Liposome Encapsulation of the Palmitoyl-KTTKS Peptide: Structural and Functional Characterization.

In this study, the amphiphilic N-palmitoyl-KTTKS peptide was integrated in the bilayer of egg-derived phosphatidylcholine (PC) vesicles using two different preparation methods, namely thin-film evaporation (TLE) and reverse-phase evaporation (REV). Both the REV and TLE methods allowed for the formation of homogeneous liposome dispersions (PdI < 0.20) with mean hydrodynamic diameters of <100 nm and <200 nm, respectively, a net negative surface charge and a percentage of structured phospholipids higher than 90%. The inclusion of the amphiphilic N-palmitoyl-KTTKS peptide within phospholipid-based vesicles could improve peptide stability and skin delivery. Therefore, the obtained liposomes were evaluated via experiments assessing the synthesis of collagen and the ECM in 3T3-NIH fibroblasts. The obtained results showed that, when delivered with PC liposomes, pal-KTTKS stimulated collagen production more than free pentapeptide and 1 mM ascorbic acid, used as a positive control.

Long-Term Stability of Lavandula x intermedia Essential Oil Nanoemulsions: Can the Addition of the Ripening Inhibitor Impact the Biocidal Activity of the Nanoformulations?

In this work, Lavandula x intermedia essential oil (LEO) was encapsulated in lipid-based nanoemulsions (NanoLEO) using the solvent-displacement technique. In order to preserve the colloidal stability of the formulation, LEO was appropriately doped with the incorporation of different levels of a water-insoluble oil used as a ripening inhibitor. All the nanoemulsion samples were evaluated in terms of the impact of the water-insoluble oil on the nanoemulsion formation, physical-chemical properties, and antibacterial effectiveness against E. coli (Gram-negative) and B. cereus (Gram-positive). The presence of the inert oil added benefits to the formulations in terms of appearance, colloidal stability, and loss of volatile components. However, the antimicrobial activity of the nanoemulsions dramatically decreased with the ripening inhibitor addition, probably because it hampered the internalization of the antimicrobial components of LEO within the bacterial cell membranes, thus nullifying the delivery ability of the nanoemulsion formulation. On the contrary, the undoped NanoLEO formulation showed unaltered antibacterial activity in both E. coli and B. cereus up to 40 weeks from the preparation.

Cryogel Scaffolds for Tissue-Engineering: Advances and Challenges for Effective Bone and Cartilage Regeneration.

Critical-sized bone defects and articular cartilage injuries resulting from trauma, osteonecrosis, or age-related degeneration can be often non-healed by physiological repairing mechanisms, thus representing a relevant clinical issue due to a high epidemiological incidence rate. Novel tissue-engineering approaches have been proposed as an alternative to common clinical practices. This cutting-edge technology is based on the combination of three fundamental components, generally referred to as the tissue-engineering triad: autologous or allogenic cells, growth-stimulating factors, and a scaffold. Three-dimensional polymer networks are frequently used as scaffolds to allow cell proliferation and tissue regeneration. In particular, cryogels give promising results for this purpose, thanks to their peculiar properties. Cryogels are indeed characterized by an interconnected porous structure and a typical sponge-like behavior, which facilitate cellular infiltration and ingrowth. Their composition and the fabrication procedure can be appropriately tuned to obtain scaffolds that match the requirements of a specific tissue or organ to be regenerated. These features make cryogels interesting and promising scaffolds for the regeneration of different tissues, including those characterized by very complex mechanical and physical properties, such as bones and joints. In this review, state-of-the-art fabrication and employment of cryogels for supporting effective osteogenic or chondrogenic differentiation to allow for the regeneration of functional tissues is reported. Current progress and challenges for the implementation of this technology in clinical practice are also highlighted.

Effects of Processing on Chemical Composition of Extracts from Sour Cherry Fruits, a Neglected Functional Food.

