Elaboration of a structured biomaterial to be used as a substrate for animal cell culture

Abstract

The global scope of this work was the study of adhesion and proliferation of fibroblasts L-929 and osteoblasts MG-63 on untreated polystyrene, tissue culture polystyrene, untreated polylactide, hydrolyzed polylactide, and aminolyzed polylactide. A state of the art and techniques was first realized. Substrates were then produced. Untreated and treated polystyrene were cut into disks while polylactide powder was pressed into disks, after optimization of the production process,. The surface of disks was chemically modified by hydrolysis and aminolysis surface treatment. Substrates were characterized by optical microscopy, SEM, and water contact angle. Finally, cell adhesion, proliferation, and viability on these substrates were assessed by cell counting and MTT assay after 1, 4, and 8 days of cell culture. Results from surface characterization showed an increase of roughness for hydrolyzed polylactide. Regarding water contact angles measurements, values were smaller on treated substrates even though the difference with untreated substrates appeared smaller than expected. Cell counting and MTT assay showed an increase of cell proliferation and cell viability for treated polylactide substrates in the case of fibroblasts and a decrease of cell proliferation for aminolyzed polylactide substrates in the case of osteoblasts. After comparison with a commercial membrane called Alvetex, it was hypothesized that the Alvatex was a suitable substrate for fibroblast culture. Future developments were also proposed such as the optimization of temperature for press molding process during PLA plates production, a deeper characterization of surface chemistry and roughness, and the investigation of the effect of floating substrates on cell proliferation.