-PTX on MCF-7 and MDA MB 231 cell lines in 2D model. Notes: Cell viability was evaluated 72 h after exposure to increasing concentrations of PTX (0.00002, 0.02, and 5 µg/mL) by APH assay. Statistical significance versus untreated cells (100%, represented by a dotted line): ** P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Activity Assay, Multiple Displacement Amplification

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: BAX and BCL-2 gene expression analyses in MCF-7 and MDA MB 231 cells in p3D cultures following 24 h treatments. Notes: Cells were incubated for 24 h with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX (PTX, 5 µg/mL). GAPDH was used as reference gene to normalize data. The effects of each treatment were compared with gene expression detectable in untreated cells (=1) as indicated by the dotted line. Statistically significant difference versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Expressing, Multiple Displacement Amplification, Incubation

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Representative H E staining of scaffold sections from p3D MCF-7 and MDA MB 231 cultures after treatments. Notes: MCF-7 (upper panels) and MDA MB 231 (lower panels) cells were initially cultured for 7 and 4 days, respectively, and then left untreated ( A ) or treated for 48 h with PTX in a free form ( B ), HSA-NPs-PTX ( C ), and KER-NPs-PTX ( D ) (PTX, 5 µg/mL). Scale bar: 100 µm. Abbreviations: H E, hematoxylin and eosin; HSA-NPs-PTX, PTX loaded in albumin nanoparticles; KER-NPs-PTX, PTX loaded in keratin nanoparticles; p3D, three-dimensional model with perfused bioreactor; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Staining, Multiple Displacement Amplification, Cell Culture

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: BAX and BCL-2 gene expression analyses in MCF-7 and MDA MB 231 cells cultured in 2D model upon 12 h treatment. Notes: Cells were incubated for 12 h with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX (PTX, 5 µg/mL). GAPDH was used as reference gene to normalize data. Effects of each treatment on gene expression levels were compared with those detectable in untreated cells (n=1) as indicated by the dotted line. Statistically significant difference versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Expressing, Multiple Displacement Amplification, Cell Culture, Incubation

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Effects of PTX in a free form or vehiculated by HSA-NPs or by KER-NPs on cell death in p3D cultures. Notes: MCF-7 and MDA MB 231 cells were exposed to PTX in a free form, HSA-NPs-PTX, and KER-NPs-PTX (PTX, 5 µg/mL) for 24 and 48 h. Apoptosis assays were carried out by flow cytometry, following APC-Annexin V and PI staining. Representative dot plots of MCF-7 ( A ) and MDA MB 231 ( B ) cell lines after each treatment. Percentages of early apoptotic cells (positive to APC-Annexin V and negative to PI) and late apoptotic cells (positive to APC-Annexin V and PI) were evaluated 24 and 48 h after treatment ( C ). Statistical significance versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Multiple Displacement Amplification, Flow Cytometry, Cytometry, Staining

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: KER-NPs-PTX analysis. Notes: Transmission electron microscopy images of fixed KER-NPs-PTX at two magnifications: scale bar 2 µm ( A ) and scale bar 200 nm ( B ). Morphological analysis showed particles spherical in shape, smooth surface, and an average dry diameter of ~80 nm. Stability of KER-NPs-PTX in physiological conditions ( C ). Fluorescence spectra of KER-NPs-PTX at different PTX loading percentages ( D ). Abbreviations: KER-NPs-PTX, PTX loaded in keratin nanoparticles; PDI, polydispersity index; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Transmission Assay, Electron Microscopy, Fluorescence

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Profiles and mechanisms of PTX release from KER-NPs in vitro. Notes: Profile of PTX release from KER-NPs ( A ). Correlation coefficient values ( R 2 ) used to determine the best fitting model among zero-order, first-order, and Higuchi modes ( B ). Abbreviations: KER-NPs, keratin nanoparticles; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: In Vitro