Sour cherries fruits (Prunus cerasus L., syn P. cerasus var. austera) are locally known as "visciola di Sezze", due to the name of the city where they are traditionally cultivated in Lazio Region, Italy. Fruit samples from three harvesting years (June 2019, 2020 and 2021), were submitted to a protocol of analyses to detect the bioactive content based on year of harvest, freezing, homogenization and thermic treatments. Polyphenolic components, particularly anthocyanin compounds, were extracted, purified and analyzed by HPLC-DAD and DI-ESI-MS. An anthocyanin content between 0.24 and 21 mg/g fresh weight and a flavonols content between 0.04 and 0.2 mg/g fresh weight were found, depending on the harvest year and the applied procedures. Anthocyanins, besides being the principal components, were mainly represented by cyanidin-3-glucosyl-rutinoside (about 80%), a not particularly widespread molecule, mostly accounting for polyphenolic content. Color analysis and anti-radical activity of the different obtained extracts were performed with the aim to correlate organoleptic characters and health potential to the detected anthocyanins and flavanols content. Results show that immediate post-harvest freezing is the best way to preserve the bioactive content, the correlated color expression and anti-radical activity.

Insights into the reaction of chondroitin sulfate with glycidyl methacrylate: 1D and 2D NMR investigation.

Chondroitin sulfate methacrylate (CS-MA) is a semisynthetic biopolymer increasingly used for the fabrication of chemical hydrogels. In this study, the methacrylation reaction of native CS was carried out with glycidyl methacrylate in dimethyl sulfoxide and optimized to obtain tunable and reproducible methacrylation degrees in a short reaction time. The methacrylation reaction was deeply characterized by mono- and bi-dimensional (1D, 2D) NMR spectroscopy of CS-MA derivatives with different methacrylation degrees. In contrast to what previously reported in the literature, HSQC, HMBC and TOCSY analyses revealed that the methacrylation reaction proceeds via both epoxy ring-opening and transesterification, involving predominantly the primary hydroxyl groups of CS, while preserving sulfate and carboxyl groups of the biopolymer. These findings are of fundamental importance for appropriate and rational design of CS-MA-based biomaterials.

Solvent Casting and UV Photocuring for Easy and Safe Fabrication of Nanocomposite Film Dressings.

The aim of this work was to optimize and characterize nanocomposite films based on gellan gum methacrylate (GG-MA) and silver nanoparticles (AgNPs) for application in the field of wound dressing. The films were produced using the solvent casting technique coupled with a photocuring process. The UV irradiation of GG-MA solutions containing glycerol as a plasticizer and different amounts of silver nitrate resulted in the concurrent crosslinking of the photocurable polymer and a reduction of Ag ions with consequent in situ generation of AgNPs. In the first part of the work, the composition of the films was optimized, varying the concentration of the different components, the GG-MA/glycerol and GG-MA/silver nitrate weight ratios as well as the volume of the film-forming mixture. Rheological analyses were performed on the starting solutions, whereas the obtained films were characterized for their mechanical properties. Colorimetric analyses and swelling studies were also performed in order to determine the AgNPs release and the water uptake capacity of the films. Finally, microbiological tests were carried out to evaluate the antimicrobial efficacy of the optimized films, in order to demonstrate their possible application as dressings for the treatment of infected hard-to-heal wounds, which is a demanding task for public healthcare.

Valorization of Kiwi Peels: Fractionation, Bioactives Analyses and Hypotheses on Complete Peels Recycle.

Kiwi fruit samples (Actinidia deliciosa Planch, cv. Hayward) represent a suitable and good source for fibers obtainment as well as for polyphenolic and carotenoid extraction. With this aim, in this study they were submitted to a double phase extraction to separate insoluble fibers by an organic phase containing lipophilic substances and an hydroalcoholic phase containing polyphenols and soluble fibers. Insoluble fibers could be separated by filtration and sent to be micronized and reused. Hydroalcoholic fractions were then furtherly fractionated by solid-phase extraction. Data coming from the color CIEL*a*b* and the HPLC-DAD analyses of the extracts were compared and correlate with those coming from the SPME-GC/MS analysis of either the finely shredded peels or of the extracts. The obtained extracts were also submitted to anti-radical activity evaluation and anti-Candida activity. Results show that all of the obtained residues are value added products. Hypotheses were also made about the nature and the possible recycle of the obtained purified solid residue.

The Impact of Bilayer Rigidity on the Release from Magnetoliposomes Vesicles Controlled by PEMFs.

