Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: Effects of SMA NPs on viable cell density and toxicity of NIH 3T3 fibroblasts and bEnd3 brain endothelial cells assessed by alamarBlue assay: (a–c) NIH 3T3 fibroblasts; (d–f) bEnd3 cells. (a) Fluorescence intensity of reduced resazurin by NIH 3T3 fibroblasts on day 5 of culture corresponding to 3 days of incubation with SMA NPs. (b) Relative cell density of NIH 3T3 fibroblasts on day 5. (c) Cytotoxicity of SMA NPs on NIH 3T3 fibroblasts. (d) Fluorescence intensity of reduced resazurin by bEnd3 cells on day 5 of culture corresponding to 3 days of incubation with SMA NPs. (e) Relative cell density of bEnd3 cells on day 5. (f) Cytotoxicity of SMA NPs on bEnd3 cells. In all panels, the fluorescence intensity is compared to the negative control (brown bars). **** p < 0.0001. n = 4.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques: Alamar Blue Assay, Fluorescence, Incubation, Negative Control

Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: ECIS measurements of cellular response to the introduction of fluorescent SMA NPs. (a) Normalized five-day real-time ECIS measurement of NIH 3T3 fibroblasts, showing real-time data of cell attachment, proliferation, and response to fluorescent SMA NPs at varying concentrations. Capacitance was recorded at 64 kHz. The dotted blue line indicates the time when the fluorescent SMA NPs were added to each well. (b) Capacitance values from NIH 3T3 cells on day five at 110 h in culture. (c) Cytotoxicity percentage of NIH 3T3 fibroblasts in response to contact with SMA NPs. (d) Normalized five-day real-time ECIS measurement of bEnd3brain epithelial cells, showing real-time data of attachment, proliferation, and response to fluorescent SMA NPs. Capacitance was recorded at 64 kHz. The dotted blue line indicates the time when the fluorescent SMA NPs were added to each well. (e) Capacitance values of bEnd3 cells on day five at 110 h in culture. (f) Cytotoxicity percentage of brain epithelial bEnd3 cells in response to contact with SMA NPs. In panels b, c, e, and f, the capacitances and cytotoxicities are compared to the negative control (brown bars). **** p < 0.0001. n = 4.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques: Cell Attachment Assay, Negative Control

Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: Barrier condition analysis of bENd3 brain epithelial cells after addition of SMA NPs at different concentrations. (a) Real-time ECIS resistance measurements at 4 kHz. (b) Comparison of resistance values between different experimental conditions, before adding SMA NPs on day 4, after SMA NPs addition on day 8, and after media replacement on day 10. (c) Real-time ECIS capacitance measurements at 64 kHz. (d) Comparison of normalized capacitance between different experimental conditions, before adding SMA NPs, after SMA NPs addition, and after media replacement. In panels b and d, the capacitance is compared to the negative control (brown bars). ** p < 0.0001. n = 5.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques: Comparison, Negative Control

Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: Fluorescence-phase contrast micrographs of randomly selected samples. Micrographs show NIH 3T3 fibroblasts (top panel) and bEnd3 brain endothelial cells (bottom panel) containing fluorescent SMA NPs (green) on day 7 at multiple concentrations. Scale bar = 275 μm.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques: Fluorescence

Journal: ACS Applied Bio Materials

Article Title: Real-Time Monitoring of the Cytotoxic Effect of Oxygen-Sensitive Fluorescent Poly(styrene-maleic anhydride) Nanoparticles Using Electrical-Substrate Impedance Sensing

doi: 10.1021/acsabm.5c01443

Figure Lengend Snippet: SEM images showing the cell–NP interaction after 7 days of culture with fluorescent SMA NPs. (Top panel) NIH 3T3 fibroblasts. (Bottom panel) bEnd3 brain endothelial cells. Turquoise hexagons point to SMA NPs attached to cells and intercalated into the extracellular matrix (ECM). The left and right panels are replicate samples. Scale bar = 10 μm. Zoomed micrograph scale bar = 400 nm.

Article Snippet: bEnd3 murine brain endothelial cells, obtained from the American Type Culture Collection (ATCC, CRL-2299, VA), were cultured in Gibco DMEM (low glucose) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 0.1% gentamicin (Thermo Fisher Scientific).

Techniques: