• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1292-1300
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• 2006

Aeromonas encheleia, a potential human intestinal pathogen, was shown to infect a human intestinal epithelial cell line (Caco-2) in a noninvasive manner. The transcriptional profile of the Caco-2 cells after infection with the bacteria revealed an upregulated expression of genes involved in chloride secretion, including that of phospholipase A2 (PLA2) and platelet-activating factor (PAF) acetylhydrolase (PAFAH2). This was also confirmed by a real-time RT-PCR analysis. As expected from PLA2 induction, PAF was produced when the Caco-2 cells were infected with the bacteria, and PAF was also produced when the cells were treated with a bacterial culture supernatant including bacterial extracellular proteins, yet lacking lipopolysaccharides. Bacterial aerolysin was shown to induce the production of PAF.

• Korean Journal of Microbiology
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• v.37 no.2
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• pp.151-157
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• 2001

This study was conducted to select lactic acid bacteria which possess potential inhibitory effect on Helicobacter pylori, and to make feasibility test of fermented milk products using them. In order to select lactic acid bacteria specifically inhibiting the growth of H. pylori, antibacterial activity using paper disk method, adherence ability to Caco-2 cell inhibitory effect on urease activity of H. pylori, and milk fermentation feasibility were measured. Among 45 strains of lactic acid bacteria tested, 28 strains showed clear zone and Lactobacillus gasseri MK-03 showed the largest clear zone. Caco-2 cell adherence by lactic acid bacteria and inhibitory effect of them on H. pylori adherence were also evaluated. Of 28 strains tested, 18 strains appeared to be effective on adherence to Caco-2 cell, and especially Bifidobacterium longum MK-26 was found to be superior to others. When Bif. longum MK-26 and H. pylori were reacted with Caco-2 cell 2hrs before, adherence percentage of H. pylori decreased from 0.105% to 0.004%. To investigate inhibitory effect of lactic acid bacteria-derived supernatant on urease activity of H. pylori, pH-adjusted fermented supernatant(pH-4.4) was assessed by co-cultivation method. There of Lb. acidophilus MK-07-derived supernatant showed the most inhibitory effect on urease activity of H. pylori. Considering milk fermentation ability of selected 3 strains, they were comparably feasible to fermented milk products. Consequently, Lb. gasseri MK-03, Lb. acidophilus MK-07, and Bif. longum MK-26 were selected to specifically inhibit the growth of H. pylori, by antibacterial activity, inhibition of urease activity, and inhibition of Caco-2 cell adherence, respectively.

• Preventive Nutrition and Food Science
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• v.16 no.2
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• pp.174-178
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• 2011

The cell permeability and cytotoxic effects of different-sized zinc oxide (ZnO) particles were investigated using a human colorectal adenocarcinoma cell line called Caco-2. Morphological observation by scanning electron microscopy revealed that three zinc oxides with different mean particle sizes (ZnO-1, 20 nm; ZnO-2, 90~200 nm; ZnO-3, $1\sim5\;{\mu}m$) tended to aggregate, particularly in the case of ZnO-1. When cytotoxicities of all three sizes of zinc oxide particles were measured at concentration ranges of $1\sim1000\;{\mu}g$/mL, significant decreases in cell viability were observed at concentrations of $50\;{\mu}g$/mL and higher. Among the three zinc oxides, ZnO-1 showed the lowest viability at $50\;{\mu}g$/mL in Caco-2 cells, followed by ZnO-2 and ZnO-3. The permeate concentration of ZnO-1 from the apical to the basolateral side in the Caco-2 model system after four hours was about three-fold higher than that of either ZnO-2 or ZnO-3. These results demonstrated that ZnO-1, with a 20 nm mean particle size, had poorer viability and better permeability in Caco-2 cells than ZnO-2 and ZnO-3.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.419.2-420
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• 2002

Caco-2 is a cell line derived from the human colon adenocarcinoma and often used as a model for studying intestinal drug absorption. It has been well-known that a strong correlation holds between in vitro permeability across Caco-2 cell monolayers and in vivo bioavailability for various drugs. but the correlation curves varied depending on laboratories. The permeabilities of drugs across Caco-2 cell monolayers have been measured under different agitation conditions. (omitted)

• Journal of Pharmaceutical Investigation
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• v.38 no.3
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• pp.183-189
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• 2008

