Estimation of Biocompatibility of Polymeric Materials Using RT-PCR Method

Abstract

One of the most important factors in determining biocompatibility is the cellular response at the tissue-biomaterial interface. In this study, we would introduce a novel research method for the estimation of biocompatibility of polymeric materials, which is the evaluation of mRNA expression of the cells contacting with polymer surfaces using reverse transcription-polymerase chain reaction(RT-PCR) method. Interleukin-1β (IL-1β) were selected to estimation of the extent of inflammation. The various polymer films were put into the 6weli tissue culture plate. Human premyelocytic leukemia cell line(HL-60) were added to each well in containing 10% serum and with or without iipopolysaccharide(LPS) stimulation, or without serum, and were differentiated to macrophage-like cells by phorbol 12-myristate 13-acetate. After predetermined time, the total RNA were isolated from HL-60 cells and mRNA expression were quantitated by RT-PCR method. As a result, in the presence of serum and LPS stimulation, the expression of IL-1β mRNA increased with increasing the contact angle of films and reached to maximum at 70–80°. In the case of without LPS stimulation, IL-1β mRNA expression was decreased compared to the condition of presence of serum and LPS stimulation. From other results, IL-1β mRNA expression was influenced by the kind of substrate, LPS stimulation, serum presence and their combination. We assumed that RT-PCR method is one of the powerful methods to study the biocompatibility of materials.