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: KER-NPs-PTX preparation. Notes: ( A ) KER-NPs-PTX preparation by aggregation method, starting from a solution of pure keratin powder. ( B ) Correlation between PTX loading and KER-NPs-PTX size. Abbreviations: KER-NPs-PTX, PTX loaded in keratin nanoparticles; PDI, polydispersity index; PTX, paclitaxel; RT, room temperature.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Immunofluorescence staining of CC3 in MCF-7 (upper panels) and MDA MB 231 (lower panels) scaffold sections. Notes: Images show DAPI (blue) and CC3 (green) staining in MCF-7 ( A ) and MDA MB 231 ( B ) p3D scaffold sections upon treatment with PTX in a free form (b), HSA-NPs-PTX (c), and KER-NPs-PTX (d) (PTX, 5 µg/mL) for 48 h or untreated (a). Scale bar: 100 µm. Abbreviations: CC3, cleaved caspase 3; DAPI, 4′,6-diamidino-2-phenylindole; HSA-NPs-PTX, PTX loaded in albumin nanoparticles; KER-NPs-PTX, PTX loaded in keratin nanoparticles; p3D, three-dimensional model with perfused bioreactor; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Immunofluorescence, Staining, Multiple Displacement Amplification

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Induction of apoptosis in tumor cells cultured in 2D by PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX in 2D model. Notes: MCF-7 and MDA MB 231 cells were exposed to PTX, HSA-NPs-PTX, and KER-NPs-PTX (PTX, 5 µg/mL) for 24 and 48 h. Apoptosis assays were carried out by flow cytometry, following APC-Annexin V and PI staining. Representative dot plots of MCF-7 ( A ) and MDA MB 231 ( B ) cell lines after each treatment. Percentage of early apoptotic cells (positive to APC-Annexin V and negative to PI) and late apoptotic cells (positive to APC-Annexin V and PI) was evaluated 24 and 48 h after treatment ( C ). Statistical significance versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Cell Culture, Multiple Displacement Amplification, Flow Cytometry, Cytometry, Staining

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Antiproliferative activity of PTX in a free form, HSA-NPs-PTX, and KER-NPs-PTX on MCF-7 and MDA MB 231 cell lines in 2D model. Notes: Cell viability was evaluated 72 h after exposure to increasing concentrations of PTX (0.00002, 0.02, and 5 µg/mL) by APH assay. Statistical significance versus untreated cells (100%, represented by a dotted line): ** P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Activity Assay, Multiple Displacement Amplification

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Representative H E staining of scaffold sections from p3D MCF-7 and MDA MB 231 cultures after treatments. Notes: MCF-7 (upper panels) and MDA MB 231 (lower panels) cells were initially cultured for 7 and 4 days, respectively, and then left untreated ( A ) or treated for 48 h with PTX in a free form ( B ), HSA-NPs-PTX ( C ), and KER-NPs-PTX ( D ) (PTX, 5 µg/mL). Scale bar: 100 µm. Abbreviations: H E, hematoxylin and eosin; HSA-NPs-PTX, PTX loaded in albumin nanoparticles; KER-NPs-PTX, PTX loaded in keratin nanoparticles; p3D, three-dimensional model with perfused bioreactor; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Staining, Multiple Displacement Amplification, Cell Culture

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: BAX and BCL-2 gene expression analyses in MCF-7 and MDA MB 231 cells cultured in 2D model upon 12 h treatment. Notes: Cells were incubated for 12 h with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX (PTX, 5 µg/mL). GAPDH was used as reference gene to normalize data. Effects of each treatment on gene expression levels were compared with those detectable in untreated cells (n=1) as indicated by the dotted line. Statistically significant difference versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Expressing, Multiple Displacement Amplification, Cell Culture, Incubation

