Supercritical impregnation of starch aerogels with quercetin: Fungistatic effect and release modelling with a compartmental model.

This work proposes the use of supercritical CO2 to impregnate starch (potato and corn) aerogels with quercetin for a potential fungistatic application. Starch aerogels were successfully produced with supercritical drying, but different results were found depending on the amylose/amylopectin ratio. A higher amount of amylose increases aerogels' specific surface area (with a structure with nanofibrils and nodes) due to the linear and amorphous character of this polymer, whereas a higher amount of amylopectin decreases this property until values of only 25 m2·g-1, obtaining an aerogel with a rough surface. These results were explained with XRD, thermogravimetric, and rheological results (triple step with two temperature sweeps and a time sweep and steady state analysis) concerning hydrogel formation. In fact, retrogradation step plays a more important role in hydrogel formation for a starch source with a higher amount of amylopectin due to an increase in the different polymers' interactions. Supercritical impregnation of quercetin on the aerogels was successfully performed (a loading around 0.30 % with respect to the amount of polymer), and in vitro results indicated that the aerogels produced a fungistatic effect on different types of fungi, but only in the first 12 h because the microorganisms adapted to the surrounding environment. Finally, a compartmental model was used to fit the drug release, which is controlled by quercetin aqueous solubility, indicating the main mass transfer resistances (mass transfer through aerogels was always around 500 min-1 and dissolution process mass transfer from 5·10-3 to 1.65·10-3 s-1) and how an increase in the specific surface area of the aerogels (in the case of corn aerogel) provided a stronger initial burst (70-80 % in 20 min). In fact, this initial burst release was mathematically related to a parameter, that varies from 0.178 to 0.036 depending on the aerogel composition. This study shows that starch aerogels can be impregnated with a hydrophobic compound with fungistatic effect by using supercritical CO2, modifying in addition the drug release by changing the native starch.

Development of a thermosensitive hydrogel based on Polaxamer 407 and gellan gum with inclusion complexes (Sulfobutylated-ß-cyclodextrin-Farnesol) as a local drug delivery system.

This work proposes the development of a thermosensitive local drug release system based on Polaxamer 407, also known as Pluronic® F-127 (PF-127), Gellan Gum (GG) and the inclusion complex Sulfobutylated-ß-cyclodextrin (CD) with Farnesol (FOH). Rheological properties of the hydrogels and their degradation were studied. According to the rheological results, a solution of 20% w/v of PF-127 forms a strong gel with a gelling temperature of about 25 °C (storage modulus of 15,000 Pa). The addition of the GG increased the storage modulus (optimal concentration of 0.5 % w/v) twofold without modifying the gelling temperature. Moreover, including 0.5% w/v of GG also increased 6 times the degradation time of the hydrogel. Regarding the inclusion complex, the addition of free CD decreased the viscosity and the gel strength since polymer chains were included in CD cavity without affecting the gelling temperature. Contrarily, the inclusion complex CD-FOH did not significantly modify any property of the formulation because the FOH was hosted in the CD. Furthermore, a mathematical model was developed to adjust the degradation time. This model highlights that the addition of the GG decreases the number of released chains from the polymeric network (which coincides with an increase in the storage modulus) and that the free CD reduces the degradation rate, protecting the polymeric chains. Finally, FOH release was quantified with a specific device, that was designed and printed for this type of system, observing a sustainable drug release (similar to FOH aqueous solubility, 8 µM) dependent on polymer degradation.

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.

Engineering Alginate-Based Dry Powder Microparticles to a Size Suitable for the Direct Pulmonary Delivery of Antibiotics.

The inhaled route is regarded as one of the most promising strategies as a treatment against pulmonary infections. However, the delivery of drugs in a dry powder form remains challenging. In this work, we have used alginate to form microparticles containing an antibiotic model (colistin sulfate). The alginate microparticles were generated by atomization technique, and they were characterized by antimicrobial in vitro studies against Pseudomonas aeruginosa. Optimization of different parameters allowed us to obtain microparticles as a dry powder with a mean size (Feret diameter) of 4.45 ± 1.40 µm and drug loading of 8.5 ± 1.50%. The process developed was able to concentrate most of the colistin deposits on the surface of the microparticles, which could be observed by SEM and a Dual-Beam microscope. This produces a fast in vitro release of the drug, with a 100% release achieved in 4 h. Physicochemical characterization using the FTIR, EDX and PXRD techniques revealed information about the change that occurs from the amorphous to a crystalline form of colistin. Finally, the cytotoxicity of microparticles was tested using lung cell lines (A549 and Calu-3). Results of the study showed that alginate microparticles were able to inhibit bacterial growth while displaying non-toxicity toward lung cells.

