Superior Drug Delivery Performance of Multifunctional Bilosomes: Innovative Strategy to Kill Skin Cancer Cells for Nanomedicine Application.

Purpose: Numerous failures in melanoma treatment as a highly aggressive form of skin cancer with an unfavorable prognosis and excessive resistance to conventional therapies are prompting an urgent search for more effective therapeutic tools. Consequently, to increase the treatment efficiency and to reduce the side effects of traditional administration ways, herein, it has become crucial to combine photodynamic therapy as a promising therapeutic approach with the selectivity and biocompatibility of a novel colloidal transdermal nanoplatform for effective delivery of hybrid cargo with synergistic effects on melanoma cells. Methods: The self-assembled bilosomes, co-stabilized with L-α-phosphatidylcholine, sodium cholate, Pluronic® P123, and cholesterol, were designated, and the stability of colloidal vesicles was studied using dynamic and electrophoretic light scattering, also provided in cell culture medium (Dulbecco's Modified Eagle's Medium). The hybrid compounds - a classical photosensitizer (Methylene Blue) along with a complementary natural polyphenolic agent (curcumin), were successfully co-loaded, as confirmed by UV-Vis, ATR-FTIR, and fluorescent spectroscopies. The biocompatibility and usefulness of the polymer functionalized bilosome with loaded double cargo were demonstrated in vitro cyto- and phototoxicity experiments using normal keratinocytes and melanoma cancer cells. Results: The in vitro bioimaging and immunofluorescence study upon human skin epithelial (A375) and malignant (Me45) melanoma cell lines established the protective effect of the PEGylated bilosome surface. This effect was confirmed in cytotoxicity experiments, also determined on human cutaneous (HaCaT) keratinocytes. The flow cytometry experiments indicated the enhanced uptake of the encapsulated hybrid cargo compared to the non-loaded MB and CUR molecules, as well as a selectivity of the obtained nanocarriers upon tumor cell lines. The phyto-photodynamic action provided 24h-post irradiation revealed a more significant influence of the nanoplatform on Me45 cells in contrast to the A375 cell line, causing the cell viability rate below 20% of the control. Conclusion: As a result, we established an innovative and effective strategy for potential metastatic melanoma treatment through the synergism of phyto-photodynamic therapy and novel bilosomal-origin nanophotosensitizers.

Antimicrobial phyto-photodynamic activity inducing by polyphenol-supported Methylene Blue co-loaded into multifunctional bilosomes: Advanced hybrid nanoplatform in the skin infections treatment?

Widespread skin infections caused primarily by bacteria and yeast, pose a growing threat to healthcare systems. Phyto-photodynamic antimicrobial therapy is a promising treatment strategy with a few mild side effects for both superficial and deeper skin infections. The combination of natural plant products (polyphenols) with conventional photosensitizers makes it possible to improve the outcome of skin infections. In the present study, nanoengineered self-assembling bilosomes were used as a nanoplatform to deliver two compounds with different solubility, i.e., curcumin applied as a hydrophobic phytochemical compound and Methylene Blue used as a hydrophilic photosensitizer. Compared with the encapsulation of Methylene Blue alone, the double-loaded bilosomes (curcumin-supported Methylene Blue) showed higher efficiency in generating reactive oxygen species. Importantly, in our study, we also confirmed that bioinspired bilosomes prevent the rapid photobleaching of Methylene Blue, thereby enhancing its photoactivity. The post-irradiation antimicrobial action was tested against two pathogens - the Gram-positive bacterium (Staphylococcus aureus) and yeast (Candida albicans). The irradiation was provided after 10, 20, and 30 min, at a specific wavelength (λ = 640 nm) corresponding to 63, 126, and 189 J cm-2 energy fluences. The most effective reduction in the microbial cells number was found 30 min post-irradiation and was 99.994% for double-loaded bilosomes compared to 99.989% killing S. aureus for bilosomes with Methylene Blue alone. For C. albicans fungal cells, the mortality was 99.669% in the presence of a Methylene Blue and curcumin mixture compared to 98.229% of those killed without the addition of curcumin. The overall results of our contribution provide evidence that curcumin in combination with MB enhances the photo-eradication efficiency of S. aureus and C. albicans planktonic cultures. Thus, the mixture of the phytochemicals with photosensitizers and their encapsulation in multifunctional bilosomes may contribute to the development of innovative antimicrobial phyto-photodynamic therapy in the future.

Self-assembled bilosomes with stimuli-responsive properties as bioinspired dual-tunable nanoplatform for pH/temperature-triggered release of hybrid cargo.

