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.

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%.

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.

Porous Aerogels and Adsorption of Pollutants from Water and Air: A Review.

Aerogels are open, three-dimensional, porous materials characterized by outstanding properties, such as low density, high porosity, and high surface area. They have been used in various fields as adsorbents, catalysts, materials for thermal insulation, or matrices for drug delivery. Aerogels have been successfully used for environmental applications to eliminate toxic and harmful substances-such as metal ions or organic dyes-contained in wastewater, and pollutants-including aromatic or oxygenated volatile organic compounds (VOCs)-contained in the air. This updated review on the use of different aerogels-for instance, graphene oxide-, cellulose-, chitosan-, and silica-based aerogels-provides information on their various applications in removing pollutants, the results obtained, and potential future developments.

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.

Microbial Exopolysaccharides as Drug Carriers.

Microbial exopolysaccharides are peculiar polymers that are produced by living organisms and protect them against environmental factors. These polymers are industrially recovered from the medium culture after performing a fermentative process. These materials are biocompatible and biodegradable, possessing specific and beneficial properties for biomedical drug delivery systems. They can have antitumor activity, they can produce hydrogels with different characteristics due to their molecular structure and functional groups, and they can even produce nanoparticles via a self-assembly phenomenon. This review studies the potential use of exopolysaccharides as carriers for drug delivery systems, covering their versatility and their vast possibilities to produce particles, fibers, scaffolds, hydrogels, and aerogels with different strategies and methodologies. Moreover, the main properties of exopolysaccharides are explained, providing information to achieve an adequate carrier selection depending on the final application.

Supercritical carbon dioxide techniques for processing microbial exopolysaccharides used in biomedical applications.

Microbial exopolysaccharides are polymers that show a great potential for biomedical applications, such as tissue engineering applications and drug delivery, due to their biocompatibility, biodegradability and their gelling properties. These polysaccharides are obtained from a microorganism culture with a relatively straightforward downstream process thanks to their extracellular character, and can be processed to obtain aerogels, fibers and micro- or nano-particles with conventional techniques. However, these techniques present several disadvantages in that they involve time-consuming processes and the use of toxic solvents. Supercritical carbon dioxide techniques can overcome these drawbacks, but their use for processing microbial exopolysaccharides is not extended in the scientific community. This review describes the most frequently used exopolysaccharides in biomedical applications and how they can be obtained, as well as the different supercritical carbon dioxide techniques that can be used for processing them and their challenges. Specifically, high pressure shows a great potential to process and sterilize exopolysaccharide biomaterials for biomedical applications (e.g. tissue engineering or drug delivery systems) in spite of the disadvantage concerning the hydrophilicity of this type of polymers.

Supercritical Phase Inversion: A Powerful Tool for Generating Cellulose Acetate-AgNO3 Antimicrobial Membranes.

Antimicrobial composite membranes, formed by cellulose acetate loaded with AgNO3 particles, were produced by supercritical phase inversion. Different cellulose acetate concentrations were tested (15%, 20%, 30%(w/w)), whereas the active agent (i.e., silver nitrate) concentration was fixed at 0.1%(w/w) with respect to the quantity of polymer used. To determine the influence of the process parameters on membranes morphology, the pressure and temperature were varied from 150 to 250 bar and from 55 to 35 °C, respectively. In all cases, regularly porous membranes were produced with a uniform AgNO3 distribution in the membrane matrix. Silver release rate depended on membrane pore size, covering a time interval from 8 to 75 h.

Role of rheological properties on physical chitosan aerogels obtained by supercritical drying.

Chitosan aerogels were obtained after using supercritical carbon dioxide to dry physical hydrogels, studying the effect of the rheological behavior of hydrogels and solutions on the final aerogels properties. An increase on the solutions pseudoplasticity increased the subsequent hydrogels physical entanglement, without showing a significant effect on aerogels morphology (nanoporous) and textural properties (pores of about 10 nm). However, an increase of hydrogel physical entanglement promoted the formation of aerogels with a higher compressive strength (from 0.2 to 0.80 MPa) and higher thermal decomposition range, while decreasing the porosity (from 90 % to 94 %). Aerogels stress-strain responses were also successfully fitted using a hyperelastic equation with three adjustable parameters (Yeoh), showing that this type of models must be taken into account when large stresses are studied.

Cellulose Acetate and Supercritical Carbon Dioxide: Membranes, Nanoparticles, Microparticles and Nanostructured Filaments.