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Effects of PTX in a free form or vehiculated by HSA-NPs or by KER-NPs on cell death in p3D cultures. Notes: MCF-7 and MDA MB 231 cells were exposed to PTX in a free form, HSA-NPs-PTX, and KER-NPs-PTX (PTX, 5 µg/mL) for 24 and 48 h. Apoptosis assays were carried out by flow cytometry, following APC-Annexin V and PI staining. Representative dot plots of MCF-7 ( A ) and MDA MB 231 ( B ) cell lines after each treatment. Percentages of early apoptotic cells (positive to APC-Annexin V and negative to PI) and late apoptotic cells (positive to APC-Annexin V and PI) were evaluated 24 and 48 h after treatment ( C ). Statistical significance versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Multiple Displacement Amplification, Flow Cytometry, Cytometry, Staining

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: KER-NPs-PTX analysis. Notes: Transmission electron microscopy images of fixed KER-NPs-PTX at two magnifications: scale bar 2 µm ( A ) and scale bar 200 nm ( B ). Morphological analysis showed particles spherical in shape, smooth surface, and an average dry diameter of ~80 nm. Stability of KER-NPs-PTX in physiological conditions ( C ). Fluorescence spectra of KER-NPs-PTX at different PTX loading percentages ( D ). Abbreviations: KER-NPs-PTX, PTX loaded in keratin nanoparticles; PDI, polydispersity index; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Transmission Assay, Electron Microscopy, Fluorescence

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Profiles and mechanisms of PTX release from KER-NPs in vitro. Notes: Profile of PTX release from KER-NPs ( A ). Correlation coefficient values ( R 2 ) used to determine the best fitting model among zero-order, first-order, and Higuchi modes ( B ). Abbreviations: KER-NPs, keratin nanoparticles; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: In Vitro

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: KER-NPs-PTX preparation. Notes: ( A ) KER-NPs-PTX preparation by aggregation method, starting from a solution of pure keratin powder. ( B ) Correlation between PTX loading and KER-NPs-PTX size. Abbreviations: KER-NPs-PTX, PTX loaded in keratin nanoparticles; PDI, polydispersity index; PTX, paclitaxel; RT, room temperature.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Immunofluorescence staining of CC3 in MCF-7 (upper panels) and MDA MB 231 (lower panels) scaffold sections. Notes: Images show DAPI (blue) and CC3 (green) staining in MCF-7 ( A ) and MDA MB 231 ( B ) p3D scaffold sections upon treatment with PTX in a free form (b), HSA-NPs-PTX (c), and KER-NPs-PTX (d) (PTX, 5 µg/mL) for 48 h or untreated (a). Scale bar: 100 µm. Abbreviations: CC3, cleaved caspase 3; DAPI, 4′,6-diamidino-2-phenylindole; HSA-NPs-PTX, PTX loaded in albumin nanoparticles; KER-NPs-PTX, PTX loaded in keratin nanoparticles; p3D, three-dimensional model with perfused bioreactor; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Immunofluorescence, Staining, Multiple Displacement Amplification

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Induction of apoptosis in tumor cells cultured in 2D by PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX in 2D model. Notes: MCF-7 and MDA MB 231 cells were exposed to PTX, HSA-NPs-PTX, and KER-NPs-PTX (PTX, 5 µg/mL) for 24 and 48 h. Apoptosis assays were carried out by flow cytometry, following APC-Annexin V and PI staining. Representative dot plots of MCF-7 ( A ) and MDA MB 231 ( B ) cell lines after each treatment. Percentage of early apoptotic cells (positive to APC-Annexin V and negative to PI) and late apoptotic cells (positive to APC-Annexin V and PI) was evaluated 24 and 48 h after treatment ( C ). Statistical significance versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Cell Culture, Multiple Displacement Amplification, Flow Cytometry, Cytometry, Staining

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Antiproliferative activity of PTX in a free form, HSA-NPs-PTX, and KER-NPs-PTX on MCF-7 and MDA MB 231 cell lines in 2D model. Notes: Cell viability was evaluated 72 h after exposure to increasing concentrations of PTX (0.00002, 0.02, and 5 µg/mL) by APH assay. Statistical significance versus untreated cells (100%, represented by a dotted line): ** P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Activity Assay, Multiple Displacement Amplification

