Transepithelial electrical resistance is not a reliable measurement of the Caco-2 monolayer integrity in Transwell.
The significance of monitoring transepithelial electrical resistance (TEER) value during the study on drug absorption through Caco-2 monolayers in Transwells was re-evaluated. TEER value was monitored before, during, and after the absorption of Streptokinase (45 KD). Four enhancers--disodium ethylenediaminetetracetate (disodium EDTA), sodium cholate (NaC), sodium taurocholate (NaTC), and sodium caprate along with alpha-hemolysin (a cell membrane pore-forming toxin)--were used to signify the outcome of this study. Modified trypan blue exclusion technique was used to examine the Caco-2 cell viability throughout the absorption studies. The enhancers at the used concentration exhibited toxic effect on the Caco-2 cells as evident from the trypan blue exclusion studies. This toxic effect was not reflected by the TEER profile because TEER value dropped after the addition of the absorption enhancers. But it came back to its initial value after the cell culture media was replaced by enhancer-free media. This toxic effect was confirmed by the antiproliferation studies on the four enhancers and alpha-hemolysin against Caco-2 cells. Therefore, we concluded that the measurement of TEER is not a reliable method to determine the absorption enhancers toxicity or integrity of the Caco-2 monolayers in the Transwells.
Mukherjee
Tusharmouli
T
Biotechnology and Drug Delivery Laboratory, Department of Pharmacy and Administrative Sciences, College of Pharmacy and Allied Health Professions, St. John's University, Jamaica, New York 11439, USA.
Squillantea
Emilio
E
Gillespieb
Marc
M
Shao
Jun
J
D000277:Adjuvants, Pharmaceutic / Q000008:administration & dosage / Q000493:pharmacokinetics; D001427:Bacterial Toxins / Q000008:administration & dosage / Q000493:pharmacokinetics; D018938:Caco-2 Cells / Q000166:cytology*; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D003470:Culture Media; D003601:Cytotoxicity Tests, Immunologic; D003652:Decanoic Acids / Q000008:administration & dosage / Q000493:pharmacokinetics; D015194:Diffusion Chambers, Culture; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000008:administration & dosage / Q000493:pharmacokinetics; D017097:Electric Impedance; D004847:Epithelial Cells / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006460:Hemolysin Proteins / Q000008:administration & dosage; D006801:Humans; D012869:Skin Absorption / Q000187:drug effects / Q000502:physiology; D020358:Sodium Cholate / Q000008:administration & dosage / Q000493:pharmacokinetics; D013300:Streptokinase / Q000378:metabolism / Q000493:pharmacokinetics; D013656:Taurocholic Acid / Q000008:administration & dosage / Q000493:pharmacokinetics; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism; D014343:Trypan Blue / Q000633:toxicity
D000277:Adjuvants, Pharmaceutic; D001427:Bacterial Toxins; D003470:Culture Media; D003652:Decanoic Acids; D006460:Hemolysin Proteins; C021091:staphylococcal alpha-toxin; C031071:decanoic acid; D013656:Taurocholic Acid; D004492:Edetic Acid; D013300:Streptokinase; D014343:Trypan Blue; D020358:Sodium Cholate