title: Role of appendages in skin resistance and iontophoretic peptide flux: human versus snake skin.
abstract: OBJECTIVE
  1. The assessment of the role of hair follicles and sweat glands in skin resistance and percutaneous iontophoretic flux of 9-desglycinamide, 8-arginine vasopressin (DGAVP) by comparing two skin species: human stratum corneum which contained hair follicles, sweat and sebaceous glands, and shed snake skin which lacked all appendages. 2. The effect of 1-dodecylazacycloheptan-2-one (dodecyl-Azone, a lipid perturbing agent) on the iontophoretic DGAVP flux.
  
  
  METHODS
  Iontophoresis in vitro was performed in a transport cell (0.79 cm2 area available for percutaneous transport) by 8-hours application of a pulsed constant current of 100 Hz, 50% duty cycle and 0.26 mA.cm-2 current density delivered by a pair of Ag/AgCl electrodes, of which the anode was facing the anatomical surface of the skin samples.
  
  
  RESULTS
  The initial resistances of human stratum corneum and shed snake skin samples were of the same order of magnitude (20-24 k omega.cm2) and both skin species showed a comparable resistance-decrease profile during 8-hours iontophoresis, indicating that the resistances were mainly determined by the stratum corneum and not greatly influenced by the appendageal structures. The initial resistances of the skin samples pretreated with dodecyl-azone were less than 50% of the values of untreated samples. Because dodecylazone is known to perturb the ordering of the intercellular lipids, the effect of azone on the resistance confirms that the resistance mainly resides within the intercellular lipids of the stratum corneum. No correlation was found between the iontophoretic DGAVP-flux and the conductance of human skin. For shed snake skin, however, a good correlation was found, indicating that the iontophoretic permeability of human skin in vitro for a peptide such as DGAVP is, unlike shed snake skin, not related to its overall permeability to ions. While the initial resistances of both human and snake skin were in the same order of magnitude and showed the same declining profile during iontophoresis, the steady state iontophoretic DGAVP flux across human stratum corneum was approximately 140 times larger than through shed snake skin. These findings suggest that small ions follow pathways common to both skin types, presumably the intercellular route, while the peptide on the other hand is transported differently: across snake skin presumably along intercellular pathways only, but across human stratum corneum along additional pathways (most likely of appendageal origin) as well. This interpretation is supported by the observations made of the effects of dodecyl-azone on DGAVP-iontophoresis. Pretreatment with dodecyl-azone did not significantly change steady state fluxes and lag times of DGAVP-iontophoresis across human stratum corneum, but resulted in a significant 3-fold lag time decrease and a 3-fold flux increase of DGAVP-iontophoresis across snake skin.
  
  
  CONCLUSIONS
  The results of these in vitro studies emphasize the importance of the appendageal pathway for iontophoretic peptide transport across human stratum corneum.
authors.affiliation: Leiden/Amsterdam Center for Drug Research, Division of Pharmaceutical Technology, Leiden University, The Netherlands.
authors.forename: W H
authors.initials: WH
authors.lastname: Craane-van Hinsberg
authors.forename: J C
authors.initials: JC
authors.lastname: Verhoef
authors.forename: L J
authors.initials: LJ
authors.lastname: Bax
authors.forename: H E
authors.initials: HE
authors.lastname: Junginger
authors.forename: H E
authors.initials: HE
authors.lastname: Boddé
mesh_terms: D000328:Adult; D000818:Animals; D001381:Azepines / Q000494:pharmacology; D004560:Electricity; D005260:Female; D018859:Hair Follicle / Q000378:metabolism; D006801:Humans; D007478:Iontophoresis; D010455:Peptides / Q000378:metabolism*; D012867:Skin / Q000187:drug effects / Q000378:metabolism*; D012869:Skin Absorption / Q000187:drug effects; D012911:Snakes; D013045:Species Specificity; D013545:Sweat Glands / Q000378:metabolism; D014867:Water / Q000378:metabolism
chemical_list: D001381:Azepines; D010455:Peptides; D014867:Water; C035229:laurocapram