Eumelanin Coated PLA Electrospun Micro Fibers as Bioinspired Cradle for SH-SY5Y Neuroblastoma Cells Growth and Maturation

Abstract

Within the framework of neurodegenerative disorder therapies, the fabrication of 3D eumelanin architectures represents a novel strategy to realize tissue-engineering scaffolds for neuronal cell growth and control by providing both mechanical support and biological signals. Here, an appropriate procedure combining electrospinning, spin coating and solid-state polymerization process is established to realize the scaffolds. For biological analysis, a human derived cell line SH-SY5Y from neuroblastoma is used. Cell maturation on eumelanin microfibers, random and aligned, is evaluated by using confocal analysis and specific markers of differentiating neurons (βIII tubulin and GAP-43 expression). Cell morphology is tested by SEM analysis and immunofluorescence techniques. As results, eumelanin coated microfibers prove capable to support biological response in terms of cell survival, adhesion and spreading and to promote cell differentiation toward a more mature neuronal phenotype as confirmed by GAP-43 expression over the culture.

Exposure of the SH-SY5Y Human Neuroblastoma Cells to 50-Hz Magnetic Field: Comparison Between Two-Dimensional (2D) and Three-Dimensional (3D) In Vitro Cultures

“…We focused on such proliferation biomarkers, as residential exposure to ELF-MF has been mainly associated with increased risk for childhood leukemia and glioma; these radiations have been indeed classified as possibly carcinogenic to humans by the International Agency for Research on Cancer (IARC, 2002) [62]. We cannot exclude that the global gene expression profile of SH-SY5Y cells might be affected by growing condition in 3D Alvetex ® scaffold compared to 2D, as previously characterized-mainly in terms of neuronal morphology and neurite outgrowth-by other authors in different matrices like collagen I, Matrigel, and innovative eumelanin-coated polylactide microfibers [10,63]. These authors performed gene expression profiling of SH-SY5Y cells after 24 h of 3D culture [10] and assessed the neuronal features after 7 (and up to 21) days in culture to suggest that 3D structure was able to decrease proliferation and promote differentiation [63].…”

“…We cannot exclude that the global gene expression profile of SH-SY5Y cells might be affected by growing condition in 3D Alvetex ® scaffold compared to 2D, as previously characterized-mainly in terms of neuronal morphology and neurite outgrowth-by other authors in different matrices like collagen I, Matrigel, and innovative eumelanin-coated polylactide microfibers [10,63]. These authors performed gene expression profiling of SH-SY5Y cells after 24 h of 3D culture [10] and assessed the neuronal features after 7 (and up to 21) days in culture to suggest that 3D structure was able to decrease proliferation and promote differentiation [63]. We tested growing phenotype after 4 days in both 2D/3D cultures to characterize phenotype over a time window that might be consistent with the following ELF exposure setup and reported no significant changes.…”

Melanin and Melanin-Like Hybrid Materials in Regenerative Medicine

“…The adhesion properties of DHI-based melanins also allowed a variety of surface functionalizations aimed to fabricate bioactive substrate for stem cell culture and differentiation [ 82 , 83 ]. In particular, eumelanin-coated poly(lactic acid) (PLA) microfibers proved to be capable of supporting survival, adhesion, and differentiation toward a more mature neuronal phenotype of neuroblastoma cell type, also opening to applications in bioelectronics [ 84 ].…”

Application of blocking and immobilization of electrospun fiber in the biomedical field

“…Moreover, the brous stent can load eumelanin in the treatment of neurodegenerative disorders. 50 A single material may not be able to fulll the needs of tissue engineering scaffolds, so PLA and its copolymers can be compounded with natural materials (e.g., chitosan (CS), collagen) to optimize their properties (Table 1). For example, the multistage electrospun membrane was prepared by combining water-soluble collagen and PLA through pattern electrospinning.…”