Multiscale characterization of electrospun non-wovens for corneal regeneration: Impact of microstructure on mechanical, optical and biological properties.

The multiscale approach in designing substrates for regenerative medicine endows them with beneficial properties determining their performance in the body. Substrates for corneal regeneration should reveal the proper transparency, mechanical properties and microstructure to maintain the functionality of the regenerated tissue. In our study, series of non-wovens with different fibres orientation (random (R), aligned (A)), topography (shish-kebab (KK), core-shell (CS)) and thickness were fabricated via electrospinning. The samples were assessed for mechanical (static tensile test) and optical properties (spectroscopy UV-Vis). The research evaluated the impact of different microstructures on the viability and morphology of three cell lines (Hs 680, HaCaT and RAW 264.7). The results showed how the fibres arrangement influenced mechanical behaviour of the non-wovens. The randomly oriented fibres were more elongated (up to 50 mm) and had a lower maximum tensile force (up to 0.46 N). In turn, the aligned fibres were characterized by lower elongation (up to 19 mm) and higher force (up to 1.45 N). The conducted transparency tests showed the relation between thickness (of the non-woven and fibres) and morphology of the substrate and light transmission. To simulate the in vivo conditions, prior to the light transmission studies, samples were immersed in water. All the samples exhibited high transparency after immersion in water (>80%). The impact of various morphologies was observed in the in vitro studies. All the samples proved high cells viability. Moreover, the substrate morphology had a significant impact on the orientation and arrangement of the fibroblast cytoskeleton. The aligned fibres were oriented in exactly the same direction. The conducted research proved that, by altering the non-wovens microstructure, the properties can be adjusted so as to induce the desirable cellular reaction. This indicates the high potential of electrospun fibres in terms of modulating the corneal cell behaviour in response to the implanted substrate.