Solution-based processing for scaffold fabrication in tissue engineering applications: A brief review

The fabrication of 3D scaffolds is under wide investigation in tissue engineering (TE) because of its incessant development of new advanced technologies and the improvement of traditional processes. Currently, scientific and clinical research focuses on scaffold characterization to restore the function of missing or damaged tissues. A key for suitable scaffold production is the guarantee of an interconnected porous structure that allows the cells to grow as in native tissue. The fabrication techniques should meet the appropriate requirements, including feasible reproducibility and time-and cost-effective assets. This is necessary for easy processability, which is associated with the large range of biomaterials supporting the use of fabrication technologies. This paper presents a review of scaffold fabrication methods starting from polymer solutions that provide highly porous structures under controlled process parameters. In this review, general information of solution-based technologies, including freeze-drying, thermally or diffusion induced phase separation (TIPS or DIPS), and electrospinning, are presented, along with an overview of their technological strategies and applications. Furthermore, the differences in the fabricated constructs in terms of pore size and distribution, porosity, morphology, and mechanical and biological properties, are clarified and critically reviewed. Then, the combination of these techniques for obtaining scaffolds is described, offering the advantages of mimicking the unique architecture of tissues and organs that are intrinsically difficult to design

Molecular Hydrogen Formation on Low Temperature Surfaces in Temperature Programmed Desorption Experiments

The study of the formation of molecular hydrogen on low temperature surfaces is of interest both because it allows to explore elementary steps in the heterogeneous catalysis of a simple molecule and because of the applications in astrochemistry. Here we report results of experiments of molecular hydrogen formation on amorphous silicate surfaces using temperature-programmed desorption (TPD). In these experiments beams of H and D atoms are irradiated on the surface of an amorphous silicate sample. The desorption rate of HD molecules is monitored using a mass spectrometer during a subsequent TPD run. The results are analyzed using rate equations and the activation energies of the processes leading to molecular hydrogen formation are obtained from the TPD data. We show that a model based on a single isotope provides the correct results for the activation energies for diffusion and desorption of H atoms. These results can thus be used to evaluate the formation rate of H_2 on dust grains under the actual conditions present in interstellar clouds.Comment: 30 pages, 1 table, 6 figures. Published versio

Residence Time Distribution of Solid Particles in a High-Aspect Ratio Multiple-Impeller Stirred Vessel

Despite its importance, experimental information on the Residence Time Distribution (RTD) of solid particles in continuous-flow stirred vessels is still scant. In this work, experimental data on particle RTD in a high-aspect-ratio vessel stirred by three equally-spaced Rushton turbines, was obtained by means of a special technique named Twin System Approach (TSA). Quite surprisingly, results indicate that, among the various possibilities that could have been devised (e.g. 6, or 3, or 1 ideal tanks in series), the flow model closest to reality for the particle phase, at least in the experimental range here investigated, is that of a single perfectly stirred vessel

Pollutants removal from municipal sewage by means of microalgae

Microalgae are microorganisms able to photosynthesize, namely transforming inorganic substrates and sun light into organic compounds and chemical energy. The industry of microalgae has expanded in the last decades and several applications are now developed, making their biomass interesting under an economic perspective. Nannochlopsis gaditana is one of the most interesting species already employed in industry because of its high content in lipids that could be employed as source for biodiesel synthesis but also in other fields such as cosmetic and pharmaceutic. One of the most promising application is the exploitation of microalgal grow for bioremediating wastewaters polluted with inorganic nutrients such as nitrates and phosphates that microalgae are able to employ as nutrients. Bio-treatment of wastewaters by using microalgae has the advantage to reclassify the water and preserve it from wasting while producing a valuable biomass. In this work, a microalgal strain, Nannochloropsis gaditana, was employed for testing its performance in the bioremediation of municipal sewages. The wastewater was taken from a municipal plant, after the primary treatment, and the algae processing was aimed at replacing the secondary treatment. Algal growth in its growth medium and in the sewage was compared and algal biomass was characterized. Biochemical Oxygen Demand (BOD), Chemical Oxygen Demand (COD), total nitrogen and total phosphorous levels of the sewage before and after algae treatment were also determined in order to evaluate the efficiency of this microalgal strain on wastewater bioremediation. Our results showed that N. gaditana grows better in wastewater than in the control growth medium and it is able to efficiently remove nutrients from the sewage. However, COD and BOD values did not decrease after algal treatment. These results suggest that the use of selected bacteria and/or yeast strains (together with microalgae) could improve the efficiency of wastewater treatments decreasing BOD and COD values

A High-Throughput Mechanical Activator for Cartilage Engineering Enables Rapid Screening of in vitro Response of Tissue Models to Physiological and Supra-Physiological Loads

Articular cartilage is crucially influenced by loading during development, health, and disease. However, our knowledge of the mechanical conditions that promote engineered cartilage maturation or tissue repair is still incomplete. Current in vitro models that allow precise control of the local mechanical environment have been dramatically limited by very low throughput, usually just a few specimens per experiment. To overcome this constraint, we have developed a new device for the high throughput compressive loading of tissue constructs: the High Throughput Mechanical Activator for Cartilage Engineering (HiT-MACE), which allows the mechanoactivation of 6 times more samples than current technologies. With HiT-MACE we were able to apply cyclic loads in the physiological (e.g., equivalent to walking and normal daily activity) and supra-physiological range (e.g., injurious impacts or extensive overloading) to up to 24 samples in one single run. In this report, we compared the early response of cartilage to physiological and supra-physiological mechanical loading to the response to IL-1β exposure, a common but rudimentary in vitro model of cartilage osteoarthritis. Physiological loading rapidly upregulated gene expression of anabolic markers along the TGF-β1 pathway. Notably, TGF-β1 or serum was not included in the medium. Supra-physiological loading caused a mild catabolic response while IL-1β exposure drove a rapid anabolic shift. This aligns well with recent findings suggesting that overloading is a more realistic and biomimetic model of cartilage degeneration. Taken together, these findings showed that the application of HiT-MACE allowed the use of larger number of samples to generate higher volume of data to effectively explore cartilage mechanobiology, which will enable the design of more effective repair and rehabilitation strategies for degenerative cartilage pathologies

