Lipid-Based Nanocarriers: Bridging Diagnosis and Cancer Therapy.

Theranostics is a growing field that matches diagnostics and therapeutics. In this approach, drugs and techniques are uniquely coupled to diagnose and treat medical conditions synergically or sequentially. By integrating diagnostic and treatment functions in a single platform, the aim of theranostics is to improve precision medicine by tailoring treatments based on real-time information. In this context, lipid-based nanocarriers have attracted great scientific attention due to their biodegradability, biocompatibility, and targeting capabilities. The present review highlights the latest research advances in the field of lipid-based nanocarriers for cancer theranostics, exploring several ways of improving in vivo performance and addressing associated challenges. These nanocarriers have significant potential to create new perspectives in the field of nanomedicine and offer promise for a significant step towards more personalized and precise medicine, reducing side effects and improving clinical outcomes for patients. This review also presents the actual barriers to and the possible challenges in the use of nanoparticles in the theranostic field, such as regulatory hurdles, high costs, and technological integration. Addressing these issues through a multidisciplinary and collaborative approach among institutions could be essential for advancing lipid nanocarriers in the theranostic field. Such collaborations can leverage diverse expertise and resources, fostering innovation and overcoming the complex challenges associated with clinical translation. This approach will be crucial for realizing the full potential of lipid-based nanocarriers in precision medicine.

Production of Biopolymeric Microparticles to Improve Cannabigerol Bioavailability.

Cannabigerol's (CBG) therapeutic effects are limited by its poor water solubility and low dissolution rate. To improve these properties, supercritical CO2-assisted atomization (SAA) was applied to produce coprecipitates, i.e., CBG nanoparticles coprecipitated in polyvinylpyrrolidone (PVP) microparticles. The experiments were performed by varying the CBG/PVP mass ratio (R) and the overall concentration of solutes CBG+PVP to study the influence of these parameters on particle morphology, particle size, and size distribution. Periodic dynamic light scattering (DLS) analysis was performed at regular time intervals to measure the size of CBG nanoparticles in PVP microparticles. It showed that CBG nanoparticles down to 105 nm were successfully produced through SAA. Dissolution tests were used to verify that a reduction of CBG particle size significantly increased its dissolution rate. In the liquid medium adopted, untreated CBG powder was released in 210 min, whereas CBG nanoparticles of 105 nm were completely dissolved in only 15 min.

Production of Agarose-Hydroxyapatite Composites via Supercritical Gel Drying, for Bone Tissue Engineering.

Bone tissue engineering (BTE) is the most promising strategy to repair bones injuries and defects. It relies on the utilization of a temporary support to host the cells and promote nutrient exchange (i.e., the scaffold). Supercritical CO2 assisted drying can preserve scaffold nanostructure, crucial for cell attachment and proliferation. In this work, agarose aerogels, loaded with hydroxyapatite were produced in view of BTE applications. Different combinations of agarose concentration and hydroxyapatite loadings were tested. FESEM and EDX analyses showed that scaffold structure suffered from partial closure when increasing filler concentration; hydroxyapatite distribution was homogenous, and Young's modulus improved. Looking at BTE applications, the optimal combination of agarose and hydroxyapatite resulted to be 1% w/w and 10% w/v, respectively. Mechanical properties showed that the produced composites could be eligible as starting scaffold for BTE, with a Young's Modulus larger than 100 kPa for every blend.

Production of PEGylated Vancomycin-Loaded Niosomes by a Continuous Supercritical CO2 Assisted Process.

Niosomes are arousing significant interest thanks to their low cost, high biocompatibility, and negligible toxicity. In this work, a supercritical CO2-assisted process was performed at 100 bar and 40 °C to produce niosomes at different Span 80/Tween 80 weight ratios. The formulation of cholesterol and 80:20 Span 80/Tween 80 was selected to encapsulate vancomycin, used as a model active compound, to perform a drug release rate comparison between PEGylated and non-PEGylated niosomes. In both cases, nanometric vesicles were obtained, i.e., 214 ± 59 nm and 254 ± 73 nm for non-PEGylated and PEGylated niosomes, respectively, that were characterized by a high drug encapsulation efficiency (95% for non-PEGylated and 98% for PEGylated niosomes). However, only PEGylated niosomes were able to prolong the vancomycin release time up to 20-fold with respect to untreated drug powder, resulting in a powerful strategy to control the drug release rate.