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: BAX and BCL-2 gene expression analyses in MCF-7 and MDA MB 231 cells in p3D cultures following 24 h treatments. Notes: Cells were incubated for 24 h with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX (PTX, 5 µg/mL). GAPDH was used as reference gene to normalize data. The effects of each treatment were compared with gene expression detectable in untreated cells (=1) as indicated by the dotted line. Statistically significant difference versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Expressing, Multiple Displacement Amplification, Incubation

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Representative H E staining of scaffold sections from p3D MCF-7 and MDA MB 231 cultures after treatments. Notes: MCF-7 (upper panels) and MDA MB 231 (lower panels) cells were initially cultured for 7 and 4 days, respectively, and then left untreated ( A ) or treated for 48 h with PTX in a free form ( B ), HSA-NPs-PTX ( C ), and KER-NPs-PTX ( D ) (PTX, 5 µg/mL). Scale bar: 100 µm. Abbreviations: H E, hematoxylin and eosin; HSA-NPs-PTX, PTX loaded in albumin nanoparticles; KER-NPs-PTX, PTX loaded in keratin nanoparticles; p3D, three-dimensional model with perfused bioreactor; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Staining, Multiple Displacement Amplification, Cell Culture

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: BAX and BCL-2 gene expression analyses in MCF-7 and MDA MB 231 cells cultured in 2D model upon 12 h treatment. Notes: Cells were incubated for 12 h with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX (PTX, 5 µg/mL). GAPDH was used as reference gene to normalize data. Effects of each treatment on gene expression levels were compared with those detectable in untreated cells (n=1) as indicated by the dotted line. Statistically significant difference versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Expressing, Multiple Displacement Amplification, Cell Culture, Incubation

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Effects of PTX in a free form or vehiculated by HSA-NPs or by KER-NPs on cell death in p3D cultures. Notes: MCF-7 and MDA MB 231 cells were exposed to PTX in a free form, HSA-NPs-PTX, and KER-NPs-PTX (PTX, 5 µg/mL) for 24 and 48 h. Apoptosis assays were carried out by flow cytometry, following APC-Annexin V and PI staining. Representative dot plots of MCF-7 ( A ) and MDA MB 231 ( B ) cell lines after each treatment. Percentages of early apoptotic cells (positive to APC-Annexin V and negative to PI) and late apoptotic cells (positive to APC-Annexin V and PI) were evaluated 24 and 48 h after treatment ( C ). Statistical significance versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Multiple Displacement Amplification, Flow Cytometry, Cytometry, Staining

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: KER-NPs-PTX analysis. Notes: Transmission electron microscopy images of fixed KER-NPs-PTX at two magnifications: scale bar 2 µm ( A ) and scale bar 200 nm ( B ). Morphological analysis showed particles spherical in shape, smooth surface, and an average dry diameter of ~80 nm. Stability of KER-NPs-PTX in physiological conditions ( C ). Fluorescence spectra of KER-NPs-PTX at different PTX loading percentages ( D ). Abbreviations: KER-NPs-PTX, PTX loaded in keratin nanoparticles; PDI, polydispersity index; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Transmission Assay, Electron Microscopy, Fluorescence

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Profiles and mechanisms of PTX release from KER-NPs in vitro. Notes: Profile of PTX release from KER-NPs ( A ). Correlation coefficient values ( R 2 ) used to determine the best fitting model among zero-order, first-order, and Higuchi modes ( B ). Abbreviations: KER-NPs, keratin nanoparticles; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: In Vitro