Stimuli-sensitive nanocarriers have recently been developed as a powerful tool in biomedical applications such as drug delivery, detection, and gene transfer techniques. Among the external triggers investigated, low intensity magnetic fields represent a non-invasive way to remotely control the release of compounds from a magneto-sensitive carrier. Magnetoliposomes (MLs), i.e., liposomes entrapping magnetic nanoparticles (MNPs), are studied due to their capacity to transport hydrophobic and hydrophilic agents, their easy production, and due to the ability of MNPs to respond to a magnetic actuation determining the triggered release of the encapsulated compounds. Here we investigated the design and optimization of the MLs to obtain an efficient on-demand release of the transported compounds, due to the magneto-mechanical actuation induced by applying low-intensity pulsed electromagnetic fields (PEMFs). In particular we studied the effect of the bilayer packing on the ability of MLs, with oleic acid-coated MNPs encapsulated in the bilayer, to respond to PEMFs application. Three kinds of MLs are produced with an increasing rigidity of the bilayer, defined as Liquid Disorder, Liquid Order, and Gel MLs and the delivery of a hydrophilic dye (as a model drug) is investigated. Results demonstrate the efficacy of the magnetic trigger on high-ordered bilayers, which are unable to dampen the perturbation produced by MNPs motion.

Injectable and In Situ Gelling Dextran Derivatives Containing Hydrolyzable Groups for the Delivery of Large Molecules.

Recently, we reported the synthesis and characterization of a new dextran derivative obtained by grafting polyethylene glycol methacrylate to a polysaccharide backbone through a carbonate bond. This moiety was introduced because it allows for the fabrication, through a photo-induced crosslinking reaction, of biodegradable hydrogels particularly suitable for the release of high molecular weight molecules. Here, we investigate the influence of the oxyethylene chain length and the molecular weight of the starting dextran on the main properties of the polymeric solutions as well as those of the corresponding hydrogels. All synthesized polymeric derivatives were characterized by FTIR, NMR, and rheological analyses. The photo-crosslinking reaction of the polymers allowed us to obtain biodegradable networks tested for their mechanical properties, swelling, and degradation behavior. The results showed that both the oxyethylene chain length as well as the molecular weight of the starting dextran influenced swelling and degradation of the hydrogel network. As a consequence, the different behaviors in terms of swelling and degradability were able to affect the release of a large model molecule over time, making these matrices suitable candidates for the delivery of high molecular weight drug substances.

Health Potential of Clery Strawberries: Enzymatic Inhibition and Anti-Candida Activity Evaluation.

Strawberries, belonging to cultivar Clery (Fragaria × ananassa Duchesne ex Weston) and to a graft obtained by crossing Clery and Fragaria vesca L., were chosen for a study on their health potential, with regard to the prevention of chronic and degenerative diseases. Selected samples, coming from fresh and defrosted berries, submitted to different homogenization techniques combined with thermal and microwave treatments, had been previously analyzed in their polyphenolic content and antioxidant capacity. In the present work, these homogenates were evaluated in relation to their enzymatic inhibition activity towards acetylcholinesterase and butyrylcholinesterase, α-amylase, α-glucosidase and tyrosinase. All these enzymes, involved in the onset of diabetes, and neurodegenerative and other chronic diseases, were modulated by the tested samples. The inhibitory effect on tyrosinase and cholinesterase was the most valuable. Antifungal activity against Candida albicans, recently shown to play a crucial role in human gut diseases as well as diabetes, rheumatoid arthritis and Alzheimer's disease, was also shown in vitro and confirmed by the in vivo text on Galleria mellonella. Overall, the obtained results confirm once again the health potential of strawberries; however, the efficacy is dependent on high quality products submitted to correct processing flow charts.

Dextran-polyethylene glycol cryogels as spongy scaffolds for drug delivery.

Cryogels are a particular type of hydrogels that possess great potential in both fields of drug delivery and tissue engineering. Based on these premises, the goal of this work was to develop a cytocompatible polymeric cryogel, which could be used as a spongy scaffold to promote the delivery of biomolecules. Precisely, the novel formulation was fabricated by combining dextran methacrylate (DEX-MA) and polyethylene glycol dimethacrylate (PEG-DMA) through radical polymerization at a temperature of -15 °C. The swelling, porosity, mechanical properties, and the drug release profile of vitamin B12 from the optimized cryogel were evaluated and compared to hydrogels fabricated at room temperature. The use of the cryo-gelation technique enabled the formation of scaffolds with improved swelling, increased interconnected porosity, and higher mechanical resistance than conventional hydrogels. The cryogels proved to be non-toxic and suitable carriers for the delivery of water-soluble biomolecules. Overall, the novel cytocompatible cryogel formulation could be used for biomedical applications that require the need of a macroporous scaffold for localized delivery of bioactive molecules.

Chemical Investigation and Screening of Anti-Proliferative Activity on Human Cell Lines of Pure and Nano-Formulated Lavandin Essential Oil.