Cellulose acetate (CA) is a very versatile biocompatible polymer used in various industrial sectors. Therefore, depending on the application, different morphologies are required. Different processes at industrial scale are commonly employed to obtain CA micro or nanoparticles (discontinuous structures) or CA membranes (continuous structures with discontinuities). In this work, two supercritical carbon dioxide (scCO2) based techniques, such as the semi-continuous supercritical antisolvent process (SAS) and the supercritical fluid phase inversion process, in which scCO2 plays the role of antisolvent, were employed. Varying the kind of organic solvent used to prepare the polymeric solution, the polymer concentration, and operating pressure and temperature, it was possible to tune the characteristics of the obtained material. In particular, using acetone as the organic solvent, filaments constituted by nanoparticles, expanded microparticles, nanoparticles with a mean diameter lower than 80 nm, and microporous membranes were obtained, varying the operating conditions. The attainment of spherical micron-sized particles was instead achieved using a mixture of acetone and DMSO as the organic solvent. Therefore, the versatility of the supercritical carbon dioxide-based techniques has been confirmed, and it was possible to obtain, using a single experimental plant, various morphologies of cellulose acetate (with controllable particles' or pores' diameters) by varying the operating conditions.

Supercritical Fluid Processing of Polymers.

The use of supercritical fluids instead of organic solvents has attracted the interest of numerous researchers, due to the unique peculiarities of supercritical fluids which are characterized by solvent powers comparable to those of liquid organic solvents, diffusivity comparable to those of gaseous substances and quasi-zero surface tension [...].

Supercritical Assisted Electrospray: An Improved Micronization Process.

A new process is proposed that can largely improve classical electrospray (ESPR) atomization, thanks to the addition of supercritical CO2 (SC-CO2) to the liquid mixture, in which a polymer is dissolved, forming an expanded liquid. The consequent reduction of surface tension and viscosity allows the production of micrometric or nanometric particles of controlled size and distribution at a production rate up to one hundred times that of the traditional process. The new process was applied to particle generation from a very high molecular weight polyvinylpyrrolidone (PVP) and tested at different polymer percentages by weight and at different pressures. Repeatable microparticle diameters and distributions were obtained, ranging between 0.55 and 2.25 µm at PVP concentrations from 1 to 5% w/w and pressures between 80 and 120 bar.

A Phenomenological Approach to Study Mechanical Properties of Polymeric Porous Structures Processed Using Supercritical CO2.

This work proposes a modeling of the mechanical properties of porous polymers processed by scCO2, using a phenomenological approach. Tensile and compression tests of alginate/gelatin and cellulose acetate/graphene oxide were modeled using three hyperelastic equations, derived from strain energy functions. The proposed hyperelastic equations provide a fair good fit for mechanical behavior of the nanofibrous system alginate/gelatin (deviations lower than 10%); whereas, due to the presence of the solid in the polymer network, a four-parameter model must be used to fit the composite cellulose acetate/graphene oxide behavior. Larger deviations from the experimental data were observed for the system cellulose acetate/graphene oxide because of its microporous structure. A finite element method was, then, proposed to model both systems; it allowed a realistic description of observable displacements and effective stresses. The results indicate that materials processed using scCO2, when submitted to large stresses, do not obey Hooke´s law and must be considered as hyperelastic.

SC-CO2-assisted process for a high energy density aerogel supercapacitor: the effect of GO loading.

Energy density, safety, and simple and environmentally friendly preparation methods are very significant aspects in the realization of a compact supercapacitor. Herein we report the use of a supercritical CO2-assisted gel drying process (SC-CO2) for the preparation of porous electrodes containing dispersed graphene in a poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) binder membrane to sandwich in a new portable supercapacitor based on graphene oxide (GO). A GO loading of 60 wt.% was found to give the best combination of factors (porosity, wettability, mechanical and electrochemical properties). Cycling voltammetry and charge/discharge studies showed an excellent capacitance behaviour and stability in an ionic liquid electrolyte, suggesting SC-CO2 processing as a promising platform to produce highly bulky and porous films for supercapacitors. The supercapacitor device delivers a very high energy density of 79.2 Wh kg-1 at a power density of 0.23 KW kg-1 (current density 0.5 A g-1, specific capacitance 36.2 F g-1) while that of steel remains at 50.3 Wh kg-1 at a power density of 2.8 KW kg-1 (current density 6 A g-1, specific capacitance 23.5 F g-1).

Regeneration techniques for bone-to-tendon and muscle-to-tendon interfaces reconstruction.