Liposomes, transfersomes and niosomes: production methods and their applications in the vaccinal field.

One of the most effective strategies to fight viruses and handle health diseases is vaccination. Recent studies and current applications are moving on antigen, DNA and RNA-based vaccines to overcome the limitations related to the conventional vaccination strategies, such as low safety, necessity of multiple injection, and side effects. However, due to the instability of pristine antigen, RNA and DNA molecules, the use of nanocarriers is required. Among the different nanocarriers proposed for vaccinal applications, three types of nanovesicles were selected and analysed in this review: liposomes, transfersomes and niosomes. PubMed, Scopus and Google Scholar databases were used for searching recent papers on the most frequently used conventional and innovative methods of production of these nanovesicles. Weaknesses and limitations of conventional methods (i.e., multiple post-processing, solvent residue, batch-mode processes) can be overcome using innovative methods, in particular, the ones assisted by supercritical carbon dioxide. SuperSomes process emerged as a promising production technique of solvent-free nanovesicles, since it can be easily scaled-up, works in continuous-mode, and does not require further post-processing steps to obtain the desired products. As a result of the literature analysis, supercritical carbon dioxide assisted methods attracted a lot of interest for nanovesicles production in the vaccinal field. However, despite their numerous advantages, supercritical processes require further studies for the production of liposomes, transfersomes and niosomes with the aim of reaching well-defined technologies suitable for industrial applications and mass production of vaccines.

Production of Exopolysaccharide-Based Porous Structures for Biomedical Applications: A Review.

Exopolysaccharides, obtained from microorganisms as fermentation products, are interesting candidates for biomedical applications as scaffolds: they are biocompatible, nontoxic, antimicrobial, antitumor materials. To produce exopolysaccharide-based scaffolds, sol-gel technology could be used, which ends with the removal of the liquid phase from the polymeric network (i.e., the drying step). The aim of this review is to point out the most relevant strengths and weaknesses of the different drying techniques, focusing attention on the production of exopolysaccharide-based porous structures. Among these drying processes, supercritical carbon dioxide-assisted drying is the most promising strategy to obtain dried gels to use in the biomedical field: it produces highly porous and lightweight devices with outstanding surface areas and regular microstructure and nanostructure (i.e., aerogels). As a result of the analysis carried out in the present work, it emerged that supercritical technologies should be further explored and applied to the production of exopolysaccharide-based nanostructured scaffolds. Moving research towards this direction, exopolysaccharide utilization could be intensified and extended to the production of high added-value devices.

Production of Antioxidant Transfersomes by a Supercritical CO2 Assisted Process for Transdermal Delivery Applications.

Transfersomes are deformable vesicles that can transport drugs across difficult-to-permeate barriers in human tissues. In this work, nano-transfersomes were produced for the first time by a supercritical CO2 assisted process. Operating at 100 bar and 40 °C, different amounts of phosphatidylcholine (2000 and 3000 mg), kinds of edge activators (Span® 80 and Tween® 80), and phosphatidylcholine to edge activator weight ratio (95:5, 90:10, 80:20) were tested. Formulations prepared using Span® 80 and phosphatidylcholine at an 80:20 weight ratio produced stable transfersomes (-30.4 ± 2.4 mV ζ-potential) that were characterized by a mean diameter of 138 ± 55 nm. A prolonged ascorbic acid release of up to 5 h was recorded when the largest amount of phosphatidylcholine (3000 mg) was used. Moreover, a 96% ascorbic acid encapsulation efficiency and a quasi-100% DPPH radical scavenging activity of transfersomes were measured after supercritical processing.

Current Applications of Liposomes for the Delivery of Vitamins: A Systematic Review.

