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. 2023 Nov;13(11):2847-2868.
doi: 10.1007/s13346-023-01353-4. Epub 2023 May 15.

Atorvastatin liposomes in a 3D-printed polymer film: a repurposing approach for local treatment of oral candidiasis

Eman M Nour  1 , Salma E El-Habashy  2 , Michael G Shehat  3 , Marwa M Essawy  4   5 , Riham M El-Moslemany  1 , Nawal M Khalafallah  1
Affiliations

Atorvastatin liposomes in a 3D-printed polymer film: a repurposing approach for local treatment of oral candidiasis

Eman M Nour et al. Drug Deliv Transl Res. 2023 Nov.

Abstract

Oral candidiasis (OC) is an opportunistic fungal infection, common amongst the elderly and the immunocompromised. Unfortunately, the therapeutic efficacy of common antifungals is imperiled by the rise of antifungal drug resistance. An alternative promising therapeutic option possibly contributing to antifungal therapy is drug repurposing. Herein, we aimed to employ novel pharmaceutical drug delivery for enhancing the emerging antifungal potential of the hypocholesterolemic drug atorvastatin (ATV). ATV-propylene-glycol-liposomes (ATV/PG-Lip) were prepared then integrated in 3D-printed (3DP) mucoadhesive films comprising chitosan, polyvinyl-alcohol and hydroxypropyl methylcellulose, as an innovative blend, for the management of OC. ATV/PG-Lip demonstrated good colloidal properties of particle size (223.3 ± 2.1 nm), PDI (0.12 ± 0.001) and zeta potential (-18.2 ± 0.3 mV) with high entrapment efficiency (81.15 ± 1.88%) and sustained drug release. Also, ATV/PG-Lip showed acceptable three-month colloidal stability and in vitro cytocompatibility on human gingival fibroblasts. The developed 3DP-films exhibited controlled ATV release (79.4 ± 1.4% over 24 h), reasonable swelling and mucoadhesion (2388.4 ± 18.4 dyne/cm2). In vitro antifungal activity of ATV/PG-Lip was confirmed against fluconazole-resistant Candida albicans via minimum inhibitory concentration determination, time-dependent antifungal activity, agar diffusion and scanning electron microscopy. Further, ATV/PG-Lip@3DP-film exceeded ATV@3DP-film in amelioration of infection and associated inflammation in an in vivo oral candidiasis rabbit model. Accordingly, the results confirm the superiority of the fabricated ATV/PG-Lip@3DP-film for the management of oral candidiasis and tackling antifungal resistance.

Keywords: Additive manufacturing; Antifungal resistance; Buccal drug delivery; Drug repurposing; Mucoadhesive dosage forms; Oral thrush.

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Conflict of interest statement

The authors have no relevant financial or non-financial interests to disclose.

