Encapsulation of Olive (Olea europaea L.) Pruning Waste Particles by Supercritical CO2 Technology.

Olive leaves (Olea europaea L.) contain a multitude of bioactive compounds such as sterols, carotenes, triterpenic alcohols and phenolic compounds. These compounds have been shown to exhibit antiviral, antioxidant, candida-growth-inhibitory, anticancer, antifungal, anti-inflammatory and antibacterial activities. In this sense, submicron particles from olive leaves with antioxidant activity were precipitated by supercritical antisolvent extraction in a previous work. Moreover, encapsulation enables the delayed release of compounds and avoids the first-step effect in medical therapies. Therefore, this work focused on encapsulation of particles with a certain antioxidant capacity from olive pruning waste using supercritical technology. A variety of experiments were carried out to test how the different operating variables (pressure, temperature and extract-polymer ratio) affect. Morphology was analyzed by SEM microscopy, obtaining encapsulates between 1 and 5 microns in size. The antioxidant capacity was determined by the DPPH assay, with most of the encapsulates having AAI values between 0.5 and 1 (moderate antioxidant capacity). An increase in polyphenol content was observed in the 1:3 ratio tests. The release of the compounds in gastric simulated medium was retarded by the polymeric encapsulation, while in intestinal fluid, the solubility was improved compared to the unencapsulated particles. Overall, the supercritical encapsulation process for the natural extract of olive pruning residues has proven to be effective in obtaining antioxidant particles with different release profiles.

Formation of PLGA-PEDOT: PSS Conductive Scaffolds by Supercritical Foaming.

The usage of conjugated materials for the fabrication of foams intended to be used as therapeutic scaffolds is gaining relevance these days, as they hold certain properties that are not exhibited by other polymer types that have been regularly used until the present. Hence, this work aims to design a specific supercritical CO2 foaming process that would allow the production of porous polymeric devices with improved conductive properties, which would better simulate matrix extracellular conditions when used as therapeutic scaffolds (PLGA-PEDOT:PSS) systems. The effects of pressure, temperature, and contact time on the expansion factor, porosity, mechanical properties, and conductivity of the foam have been evaluated. The foams have been characterized by scanning electron and atomic force microscopies, liquid displacement, PBS degradation test, compression, and resistance to conductivity techniques. Values close to 40% porosity were obtained, with a uniform distribution of polymers on the surface and in the interior, expansion factors of up to 10 orders, and a wide range of conductivity values (2.2 × 10-7 to 1.0 × 10-5 S/cm) and mechanical properties (0.8 to 13.6 MPa Young's modulus in compression test). The conductive and porous scaffolds that have been produced by supercritical CO2 in this study show an interesting potential for tissue engineering and for neural or cardiac tissue regeneration purposes due to the fact that electrical conductivity is a crucial factor for proper cell function and tissue development.

Generation of Highly Antioxidant Submicron Particles from Myrtus communis Leaf Extract by Supercritical Antisolvent Extraction Process.

Submicron particles have been produced from an ethanolic extract of Myrtus communnis leaves using supercritical carbon dioxide technology, hereinafter referred to as Supercritical Antisolvent Extraction (SAE). The influence of pressure (9-20 MPa), temperature (308 and 328 K) and injection rate (3 and 8 mL/min) on the particles' precipitation has been investigated, and it has been confirmed that increases in pressure and temperature led to smaller particle sizes. The obtained particles had a quasi-spherical shape with sizes ranging from 0.42 to 1.32 µm. Moreover, the bioactivity of the generated particles was assessed and large contents of phenolic compounds with a high antioxidant activity were measured. The particles were also subjected to in vitro studies against oxidative stress. The myrtle particles demonstrated cytoprotective properties when applied at low concentrations (1 µM) to macrophage cell lines.

Supercritical Impregnation of Mangifera indica Leaves Extracts into Porous Conductive PLGA-PEDOT Scaffolds.

