Cytotoxicity evaluation of enzyme inhibitors and absorption enhancers in Caco-2 cells for oral delivery of salmon calcitonin.
The usefulness of enzyme inhibitors and absorption enhancers with least mucosal cell cytotoxicity was evaluated on Caco-2 cell monolayers. The temporal cytotoxicity of several protease inhibitors at 500 microg/mL (e.g., turkey and chicken ovomucoids, aprotinin, and Protease Inhibitor Cocktail) and absorption enhancers [e.g., cholate (3%), glycocholate (3%), glycosursodeoxycholate (3%), ethylenediaminetetraacetic acid (EDTA, 0.1%), hydroxypropyl-beta-cyclodextrin (HP-beta-CD, 5%), hydroxypropyl-gamma-cylcodextrin (HP-gamma-CD, 5%), gamma-cylcodextrin (gamma-CD, 5%), tetradecyl-beta-D-maltoside (0.25%), octylglucoside (0.25%), citric acid (10%), glycyrrhetinic acid (0.34 mM), and Tween-80 (0.1%)] was measured by monitoring their effect on Caco-2 cell viability. Cell viability was measured by mannitol permeability measurements, transepithelial electrical resistance (TEER) measurements, DNA-propidium iodide staining assay, and WST-1 assay (tetrazolium salt based assay). Sodium dodecyl sulfate (0.1%), a potent surfactant, was used as a positive control. Chicken and turkey ovomucoids were nontoxic to cells as evaluated by all the methods used. Aprotinin decreased the TEER, whereas plasma membrane damage was seen with Protease Inhibitor Cocktail after a 24-h period. With respect to the absorption enhancers, the toxicity increased directly as a result of an increase in the time of incubation. The enhancers EDTA and HP-beta-CD can be used safely for a short period of time, whereas glycosursodeoxycholate, glycyrrhetinic acid, octylglucoside, HP-gamma-CD, and gamma-CD can be used for a longer period.
School of Pharmacy, Texas Tech University Health Sciences Center, 1300 Coulter, Amarillo, Texas 79106, USA.
Rakhi B
RB
Shah
Anitha
A
Palamakula
Mansoor A
MA
Khan
D000042:Absorption / Q000187:drug effects; D018938:Caco-2 Cells; D002116:Calcitonin / Q000008:administration & dosage*; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents; D004247:DNA / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004553:Electric Conductivity; D004791:Enzyme Inhibitors / Q000633:toxicity*; D005079:Excipients / Q000633:toxicity*; D005562:Formazans / Q000737:chemistry; D006801:Humans; D008353:Mannitol / Q000737:chemistry; D009685:Nuclear Envelope / Q000187:drug effects; D010088:Oxidoreductases / Q000737:chemistry; D011419:Propidium; D013778:Tetrazolium Salts; D013936:Thymidine / Q000378:metabolism
C110656:2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; D004396:Coloring Agents; D004791:Enzyme Inhibitors; D005079:Excipients; D005562:Formazans; D013778:Tetrazolium Salts; D011419:Propidium; D008353:Mannitol; C028815:salmon calcitonin; D002116:Calcitonin; D004247:DNA; D010088:Oxidoreductases; D013936:Thymidine