Nanofibrous materials affect the reaction of cytotoxicity assays

Abstract

Nanofibrous materials are widely investigated as a replacement for the extracellular matrix, the 3D foundation for cells in all tissues. However, as with every medical material, nanofibers too must pass all safety evaluations like in vitro cytotoxicity assays or in vivo animal tests. Our literature research showed that differences in results of widely used cytotoxicity assays applied to evaluate nanofibrous materials are poorly understood. To better explore this issue, we prepared three nanofibrous materials with similar physical properties made of poly-L-lactic acid, polyurethane, and polycaprolactone. We tested five metabolic cytotoxicity assays (MTT, XTT, CCK-8, alamarBlue, PrestoBlue) and obtained different viability results for the same nanofibrous materials. Further, the study revealed that nanofibrous materials affect the reaction of cytotoxicity assays. Considering the results of both described experiments, it is evident that validating all available cytotoxicity assays for nanofibrous materials and possibly other highly porous materials should be carefully planned and verified using an additional analytical tool, like scanning electron microscopy or, more preferably, confocal microscopy.

Minimally Invasive Intradiscal Delivery of BM-MSCs via Fibrous Microscaffold Carriers

“…We used two different tests to minimize the influence of polymer fibers on the measurement result. 49 There were no statistically significant differences in the proliferative activity of cells cultured on the PLLA, PLLA/CH, and PLLA/CS on days 3 and 6 of culture relative to the control group. In the case of PLLA/CH, there was a decrease in the proliferative activity of cells between 6 and 9 days of culture (Figure S9, Supporting Information).…”

“…The BrdU test quantitatively measures DNA synthesis, thereby illustrating the degree of cell proliferation. We used two different tests to minimize the influence of polymer fibers on the measurement result . There were no statistically significant differences in the proliferative activity of cells cultured on the PLLA, PLLA/CH, and PLLA/CS on days 3 and 6 of culture relative to the control group.…”

Tunable electrospun scaffolds of polyacrylonitrile loaded with carbon nanotubes: from synthesis to biological applications

“…9B, and other maps can be found in Supplementary Information SI2). For all scaffolds with different concentrations of MWCNTs and alignments, we found a minimum of ~80-85 % of living cells after 5 days (Figure 9A, C, D) implying very satisfying biocompatibility of the scaffolds, [27] whereby cells were found throughout the whole fiber mat (data not shown). To summarize, with these new evidences of the biocompatible character of three cell lines belonging to muscular tissues, we can now assess that the PAN/MWNCT electrospun scaffolds present a strong interest for the in vitro study of a variety of cells ranging from brain to muscular tissues (including cardiac tissues).…”

Navigating α‐Synuclein Aggregation Inhibition: Methods, Mechanisms, and Molecular Targets

“…However, in the case of αSyn aggregation, cell viability decline can reach only up to 40 % [89] which is hardly suitable for quantitative inhibitor assessment. Moreover, the commonly used MTT cell viability assay may yield false-positive results [76,90] and can show inhibition activity for not active compounds. Finally, the exact mechanism of toxicity is not yet fully understood making the approach poorly reliable.…”