Liposomes have been used for several decades for the encapsulation of drugs and bioactives in cosmetics and cosmeceuticals. On the other hand, the use of these phospholipid vesicles in food applications is more recent and is increasing significantly in the last ten years. Although in different stages of technological maturity-in the case of cosmetics, many products are on the market-processes to obtain liposomes suitable for the encapsulation and delivery of bioactives are highly expensive, especially those aiming at scaling up. Among the bioactives proposed for cosmetics and food applications, vitamins are the most frequently used. Despite the differences between the administration routes (oral for food and mainly dermal for cosmetics), some challenges are very similar (e.g., stability, bioactive load, average size, increase in drug bioaccessibility and bioavailability). In the present work, a systematic review of the technological advancements in the nanoencapsulation of vitamins using liposomes and related processes was performed; challenges and future perspectives were also discussed in order to underline the advantages of these drug-loaded biocompatible nanocarriers for cosmetics and food applications.

Supercritical Fluid Extraction of Essential Oil and Sclareol from a Clary Sage Concrete.

Clary Sage extracts are of industrial interest: in particular, sclareol shows a strong pharmaceutical potential. Supercritical fluid extraction was used to recover compounds of interest from a Salvia sclarea L. waxy n-hexane extract ("concrete"), using semi-continuous fractionation and a multi-step extraction strategy. Multi-step extraction experiments were carried out in two phases: the first one operated at 90 bar and 50 °C; the second one at 100 bar and 40 °C. GC-MS traces showed that during the first extraction step, only lighter compounds (e.g., monoterpenes, sesquiterpenes, and derivatives) were collected, whereas, in the second step, only sclareol and related compounds were recovered. By adjusting operating conditions (temperature and pressure), selective extraction of different families of compounds was accomplished, with no further need for post-processing of the products. Moreover, using two separators in series, the compounds of interest were fractionated from paraffins and, by changing the operating conditions, the extraction yield increased from about 6.0% to 9.3% w/w as CO2 density increased.

Validation of a compartmental model to predict drug release from porous structures produced by ScCO2 techniques.

A global release model is proposed to study the drug release from porous materials for pharmaceutical applications. This model is defined by implementing a compartmental model where the release profile could be explained as the combination of mass transfer phenomena through three compartments as well as a desorption process or dissolution process from the support. This model was validated with five different systems produced with supercritical CO2 (aerogels, membranes, and fibers), showing different release processes. Numerical results indicate that this compartmental approach can be useful to determine adsorption and desorption constants as well as mass transfer resistances within the material. Likewise, this model can predict lag phases and imbibition phenomena. Therefore, the development of compartmental models can be an alternative to traditional models to successfully predict the drug profile of porous materials, achieving a complete understanding of the involved phenomena regardless of the material characteristics.

Supercritical Phase Inversion to Produce Photocatalytic Active PVDF-coHFP_TiO2 Composites for the Degradation of Sudan Blue II Dye.

TiO2-loaded poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-coHFP) membranes were produced by supercritical CO2-assisted phase inversion. Three different TiO2 loadings were tested: 10, 20, and 30 wt% with respect to the polymer. Increasing the TiO2 amount from 10 wt% to 20 wt% in the starting solution, the transition from leafy-like to leafy-cellular morphology was observed in the section of the membrane. When 30 wt% TiO2 was used, the entire membrane section showed agglomerates of TiO2 nanoparticles. These polymeric membranes were tested to remove Sudan Blue II (SB) dye from aqueous solutions. The adsorption/photocatalytic processes revealed that membrane morphology and TiO2 cluster size were the parameters that mainly affected the dye removal efficiency. Moreover, after five cycles of exposure of these membranes to UV light, SB removal was higher than 85%.

Salicylic Acid Co-Precipitation with Alginate via Supercritical Atomization for Cosmetic Applications.

Alginate-based microparticles were produced via supercritical assisted atomization (SAA) with the aim of obtaining a biocompatible and low-cost carrier for the delivery of active compounds in cosmetic applications. Salicylic acid was selected as an active model compound, and it was co-precipitated with alginate via SAA, operating at 82 bar and 80 °C. In particular, the drug-to-polymer weight ratio was fixed at 1/4, whereas polymer concentration was varied from 5 to 20 mg/mL in the starting aqueous solution. Operating in this way, alginate-salicylic acid microparticles were characterized by a mean diameter of 0.72 ± 0.25 µm, and the active compound became amorphous after processing. A salicylic acid encapsulation efficiency close to 100% was reached, and the drug release time from the biopolymeric microparticles was prolonged up to nine times with respect to untreated salicylic acid powder.