Lavandin essential oil (LEO), a natural sterile hybrid obtained by crossbreeding L. angustifolia × L. latifolia, is mainly composed by active components belonging to the family of terpenes endowed with relevant anti-proliferative activity, which can be enhanced by proper application of nanotechnology. In particular, this study reports the chemical characterization and the screening of the anti-proliferative activity on different human cell lines of pure and nano-formulated lavandin essential oil (EO). LEO and its formulation (NanoLEO) were analyzed by HS/GC-MS (Headspace/Gas Chromatography-Mass Spectrometry) to describe and compare their chemical volatile composition. The most abundant compounds were linalool and 1,8-cineole (LEO: 28.6%; 27.4%) (NanoLEO: 60.4%; 12.6%) followed by α-pinene (LEO: 9.6%; NanoLEO: 4.5%), camphor (LEO: 6.5%; NanoLEO: 7.0%) and linalyl acetate (LEO: 6.5%; NanoLEO: 3.6%). The cytotoxic effects of LEO and NanoLEO were investigated on human neuroblastoma cells (SHSY5Y), human breast adenocarcinoma cells (MCF-7), human lymphoblastic leukemia cells (CCRF CEM), human colorectal adenocarcinoma cells (Caco-2) and one normal breast epithelial cell (MCF10A) by the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide)-assay. Caco-2, MCF7 and MCF10A normal cells resulted more resistant to the treatment with LEO, while CCRF-CEM and SHSY5Y cells were more sensitive. The antiproliferative effect of LEO resulted amplified when the essential oil was supplied as nanoformulation, mainly in Caco-2 cells. Scanning and transmission electron microscopy investigations were carried out on Caco-2 cells to outline at ultrastructural level possible affections induced by LEO and NanoLEO treatments.

Enhanced Loading Efficiency and Mucoadhesion Properties of Gellan Gum Thin Films by Complexation with Hydroxypropyl-ß-Cyclodextrin.

Polymeric oral thin films (OTFs) were prepared by the casting method, combining gellan gum (GG), a water-soluble polysaccharide, and glycerol (Gly) as a plasticizing agent. GG-Gly films were investigated as potential systems for buccal drug delivery using fluconazole (Class I of the Biopharmaceutical Classification System) as a model drug. At a low concentration of Gly drug precipitation occurred while, for higher concentrations of Gly, a significant deterioration of mucoadhesive and mechanical properties was observed. One possible way to overcome all these problems could be the addition of hydroxypropyl-ß-cyclodextrin (HP-ß-CD) to the GG-Gly formulation as a drug-precipitation inhibitor. In this work the effect of cyclodextrin addition on the mechanical, mucoadhesive, swelling and release properties of GG-Gly films was investigated. In-vitro drug release studies were carried out using the paddle type dissolution apparatus (USP II) and the millifluidic flow-through device (MFTD). A moving-boundary model for swelling dynamics and release in USP II is proposed to estimate the effective diffusivity of the solvent, HP-ß-CD, fluconazole and complex fluconazole/HP-ß-CD in the swelling film. Experimental results, supported by theoretical modeling, confirmed that gellan gum-low glycerol thin films including HP-ß-CD represent a suitable formulation for fluconazole drug delivery. A sustained release was observed when GG-Gly film is loaded with a preformed complex fluconazole/HP-ß-CD.

Gelation of the internal core of liposomes as a strategy for stabilization and modified drug delivery II. Theoretical analysis and modelling of in-vitro release experiments.

PEG-DMA was incorporated in unilamellar liposomes. PEG-DMA crosslinking by photo-induced radical reaction transforms the liquid aqueous core of the liposome into a hydrogel. The molecular weight of PEG-DMA significantly influences both structural and release properties of these hybrid nanosystems, by affecting both membrane permeability and diffusional properties of the inner core. Release studies of 5-(6) carboxyfluorescein from Conventional Liposomes (CL) and Gel-in-Liposome (GiL) systems were carried out in a vertical Franz Diffusion Cell. A detailed transport model is proposed, aimed at describing the entire drug diffusive pathway from the vesicles' inner core, through the double-layer membrane, into the buffer solution in the donor chamber of the Franz Cell and from there to the receptor chamber, where withdrawals are performed to evaluate the released drug concentration. The model permits to give a quantitative estimate of the diffusional resistances offered by the inner core (liquid or gelled) and by the double-layer membrane for CLs and different GiLs systems. The theoretical analysis of experimental release data strongly supports the basic assumption that, by varying the molecular weight of PEG-DMA, a different arrangement of the polymer within the liposomal structure and a different interaction with the membrane occur. PEG750-DMA decreases the transport resistance of the double layer membrane with respect to CLs, while PEG4000-DMA plays the opposite role. After gelation of the internal core, the diffusional resistance to drug transport inside GiLs becomes controlling, thus significantly slowing down drug release from these systems. Therefore, the combination of PEG-DMA with phospholipid vesicles appears an interesting strategy to develop sustained drug delivery systems.