Plant leaves, such as those from Mangifera indica, represent a potential utilization of waste due to their richness in bioactive compounds. Supercritical CO2 allows these compounds to be incorporated into various matrices by impregnation. Combined with its ability to generate polymeric scaffolds, it represents an attractive strategy for the production of biomedical devices. For this purpose, conjugated polymeric scaffolds of biodegradable PLGA (poly(lactic-co-glycolic acid)) and PEDOT:PSS (poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate)), generated in situ by foaming, were employed for the supercritical impregnation of ethanolic mango leaves extract (MLE) in tissue engineering as a potential application. The extraction of MLE was performed by Enhanced Solvent Extraction. The effects of pressure (120-300 bar), temperature (35-55 °C), and depressurization rate (1-50 bar/min) on the physical/conductive properties and the impregnation of MLE were studied. The scaffolds have been characterized by liquid displacement, scanning electron microscope, resistance to conductivity techniques, measurements of impregnated load, antioxidant capacity and antimicrobial activity. Porosity values ranging 9-46% and conductivity values between 10-4-10-5 S/cm were obtained. High pressures, low temperatures and rapid depressurization favored the impregnation of bioactive compounds. Scaffolds with remarkable antioxidant activity were obtained (75.2-87.3% oxidation inhibition), demonstrating the ability to inhibit S. aureus bacterial growth (60.1 to 71.4%).

Inclusion of Natural Antioxidants of Mango Leaves in Porous Ceramic Matrices by Supercritical CO2 Impregnation.

Mango is one of the most important, medicinal tropical plants in the world from an economic point of view due to the presence of effective bioactive substances as co-products in its leaves. The aim of this work was to enhance the impregnation of natural antioxidants from mango leaves into a porous ceramic matrix. The effects of pressure, temperature, impregnation time, concentration of the extract and different porous silica on impregnation of phenolic compounds and antioxidant activity were analyzed. The volume of the pressurized fluid extract and amount of porous ceramic matrix remained constant. The best impregnation conditions were obtained at 6 h, 300 bar, 60 mg/mL, 35 °C and with MSU-H porous silica. The results indicated that increasing the pressure, concentration of the extract and temperature during impregnation with phenolic compounds such as gallic acid and iriflophenone 3-C (2-O-p-hydroxybenzolyl)-ß-D-glucoside increased the antioxidant activity and the amount of total phenols.

Development of Porous Polyvinyl Acetate/Polypyrrole/Gallic Acid Scaffolds Using Supercritical CO2 as Tissue Regenerative Agents.

Scaffolds are advanced devices employed in tissue engineering, as they are intended to mimic the characteristics of extracellular matrices. In this respect, conjugated materials are gaining relevance in the manufacturing of the foams used for therapeutic scaffolds, since they can provide certain properties that are missing in the other polymers used to form the scaffolds. This work has, therefore, focused on the development of functional scaffolds formed by conjugated-non-conjugated polymers such as polyvinyl acetate and polypyrrole, impregnated with gallic acid as the model drug and produced by means of a supercritical CO2 foaming/impregnation process. The effects from a series of parameters such as pressure, temperature, depressurization rate, and contact time of the scaffold production process have been determined. The impregnated foams have been characterized according to their morphology, including their porosity and expansion factor, their drug loading and delivering capabilities, and their mechanical and electrical properties. The characterization of the experiments was carried out using scanning electron microscopy, liquid displacement, in vitro release, electrochemical impedance spectroscopy, and compression techniques. The results from our tests have revealed a considerable influence of all the input variables studied, as well as relevant interactions between them. Values close to 35% porosity were obtained, with a drug release of up to 10 h with a fast initial release. The best operating conditions were 353 K, 30 MPa, 0.5 MPa/min depressurization rate, and 1 h contact time. By means of the supercritical foaming/impregnation technique, scaffolds with potential in tissue engineering due to their studied properties were obtained.

An Attempt to Optimize Supercritical CO2 Polyaniline-Polycaprolactone Foaming Processes to Produce Tissue Engineering Scaffolds.

