Cyquant mtt cell viability assay

The biocompatibility of the materials was tested on HeLa cells. Cells were cultivated at a density of 10⁵ cells/ well in Dulbecco’s Modified Eagle Medium with 10% fetal bovine serum. The materials were UV sterilized and added on top of the cells for 24 h. Cytotoxicity was measured using the LDH Cytotoxicity kit (Sigma) following the manufacturer’s instructions. Absorbance was read at λ = 490 nm using a NanoQuant Infinite M200 Pro instrument. The viability of the cells was analyzed using the Live/Dead assay (cat. No. L3224). Imaging was performed at λ = 494/517 (live cells) and at λ = 517/617 (dead cells) using a Zeiss fluorescence microscope.
Cell proliferation was quantified using the CyQUANT™ MTT Cell Viability Assay (Thermos Scientific, Zeiss Oberkochen, Baden-Wurttenberg, Germany) following the manufacturer’s instructions.

BEAS-2B cells were plated in 96 well plates at 70% confluency and treated for 48 h with each drug at indicated doses. Cell mitochondrial activity was profiled using CyQuant MTT Cell Viability Assay (Thermo Fisher, Catalog Number V13154) following manufacturer instructions. The absorbance at 590 nm was quantified using Spectramax i3 (Molecular Devices). Cytotoxicity was quantified using CyQuant LDH Cytotoxicity Assay (Thermo Fisher Catalog Number C20301) following manufacturer instructions. Briefly, 50 μl of media supernatant from each well was used to quantify cell toxicity and was normalized to cells lysed with 10x cell lysis buffer as 100% cell death. Absorbance was measured at 490 nm and 680 nm with the 680 nm absorbance used to determine background plate absorbance. Mitochondrial Super Oxide production was quantified using MitoSOX™ Red Mitochondrial Superoxide Indicator (Thermo Fisher Catalog Number M36008) using manufacturer instructions. Hoescht 33342 was used as a nuclear counterstain. Sixteen images were captured per well using 20x objective of the Image Express Pico and analyzed using 2 color cell scoring system to determine average Mitochondrial Superoxide Intensity per cell.

CyQuant™ MTT Cell Viability Assay (ThermoFisher) was used according to the manufacturers’ quick protocol instructions. Cells were plated in black 96-well plates (Corning or ThermoFisher). NSCs were differentiated for 7 days prior to treatment with MNPLs. Cells were treated with non-fluorescent microspheres (Polybead^®) 0–1000 µg/mL for 24 h or 7 days, with new microbeads in fresh media every 2–3 days. Absorbance was measured at 540 nm (Cytation 3; BioTek, Winooski, VT, USA). Viability of the control cells (not exposed to microspheres) was set to 100%.