Evaluation of biological responses to polymeric biomaterials by RT-PCR analysis. I. Study of IL-1 beta mRNA expression.
In this study, we introduce a novel research methodology, the evaluation of mRNA expression of cells contacting with polymeric materials using reverse transcription-polymerase chain reaction (RT-PCR) analysis HL-60 was used as a model of the macrophages. The expression of interleukin-1 beta (IL-1 beta) mRNA, a cytokine secreted by macrophages, was selected to estimate the extent of inflammation. The expression of IL-1 beta mRNA in the HL60 cells cultured on various substrates and in various conditions was studied. Expression of IL-1 beta could be successfully determined by RT-PCR analysis. A 48 h incubation period was necessary to clarify the expression of IL-1 beta mRNA. It became clear that lipopolysaccharide stimulation was not necessary in this analysis because of the high sensitivity of RT-PCR analysis. It is concluded that RT-PCR analysis is a powerful tool for studying cell-polymer interaction, and is a complementary method for ELISA.
Kishida
A
A
Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering, Kagoshima University, Japan.
Kato
S
S
Ohmura
K
K
Sugimura
K
K
Akashi
M
M
D001483:Base Sequence; D001672:Biocompatible Materials / Q000633:toxicity*; D017931:DNA Primers / Q000235:genetics; D005069:Evaluation Studies as Topic; D015870:Gene Expression; D018922:HL-60 Cells; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics*; D008422:Materials Testing / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011108:Polymers / Q000633:toxicity*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*
D001672:Biocompatible Materials; D017931:DNA Primers; D007375:Interleukin-1; D011108:Polymers; D012333:RNA, Messenger