Investigations on inducible nitric oxide synthase (iNOS) gene expression and SEM analysis during the interaction of a novel tissue adhesive with J774A.1 mouse macrophage cells
Note: [] Permanent address: School of Biotechnology and Chemical Engineering, Vellore Institute of Technology, Vellore-632014 Tamil Nadu, India.
Abstract: A novel tissue adhesive consisting of human serum albumin (HSA) and tartaric acid derivative (TAD) was developed by us. Four different concentrations of TAD namely, 0.05 mmol, 0.1 mmol, 0.2 mmol and 0.3 mmol 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. Nitric oxide activity was studied using nitrate/nitrite colorimetric assay method. It was observed that the total NO production is almost the same for 40% and 42% HSA for the respective concentrations of 0.05 mmol to 0.3 mmol TAD at all incubation times from 1 d to 3 d. On the other hand, the NO activity considerably decreases for 44% HSA at all concentrations from 0.05 to 0.3 mmol TAD. This trend is observed for all the three days of incubation. From the iNOS mRNA RT-PCR experiments, it was observed that no tendency of the gene expression of iNOS was significantly found and the expression level of this gene is not affected by the concentrations of both HSA and TAD. iNOS may be up-regulated by TAD in the early stage of culture (day 1) but no significant change was observed on day 2 and 3. On the whole 0.3 mmol TAD with 44% HSA could be considered as a challenging material as a tissue adhesive for use in the field of tissue engineering. Ours is the first report on iNOS gene expression on a tissue adhesive.
