Fabrication and characterization of PVA/CS-PCL/gel multi-scale electrospun scaffold: simulating extracellular matrix for enhanced cellular infiltration and proliferation

A new bi-component poly(vinylalcohol)(PVA)/chitosan(CS)-poly(e-caprolactone)(PCL)/gelatin(Gel) multiscale electrospun scaffold was developed and analyzed in comparison with several other single scale systems. To mimic the native extracellular matrix in composition and structure and promote the migration of cells inside the scaffold, PVA/CS composite nanofibers (102 +/- 52 nm) and PCL/Gelcomposite microfiber (2.5 +/- 1.0 mu m) were simultaneously electrospun from the two opposite syringes and mixed on a rotating mandrel to generate a bi-component multi-scale membrane. The bi-component membrane was crosslinked by glutaraldehyde vapor to maintain its fiber morphology in the wet stage. Morphology, shrinkage and spectroscopic of the electrospun membranes were characterized. To test the newly developed multiscale membrane, we seeded mesenchymal stem cells (MSCs) derived from rabbit onto five different fiber scaffolds (PVA, PVA/CS, PCL, PCL/Gel and PVA/CS-PCL/Gel) and compared cell adhesion and proliferation between different groups for 3 days using scanning electron microscopy, inverted microscope observations assay and MTT colorimetric. Cell culture results suggest that the incorporation of chitosan and gelatin could enhance cell adhesion and cell spreading in comparison to the performance of single component scaffolds of PVA and PCL. The multiscale PVA/CS-PCL/Gel membrane scaffolds provide a better environment to increase the growth, adhesion, and proliferation of cells. Scanning electron microscopy (SEM) observations showed that the cells were not only adhered well and proliferated on the surface of the scaffolds, but were also able to infiltrate inside the scaffold within 3 days of culture. MTT assay and inverted microscope observations also showed that the PVA/CS-PCL/Gel complex fibrous membrane exhibited better activity than other single component/scale systems scaffolds. Our results provide the underlying insights needed to guide the design of the native extracellular matrix.