Fosfolipidne formulacije s timolom - ispitivanje biokompatibilnosti na humanim keratinocitima

master's thesis

• Title (english): Phospholipid formulations with thymol - biocompatibility testing on human keratinocytes
• Author: Kristina Kranjčec
• Mentor: Zora Rukavina (mentor)
• Committee member: Zora Rukavina (predsjednik povjerenstva)
• Committee member: Željka Vanić (član povjerenstva)
• Committee member: Emerik Galić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical technology); Zagreb
• Defense date and country: 2024-06-27, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Timol pokazuje niz bioloških učinaka, koji mogu biti veoma korisni u lokalnoj terapiji kožnih oboljenja. Međutim, zbog mogućih citotoksičnih učinaka timola na stanice kože, njegova topikalna dermalna primjena može biti ograničena. Cilj ovog istraživanja bio je ispitati biokompatibilnost timola in vitro na HaCaT staničnoj liniji nakon uklapanja u fosfolipidne formulacije: vezikularne fosfolipidne gelove (VFG-ovi) i liposome analognog fosfolipidnog sastava. VFG-ovi su priređeni visokotlačnom homogenizacijom, a liposomske formulacije film metodom, nakon čega su homogenizirane ekstruzijom kroz polikarbonatne membrane. Za VFG-ove je utvrđen srednji promjer vezikula od 140 do 151 nm (indeks polidisperznosti ˂ 0,27), dok su ekstrudirani liposomi bili neznatno veći (144 − 164 nm) uz indeks polidisperznosti ˂ 0,12. Zeta potencijali VFG-ova (od -2,1 do -3,8 mV) bili su manje negativni u odnosu na liposome sa zeta potencijalom od -19,4 do -21,7 mV. Prisutnost kolesterola u formulacijama utjecala je na blago povećanje srednjeg promjera vezikula i indeksa polidisperznosti, te postizanje negativnijeg zeta potencijala u odnosu na formulacije bez kolesterola. Biokompatibilnost formulacija s timolom ispitana je u in vitro uvjetima na HaCaT staničnoj liniji pomoću MTT testa 24 h nakon tretiranja stanica. Dobiveni rezultati potvrdili su citotoksični potencijal slobodnog timola pri koncentracijama  125 μg/ml. Također, rezultati potvrđuju da se citotoksični učinci timola eliminiraju uklapanjem u fosfolipidne formulacije, budući da su testirane formulacije s timolom bile biokompatibilne s HaCaT stanicama u čitavom rasponu testiranih koncentracija (7,82 – 4000 μg/ml).

Abstract (english)

Thymol exhibits a range of biological effects, which can be highly beneficial in the local treatment of skin diseases. However, due to the possible cytotoxic effects of thymol on skin cells, its topical dermal application may be limited. This study aimed to examine the biocompatibility of thymol in vitro on the HaCaT cell line, after being incorporated into phospholipid formulations: vesicular phospholipid gels (VPGs) and liposomes of analogous phospholipid composition. VPGs were prepared by high-pressure homogenization, while liposomal formulations were produced by the film method, followed by homogenization through extrusion using polycarbonate membranes. The VPGs exhibited an average vesicle diameter ranging from 140 to 151 nm (polydispersity index < 0.27), while the extruded liposomes were slightly larger (144 - 164 nm) with a polydispersity index < 0.12. The zeta potential of the VPGs ranged from -2.1 to -3.8 mV, compared to the liposomes with zeta potential ranging from -19.4 to -21.7 mV. The presence of cholesterol in the formulations resulted in a modest increase in the average vesicle diameter and polydispersity index, as well as slightly more negative zeta potential compared to the formulations without cholesterol. The biocompatibility of the formulations containing thymol was tested in vitro on the HaCaT cell line using the MTT assay 24 hours after treating the cells. The results confirmed the cytotoxic potential of free thymol at concentrations  125 μg/ml. Additionally, the results demonstrate that the cytotoxic effects of thymol are eliminated when incorporated into the phospholipid formulations, as the tested formulations containing thymol were biocompatible with HaCaT cells across the entire range of tested concentrations (7.82 – 4000 μg/ml).

• Keywords: timol; vezikularni fosfolipidni gelovi; liposomi; biokompatibilnost; keratinociti
• Keywords (english): thymol; vesicular phospholipid gels; liposomes; biocompatibility; keratinocytes
• Language: croatian
• URN:NBN: urn:nbn:hr:163:832860
• Study programme: Title: Pharmacy - integrated undergraduate and graduate - university study; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: sveučilišni magistar farmacije (sveučilišni magistar farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-09-27 12:47:15