Supercritical Antisolvent Technique for the Production of Breathable Naringin Powder.

Flavonoids are polyphenolic compounds largely present in fruits and vegetables possessing antioxidant properties, anti-inflammatory and antibacterial activities. Their use in clinical practice is very poor due to their low bioavailability, susceptibility to oxidation and degradation. Moreover, their slight solubility in biological fluids and a consequent low dissolution rate leads to an irregular absorption from solid dosage forms, even though, anti-inflammatory formulations could be used as support for several disease treatment, i.e. the COVID-19 syndrome. To improve flavonoid bioavailability particle size of the powder can be reduced to make it breathable and to promote the absorption in the lung tissues. Supercritical fluid based antisolvent technique has been used to produce naringin particles, with size, shape and density as well as free flowing properties able to fit inhalation needs. The dried particles are produced with the removal of the solvent at lower temperatures compared to the most used traditional micronization processes, such as spray drying. The best breathable fraction for naringin particles is obtained for particles with a d50~7 µm manufactured at 35 °C-150 bar and at 60 °C-130 bar, corresponding to 32.6% and 36.7% respectively. The powder is produced using a high CO2 molar fraction (0.99) that assure a better removal of the solvent. NuLi-1 cell line of immortalised bronchial epithelial cells adopted to evaluate powder cytotoxicity indicated after 24 h absence of toxicity at concentration of 25 µM.

The Nanostructure of Polymer-Active Principle Microparticles Produced by Supercritical CO2 Assisted Processing.

Traditional and supercritical CO2 assisted processes are frequently used to produce microparticles formed by a biopolymer containing an active principle to improve the bioavailability of the active principle. However, information about the internal organization of these microparticles is still scarce. In this work, a suspension of dextran + Fe3O4 nanoparticles (model system) and a solution of polyvinylpyrrolidone (PVP) + curcumin were used to produce spherical microparticles by supercritical CO2 processing. Periodic dynamic light scattering measurements were used to analyze the evolution of the microparticles dissolution, size, and size distribution of the guest active principle in the polymeric matrix. It was found that curcumin was dispersed in the form of nanoparticles in the PVP microparticles, whose size largely depended on its relative concentration. These results were validated by transmission electron microscopy and scanning electron microscopy of the PVP microparticles and curcumin nanoparticles, before and after the dissolution tests.

Production of Porous Agarose-Based Structures: Freeze-Drying vs. Supercritical CO2 Drying.

In this work, the effect of two processes, i.e., freeze-drying and supercritical CO2 (SC-CO2) drying, on the final morphology of agarose-based porous structures, was investigated. The agarose concentration in water was varied from 1 wt% up to 8 wt%. Agarose cryogels were prepared by freeze-drying using two cooling rates: 2.5 °C/min and 0.1 °C/min. A more uniform macroporous structure and a decrease in average pore size were achieved when a fast cooling rate was adopted. When a slower cooling rate was performed instead, cryogels were characterized by a macroporous and heterogenous structure at all of the values of the biopolymer concentration investigated. SC-CO2 drying led to the production of aerogels characterized by a mesoporous structure, with a specific surface area up to 170 m2/g. Moreover, agarose-based aerogels were solvent-free, and no thermal changes were detected in the samples after processing.

PVDF HFP_RuO2 Nanocomposite Aerogels Produced by Supercritical Drying for Electrochemical Oxidation of Model Tannery Wastewaters.

A supercritical CO2 drying process was used to prepare an innovative nanocomposite, formed by a poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF HFP) aerogel loaded with RuO2 nanoparticles. The produced nanocomposites, at 10% and 60% w/w of RuO2, were tested for the electrochemical oxidation of model tannery wastewaters. The effect of the electrochemical oxidation parameters, like pH, temperature, and current density, on tannic acid, intermediates, and chemical oxygen demand (COD) removal, was investigated. In particular, the electrolysis of a simulated real tannery wastewater, using PVDF HFP_RuO2 60, was optimized working at pH 10, 40 °C, and setting the current density at 600 A/m2. Operating in this way, surfactants, sulfides, and tannins oxidation was achieved in about 2.5 h, ammonium nitrogen oxidation in 3 h, and COD removal in 5 h. When chloride-containing solutions were tested, the purification was due to indirect electrolysis, related to surface redox reactions generating active chlorine. Moreover, sulfide ions were converted into sulfates and ammonium nitrogen in gaseous N2.

