Investigation Of The Cytotoxicity Of Cassava Microfiber-Gelatin Composite Scaffold

Abstract

Cellulose fiber-reinforced composite scaffolds have recently become an interesting target for Biomedical Engineers and Biomaterials Researchers for Tissue Engineering (TE) applications. This has led to the exploration of cellulose from diverse sources such as tunicates, bacteria and especially from plants. Cassava bagasse, which is a fibrous solid residue obtained after the extraction of cassava and soluble sugars, has recently been explored as a potential source of cellulose, and has been successfully used to enhance the mechanical properties of gelatin scaffold for TE purposes. However, there is lack of knowledge on the biocompatibility of this fabricated scaffold, limiting its potential to be considered at the research level as a biomaterial for TE purposes. This study provided knowledge on the cytotoxicity of the scaffold by using HEK 293 and MDA MB 231 cell lines. The tests performed were according to ISO standards for checking in vitro cytotoxicity of medical devices- extraction and direct contact tests. Scaffold prepared with gelatin only, and cells cultured on well plates with no scaffold were used as the controls. Extracts obtained from the samples were exposed to the cells and analyzed after 24 h and 48 h of incubation, using optical and fluorescence microscopy. Additionally, cells were seeded directly on the samples and analyzed after day 1, 3 and 5 using tetrazolium-based colorimetric assay. Results obtained for HEK 293 cells demonstrated enhanced cell viability and little/no changes in cell morphology. However, there was a decline in cell viability and changes in cell morphology for MDA MB 231 cells. These results suggest that the presence of the fiber in gelatin is not cytotoxic to HEK 293 cells and can be considered for TE purposes when using normal cells. On the contrary, the presence of the fiber in gelatin is cytotoxic to MDA MB 231 cells and may not be considered for TE purposes such as 3D tumor cell studies that require the growth of cancer cells. However, further studies are required to explore the use of the cassava bagasse for its anti-cancer cell properties as demonstrated in this study.