Razvoj in vrednotenje nanovlaken s klotrimazolom za lokalno zdravljenje vulvovaginalne kandidiaze

Gorenčič, Luka (avtor), Zupančič, Špela (mentor), Grilc, Nina Katarina (komentor)

Standardno zdravljenje vulvovaginalne kandidiaze temelji na uporabi antimikotikov, kot sta klotrimazol in mikonazol. Kljub razpoložljivim terapijam pa se ženske pogosto soočajo s ponovitvami okužbe, kar kaže na potrebo po učinkovitejših in trajnejših pristopih. Namen magistrskega dela je bil razviti nanovlakna z vgrajenim antimikotikom in raziskati potencialno kombinirano uporabo antimikotika z modelnim potencialno probiotičnim sevom, ki bi lahko dodatno prispeval k obnovi vaginalne mikrobiote. Izdelovali smo hidrofilna nanovlakna iz poliakrilne kisline (PAA) in hidrofobna nanovlakna iz polikaprolaktona (PCL). V okviru raziskave smo najprej ocenili bakteriostatični vpliv klotrimazola in mikonazola na modelni probiotični sev Lactobacillus jensenii ATCC 25258, od katerih noben ni povsem inhibiral rasti. Za nadaljnjo vgradnjo učinkovine v nanovlakna PAA in PCL smo tako kot terapijo prvega izbora za vulvovaginalno kandidiazo izbrali klotrimazol. V primeru nanovlaken PAA smo preverili tudi preživetje modelnega probiotika v disperziji polimera in klotrimazola, ki bi bila primerna za sočasno vgradnjo tako antimikotika kot probiotikov v nanovlakna, in ugotovili, da je PAA baktericidna, zaradi česar sočasna vgradnja v isti dostavni sistem na osnovi PAA ni mogoča in je smiseln razvoj nanovlaken le s klotrimazolom. V sklopu razvoja nanovlaken s 5 % (m/m) klotrimazola smo določili ustrezne pogoje in najboljši sestavi disperzije za elektrostatsko sukanje, ki sta bili 7,5 % (m/V) PAA v vodni raztopini 0,1 % (m/V) Tween 80 oziroma 13,5 % (m/V) PCL v zmesi mravljične in ocetne kisline v razmerju 3:1 (V/V). Ustrezno morfologijo nanovlaken smo potrdili z vrstično elektronsko mikroskopijo. Nanovlakna PAA smo za izboljšanje stabilnosti v vodnem mediju dodatno uspešno premrežili s 100 mM železovim (III) kloridom. Stabilnost premreženih nanovlaken PAA in nanovlaken PCL v vodi smo preverili z vrednotenjem izgube mase, pri čemer so nanovlakna PAA po 24 h izgubila do 36 % mase, PCL pa le 8 %. Samo uspešnost vgrajevanja klotrimazola v nanovlakna in njegovega sproščanja v modificirano simulirano vaginalno tekočino smo želeli ovrednotiti z metodo na osnovi tekočinske kromatografije visoke ločljivosti. V ta namen smo metodo razvili, vendar smo ugotovili, da klotrimazol v modificirani simulirani vagnialni tekočini ni stabilen in razpada, zato sproščanja v njej ni bilo mogoče kvantitativno ovrednotiti. Razvita nanovlakna PAA in PCL s klotrimazolom ob ustreznem nadaljnjem analitskem razvoju za ugotavljanje kinetike sproščanja predstavljajo obetaven sistem za zdravljenje vulvovaginalne kandidiaze.

• Jezik: Slovenski jezik
• Ključne besede: nanovlakna, elektrostatsko sukanje, klotrimazol, PAA, PCL
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2025
• PID: 20.500.12556/RUL-177562
• Datum objave v RUL: 24.12.2025

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Development and evaluation of clotrimazole-loaded nanofibers for local treatment of vulvovaginal candidiasis

Izvleček

Standard treatment of vulvovaginal candidiasis relies on antimycotics such as clotrimazole and miconazole, yet recurrences remain common, highlighting the need for more durable therapies. This master’s thesis aimed to develop antimycotic-loaded nanofibers and explore their potential combined use with a model probiotic strain to support restoration of the vaginal microbiota. We prepared hydrophilic nanofibers from poly(acrylic acid) (PAA) and hydrophobic nanofibers from poly(caprolactone) (PCL). First, we evaluated the bacteriostatic effects of clotrimazole and miconazole on the model probiotic strain Lactobacillus jensenii ATCC 25258, neither of which fully inhibited bacterial growth. Based on these findings, clotrimazole was selected for incorporation into PAA and PCL nanofibers. In the case of PAA nanofibers, we also evaluated probiotic viability in polymer–clotrimazole dispersions suitable for potential co-incorporation. PAA showed bactericidal activity, making simultaneous incorporation infeasible; therefore, only clotrimazole-loaded PAA nanofibers were developed. For the development of nanofibers containing 5% (m/m) clotrimazole, we optimized the formulation and processing parameters for electrospinning. The optimal dispersions were 7,5% (m/V) PAA in an aqueous solution containing 0,1% (m/V) Tween 80, and 13,5% (m/V) PCL dissolved in a 3:1 (V/V) mixture of formic and acetic acid. The morphology of the resulting nanofibers was evaluated by scanning electron microscopy. To improve their stability in aqueous media, PAA nanofibers were additionally crosslinked with 100 mM iron (III) chloride. The stability of crosslinked PAA and PCL nanofibers in water was assessed by mass loss measurements, where PAA nanofibers lost up to 36% of their mass after 24 h, whereas PCL nanofibers lost only 8%. We aimed to quantify the incorporation efficiency of clotrimazole and its release into modified simulated vaginal fluid using high-performance liquid chromatography. For this purpose, we developed the method; however, we found that clotrimazole is not stable in modified simulated vaginal fluid and undergoes degradation, preventing quantitative evaluation of its release in this medium. Nevertheless, the developed PAA and PCL nanofibers loaded with clotrimazole represent a promising system for the local treatment of vulvovaginal candidiasis, provided that suitable analytical approaches for evaluating release kinetics are further optimized.

• Ključne besede: nanofibers, electrospinning, clotrimazole, PAA, PCL