Agarose, Alginate and Chitosan Nanostructured Aerogels for Pharmaceutical Applications: A Short Review.

In this short review, drug delivery systems, formed by polysaccharide-based (i.e., agarose, alginate, and chitosan) aerogels, are analyzed. In particular, the main papers, published in the period 2011-2020 in this research field, have been investigated and critically discussed, in order to highlight strengths and weaknesses of the traditional production techniques (e.g., freeze-drying and air evaporation) of bio-aerogels with respect to supercritical CO2 assisted drying. Supercritical CO2 assisted drying demonstrated to be a promising technique to produce nanostructured bio-aerogels that maintain the starting gel volume and shape, when the solvent removal occurs at negligible surface tension. This characteristic, coupled with the possibility of removing also cross-linking agent residues from the aerogels, makes these advanced devices safe and suitable as carriers for controlled drug delivery applications.

Polysaccharide-Based Aerogel Production for Biomedical Applications: A Comparative Review.

A comparative analysis concerning bio-based gels production, to be used for tissue regeneration, has been performed in this review. These gels are generally applied as scaffolds in the biomedical field, thanks to their morphology, low cytotoxicity, and high biocompatibility. Focusing on the time interval 2015-2020, the production of 3D scaffolds of alginate, chitosan and agarose, for skin and bone regeneration, has mainly been investigated. Traditional techniques are critically reviewed to understand their limitations and how supercritical CO2-assisted processes could overcome these drawbacks. In particular, even if freeze-drying represents the most widespread drying technique used to produce polysaccharide-based cryogels, supercritical CO2-assisted drying effectively allows preservation of the nanoporous aerogel structure and removes the organic solvent used for gel preparation. These characteristics are essential for cell adhesion and proliferation.

Supercritical CO2 Extraction of Organic Solvents from Flunisolide and Fluticasone Propionate.

In this work, Class 2 and Class 3 solvents contained in two corticosteroids, flunisolide (Fluni) and fluticasone propionate (Fluti), were reduced to a few ppm by supercritical CO2 extraction. The process was carried out at pressures from 80 to 200 bar, temperatures of 40 °C and 80 °C, and at a fixed CO2 flow rate of 0.7 kg/h. The results demonstrated that CO2 density is the key parameter influencing the extraction kinetics and the solvent final residue. In particular, in the range investigated, optimal pressure and temperature conditions for the extraction of residual organic solvents were found working at 200 bar and 40 °C, which corresponds to a CO2 density of 0.840 g/cm3. Operating in this way, total organic solvent residues were reduced from 13,671 ppm and 326 ppm to 12 ppm and 10 ppm for Fluni and Fluti, respectively.

Supercritical emulsion extraction fabricated PLA/PLGA micro/nano carriers for growth factor delivery: Release profiles and cytotoxicity.

Controlled delivery of human growth factors is still a challenge in tissue engineering protocols, and poly-lactic acid and poly-lactic-co-glycolic acid carriers have been recently proposed for this purpose. Here, the microencapsulation of two human growth factors, namely Growth Differentiation Factor -5 (hGDF-5) and Transforming Growth Factor ß1 (hTGF-ß1) was tested, by processing different emulsions with Supercritical Emulsion Extraction (SEE) technology. Polymer molecular weight, co-polymer ratio and surfactant amount in aqueous phases as well as phases mixing rate were varied to fabricate carriers with suitable size and loadings. Carriers with different mean sizes from 0.4 ± 0.09 µm up to 3 ± 0.9 µm were obtained by SEE technology when processing emulsions with different formulations; carriers were loaded with 3 µg/g and 7 µg/g for hGDF-5 and hTGF-ß1 with controlled growth factor release over 25 days. Carriers displayed extremely low cytotoxicity when evaluated in Chinese Hamster Ovary cells (CHO-K1). Further, they also exhibited reduced cytotoxicity with respect to carriers obtained by conventional evaporation techniques, and low reactivity on human peripheral blood mononuclear cells (hPBMCs), suggesting their safety and potential use in tissue engineering protocols.