The present investigation is aimed at preparing chitosan films capable of simulating the flux of modal drugs, 5-fluorouracil (5-FU) and indomethacin (INDO), across rat, rabbit and human cadaver epidermal sheets. Application of statistical design revealed that the concentration of chitosan, crosslinking time and concentration of crosslinking agent significantly influenced the in vitro flux of 5-FU and INDO across chitosan films. Multiple linear regression revealed a linear influence of all these active variables on 5-FU and INDO flux. It was deduced from atomic absorption spectroscopic analyses, DSC and IR spectroscopic data that 5% (m/V) sodium tripolyphosphate (NaTPP) produced optimum crosslinking of chitosan films. The in vitro permeation of both 5-FU and INDO across optimized film formulations was found to be comparable to that obtained across rat, rabbit and human epidermal sheets. These results indicate that optimized chitosan films have a potential to be developed as a substitute for animal and human cadaver epidermal sheets for preliminary in vitro permeation studies.U radu je opisana priprava kitosanskih filmova pogodnih za simulaciju prijelaza modelnih lijekova, 5-fluorouracila (5-FU) i indometacina (INDO), kroz epidermalne slojeve štakora, zeca i čovjeka. Koncentracija kitosana, vrijeme umrežavanja i koncentracija reagensa za umrežavanje značajno su utjecale na in vitro prolaz 5-FU i INDO kroz kitosanske filmove. Multiplom linearnom regresijom pokazano je da sve navedene varijable imaju linearni utjecaj na prolaz 5-FU i INDO. Uz pomoć atomske apsorpcijske spektralne analize, DSC i IR spektroskopskih podataka zaključeno je da je 5%-tna (m/V) otopina natrijevog tripolifosfata (NaTPP) najoptimalnija za umrežavanje kitosanskih filmova. Pronađeno je da je in vitro permeacija 5-FU i INDO kroz optimiziranu formulaciju kitozanskog filma usporediva s permeacijom kroz epidermalne slojeve štakora, zeca i čovjeka. Rezultati upućuju da se optimizirani kitozanski filmovi mogu upotrijebiti kao nadomjestak animalne i humane epiderme u preliminarnim in vitro permeacijskim istraživanjima
