Nanostructured Biomaterials designed for chondral Tissue Engineering

LASTRA, M.L.; MOLINUEVO, M.S.; MIJANGOS, C.; CORTIZO, A.M.; BLASZCZYK-LEZAK, I. ; CORTIZO, M.S.

Maresias, Sao Paulo

Congreso; 14° Congresso da Sociedade Latino Americana de Biomateriais, Orgãos Artificiais e Engenharia de Tecidos ? SLABO; 2017

SLABO

The hypothesis of the present work is that the imitation of the extracellular matrix (ECM) using nanostructured materials based on natural polymers could restore the damaged osteochondral tissue. To prove our hypothesis we designed nanostructured matrices based on fumaric copolymer (diisopropyl fumarate-vinyl acetate copolymer) and chitosan crosslinked with borax, using anodized aluminum oxide (AAO) templates of two different pore diameter (170 nm and 300 nm). The obtained nanorods presented a highly homogenous diameter and length pattern as demonstrated by SEM. Since, we have previously demonstrated that this nanostructured scaffolds is biocompatible, improves the growth and osteogenicdevelopment of bone marrow progenitor cells. Here, we extended our studies evaluating the development of the chondroblastic EMC employing primary chondrocyte cells. We found that primary chondrocytes were able to proliferate on the nanostructured matrix. Moreover, the production the sulfated glycosaminoglycans (GAGs, evaluated by the alcian blue colorimetric method) were promoted on condrocytes cultured on the nanostructured scaffolds. We also evaluated the cytotoxicity of the nanomaterial growing murine macrophages on the scaffolds, and evaluating the release of interleukin-1? (ELISA Kit IL1?) to the culture medium at different times. In addition, we also measured the expression of cytotoxicity markers (IL1?, tumor necrosis factor ? (TNF?) and Nitric oxide synthase (iNOS)) using the semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) technique. Our results demonstrated that the nanostructured scaffold caused low cytotoxicity. In conclusion, we obtained a nanostructured scaffold of diisopropylfumarate-vinyl acetate copolymer crosslinked with chitosan by the infiltration in AAO template methodology, where nanofibers exhibited a homogeneous morphology. On the other hand, our scaffoldspromoted the development of the chondroblastic EMC with low inflammatory response in vitro. Our results on cell differentiation or cytotoxicity were independent of the nanofibers-diameter studied. Altogether our results suggest that nanostructuration of the polymers would improve chondrogenic growth and development.