Aligned Nanofibrous Net Deposited Perpendicularly on Microridges Supports Endothelium Formation and Promotes the Structural Maturation of hiPSC-Derived Cardiomyocytes

Abstract

Cell alignment widely exists in various in vivo tissues and also plays an essential role in the construction of in vitro models, such as vascular endothelial and myocardial models. Recently, microscale and nanoscale hierarchical topographical structures have been drawing increasing attention for engineering in vitro cell alignment. In the present study, we fabricated a micro-/nanohierarchical substrate based on soft lithography and electrospinning to assess the synergetic effect of both the aligned nanofibrous topographical guidance and the off-ground culture environment provided by the substrate on the endothelium formation and the maturation of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The morphology, proliferation, and barrier formation of human umbilical vein endothelial cells (HUVECs) as well as the alignment, cardiac-specific proteins, and maturity-related gene expression of hiPSC-CMs on the aligned-nanofiber/microridge (AN-MR) substrate were studied. Compared with the glass slide and the single-aligned nanofiber substrate, the AN-MR substrate enhanced the proliferation, alignment, and cell−cell interaction of HUVECs and improved the length of the sarcomere and maturation-related gene expression of hiPSC-CMs. Finally, the response of hiPSC-CMs on different substrates to two typical cardiac drugs (isoproterenol and E-4031) was tested and analyzed, showing that the hiPSC-CMs on AN-MR substrates were more resistant to drugs than those in other groups, which was related to the higher maturity of the cells. Overall, the proposed micro-/nanohierarchical substrate supports the in vitro endothelium formation and enhances the maturation of hiPSC-CMs, which show great potential to be applied in the construction of in vitro models and tissue engineering.

Maturation of human cardiomyocytes derived from induced pluripotent stem cells (iPSC-CMs) on polycaprolactone and polyurethane nanofibrous mats

“…In many literature sources, cardiomyocytes differentiated from induced stem cells are cultured on different types of nanofibrous scaffolds, but there is a lack of studies evaluating the effect of seeding density on cell viability and morphology, and thus on their maturation 14 , 37 , 38 , 40 . Additionally, there are reports in which iPSC-CMs seeded at a much higher density (10 6 –4 × 10 7 cells/cm 2 ) 44 – 46 were used for studies.…”

3D Aligned Nanofiber Scaffold Fabrication with Trench-Guided Electrospinning for Cardiac Tissue Engineering

“…19 Tang et al fabricated a micro/ nanohierarchical substrate based on soft lithography and electrospinning to assess the synergetic effect of the off-ground culture environment on the maturation of CMs. 20 These composite printing strategies provide important references for the efficient fabrication of 3D aligned scaffolds; however, the scaffold morphology cannot be well defined due to the complex electric field modulation for electrospun nanofibers. 21 Here, we report a trench-guided electrospinning (ES) strategy that enables the direct fabrication of 3D aligned nanofiber scaffolds with alternative ES and a direct ink writing (DIW) process.…”

“…Dual-scale oriented scaffolds with low density were prepared by alternative 3D printing and ES to promote cell alignment and infiltration . Tang et al fabricated a micro/nanohierarchical substrate based on soft lithography and electrospinning to assess the synergetic effect of the off-ground culture environment on the maturation of CMs . These composite printing strategies provide important references for the efficient fabrication of 3D aligned scaffolds; however, the scaffold morphology cannot be well defined due to the complex electric field modulation for electrospun nanofibers …”