Investigations on the interaction of tartaric acid derivative/human serum albumin tissue adhesive with J774A.1 mouse macrophage cells through SEM, IL-6 cytokine and gene expression techniques.
ABSTRACT We developed a novel tissue adhesive consisting of human serum albumin (HSA) and tartaric acid derivative (TAD). Four different concentrations of TAD namely, 0.05 mM, 0.1 mM, 0.2 mM and 0.3 mM were mixed with 40%, 42% and 44% HSA individually and were made in the form of disks. J774A.1 mouse macrophage cells were seeded on top of these disks. The disks were pre-treated with sterile water and Eagle's medium before every seeding. All the seeding was incubated from 1 day to 3 days before making any investigations on it. SEM images were recorded and it was observed that these cells adhered to these materials very well. Mouse IL-6 cytokine expressions were studied using ELISA. It was seen from the cytokine expression results that the release of IL-6 was minimum at 0.3 mM TAD concentrations with 44% HSA disks. No significant difference was observed in the cytokine expressions of IL-6 at 42% and 44% HSA at all concentrations of TAD studied in this work. mRNA gene expressions of IL-6 were investigated using RT-PCR technique. In 40% HSA, the gene expression level of IL-6 gene did not change during 3-day-culture in the range of TAD concentration of 0.05 mmol to 0.2 mmol. However, 0.3 mM TAD suppressed the gene expression at all concentration of HSA. In 42% HSA, although 0.05 mM and 0.1 mM TAD did not affect the gene expression, 0.2 mM and 0.3 mM TAD induced the expression level with incubation time. In 44% HSA, all the concentration of TAD increased the expression level even though the cytokine expression levels were quite low. Hence it could be thought that the expression at the cytokine level is quite insignificant where as it is to be considered at the gene expression level. On the whole, 0.3 mM TAD with 44% HSA could be considered as a challenging material as a tissue adhesive material for use in the field of tissue engineering.