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Accessing the Cytotoxicity and Cell Response to Biomaterials

2021; MyJOVE; Issue: 173 Linguagem: Inglês

10.3791/61512

1940-087X

Anabela Paula, Mafalda Laranjo, Ana Coelho, Ana Margarida Abrantes, Ana Cristina Gonçalves, Ana Bela Sarmento‐Ribeiro, Manuel Marques Ferreira, Maria Filomena Botelho, Carlos Miguel Marto, Eunice Carrilho,

3D Printing in Biomedical Research

Biomaterials contact directly or indirectly with the human tissues, making it important to evaluate its cytotoxicity. This evaluation can be performed by several methods, but a high discrepancy exists between the approaches used, compromising the reproducibility and the comparison among the obtained results. In this paper, we propose a protocol to evaluate biomaterials cytotoxicity using soluble extracts, which we use for dental biomaterials. The extracts preparation is detailed, from pellets production to its extraction in a culture medium. The biomaterials cytotoxicity evaluation is based on metabolic activity using the MTT assay, cell viability using the Sulphorhodamine B (SBR) assay, cell death profile by flow cytometry, and cell morphology using May-Grünwald Giemsa. Additional to cytotoxicity evaluation, a protocol to evaluate cell function is described based on the expression of specific markers assessed by immunocytochemistry and PCR. This protocol provides a comprehensive guide for biomaterials cytotoxicity and cellular effects evaluation, using the extracts methodology, in a reproducible and robust manner.