Gelation of the internal core of liposomes as a strategy for stabilization and modified drug delivery I. Physico-chemistry study.

Since the application of nanotechnology to drug delivery, both polymer-based and lipid-based nanocarriers have demonstrated clinical benefits, improving both drug efficacy and safety. However, to further address the challenges of the drug delivery field, hybrid lipid-polymer nanocomposites have been designed to merge the beneficial features of both polymer-based and lipid-based delivery systems in a single nanocarrier. Within this scenario, this work is aimed at developing novel hybrid vesicles following the recent strategy of modifying the internal structure of liposomes. Specifically, polyethylene glycol-dimethacrylate (PEG-DMA, molecular weight 750 or 4000), was entrapped within unilamellar liposomes made of hydrogenated soybean phosphatidylcholine/cholesterol, and photo-crosslinked, in order to transform the aqueous inner core of liposomes into a soft and elastic hydrogel. After appropriate optimization of the preparation and gelation procedures, the primary objective of this work was to analyze the effect of the molecular weight of PEG-DMA on the main properties of these Gel-in-Liposome (GiL) systems. Indeed, by varying the molecular weight of PEG-DMA also its hydrophilic/lipophilic balance was modified and different arrangements of the polymer within the structure of liposomes as well as different interaction with their membrane were obtained. Both polymers were found in the inner core of the liposomes, however, the more hydrophobic PEG750-DMA also formed localized clusters within the liposome membrane, whereas the more hydrophilic PEG4000-DMA formed a polymeric corona on the vesicle surface. Preliminary cytotoxicity studies were also performed to evaluate the biological safety of these GiL systems and their suitability as innovative materials drug delivery application.

Investigating the Role of Polydopamine to Modulate Stem Cell Adhesion and Proliferation on Gellan Gum-Based Hydrogels.

Gellan gum-based hydrogels display limited cell adhesion ability due to the absence of cell-anchorage points usually present in proteins found in the extracellular matrix (ECM). This issue limits their use in the biomedical field as scaffolds to promote tissue repair. Our work addresses this challenge by investigating the use of polydopamine (pDA) as a bioactive layer to improve the surface and biological properties of gellan gum-based hydrogels cross-linked using carbodiimide chemistry. Upon treatment with a pDA layer, the hydrogel displayed an increase in wettability and swelling properties. This change in physical properties had a direct impact on the biological properties of the scaffolds. Precisely, human adipose-derived stem cells (hASCs) seeded on the pDA coated gellan gum hydrogels displayed larger cell area, increased proliferation rate, and enhanced gene expression of focal adhesion and cytoskeletal proteins. Overall, the findings of this research support the use of pDA coating as a possible approach to improve the biological features of gellan gum-based scaffolds and modulate stem cell morphology and proliferation.

Gellan Gum/Laponite Beads for the Modified Release of Drugs: Experimental and Modeling Study of Gastrointestinal Release.

In this study, gellan gum (GG), a natural polysaccharide, was used to fabricate spherical porous beads suitable as sustained drug delivery systems for oral administration. GG was cross-linked with calcium ions to prepare polymeric beads. Rheological studies and preliminary experiments of beads preparation allowed to identify the GG and the CaCl2 concentrations suitable for obtaining stable and spherical particles. GG beads were formed, through ionotropic gelation technique, with and without the presence of the synthetic clay laponite. The resultant beads were analyzed for dimensions (before and after freeze-drying), morphological aspects and ability to swell in different media miming biological fluids, namely SGF (Simulated Gastric Fluid, HCl 0.1 M) and SIF (Simulated Intestinal Fluid, phosphate buffer, 0.044 M, pH 7.4). The swelling degree was lower in SGF than in SIF and further reduced in the presence of laponite. The GG and GG-layered silicate composite beads were loaded with two model drugs having different molecular weight, namely theophylline and cyanocobalamin (vitamin B12) and subjected to in-vitro release studies in SGF and SIF. The presence of laponite in the bead formulation increased the drug entrapment efficiency and slowed-down the release kinetics of both drugs in the gastric environment. A moving-boundary swelling model with "diffuse" glassy-rubbery interface was proposed in order to describe the swelling behavior of porous freeze-dried beads. Consistently with the swelling model adopted, two moving-boundary drug release models were developed to interpret release data from highly porous beads of different drugs: drug molecules, e.g., theophylline, that exhibit a typical Fickian behavior of release curves and drugs, such as vitamin B12, whose release curves are affected by the physical/chemical interaction of the drug with the polymer/clay complex. Theoretical results support the experimental observations, thus confirming that laponite may be an effective additive for fabricating sustained drug delivery systems.