The present study compared the feasibility of Caco-2 and MDCK cells as an efficient in-vitro model for the drug classification based on Biopharmaceutics Classification System (BCS) as well as an in-vitro model for drug interactions mediated by P-gp inhibition or P-gp induction. Thirteen model drugs were selected to cover BCS Class I{\sim}IV$ and their membrane permeability values were evaluated in both Caco-2 and MDCK cells. P-gp inhibition studies were conducted by using vinblastine and verapamil in MDCK cells. P-gp induction studies were also performed in MDCK cells using rifampin and the P-gp expression level was determined by western blot analysis. Compared to Caco-2 cells, MDCK cells required shorter period of time to culture cells before running the transport study. Both Caco-2 and MDCK cells exhibited the same rank order relationship between in-vitro permeability values and human permeability values of all tested model compounds, implying that those in-vitro models may be useful in the prediction of human permeability (rank order) of new chemical entities at the early drug discovery stage. However, in the case of BCS drug classification, Caco-2 cells appeared to be more suitable than MDCK cells. P-gp induction by rifampin was negligible in MDCK-cells while MDCK cells appeared to be feasible for P-gp inhibition studies. Taken all together, the present study suggests that Caco-2 cells might be more applicable to the BCS drug classification than MDCK-cells, although MDCK cells may provide some advantage in terms of capacity and speed in early ADME screening process.

• Journal of Pharmaceutical Investigation
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• v.35 no.2
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• pp.111-116
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• 2005

The unstirred water layer (UWL), which has been known to exist in the boundary of the intestinal lumen and intestinal wall, often behaves as an absorption barrier especially for lipophilic drugs. The intestinal absorption of drugs is often characterized using Caco-2 cell monolayers grown on Transwell polycarbonate membranes. The permeability $(P_{app})$ of drugs across the cell monolayer might be influenced by the agitation of the donor compartment, since the width of UWL on the surface of the cell monolayer would be reduced by the agitation. In this study, the effect of agitation of the donor compartment with 60 rpm on the permeability was measured for 12 drugs with a wide range of lipophilicity and permeability. The $P_{app}$ of mannitol, tributylmethyl ammonium, cimetidine, ranitidine, hydrocortisone, benzylpenicillin and loxoprofen was not influenced by the agitation, while the $P_{app}$ of theophylline, propranolol, YH439, phenylpropanolamine and testosterone was increased by the agitation. There was a significant correlation between the increase of $P_{app}$ by agitation and the lipophilicity for the compounds having $P_{app}>2{\times}10^{-5}$ cm/sec. No correlation was observed for the difference in $P_{app}$ by agitation and the molecular weight, or lipophilicity of the drugs. Therefore, the agitation rate of the donor compartment in the Caco-2 cell monolayer study should be carefully controlled in order to estimate $P_{app}$ reproducibly especially for lipophilic drugs.

• Archives of Pharmacal Research
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• v.25 no.1
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• pp.86-92
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• 2002

The enhancement of paracellular transport of heparin disaccharide using several absorption enhancers across Caco-2 cell monolayers was tested . The cytotoxicity of these enhancers was also examined. The enhancing effects by Quillaja saponin, diponin glycyrrhizinate, $18{\beta}-glycyrrhetinic$ acid, sodium caprate and taurine were determined by changes in transepithelial electrical resistance (TEER) and the amount of heparin disaccharide transported across Caco-2 cell monolayers. Among the absorption enhancers, $18{\beta}-glycyrrhetinic$ acid arid taurine decreased TEER and increased the permeability of heparin disaccharide in a dose-dependent and time-dependent manner with little or negligible cytotoxicity. Our results indicate that these absorption enhancers can widen the tight junction, which is a dominant paracellular absorption route of hydrophilic compounds . It is highly possible that these absorption enhancers can be applied as pharmaceutical excipients to improve the transport of macromolecules and hydrophilic drugs having difficulty in permeability across the intestinal epithelium.