The rapid development of colloid chemistry has raised the possibility of using nanocarriers for the targeted delivery and the controlled drug release at predictable locations to reduce side effects and enhance therapeutic efficacy. In the present work, we focused on the influence of temperature and pH upon in vitro controlled phytochemical/dye-release from a modified bilosome. Drug molecules can affect the properties of nanocarriers, so the effect of encapsulated bioactive compounds on nanoparticle structure has been investigated. The self-assembly process of bioinspired components (i.e., phospholipids, bile salts, and cholesterol), and biocompatible polymeric triblock materials, made it possible to receive structures with a size below 100 nm, demonstrated good capacity for active cargo encapsulation. Differential scanning calorimetry studies showed the possibility of the payloads' interaction with the bilosomes structure. A highly lipophilic compound, such as curcumin, can weaken hydrophobic interactions between the acyl chains of phospholipids, leading to a more flexible membrane. The in vitro release profiles have proved that both solubilities of the therapeutic substances and various environmental conditions affect the release rate of the hybrid cargo. Overall, the obtained double-loaded bilosomes represent a promising bioinspired nanoplatform for oral, intravenous, and topical drug delivery in future biomedical applications.

Biodegradable Amphoteric Surfactants in Titration-Ultrasound Formulation of Oil-in-Water Nanoemulsions: Rational Design, Development, and Kinetic Stability.

Amphoteric amphiphilic compounds, due to their unique properties, may represent a group of safe and biocompatible surface-active agents for effective colloidal stabilization of nanoformulations. For this reason, the aim of this work was to develop and characterize the oil-in-water nanoemulsions based on two betaine-derived surfactants with high biodegradability, i.e., cocamidopropyl betaine and coco-betaine. In the first step, we investigated ternary phase diagrams of surfactant-oil-water systems containing different weight ratios of surfactant and oil, as the betaine-type surfactant entity (S), linoleic acid, or oleic acid as the oil phase (O), and the aqueous phase (W) using the titration-ultrasound approach. All the received nanoemulsion systems were then characterized upon droplets size (dynamic light scattering), surface charge (electrophoretic light scattering), and morphology (transmission electron as well as atomic force microscopy). Thermal and spinning tests revealed the most stable compositions, which were subjected to further kinetic stability analysis, including turbidimetric evaluation. Finally, the backscattering profiles revealed the most promising candidate with a size <200 nm for potential delivery of active agents in the future cosmetic, pharmaceutical, and biomedical applications.

Novel Surface-Modified Bilosomes as Functional and Biocompatible Nanocarriers of Hybrid Compounds.

In the present contribution, we demonstrate a new approach for functionalization of colloidal nanomaterial consisting of phosphatidylcholine/cholesterol-based vesicular systems modified by FDA-approved biocompatible components, i.e., sodium cholate hydrate acting as a biosurfactant and Pluronic P123-a symmetric triblock copolymer comprising poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO) blocks Eight novel bilosome formulations were prepared using the thin-film hydration method followed by sonication and extrusion in combination with homogenization technique. The optimization studies involving the influence of the preparation technique on the nanocarrier size (dynamic light scattering), charge (electrophoretic light scattering), morphology (transmission electron microscopy) and kinetic stability (backscattering profiles) revealed the most promising candidate for the co-loading of model active compounds of various solubility; namely, hydrophilic methylene blue and hydrophobic curcumin. The studies of the hybrid cargo encapsulation efficiency (UV-Vis spectroscopy) exhibited significant potential of the formulated bilosomes in further biomedical and pharmaceutical applications, including drug delivery, anticancer treatment or diagnostics.

Recent Advances in the Structural Design of Photosensitive Agent Formulations Using "Soft" Colloidal Nanocarriers.

The growing demand for effective delivery of photosensitive active compounds has resulted in the development of colloid chemistry and nanotechnology. Recently, many kinds of novel formulations with outstanding pharmaceutical potential have been investigated with an expansion in the design of a wide variety of "soft" nanostructures such as simple or multiple (double) nanoemulsions and lipid formulations. The latter can then be distinguished into vesicular, including liposomes and "smart" vesicles such as transferosomes, niosomes and ethosomes, and non-vesicular nanosystems with solid lipid nanoparticles and nanostructured lipid carriers. Encapsulation of photosensitive agents such as drugs, dyes, photosensitizers or antioxidants can be specifically formulated by the self-assembly of phospholipids or other amphiphilic compounds. They are intended to match unique pharmaceutic and cosmetic requirements and to improve their delivery to the target site via the most common, i.e., transdermal, intravenous or oral administration routes. Numerous surface modifications and functionalization of the nanostructures allow increasing their effectiveness and, consequently, may contribute to the treatment of many diseases, primarily cancer. An increasing article number is evidencing significant advances in applications of the different classes of the photosensitive agents incorporated in the "soft" colloidal nanocarriers that deserved to be highlighted in the present review.