Hierarchical electrospun tendon-ligament bioinspired scaffolds induce changes in fibroblasts morphology under static and dynamic conditions

This study investigated how bioinspired, hierarchically structured electrospun tendon/ligament scaffolds influence human fibroblast morphology under static versus dynamic culture. Scaffolds were fabricated from a PLLA/collagen blend (75/25 w/w) and designed with an external nanofibrous sheath surrounding internal ring-shaped bundles of axially aligned nanofibers to mimic tendon/ligament multiscale architecture. Human fibroblasts (Hs27) were cultured for 7 days on the 3D scaffolds under static conditions or with two brief dynamic stretching sessions. Multi-modal imaging (high-resolution X-ray computed tomography,SEM,fluorescence microscopy,and histology) was used to map cell distribution and quantify cell shape and orientation on both the sheath and internal bundles. Under static culture, fibroblasts on the sheath predominantly aligned circumferentially with spread morphology, whereas dynamic stimulation promoted preferential axial alignment with thinner,more slender cell bodies on the sheath. In both static and dynamic conditions, fibroblasts infiltrated into the scaffold and aligned axially along the internal bundles’ nanofiber direction, with dynamic culture generally associated with thinner cellular morphology. Overall, the work demonstrates that hierarchical electrospun scaffold architecture combined with controlled mechanical stimulation can modulate fibroblast orientation and morphology in a load-direction-dependent manner, and it establishes an imaging workflow for assessing cell–scaffold interactions in complex nanofibrous constructs.