Figures

Fig. 1
Fig. 1
Characterization of the developed ATV/PG-Lip (a–c). Transmission electron microscopy images showing the morphology of blank PG-Lip and ATV/PG-Lip (a), scale bar = 200 nm. Physical stability data of ATV/PG-Lip based on vesicle size and polydispersity index when stored at 4 °C for 12 weeks (b), n = 3 at p ≤ 0.05. Cell cytocompatibility study for different formulations on human gingival fibroblasts (c), n = 7. Data represents mean ± SD. ns: statistically nonsignificant difference at p ≤ 0.05
Fig. 2
Fig. 2
Viscosity measurement of the developed inks for 3D printing (a, b). Initial viscosity values (at 1.2 s−1 shear rate) of tested plain polymer inks containing 3% w/v PVA and 3% w/v HPMC using different chitosan concentrations (3–7% w/v) (a). Data indicates mean ± SD, n = 3. Bars bearing different letters indicate statistically significant difference: a > b, at p ≤ 0.05. Viscosity values (at different shear rates) of tested plain inks containing 3% w/v PVA and 3% w/v HPMC using different chitosan concentrations and corresponding hysteresis loops of plain polymer inks; ink-D (7%), ink-E (5%), ink-F (4%) and ink-G (3%) (b)
Fig. 3
Fig. 3
Optimization of 3D printing parameters (a–c). Optimization of ink spreading ratio for different tested inks containing 3% w/v PVA and 3% w/v HPMC, using 4% and 5% w/v chitosan (a). Data indicates mean ± SD, n = 3, different letters indicate statistically significant difference: a > b > c, at p ≤ 0.05. Effect of glutaraldehyde concentration on percentage swelling of ATV@3DP-film (b). FTIR spectra of chitosan powder and plain 3DP-film (c), verifying efficient chitosan crosslinking by glutaraldehyde
Fig. 4
Fig. 4
3D printing of mucoadhesive buccal films (a–d). 3D computer-aided design of monophasic ATV/PG-Lip@3DP-film with dimensions 10 mm × 10 mm × 2 mm (a). Representative image of freshly prepared (b) and dried (c) ATV/PG-Lip@3DP-film. Scale bar = 10 mm. Scanning electron micrographs of the developed 3DP-films (d), illustrating top view and cross-sectional view of ATV@3DP-film, showing a relatively homogenous phase. Whereas the top view and cross-sectional view of biphasic 3DP-film show different microstructure for upper and lower phases. Scale bar = 200 µm
Fig. 5
Fig. 5
Characterization of the developed 3D printed films (a–e). Swelling (a) and loss in maximum swollen wet weight over time (b) of the developed 3DP-films. In vitro cumulative release profile for ATV solution and ATV/PG-Lip using the dialysis bag technique (c) and ATV@3DP-film, ATV/PG-Lip@3DP-film and biphasic film using the total immersion method (d). Release experiments were conducted at 37 °C in 5% ethanol phosphate buffer, pH 6.8. Mucoadhesive force and adhesiveness of different films (e). Data indicates mean ± SD, n = 3, p ≤ 0.05
Fig. 6
Fig. 6
In vitro antifungal activity (a–c). Antifungal activity time profile for MIC values of ATV (32 µg/mL) and ATV/PG-Lip (128 µg/mL) (a). Fungal inhibition zones of different formulations (b). Data indicates mean ± SD, n = 3, different letters indicate statistically significant difference: a > b > c, at p ≤ 0.05. SEM micrographs of control C. albicans ATCC 10231 cells and cells treated for 2 h with MIC values of ATV solution (32 µg/mL) and ATV/PG-Lip (128 µg/mL) in comparison to PG-Lip (c). Micrographs are taken at (× 10,000), scale bars = 1 µm
Fig. 7
Fig. 7
Level of pro-inflammatory cytokines TNF-α and IL-6 in tongue tissue for in vivo model. Data indicates mean ± SD, n = 3. Different letters indicate statistically significant difference: a > b > c > d at p ≤ 0.05
Fig. 8
Fig. 8
PAS-stained photomicrographs of the C. albicans infected rabbit buccal and tongue mucosa with/out treatment (a–e). The untreated mucosa displays the persistence of the hyphae and spores at the surface epithelium (blue arrows), inducing mild dysplastic changes in terms of pleomorphism, hyperchromatism and basilar hyperplasia in the basal epithelial third. Red arrows denote the candidal-induced intraepithelial macro abscesses (a). The PG-Lip@3DP-film group reveals cheek mucosa with acanthotic stratified squamous epithelium, showing signet ring fungal infected epithelial cells (green arrow in the inset). In both buccal and tongue mucosae, the PAS positive hyphae (blue arrows) occupy the upper 2/3 of the epithelium with evident compressed hyphae in the connective tissue (black arrows). Yellow arrows point out the mitotic activity induced by candidal infection (b). The buccal and lingual mucosae treated with ATV@3DP-film reveals candidal hyphae/spores within the epithelial (blue arrows) and subepithelial layers (black arrows), eliciting diffuse submucosal inflammation (c). Except for the sporadic spores (blue arrow) detected in the upper cornified layer of the lingual papillae, none of the candidal hyphae nor spores are present in the nonkeratinized cheek mucosa treated ATV/PG-Lip@3DP-film (d). Scale bar = 100 µm (× 200 lens magnification power) and = 50 µm (× 400 magnification). The morphometric heat-map for candida infectivity and inflammation score (e), revealing the antimycotic activity of the ATV/PG-Lip@3DP-film with restoring the score almost to the baseline healthy one. n = 6 measurements. The ** denotes p < 0.01, *** denotes p < 0.001, while non significance (ns) means p > 0.05
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References

    1. Archambault LS, Dongari-Bagtzoglou A. Probiotics for Oral Candidiasis: Critical Appraisal of the Evidence and a Path Forward. Front Oral Health. 2022;3:880746. 10.3389/froh.2022.880746. - PMC - PubMed
    1. Patil S, Rao RS, Majumdar B, Anil S. Clinical appearance of oral Candida infection and therapeutic strategies. Front Microbiol. 2015;6:1391. 10.3389/fmicb.2015.01391. - PMC - PubMed
    1. Alkhalidi HM, Hosny KM, Rizg WY. Oral gel loaded by fluconazole‒sesame oil nanotransfersomes: development, optimization, and assessment of antifungal activity. Pharmaceutics. 2020;13(1):27. 10.3390/pharmaceutics13010027. - PMC - PubMed
    1. Jia D, Sun W. Silver nanoparticles offer a synergistic effect with fluconazole against fluconazole-resistant Candida albicans by abrogating drug efflux pumps and increasing endogenous ROS. Infect Genet Evol. 2021;93:104937. 10.1016/j.meegid.2021.104937. - PubMed
    1. Li X-N, Zhang L-M, Wang Y-Y, Zhang Y, Jin Z-H, Li J, Wang R-R, Xiao W-L. SWL-1 Reverses Fluconazole Resistance in Candida albicans by Regulating the Glycolytic Pathway. Front Microbiol. 2020;11:572608. 10.3389/fmicb.2020.572608. - PMC - PubMed

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