SPC Liposomes as Possible Delivery Systems for Improving Bioavailability of the Natural Sesquiterpene ß-Caryophyllene: Lamellarity and Drug-Loading as Key Features for a Rational Drug Delivery Design.

The natural sesquiterpene ß-caryophyllene (CRY) has been highlighted to possess interesting pharmacological potentials, particularly due to its chemopreventive and analgesic properties. However, the poor solubility of this sesquiterpene in aqueous fluids can hinder its uptake into cells, resulting in inconstant responses of biological systems, thus limiting its application. Therefore, identifying a suitable pharmaceutical form for increasing CRY bioavailability represents an important requirement for exploiting its pharmacological potential. In the present study, the ability of soybean phosphatidylcholine (SPC) liposomes to improve bioavailability and absorption of CRY in cancer cells has been evaluated. Liposomal formulations of CRY, differing for lamellarity (i.e., unilamellar and multilamellar vesicles or ULV and MLV) and for the drug loading (i.e., 1:0.1, 1:0.3 and 1:0.5 mol/mol between SPC and CRY) were designed with the aim of maximizing CRY amount in the liposome bilayer, while avoiding its leakage during storage. The low-loaded formulations significantly potentiated the antiproliferative activity of CRY in both HepG2 and MDA-MB-468 cells, reaching a maximum IC50 lowering (from two to five folds) with 1:0.3 and 1:0.1 SPC/CRY MLV. Conversely, increasing liposome drug-loading reduced the ability for CRY release, likely due to a possible interaction between SPC and CRY that affects the membrane properties, as confirmed by physical measures.

Experimental and Modeling Study of Drug Release from HPMC-Based Erodible Oral Thin Films.

In this work hydroxypropyl methylcellulose (HPMC) fast-dissolving thin films for oral administration are investigated. Furosemide (Class IV of the Biopharmaceutical Classification System) has been used as a model drug for in vitro release tests using three different set-ups: the Franz cell, the millifluidic flow-through device, and the paddle type dissolution apparatus (USP II). In order to enable drug incorporation within HPMC films, a multifunctional excipient, hydroxypropyl- ß -cyclodextrin (HP- ß -CD) has been included in the formulation, and the influence of HP- ß -CD on film swelling, erosion, and release properties has been investigated. Mathematical models capable of describing the swelling and release processes from HPMC erodible thin films in different apparatuses have been developed. In particular, we propose a new model for the description of drug transport and release in a Franz cell that accounts for the effect of the unavoidable imperfect mixing of the receptor chamber.

Effect of glycerol on the physical and mechanical properties of thin gellan gum films for oral drug delivery.

In this work, deacylated gellan gum and the plasticizer glycerol were used as primary components for the preparation of thin films intended for the oral delivery of therapeutic molecules. The samples were prepared by a solvent casting method and characterized for their thickness, tensile properties, swelling ability, mucoadhesion capacity and uniform drug distribution. The amount of glycerol was varied from 20% to 75% w/w in order to obtain films with tunable mechanical properties and high drug loading efficiency. The addition of glycerol was able to positively influence the mechanical characteristics of gellan gum thin film overcoming the brittleness caused by the rigid interconnection among the polymeric chains. Plasticized gellan gum films containing 50% w/w of glycerol showed optimal mechanical resistance and mucoadhesion capacity, which were adversely affected by the inclusion of higher concentrations of glycerol. On the contrary, only high amounts of the plasticizer (≥70% w/w) enabled a homogeneous distribution of the model drug fluconazole within the polymeric matrix. Overall, these results indicate that gellan gum-based thin films can be potentially used for buccal drug delivery upon precise selection of the appropriate concentration of glycerol used as a plasticizer.