Tuning Alginate Microparticle Size via Atomization of Non-Newtonian Fluids.

A new approach based on the atomization of non-Newtonian fluids has been proposed to produce microparticles for a potential inhalation route. In particular, different solutions of alginate were atomized on baths of different crosslinkers, piperazine and barium chloride, obtaining microparticles around 5 and 40 microns, respectively. These results were explained as a consequence of the different viscoelastic properties, since oscillatory analysis indicated that the formed hydrogel beads with barium chloride had a higher storage modulus (1000 Pa) than the piperazine ones (20 Pa). Pressure ratio (polymer solution-air) was identified as a key factor, and it should be from 0.85 to 1.00 to ensure a successful atomization, obtaining the smallest particle size at intermediate pressures. Finally, a numerical study based on dimensionless numbers was performed to predict particle size depending on the conditions. These results highlight that it is possible to control the microparticles size by modifying either the viscoelasticity of the hydrogel or the experimental conditions of atomization. Some experimental conditions (using piperazine) reduce the particle size up to 5 microns and therefore allow their use by aerosol inhalation.

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.

Biopesticide Encapsulation Using Supercritical CO2: A Comprehensive Review and Potential Applications.

As an alternative to synthetic pesticides, natural chemistries from living organisms, are not harmful to nontarget organisms and the environment, can be used as biopesticides, nontarget. However, to reduce the reactivity of active ingredients, avoid undesired reactions, protect from physical stress, and control or lower the release rate, encapsulation processes can be applied to biopesticides. In this review, the advantages and disadvantages of the most common encapsulation processes for biopesticides are discussed. The use of supercritical fluid technology (SFT), mainly carbon dioxide (CO2), to encapsulate biopesticides is highlighted, as they reduce the use of organic solvents, have simpler separation processes, and achieve high-purity particles. This review also presents challenges to be surpassed and the lack of application of SFT for biopesticides in the published literature is discussed to evaluate its potential and prospects.

Steady and Oscillatory Shear Flow Behavior of Different Polysaccharides with Laponite®.

The rheological behavior, in terms of steady and oscillatory shear flow, of Laponite® with different polysaccharides (alginate, chitosan, xanthan gum and levan) in salt-free solutions was studied. Results showed that a higher polymer concentration increased the zero-rate viscosity and decreased the critical strain rate (Cross model fit) as well as increasing the elastic and viscous moduli. Those properties (zero-rate viscosity and critical strain rate) can be a suitable indicator of the effect of the Laponite® on the shear flow behavior for the different solutions. Specifically, the effect of the Laponite® predominates for solutions with large critical strain rate and low zero-rate viscosity, modifying significantly the previous parameters and even the yield stress (if existing). On the other hand, larger higher polymeric concentration hinders the formation of the platelet structure, and polymer entanglement becomes predominant. Furthermore, the addition of high concentrations of Laponite® increases the elastic nature, but without modifying the typical mechanical spectra for polymeric solutions. Finally, Laponite® was added to (previously crosslinked) gels of alginate and chitosan, obtaining different results depending on the material. These results highlight the possibility of predicting qualitatively the impact of the Laponite® on different polymeric solutions depending on the solutions properties.

Comparative anatomy and 3D geometric morphometrics of the El Sidrón atlases (C1).

