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The viability of keratinocytes in adherent and non-adherent hydrogels


Introduction: Biopolymers, such as alginate and gelatin, have been widely used as hydrogel scaffolds for tissue engineering applications. Alginate is a non-adherent polymer, because it does not contain integrin-binding motifs. Gelatin is obtained by the hydrolysis of collagen which is the principal protein found in the skin.

Objetivos - Metodologia - Resultados - Discussão dos Resultados/Objectives - Methodology - Results - Discussion of Results/Objetivos - Metodología - Resultados - Discusión de los resultados

Objectives - Methodology - Results - Discussion of Results: With the aim of producing tissue-engineered skin substitutes, in this study, gelatin and alginate (an adherent and a non-adherent hydrogel) were investigated regarding their influence on the viability of human keratinocyte cells. Cells from the immortalized keratinocyte line (HaCaT) were seeded directly at a 220,000/well density, in 48 well tissue culture plates (TCPs), a two dimensional culture (2D), and in three-dimensional (3D) scaffolds composed of 0.1mL of 1% alginate, 1.5% gelatin or 1% alginate associated with 1.5% of gelatin. Cell viability was evaluated by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay after one week of cultivation. The results demonstrated that the cells remained viable in all treatments after seven days of cultivation. The average ± standard deviation (SD) values of absorbance obtained related to cell viability were 0.38 ± 0.06 for the TCP group (control); 0.52 ± 0.11 (p≤0.01) for the 1% alginate only; 0.43 ± 0.09 (p≤0.01) for the 1.5% gelatin only and 0.38 ± 0.08 (ns) for the combination of 1% alginate and 1.5% gelatin. These results indicate that cell viability was better in the alginate and gelatin when compared with the control. The lower cell viability in the TCPs can be attributed to the reduced space for cell proliferation at 2D. A statistical difference was not found between the alginate and gelatin tested, thus demonstrating that cell viability was not influenced by adherence to the proposed systems. The cells cultivated in 1% alginate associated with 1.5% gelatin showed lower cell viability than alginate only and gelatin only. The lower viability may be caused by the higher stiffness promoted by the combination of alginate and gelatin.

Considerações Finais/Final considerations/Consideraciones finales

Final considerations: The obtained scaffolds can be combined with other biomaterials, such as synthetic polymers obtained by electrospinning, with the aim of further use in regenerative medicine of soft tissues, such as skin.
Acknowledgments: MCTIC, FINEP, CNPq and Stem Cell Research Institute (IPCT).

Palavras-chave/Key words/Palabras clave

Key words: Natural polymers, biomaterials.




PAULA MARTINS FERNANDES, Marina Moraes Mattarredona Brião, Natasha Maurmann, Patricia Pranke