Introduction: The vast majority of in vitro research on microglia are based on cells isolated from neonatal animals (3-5 days of age). Studying microglia of adults has been limited by the lack of a suitable culture system that supports their growth. In this study, we describe a protocol for growing microglia of adults based on modifications of the technique for culturing microglia isolated from neonatal rats. Methods: Mixed glia isolated from adult rats (age range of 1 month to 3 years old) were seeded in culture flasks coated with poly-L-lysine. Cells were maintained in DMEM media supplemented with insulin-transferrin-selenium (ITS) and recombinant human macrophage colony-stimulating factor (M-CSF). Mild trypsinisation was carried out to isolate microglia from mixed glia culture. Results: Microglia cells of adult rats were successfully grown in vitro. For the expansion of adult microglia, it was observed that coating the cell culture flasks with poly-L-lysine was crucial to encourage cell adherence. The substitution of insulin in culture media with ITS was found to improve cell yield and reduced the number of days required for culture from 28 days to 14 days. Addition of M-CSF to cell culture medium, along with the improvisations described above provided the best adult microglia cell yield (2.91 ± 0.56 x 10⁶ cells) compared to the technique of replating cells (0.91 ± 0.65 x 10⁶ cells; p<0.05). Conclusion: Optimisation of primary cell culture technique by coating culture flasks with poly-L-lysine,supplementation of culture medium with ITS and M-CSF allowed microglia of adult rats to be successfully cultured in vitro

Objective: We aimed to evaluate the cell viability and antimicrobial effects of orthodontic bands coated with silver or zinc oxide nanoparticles (nanoAg and nano-ZnO, respectively). Methods: In this experimental study, 30 orthodontic bands were divided into three groups (n = 10 each): control (uncoated band), Ag (silver-coated band), and ZnO (zinc oxide-coated band). The electrostatic spray-assisted vapor deposition method was used to coat orthodontic bands with nano-Ag or nano-ZnO. The biofilm inhibition test was used to assess the antimicrobial effectiveness of nano-Ag and nano-ZnO against Streptococcus mutans, Lactobacillus acidophilus, and Candida albicans. Biocompatibility tests were conducted using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. The groups were compared using oneway analysis of variance with a post-hoc test. Results: The Ag group showed a significantly higher reduction in the number of L. acidophilus, C. albicans, and S.mutans colonies than the ZnO group (p = 0.015, 0.003, and 0.005, respectively). Compared with the control group, the Ag group showed a 2-log 10 reduction in all the microorganisms' replication ability, but only S. mutants showed a 2-log10 reduction in replication ability in the ZnO group. The lowest mean cell viability was observed in the Ag group, but the difference between the groups was insignificant (p > 0.05). Conclusions Coating orthodontic bands with nanoZnO or nano-Ag induced antimicrobial effects against oral pathogens. Among the nanoparticles, nano-Ag showed the best antimicrobial activity and nanoZnO showed the highest biocompatibility.