The first cervical vertebra (atlas, C1) is an important element of the vertebral column because it connects the cranial base with the cervical column, thus helping to maintain head posture and contributing to neck mobility. However, few atlases are preserved in the fossil record because of the fragility of this vertebra. Consequently, only eight well-preserved atlases from adult Neandertals have been recovered and described. Here, we present nine new atlas remains from the El Sidrón Neandertal site (Asturias, Spain), two of which (SD-1643 and SD-1605/1595) are sufficiently well preserved to allow for a detailed comparative and three-dimensional geometric morphometric analysis. We compared standard linear measurements of SD-1643 and SD-1605/1595 with those of other Neandertal atlases and carried out three-dimensional geometric morphometric analyses to compare size and shape of SD-1643 and SD-1605/1595 with those of 28 Pan (Pan troglodytes and Pan paniscus), a broad comparative sample of 55 anatomically modern humans from African and European populations, and other fossil hominins (Neandertals, Homo antecessor, Paranthropus boisei). The El Sidrón atlas fossils show typical features of the Neandertal atlas morphology, such as caudal projection of the anterior tubercle, gracility of both the posterior tubercle and the tuberosity for the insertion of the transverse ligament, and an anteroposteriorly elongated neural canal. Furthermore, when compared with atlases from the other taxa, Neandertals exhibit species-specific features of atlas morphology including a relatively lower lateral mass height, relatively narrower transverse foramina, and flatter and more horizontally oriented articular facets. Some of these features fit with previous suggestions of shorter overall length of the cervical spine and potential differences in craniocervical posture and mobility. Our results may support a different spinopelvic alignment in this species, as the atlas morphology suggests reduced cervical lordosis.

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.

The study of the lower limb entheses in the Neanderthal sample from El Sidrón (Asturias, Spain): How much musculoskeletal variability did Neanderthals accumulate?

Entheses have rarely been systematically studied in the field of human evolution. However, the investigation of their morphological variability (e.g., robusticity) could provide new insight into their evolutionary significance in the European Neanderthal populations. The aim of this work is to study the entheses and joint features of the lower limbs of El Sidrón Neanderthals (Spain; 49 ka), using standardized scoring methods developed on modern samples. Paleobiology, growth, and development of both juveniles and adults from El Sidrón are studied and compared with those of Krapina Neanderthals (Croatia, 130 ka) and extant humans. The morphological patterns of the gluteus maximus and vastus intermedius entheses in El Sidrón, Krapina, and modern humans differ from one another. Both Neanderthal groups show a definite enthesis design for the gluteus maximus, with little intrapopulation variability with respect to modern humans, who are characterized by a wider range of morphological variability. The gluteus maximus enthesis in the El Sidrón sample shows the osseous features of fibrous entheses, as in modern humans, whereas the Krapina sample shows the aspects of fibrocartilaginous ones. The morphology and anatomical pattern of this enthesis has already been established during growth in all three human groups. One of two and three of five adult femurs from El Sidrón and from Krapina, respectively, show the imprint of the vastus intermedius, which is absent among juveniles from those Neanderthal samples and in modern samples. The scant intrapopulation and the high interpopulation variability in the two Neanderthal samples is likely due to a long-term history of small, isolated populations with high levels of inbreeding, who also lived in different ecological conditions. The comparison of different anatomical entheseal patterns (fibrous vs. fibrocartilaginous) in the Neanderthals and modern humans provides additional elements in the discussion of their functional and genetic origin.

Analyses of the neandertal patellae from El Sidrón (Asturias, Spain) with implications for the evolution of body form in Homo.

The evolution of the body form in Homo and its potential morphological connection to the arrangement of different skeletal systems is of major interest in human evolution. Patella morphology as part of the knee is potentially influenced by body form. Here, we describe for the first time the patellae remains recovered at El Sidrón Neandertal site and analyze them in a comparative evolutionary framework. We aim to clarify whether morphometric features frequently observed in Neandertal and modern human patellae are retained from a primitive anatomical arrangement or whether they represent derived features (apomorphies). For this purpose, we combine analyses of discrete features, classic anthropological measurements, and 3D geometric morphometrics based on generalized Procrustes analysis, mean size and shape comparisons, and principal components analysis. We found a size increment of the patella in hominin evolution, with large species showing a larger patella. Modern humans and Neandertals exhibit overall larger patellae, with maximum values observed in the latter, likely as a consequence of their broader body shape. Also, some Neandertals display a thicker patella, which has been linked to larger quadriceps muscles. However, Neandertals retain a primitive morphology in their patellar articular surfaces, with similar-sized lateral and medial articular facets, leading to a more symmetrical internal face. This feature is inherited from a primitive Homo ancestor and suggests a different configuration of the knee in Neandertals. Conversely, Homo sapiens exhibits an autoapomorphic patellar anatomy with expanded lateral articular facets. We propose that these distinct configurations of the patella within Homo may be a consequence of different body forms rather than specific functional adaptations of the knee. Thus, the slender body form of modern humans may entail a medial reorientation of the tibial tuberosity (patellar ligament), allowing lateral surface expansion. These anatomical evolutionary variations may involve subtle secondary differences in bipedalism within Homo.