INTRODUCTION: The complex structure of the bone-tendon and muscle-tendon junctions makes their reproduction for tissue engineering applications very difficult. Relatively few studies have investigated the characteristics of these regions from a tissue engineering view point. SOURCES OF DATA: PubMed, Thomson Reuters, Scopus and Google Scholar databases were searched using various combinations of the keywords 'Tendon', 'Myotendinous junction', 'Osteotendinous junction', 'Tissue engineering' and 'Scaffold'. AREAS OF AGREEMENT: The available studies can be divided according to whether the objective is to build an entire composite tissue unit or to assist the recreation of interfaces, such as improving integration of autografts with the surrounding bone or with the muscle. The most used techniques are based on the electrospinning and the self-reorganized constructs process, which were applied to both bone-to-tendon junction (BTJ) and muscle-to-tendon junction (MTJ) regeneration. The use of nanofibers that mimic the hierarchical structure of the extracellular matrix (ECM), eventually functionalized by encapsulation of bioactive components, allowed cell attachment and differentiation. AREAS OF CONTROVERSY: There have been no translational investigations. GROWING POINTS: There is a need to devise suitable techniques that allow suitable tissue engineering of BTJ and MTJ. AREAS TIMELY FOR DEVELOPING RESEARCH: Appropriately planned studies are needed to translate tissue engineering from a scientific challenge to a clinically applicable technique.

Supercritical fluid assisted process for the generation of cellulose acetate loaded structures, potentially useful for tissue engineering applications.

Supercritical CO2 phase inversion offers an alternative to obtain solvent free structures with short processing times and preservation of the morphology. We prepared cellulose acetate structures loaded with drug (ibuprofen) to perform experiments at pressures and temperatures ranging between 150 and 250 bars and 35 and 55 °C. The structures were properly characterized by SEM, EDX and DSC; drug controlled release experiments were also performed. Analyses showed that the operating conditions strongly influenced the structure morphology, porosity and drug release profiles. Indeed, connected microparticles, nanofibrous networks and cellular membranes were produced, which have generated different drug release profiles.

Interpenetration of Natural Polymer Aerogels by Supercritical Drying.

Natural polymers, such as alginate and gelatin, can be used to produce scaffolds for tissue engineering applications; but, their mechanical and biochemical performance should be improved. A possible solution to obtain this result, is the generation of multi-component scaffolds, by blending two or more polymers. One way to realize it, is the formation of an interpenetrating polymer network (IPN). In this work, the interpenetration of alginate and gelatin hydrogels has been successfully obtained and preserved by supercritical CO2 (SC-CO2) drying performed at 200 bar and 35 °C, using different blend compositions: from alginate/gelatin = 20:80 v/v to alginate/gelatin = 80:20 v/v. The process allowed modulation of morphology and mechanical properties of these blends. The overall result was made possible by the supercritical drying process that, working at zero surface tension, allows preserving the hydrogels nanostructure in the corresponding aerogels.

Tubular perfusion system culture of human mesenchymal stem cells on poly-L-lactic acid scaffolds produced using a supercritical carbon dioxide-assisted process.

In vitro human mesenchymal stem cell (hMSC) proliferation and differentiation is dependent on scaffold design parameters and specific culture conditions. In this study, we investigate how scaffold microstructure influences hMSC behavior in a perfusion bioreactor system. Poly-L-lactic acid (PLLA) scaffolds are fabricated using supercritical carbon dioxide (SC-CO(2)) gel drying. This production method results in scaffolds fabricated with nanostructure. To introduce a microporous structure, porogen leaching was used in addition to this technique to produce scaffolds of average pore size of 100, 250, and 500 µm. These scaffolds were then cultured in static culture in well plates or dynamic culture in the tubular perfusion system (TPS) bioreactor. Results indicated that hMSCs were able to attach and maintain viability on all scaffolds with higher proliferation in the 250 µm and 500 µm pore sizes of bioreactor cultured scaffolds and 100 µm pore size of statically cultured scaffolds. Osteoblastic differentiation was enhanced in TPS culture as compared to static culture with the highest alkaline phosphatase expression observed in the 250 µm pore size group. Bone morphogenetic protein-2 was also analyzed and expression levels were highest in the 250 µm and 500 µm pore size bioreactor cultured samples. These results demonstrate cellular response to pore size as well as the ability of dynamic culture to enhance these effects.

Biodegradable synthetic scaffolds for tendon regeneration.

Tissue regeneration is aimed at producing biological or synthetic scaffolds to be implanted in the body for regenerate functional tissues. Several techniques and materials have been used to obtain biodegradable synthetic scaffolds, on which adhesion, growth, migration and differentiation of human cells has been attempted. Scaffolds for tendon regeneration have been less frequently proposed, because they have a complex hierarchical structure and it is very difficult to mimic their peculiar mechanical properties. In this review, we critically analyzed the proposed materials and fabrication techniques for tendon tissue engineering and we indicated new preparation processes, based on the use of supercritical fluids, to produce scaffolds with characteristics very similar to the native tendon structure.