title: Transport of octreotide and evaluation of mechanism of opening the paracellular tight junctions using superporous hydrogel polymers in Caco-2 cell monolayers.
abstract: The purpose of this study was to investigate the mechanism of opening of tight junctions in Caco-2 cell monolayers using superporous hydrogel (SPH) and SPH composite (SPHC) polymers as permeation enhancers for peptide drug delivery. Moreover, the transport of octreotide across Caco-2 cell monolayers was assessed by application of SPH and SPHC polymers on Caco-2 cell monolayers. In these experiments, N,N,N-trimethyl chitosan chloride with 60% quaternization (TMC60) was used as a positive control for opening of tight junctions. Transepithelial electrical resistance (TEER) studies showed that all three polymers (TMC60, SPH, and SPHC) were able to decrease TEER values to approximately 30% of the initial values, indicating the ability of these polymers to open the tight junctions. Recovery TEER studies showed that the effects of the polymers on Caco-2 cell monolayers were reversible, indicating viability of the cells after incubation with polymers. Both SPH and SPHC (compared with TMC60) were able to increase the paracellular transport of octreotide by their mechanical pressures on tight junctions. The mechanistic studies showed that junctional proteins, including actin, occludin, and claudin-1, were influenced by application of SPH and SPHC polymers to the Caco-2 cell monolayers. SPH and SPHC induced clear changes in the staining pattern of all three proteins compared with the control, indicating that the expression of these proteins in the tight junctions was increased, most likely due to the mechanical pressure of the polymers on the junctional proteins.
authors.affiliation: Department of Pharmaceutical Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands. farid.dorkoosh@organon.com
authors.forename: Farid A
authors.initials: FA
authors.lastname: Dorkoosh
authors.forename: Corine A N
authors.initials: CA
authors.lastname: Broekhuizen
authors.forename: Gerrit
authors.initials: G
authors.lastname: Borchard
authors.forename: Morteza
authors.initials: M
authors.lastname: Rafiee-Tehrani
authors.forename: J Coos
authors.initials: JC
authors.lastname: Verhoef
authors.forename: Hans E
authors.initials: HE
authors.lastname: Junginger
mesh_terms: D018938:Caco-2 Cells; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006801:Humans; D020100:Hydrogels / Q000493:pharmacokinetics*; D015282:Octreotide / Q000493:pharmacokinetics*; D009995:Osmosis; D011108:Polymers / Q000493:pharmacokinetics*; D016062:Porosity / Q000187:drug effects; D021381:Protein Transport; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism*
chemical_list: D020100:Hydrogels; D011108:Polymers; D015282:Octreotide