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.

A comprehensive study on levan nanoparticles formation: Kinetics and self-assembly modeling.

Levan nanoparticles formation is a complicated phenomenon involving simultaneously polymeric reaction kinetics and nanoparticles self-assembly theory. These phenomena are studied in this work with experimental and computational methodologies. Specifically, the effect of different parameters on levan kinetics and nanoparticles production in a cell-free system environment have been studied. Results point out that 37 °C is the best temperature for synthesizing levan as well as the existence of a substrate inhibition effect for polymeric reaction. This work also highlights that raffinose can be used for producing and that an increase on the ratio enzyme-substrate increases the velocity of conversion. However, the previous experimental conditions did not produce an important effect on self-assembly formed levan nanoparticles (always 110 nm) as long as the required levan concentration (CAC) for nanoparticles reorganization is achieved. To have a better understanding of these results, a model was developed to explain numerically levan kinetics and nanoparticle self-assembly. This model was built by taking into account enzyme poisoning effect (also demonstrated experimentally) and a diffusion limited cluster model for the aggregation phenomenon. Simulation results fit properly experimental data and catalytic parameters as well as predicting accurately the value of CAC for producing its reorganization into nanoparticles by self-assembly.

Differences in levan nanoparticles depending on their synthesis route: Microbial vs cell-free systems.

Differences between the levan obtained from bacteria and from cell-free systems were studied in this work. Results showed that both polymers are non-porous solids (type II isotherm with 20 m2/g) with a main thermal decomposition at 200 °C and a negligible value of protein adsorption. Microbial levan produced nanoparticles of 90 nm in diameter whereas nanoparticles of 110 nm were obtained with the polymer obtained from a cell-free system. Both polymers behave as aggregates depending on the critical aggregation concentration. At the same time, that concentration depends on the technique used for the polymer synthesis. Cell-free system aggregation concentration is 0.24 mg/mL whereas a concentration of 0.05 mg/mL was found for the microbial system. In both cases, the average molecular weight of the aggregate is higher than 2000 kDa. These results highlight the existence of aggregation equilibrium for both polymers that has to be taken into account for future applications.

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.

Levan-Capped Silver Nanoparticles for Bactericidal Formulations: Release and Activity Modelling.

An environmentally friendly technique was used to produce levan-capped silver nanoparticles of about 30 nm (with a loading of 30%) that showed bactericide effect, for E. coli and B. subtilis. That effect was mathematically studied with a dose-response model (lethal dose of 12.4 ppm and 6.8 ppm respectively). These silver nanoparticles were subsequently introduced in a gel to create a silver release system with bacteria inhibition activity. Silver release from the gel and its bactericidal activity was theoretically studied to develop a unique model that is able to predict accurately both silver release and lethal dose for any type of bacteria. This model will be useful for performing predictions for future silver in gel applications.

Primary visual cortex in neandertals as revealed from the occipital remains from the El Sidrón site, with emphasis on the new SD-2300 specimen.

The comparative analysis of the endocranial surface of the El Sidrón new occipital fragment SD-2300 shows meaningful differences in the configuration of the occipital pole region between neandertals and anatomically modern humans (AMH). The particular asymmetries found in neandertals in the venous sinus drainage and the petalial patterns are recognizable in this new specimen as well. In addition, the supra- and infracalcarine fossae of the occipital pole region appear to deviate obliquely from the mid-line when compared with sapiens. Due to the excellent preservation conditions of SD-2300, the main sulci and gyri of the occipital pole area have been identified, this degree of detail being uncommon in a fossil specimen; in general, the gyrification pattern is similar to AMH, but with some notable differences. Particularly interesting is the description of the lunate and the calcarine sulci. The lunate sulcus is located close to the occipital pole, in a similar posterior position to in other Homo species. Regarding the calcarine sulcus, there are significant differences in the primary visual cortex, with the V1 area, or Brodmann area 17, being larger in Homo neanderthalensis than in Homo sapiens. This may lead to greater visual acuity in neandertals than in sapiens.