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: KER-NPs-PTX preparation. Notes: ( A ) KER-NPs-PTX preparation by aggregation method, starting from a solution of pure keratin powder. ( B ) Correlation between PTX loading and KER-NPs-PTX size. Abbreviations: KER-NPs-PTX, PTX loaded in keratin nanoparticles; PDI, polydispersity index; PTX, paclitaxel; RT, room temperature.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Immunofluorescence staining of CC3 in MCF-7 (upper panels) and MDA MB 231 (lower panels) scaffold sections. Notes: Images show DAPI (blue) and CC3 (green) staining in MCF-7 ( A ) and MDA MB 231 ( B ) p3D scaffold sections upon treatment with PTX in a free form (b), HSA-NPs-PTX (c), and KER-NPs-PTX (d) (PTX, 5 µg/mL) for 48 h or untreated (a). Scale bar: 100 µm. Abbreviations: CC3, cleaved caspase 3; DAPI, 4′,6-diamidino-2-phenylindole; HSA-NPs-PTX, PTX loaded in albumin nanoparticles; KER-NPs-PTX, PTX loaded in keratin nanoparticles; p3D, three-dimensional model with perfused bioreactor; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Immunofluorescence, Staining, Multiple Displacement Amplification

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Induction of apoptosis in tumor cells cultured in 2D by PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX in 2D model. Notes: MCF-7 and MDA MB 231 cells were exposed to PTX, HSA-NPs-PTX, and KER-NPs-PTX (PTX, 5 µg/mL) for 24 and 48 h. Apoptosis assays were carried out by flow cytometry, following APC-Annexin V and PI staining. Representative dot plots of MCF-7 ( A ) and MDA MB 231 ( B ) cell lines after each treatment. Percentage of early apoptotic cells (positive to APC-Annexin V and negative to PI) and late apoptotic cells (positive to APC-Annexin V and PI) was evaluated 24 and 48 h after treatment ( C ). Statistical significance versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Cell Culture, Multiple Displacement Amplification, Flow Cytometry, Cytometry, Staining

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Antiproliferative activity of PTX in a free form, HSA-NPs-PTX, and KER-NPs-PTX on MCF-7 and MDA MB 231 cell lines in 2D model. Notes: Cell viability was evaluated 72 h after exposure to increasing concentrations of PTX (0.00002, 0.02, and 5 µg/mL) by APH assay. Statistical significance versus untreated cells (100%, represented by a dotted line): ** P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Activity Assay, Multiple Displacement Amplification

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Representative H E staining of scaffold sections from p3D MCF-7 and MDA MB 231 cultures after treatments. Notes: MCF-7 (upper panels) and MDA MB 231 (lower panels) cells were initially cultured for 7 and 4 days, respectively, and then left untreated ( A ) or treated for 48 h with PTX in a free form ( B ), HSA-NPs-PTX ( C ), and KER-NPs-PTX ( D ) (PTX, 5 µg/mL). Scale bar: 100 µm. Abbreviations: H E, hematoxylin and eosin; HSA-NPs-PTX, PTX loaded in albumin nanoparticles; KER-NPs-PTX, PTX loaded in keratin nanoparticles; p3D, three-dimensional model with perfused bioreactor; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Staining, Multiple Displacement Amplification, Cell Culture

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: BAX and BCL-2 gene expression analyses in MCF-7 and MDA MB 231 cells cultured in 2D model upon 12 h treatment. Notes: Cells were incubated for 12 h with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX (PTX, 5 µg/mL). GAPDH was used as reference gene to normalize data. Effects of each treatment on gene expression levels were compared with those detectable in untreated cells (n=1) as indicated by the dotted line. Statistically significant difference versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Expressing, Multiple Displacement Amplification, Cell Culture, Incubation

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Effects of PTX in a free form or vehiculated by HSA-NPs or by KER-NPs on cell death in p3D cultures. Notes: MCF-7 and MDA MB 231 cells were exposed to PTX in a free form, HSA-NPs-PTX, and KER-NPs-PTX (PTX, 5 µg/mL) for 24 and 48 h. Apoptosis assays were carried out by flow cytometry, following APC-Annexin V and PI staining. Representative dot plots of MCF-7 ( A ) and MDA MB 231 ( B ) cell lines after each treatment. Percentages of early apoptotic cells (positive to APC-Annexin V and negative to PI) and late apoptotic cells (positive to APC-Annexin V and PI) were evaluated 24 and 48 h after treatment ( C ). Statistical significance versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Multiple Displacement Amplification, Flow Cytometry, Cytometry, Staining

