The of the basolateral to apical direction is higher than that of the apical to basolateral direction. bars represent standard error of the mean value for three determinations. Abbreviations: CyA, cyclosporine; MK-571, Sigma-MK-571 sodium salt hydrate; Tyrosol Caco-2, continuous FNDC3A line of heterogeneous human epithelial colorectal adenocarcinoma cells. Open in a separate windows Physique 7 Effect of oxophenylarsine and SDCh around the transport of Lac-NCTD. Notes: # 0.05 versus control group. Error bars represent standard error of the mean value for three determinations. Abbreviations: SDCh, sodium deoxycholate; Lac-NCTD, lactosyl-norcantharitin. Table 1 Uptake of Lac-NCTD on Caco-2 cells with different preparations (n = 3) 0.01 versus Lac-NCTD; # 0.05 versus Lac-NCTD-CS-NPs. Abbreviations: Lac-NCTD, lactosyl-norcantharitin; Caco-2, continuous line of heterogeneous human epithelial colorectal adenocarcinoma cells. Table 3 of Lac-NCTD on Caco-2 cells with different preparations (n = 3) (10?6) 0.05 versus Lac-NCTD; ** 0.01 versus Lac-NCTD; # 0.05 versus Lac-NCTD-CS-NPs. Abbreviations: Lac-NCTD, lactosyl-norcantharitin; Caco-2, continuous line of heterogeneous human epithelial colorectal adenocarcinoma cells. Results Preparation and evaluation of Lac-NCTD-TMC-NPs Lac-NCTD-TMC-NPs were prepared through an ionic crosslinking process and evaluated in terms of the NPs EE, DL, particle size, and polydispersity index (PDI). The particle size of Lac-NCTD-TMC-NPs was around 120 nm (PDI 0.3) (Physique 1). The zeta potential of Lac-NCTD-CS-NPs was 28.08 3.95 mV, whereas that of Lac-NCTD-TMC-NPs was 37.37 3.62 mV, mainly because of the high degree of TMC quaternization (65%). Open in a separate window Physique 1 Particle size distribution analysis using a Zetasizer Nano particle analyzer. The EE and DL of Lac-NCTD-TMC-NPs were 69.29% 0.76% and 9.1% 0.07%, respectively. A TEM image of the Lac-NCTD-TMC-NPs is usually shown in Physique 2. Under TEM, most of the NPs appeared to be uniform and round, with the majority Tyrosol of particles separated from one another. Open in a separate window Physique 2 TEM micrograph of Lac-NCTD-TMC-NPs. Abbreviations: TEM, transmission electron microscopy; Lac-NCTD-TMC-NPs, lactosyl-norcantharitin (Lac-NCTD)-associated 0.05). Hence, the subsequent experiments were performed using the medium at pH 7.4. The uptake amount of Lac-NCTD (250 g/mL) into Caco-2 monolayers in 4C and 37C were (1.35 0.13) and (0.80 0.15) mg/g protein, respectively. The uptake of Lac- NCTD across Caco-2 cell monolayers showed heat dependence, and was enhanced at Tyrosol low temperatures. Effect of protein inhibitor The saturation possibility of the P-glycoprotein (P-gp) and the multidrug resistance-associated protein 2 (MRP2) function was investigated in the present study. Hence, the uptake of Lac-NCTD across Caco-2 in solutions with added P-gp or MRP2 inhibitors 20 mol/L cyclosporine (CyA) or 50 mol/L MK-571 was investigated. Results are shown Tyrosol in Physique 5, which indicate that CyA and MK-571 can facilitate the uptake procedure. However, the latter had an even stronger action. Uptake of Lac-NCTD-NPs into Caco-2 monolayers The uptake of Lac-NCTD-NPs (Lac-NCTD-CS-NPs, Lac-NCTD-TMC-NPs) and Lac-NCTD answer at a dose of 250 g/mL at 37C was investigated. The results are shown in Table 1. Among the three formulations, the order of uptake amount was Lac-NCTD-TMC-NPs Lac-NCTD-CS-NPs Lac-NCTD answer. Transport of Lac-NCTD across Caco-2 monolayers Effect of concentration on the transport of Lac-NCTD The influence of concentration on the transport of Lac-NCTD across the Caco-2 cell monolayers was measured (Table 2). The bidirectional transport data for Lac-NCTD showed that this secretory (BL-AP) of Lac-NCTD was higher than its absorptive (AP-BL) at every concentration point, indicating the presence of efflux proteins.12C13 Table 2 of.