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Development of a conduit of PLGA-gelatin aligned nanofibers produced by electrospinning seeded with stem cells for nerve regeneration
A promising alternative to conventional nerve grafting is the use of artificial grafts made from biodegradable and biocompatible materials and supportive cells. In this study, a poly (lactic-co-glycolic acid) (PLGA) conduit of aligned nanofibers was produced by the electrospinning method and seeded with mouse embryonic stem cells (mESCs) that were subsequently differentiated into neural precursor cells.
Objetivos - Metodologia - Resultados - Discussão dos Resultados/Objectives - Methodology - Results - Discussion of Results/Objetivos - Metodología - Resultados - Discusión de los resultados
Scaffolds of aligned fibers with an average diameter of 0.90±0.36 µm, an alignment coefficient of 0.817±0.07 and an 112.5±0.12º contact angle were produced. A treatment with gelatin increased the fiber diameter to 1.05±0.32 µm, reduced the alignment coefficient to 0.655±0.045 and made the scaffold very hydrophilic with a contact angle of 0°. The conduits seeded with murine embryonic stem cells (mESCs) were submitted to a neural differentiation protocol and the cell proliferation, differentiation and viability were analyzed. The real time PCR analysis showed that the cells were upregulating the expression of neural markers nestin and beta3 tubulin. The MTT assay showed that the cells maintained viable and proliferated after 7 days in culture, showing better viability on gelatin treated conduits. Moreover, the Live/Dead assay indicated high cell viability, with almost no dead cells. Confocal images of phalloidin/DAPI staining showed that the cells adhered and proliferated widely, in fully adaptation with the biomaterial.
Considerações Finais/Final considerations/Consideraciones finales
In conclusion, a method is presented here for producing conduits made of aligned biocompatible and biodegradable PLGA nanofibers with a very good cell-conduit interaction as promising regenerative product for nerve grafting.
Palavras-chave/Key words/Palabras clave
biomaterial, stem cells, nerve graft, conduit, aligned nanofibers, tissue engeneering, regenerative medicine
LAURA GONÇALVES POZZOBON, LAURA ELENA SPERLING, PATRICIA LUCAS PRANKE