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: KER-NPs-PTX analysis. Notes: Transmission electron microscopy images of fixed KER-NPs-PTX at two magnifications: scale bar 2 µm ( A ) and scale bar 200 nm ( B ). Morphological analysis showed particles spherical in shape, smooth surface, and an average dry diameter of ~80 nm. Stability of KER-NPs-PTX in physiological conditions ( C ). Fluorescence spectra of KER-NPs-PTX at different PTX loading percentages ( D ). Abbreviations: KER-NPs-PTX, PTX loaded in keratin nanoparticles; PDI, polydispersity index; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Transmission Assay, Electron Microscopy, Fluorescence

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Profiles and mechanisms of PTX release from KER-NPs in vitro. Notes: Profile of PTX release from KER-NPs ( A ). Correlation coefficient values ( R 2 ) used to determine the best fitting model among zero-order, first-order, and Higuchi modes ( B ). Abbreviations: KER-NPs, keratin nanoparticles; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: In Vitro

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: KER-NPs-PTX preparation. Notes: ( A ) KER-NPs-PTX preparation by aggregation method, starting from a solution of pure keratin powder. ( B ) Correlation between PTX loading and KER-NPs-PTX size. Abbreviations: KER-NPs-PTX, PTX loaded in keratin nanoparticles; PDI, polydispersity index; PTX, paclitaxel; RT, room temperature.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Immunofluorescence staining of CC3 in MCF-7 (upper panels) and MDA MB 231 (lower panels) scaffold sections. Notes: Images show DAPI (blue) and CC3 (green) staining in MCF-7 ( A ) and MDA MB 231 ( B ) p3D scaffold sections upon treatment with PTX in a free form (b), HSA-NPs-PTX (c), and KER-NPs-PTX (d) (PTX, 5 µg/mL) for 48 h or untreated (a). Scale bar: 100 µm. Abbreviations: CC3, cleaved caspase 3; DAPI, 4′,6-diamidino-2-phenylindole; HSA-NPs-PTX, PTX loaded in albumin nanoparticles; KER-NPs-PTX, PTX loaded in keratin nanoparticles; p3D, three-dimensional model with perfused bioreactor; PTX, paclitaxel.

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Immunofluorescence, Staining, Multiple Displacement Amplification

Journal: International Journal of Nanomedicine

Article Title: Anticancer activity of paclitaxel-loaded keratin nanoparticles in two-dimensional and perfused three-dimensional breast cancer models

doi: 10.2147/IJN.S159942

Figure Lengend Snippet: Induction of apoptosis in tumor cells cultured in 2D by PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX in 2D model. Notes: MCF-7 and MDA MB 231 cells were exposed to PTX, HSA-NPs-PTX, and KER-NPs-PTX (PTX, 5 µg/mL) for 24 and 48 h. Apoptosis assays were carried out by flow cytometry, following APC-Annexin V and PI staining. Representative dot plots of MCF-7 ( A ) and MDA MB 231 ( B ) cell lines after each treatment. Percentage of early apoptotic cells (positive to APC-Annexin V and negative to PI) and late apoptotic cells (positive to APC-Annexin V and PI) was evaluated 24 and 48 h after treatment ( C ). Statistical significance versus untreated cells: * P

Article Snippet: For p3D experiments, 2×106 MCF-7 and MDA MB 231 cells were precultured for 7 and 4 days, respectively, and then treated with PTX in a free form, HSA-NPs-PTX, or KER-NPs-PTX ([PTX] at the same drug concentration [5 µg/mL] used for 2D cultures) for 48 h. Entire scaffolds then underwent collagenase IV (Worthington Biochemical Corporation, Lakewood, NJ, USA) treatment at 37°C for 40 min, and apoptosis in the cell suspensions was evaluated by APC-Annexin V and PI Apoptosis Detection Kit.

Techniques: Cell Culture, Multiple Displacement Amplification, Flow Cytometry, Cytometry, Staining