Synthetic nanofibers can degrade over time and be replaced by natural ECM, and such nanofibers, if properly designed, can stimulate the development of specific cell phenotypes 10. Nanofibrous materials can mimic collagen fibers' structural and mechanical properties in living tissues 9. Nowadays, it is also possible to fabricate nanofibrous materials by other methods, such as self-assembly, solution blow spinning, polymerization or template-based synthesis, and centrifugal spinning 6, 7, 8. The high surface-to-volume ratio and the fact that nanofibers can be obtained from a broad spectrum of polymers and additives 3 give a chance to prepare a new generation of advanced materials used in filtration techniques and catalytic chemistry, medicine or electronics 4, 5. One-dimensional, usually 10–1000 nm-thin fibers, called nanofibers, were first electrospun more than a century ago by John Francis Cooley in 1900 or Anton Formhals in 1934 1, 2. ![]() 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. Further, the study revealed that nanofibrous materials affect the reaction of cytotoxicity assays. We tested five metabolic cytotoxicity assays (MTT, XTT, CCK-8, alamarBlue, PrestoBlue) and obtained different viability results for the same nanofibrous materials. To better explore this issue, we prepared three nanofibrous materials with similar physical properties made of poly-L-lactic acid, polyurethane, and polycaprolactone. Our literature research showed that differences in results of widely used cytotoxicity assays applied to evaluate nanofibrous materials are poorly understood. However, as with every medical material, nanofibers too must pass all safety evaluations like in vitro cytotoxicity assays or in vivo animal tests. Nanofibrous materials are widely investigated as a replacement for the extracellular matrix, the 3D foundation for cells in all tissues.
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