• Food Science of Animal Resources
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• v.34 no.1
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• pp.20-25
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• 2014

This study evaluated the effects of frankfurter fat content on Listeria monocytogenes resistance to heat stress and gastric fluid, and the Caco-2 cell invasion efficiency of the pathogen. A 10-strain mixture of L. monocytogenes was inoculated on frankfurters formulated with 10%, 20%, and 30% fat content (10%: F10, 20%: F20, 30%: F30) and stored at $10^{\circ}C$ for 30 d. The samples were analyzed for L. monocytogenes resistance to heat stress and a simulated gastric fluid challenge. The total bacteria and L. monocytogenes survival rates were measured on tryptic soy agar plus 0.6% yeast extract and Palcam agar, respectively. L. monocytogenes colonies inoculated on F10, F20, and F30 samples were used for a Caco-2 cell invasion assay. In general, no obvious differences were observed between the survival rates of total bacteria and L. monocytogenes grown on different fat contents under heat stress and gastric fluid challenge. However, L. monocytogenes obtained from the F30 samples had a significantly higher Caco-2 cell invasion efficiency than those in the F10 and F20 samples (p<0.05). These results indicate that although high fat content in food may not be related to L. monocytogenes resistance to heat stress and gastric fluid, it may increase the Caco-2 cell invasion efficiency of the pathogen.

• Food Science of Animal Resources
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• v.33 no.4
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• pp.481-486
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• 2013

This study evaluates the effects of fat contents on the thermal resistance, antibiotic sensitivity, and Caco-2 cell invasion of Listeria monocytogenes. Ten strain mixture of L. monocytogenes in milk (0, 1, and 4% fat) and pork sausage patties (10, 20, and 30% fat) were exposed to $63^{\circ}C$. To evaluate effects of fat on the antibiotic sensitivity of L. monocytogenes, the L. monocytogenes strains NCCP10811 (most antibiotic resistant to streptomycin) and NCCP10943 (most antibiotic sensitive to streptomycin) were exposed to different fat contents in milk and pork sausage patties, and L. monocytogenes from the foods were used for antibiotic sensitivity assays. The most invasive L. monocytogenes strains (NCCP10943) was exposed to different fat contents in milk or pork sausage patties, and L. monocytogenes from the foods were used for the Caco-2 cell invasion assays. The reductions of L. monocytogenes populations were not generally influenced by fat contents. The L. monocytogenes subjected to milk fat had increased sensitivities (p<0.05) due to some antibiotics. In addition, Caco-2 cell invasion efficiency of L. monocytogenes NCCP10943 increased (p<0.05) as fat contents increased. These results indicated that higher fat contents may be related to L. monocytogenes invasions and heat resistances in pork sausage patties, but the relationship between fat and antibiotic sensitivity varied according to antibiotics, strains, and fat contents.

• Journal of Nutrition and Health
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• v.36 no.3
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• pp.270-279
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• 2003

Conjugated linoleic acid (CLA) is a group of positional and geometric isomers of linoleic acid (LA) and exhibits anticarcinogenic activity in a variety of animal models. We have previously observed that CLA inhibited the growth of Caco-2 cells, a human colon adenocarcinoma cell line. The present study was performed to determine whether the growth inhibitory effect of CLA is related to change in secretion of IGF- II and/or IGF-binding proteins (IGFBPs) that have been shown to regulate Caco-2 cell proliferation by an autocrine mechanism. Cells were incubated in serum-free medium with various concentrations of CLA or linoleic acid (LA). Immunoblot analysis of 24-hours, serum-free, conditioned medium using a monoclonal anti-IGF-IIantibody revealed that Caco-2 cells secreted both mature 6,500 Mr and higher Mr forms of pro IGF-II. The levels of pro IGF-II and mature IGF-IIwere decreased by 43 $\pm$ 2% and 53 $\pm$ 6%, respectively by treatment with 50 $\mu$ M CLA. LA slightly increased pro IGF- II levels. Results from Northern blot analysis showed that CLA decreased IGF-II mRNA levels at 50 $\mu$ M concentration suggesting that CLA regulation of IGF-II protein expression occurs partly at the transcriptional level. Ligand blot analysis of conditioned media using 1251-IGF-II revealed that CLA slightly decreased IGFBP-2 levels and increased IGFBP-4 levels. We confirmed our previous results that CLA inhibited cell growth in a dose-dependent manner but LA slightly increased cell growth. Exogenous IGF-II mitigated the growth inhibitory effect of CLA. These results indicate that the growth inhibitory effect of CLA may be at least in part mediated by decreasing IGF-II and IGFBP-2 secretion and increasing IGFBP-4 secretion in Caco-2 cells.