The cells were observed as single cells at the time of isolation (Figure 2A and B). Thereafter, there was an increase in their size and density of the cells. Nucleus was clearly visible by day 2 and shape of the cells changed throughout the time of observation (Figure 2C and D). Day 3 onwards the cells differentiated into

different shapes ranging from oval to round shape cells (Figure 2E and F). The cells obtained on day 5 (Figure 2G) were chosen for adherence studies as significant increase in size was attained by this time. Figure 2 Isolated murine nasal epithelial cells as observed under 40X Olympus light microscope on different days post-seeding. A) and B) unstained and stained preparation of isolated single cells seen on the day of isolation C) unstained and D) stained preparation of cultured NEC on day 2 post seeding.

CH5183284 purchase Nucleus is clearly evident in all the cells E) and F) cells as seen on day 3 post seeding of different shapes and sizes and G) Polygonal shaped NEC as seen on day 5 with significant Ro 61-8048 chemical structure increase in size as well. These cells were harvested, counted and used for adherence and invasion studies. Since bacterial adherence is an essential step in the colonisation process of an organism, hence the percentage adherence of MRSA 43300 was studied using cultured NEC. PSI-7977 bacteria was added in order to obtain bacteria: nasal epithelial cell ratio of 1:1 and 10:1. The results presented in Table 1 show that bacteria exhibited high adherence (>50%) to nasal cells. The adherence was more (73.7%) when treated with higher number of bacterial cells i.e. 10:1. However, invasion of NEC was low, with only a maximum of 30% Rolziracetam cells being invaded by the test bacteria. Similarly, cytotoxic damage inflicted by MRSA 43300 onto the cultured NEC was very low with an estimated value of just 3.6% and 9% at bacteria: NEC ratio of 1:1 and 10:1 respectively. Table 1 Effect of phage on adhesion, invasion and

cytotoxicity of NEC by S. aureus 43300 Treatments Mean percent (%)   Adherence Invasion Cytotoxicity post 24 h Control (Bacteria + NEC;1:1) 58.6 ± 7.01 25 ± 3.73 3.6 ± 1.4 Control (Bacteria + NEC;10:1) 73.77 ± 7.8 31.90 ± 1.34 11.1 ± 0.7 Phage (MOI-1) 0.41 ± 0.202 0.0307 ± 0.012 0.21 ± 0.035 Phage (MOI-10) 0.0258 ± 0.005 No invasion No cytotoxicity Effect of phage addition on bacterial adhesion, invasion and cytotoxicity of NEC To demonstrate the effect of phage on the adherence and consecutively invasion and cytotoxicity of NEC by host bacteria, cultured NEC cells were processed in the same way with bacteria added in a ratio of 10:1. Following bacterial addition, phage was added at MOI-1 and MOI-10. Cells were then incubated for allowing adherence and invasion to occur. From Table 1, it is evident that phage when added at MOI-1 and MOI-10 to S. aureus 43300, was able to significantly reduce (p < 0.05) all the three parameters as compared to untreated control. Only 0.