Biološko in fizikalno-kemijsko vrednotenje liposomov in transferosomov za dermalno dostavo betametazondipropionata

Krištofelc, Melanie (author), Zvonar Pobirk, Alenka (mentor), Vitek, Mercedes (comentor)

Vse pogostejša je enkapsulacija zdravilnih učinkovin v različne dostavne sisteme, ki pripomorejo k zmanjšanju neželenih učinkov ter doseganju večjega terapevtskega učinka. Liposomi in transferosomi so čedalje bolj priljubljeni za dermalno uporabo, saj so sestavljeni iz biokompatibilnih sestavin, ki izkazujejo visoko raven sprejemljivosti na koži. V okviru diplomske naloge smo na celični liniji keratinocitov (NCTC2544) ugotavljali potencialno toksičnost liposomov in transferosomov brez in z vgrajeno učinkovino, betametazondipropionatom. Le-ta spada v skupino kortikosteroidov in se uporablja pri kožnih boleznih z močno izraženimi vnetnimi procesi. Proučevane sisteme smo izdelali iz netoksičnih in za nanos na kožo sprejemljivih sestavin: bidestilirane vode, lecitina, betametazondipropionata ter Tween 80. Pred biološkim vrednotenjem smo sisteme proučili tudi z vidika fizikalno-kemijske stabilnosti. Citotoksičnost smo določali na dva načina, in sicer s testom metabolne aktivnosti ter s pomočjo invertnega svetlobnega mikroskopa. Proučili smo vpliv posameznih sestavin (z izjemo bidestilirane vode) na citotoksičnost. Največji vpliv je imel Tween 80, kar je bilo pričakovano, saj gre za površinsko aktivno snov z določenim iritacijskim potencialom (metabolna aktivnost celic je pri koncentraciji 0,5 mg/mL znašala le 2 %). Nadalje smo keratinocite izpostavili štirim različnim koncentracijam proučevanih sistemov v območju od 0,05 mg/mL do 0,5 mg/mL, z namenom določitve najbolj optimalne koncentracije za uporabo na zdravi kot tudi poškodovani (atopijski) koži. Pri transferosomih smo zabeležili nižjo viabilnost celic kot pri liposomih, nekompatibilnost pa je bila očitna tudi pri proučevanju celic pod invertnim svetlobnim mikroskopom, saj je bila večina celic po 24-urni izpostavitvi odlepljenih od podlage in netipične okrogle oblike brez medceličnih stikov. Na podlagi rezultatov vrednotenja posameznih sestavin lahko citotoksičen učinek transferosomov pripišemo Tween 80. Tudi po vgradnji učinkovine v proučevane sisteme je bila viabilnost celic nekoliko nižja. Izmed testiranih koncentracij lahko na podlagi dobljenih rezultatov koncentracijo 0,5 mg/mL opredelimo kot subtoksično (relativno varno) za liposome, za transferosome pa le-ta znaša 0,25 mg/mL. Zaključimo lahko, da liposomi z vidika biokompatibilnosti predstavljajo bolj obetaven dostavni sistem za betametazondipropionat kot transferosomi.

• Language: Slovenian
• Keywords: atopijski dermatitis, kortikosteroidi, liposomi, transferosomi, citotoksičnost
• Work type: Bachelor thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2023
• PID: 20.500.12556/RUL-149800
• Publication date in RUL: 10.09.2023

Secondary language

• Language: English
• Title: Biological and physicochemical evaluation of liposomes and transferosomes for dermal delivery of betamethasone dipropionate

Abstract

The encapsulation of active ingredients in various delivery systems is increasing, as it helps reduce side effects and achieve a greater therapeutic effect. Liposomes and transferosomes are common for dermal application due to their biocompatible ingredients and a high level of acceptance by the skin. As part of the diploma thesis, we determined the potential toxicity of liposomes and transferosomes with and without the active ingredient, betamethasone dipropionate, on the keratinocyte cell line (NCTC2544). Betamethasone diproponate belongs to the group of corticosteroids, used for treating skin diseases with overly expressed inflammation. The studied systems were formulated with non-toxic ingredients, acceptable for dermal application: bidistilled water, lecithin, betamethasone dipropionate and Tween 80. Before the biological evaluation, we also studied the systems physical and chemical stability. Cytotoxicity was determined in two ways, firstly with a metabolic activity assay, and secondly with an inverted light microscope. In addition, we studied the influence of individual ingredients (with the exception of bidistilled water) on cytotoxicity. Tween 80 had the greatest impact, which was expected, since it is a surfactant with a certain irritating potential (metabolic activity of the cells at a concentration of 0,5 mg/mL was merely 2 %). Furthermore, we exposed the keratinocytes to four different system concentrations between 0,05 mg/mL and 0,5 mg/mL with the aim of determining the optimal concentration for use on healthy as well as compromised (atopic) skin. A lower cell viability was noted for transferosomes in comparison to liposomes. The following incompatibility was also evident when examining the cells under an inverted light microscope, as most of the cells were detached after 24 hours of exposure and had an atypical shape without intercellular connections. Based on the results of the evaluation of each individual ingredient, the cytotoxic effect of transferosomes can be attributed to Tween 80. The cells also demonstrated lower viability in the presence of the active ingredient. In conclusion, the concentration of 0,5 mg/mL can be defined as subtoxic for liposomes, but not also transferosomes, for which the subtoxic concentration is 0,25 mg/mL. Due to their biocompatibility, liposomes represent a far more promising delivery system for betamethasone dipropionate than transferosomes.

• Keywords: atopic dermatitis, corticosteroids, liposomes, transferosomes, cytotoxicity