Conjugated polymers are biomaterials with high conductivity characteristics because of their molecular composition. However, they are too rigid and brittle for medical applications and therefore need to be combined with non-conductive polymers to overcome or lessen these drawbacks. This work has, consequently, focused on the development of three-dimensional scaffolds where conductive and non-conductive polymers have been produced by combining polycaprolactone (PCL) and polyaniline (PANI) by means of supercritical CO2 foaming techniques. To evaluate their therapeutic potential as implants, a series of experiments have been designed to determine the most influential variables in the production of the three-dimensional scaffolds, including temperature, pressure, polymer ratio and depressurization rate. Internal morphology, porosity, expansion factor, PANI loads, biodegradability, mechanical and electrical properties have been taken as the response variables. The results revealed a strong influence from all the input variables studied, as well as from their interactions. The best operating conditions tested were 70 °C, 100 bar, a ratio of 5:1 (PCL:PANI), a depressurization rate of 20 bar/min and a contact time of 1 h.

Determining the Optimal Conditions for the Production by Supercritical CO2 of Biodegradable PLGA Foams for the Controlled Release of Rutin as a Medical Treatment.

Poly(D,L,-lactide-co-glycolide) (PLGA) foam samples impregnated with rutin were successfully produced by supercritical foaming processes. A number of parameters such as pressure (80-200 bar), temperature (35-55 °C), depressurization rate (5-100 bar/min), ratio lactide:glycolide of the poly(D,L,-lactide-co-glycolide) (50:50 and 75:25) were studied to determine their effect on the expansion factor and on the glass transition temperature of the polymer foams and their consequences on the release profile of the rutin entrapped in them. The impregnated foams were characterized by scanning electron microscopy, differential scanning calorimetry, and mercury intrusion porosimetry. A greater impregnation of rutin into the polymer foam pores was observed as pressure was increased. The release of rutin in a phosphate buffer solution was investigated. The controlled release tests confirmed that the modification of certain variables would result in considerable differences in the drug release profiles. Thus, five-day drug release periods were achieved under high pressure and temperature while the depressurization rate remained low.

Foaming of Polycaprolactone and Its Impregnation with Quercetin Using Supercritical CO2.

Foamed polycaprolactone impregnated with quercetin was carried out with a batch foaming technique using supercritical CO2. The experimental design was developed to study the influence of pressure (15-30 MPa), temperature (308-333 K), and depressurization rate (0.1-20) on the foam structure, melting temperature, and release tests of composites. The characterization of the experiments was carried out using scanning electron microscopy, X-ray diffractometer, and differential scanning calorimetry techniques. It was observed that the porosity created in the polymer had a heterogeneous structure, as well as the impregnation of the quercetin during the process. On the other hand, controlled release tests showed a significant delay in the release of quercetin compared to commercial quercetin.

Micro-Raman Spectroscopy for the Determination of Local Temperature Increases in TiO2 Thin Films due to the Effect of Radiation.

This study applied a classic method involving Raman spectroscopy and the use of Stokes and anti-Stokes peaks to measure the temperature of TiO2 thin films found in dye-sensitized solar cells (DSSCs). In addition, three mathematical formulae were used and analyzed to estimate the increase in temperature generated solely by the effect of the radiation. The tests and calculations performed showed an increase in the temperature of the TiO2 film. That is, the films were heated by the radiation they were exposed to. A temperature increase of up to 30 K was detected for the sample with a single layer of TiO2, and over 40 K for the sample with three layers for the highest radiation powers used, and greater increases in temperature were observed in the thicker films.

A Study of Overheating of Thermostatically Controlled TiO2 Thin Films by Using Raman Spectroscopy.

In this study a classic Raman spectroscopy method is applied and the intensity ratio of Stokes and anti-Stokes peaks is used to measure the temperature of thermostatically controlled TiO2 thin films. In addition, three mathematical formulae are used and analyzed to estimate the temperature of the TiO2 thin films. Overheating of the samples above the thermostatically controlled temperature was observed while recording the Raman spectra, with a temperature increase of up to 30 K being detected. DFT-periodic calculations showed that the anatase (101) surface had a smaller band gap than bulk anatase. Thus, it can absorb the laser radiation with a wavelength of 532 nm that is used in the experimental setup. Part of the absorbed photon energy transfers into phonon energy, heating up the anatase phase, thus leading to the heating of the samples. Moreover, overheating of the samples indicates that the experimental method used in this study can lead to deviations in their real absolute temperature values.