A Biodegradable, Bio-Based Polymer for the Production of Tools for Aquaculture: Processing, Properties and Biodegradation in Sea Water

Bio-based, biodegradable polymers can dramatically reduce the carbon dioxide released into the environment by substituting fossil-derived polymers in some applications. In this work, prototypes of trays for aquaculture applications were produced via injection molding by using a biodegradable polymer, Mater-Bi-(R). A characterization carried out via calorimetric, rheological and mechanical tests revealed that the polymer employed shows properties suitable for the production of tools to be used in aquaculture applications. Moreover, the samples were subjected to a biodegradation test in conditions that simulate the marine environment. The as-treated samples were characterized from gravimetrical, morphological and calorimetric point of views. The obtained data showed a relatively low biodegradation rate of the thick molded samples. This behavior is of crucial importance since it implies a long life in marine water for these manufacts before their disappearing

Physical and biological properties of electrospun poly(d,l-lactide)/nanoclay and poly(d,l-lactide)/nanosilica nanofibrous scaffold for bone tissue engineering

Electrospun scaffolds exhibiting high physical performances with the ability to support cell attachment and proliferation are attracting more and more scientific interest for tissue engineering applications. The inclusion of inorganic nanoparticles such as nanosilica and nanoclay into electrospun biopolymeric matrices can meet these challenging requirements. The silica and clay incorporation into polymeric nanofibers has been reported to enhance and improve the mechanical properties as well as the osteogenic properties of the scaffolds. In this work, for the first time, the physical and biological properties of polylactic acid (PLA) electrospun mats filled with different concentrations of nanosilica and nanoclay were evaluated and compared. The inclusion of the particles was evaluated through morphological investigations and Fourier transform infrared spectroscopy. The morphology of nanofibers was differently affected by the amount and kind of fillers and it was correlated to the viscosity of the polymeric suspensions. The wettability of the scaffolds, evaluated through wet contact angle measurements, slightly increased for both the nanocomposites. The crystallinity of the systems was investigated by differential scanning calorimetry highlighting the nucleating action of both nanosilica and nanoclay on PLA. Scaffolds were mechanically characterized with tensile tests to evaluate the reinforcing action of the fillers. Finally, cell culture assays with pre-osteoblastic cells were conducted on a selected composite scaffold in order to compare the cell proliferation and morphology with that of neat PLA scaffolds. Based on the results, we can convince that nanosilica and nanoclay can be both considered great potential fillers for electrospun systems engineered for bone tissue regeneration

Tubular composite scaffolds produced via Diffusion Induced Phase Separation (DIPS) as a shaping strategy for anterior cruciate ligaments reconstruction

Injuries of tendons and ligaments are common, especially among the young population. Anterior cruciate ligament (ACL) injuries do not heal due to its limited vascularization and hence, surgical intervention is usually required. The ideal scaffold for ligament tissue engineering (TE) should be biocompatible and possess mechanical and functional characteristics comparable to the native ACL. The Diffusion Induced Phase Separation (DIPS) technique allows the preparation of homogenous porous tubular scaffold with micro-pores using a rather simple procedure. Composites based on biodegradable polymers and bioglass have attracted much attention in tissue reconstruction and repair because of their biological and physicochemical advantages. In this work a new approach in ACL TE will be proposed focussing on the development of a suitable technique for in vitro seeding of lapine ACL fibroblasts into tubular-shaped instructive Poly-lactic-acid (PLLA) scaffolds, supplemented or not with bioglass (BG) 1393, produced via DIPS. Tubular composite scaffold (diameters: 1.2 and 2 mm, +/- BG) were obtained through a dip coating around a cylindrical support followed by a DIPS. An 8%wt PLLA/dioxane solution was prepared with 5%wt of BG-1393 as filler. Preliminary in vitro cell culture trials were carried out by seeding lapine ACL fibroblasts inside the scaffolds (2 cm as length) employing different seeding strategies in order to find the best way that allows to obtain a homogeneous fibroblast distribution inside the tubes. (1) First trials consisted in the inoculating of the cell suspension inside the tubes and maintaining them in dynamical culture. (2) The second one was done by suspending the cells in a fibrin gel polymerized within the tubes by using of thrombin. (3) The third approach was carried out by using cell spheroids (three-dimensional self-assembled cell agglomerates). Cell attachment, viability and morphology were examined by live-death and Hematoxylin/Eosin stainings after 1, 7, 14 d and vimentin immunolabelings (7 d). Scanning electron microscopical analysis revealed that the internal surface of the tubes was homogeneously structured with micropores sized around 5 µm and a mean thickness of the wall of 60 µm. The results showed cell adhesion to the wall of the tubes with all seeding techniques applied even though with fibrin gel it was more homogenous. Furthermore, colonized areas expanded with culture time and the majority of cell survived irrespectively of seeding techniques. (1) In inoculation phase, many cells left the scaffold and attached on the plate. Even after the dynamic culture (rotating device) most cells covered only half the tube inner surface. (2) In the second trial, a fibrin gel was used to achieve a homogenous cell distribution during seeding. In the early stage (48 h) cells remained captured inside the fibrin, but after 7 d they become elongated and migrated from the fibrin to the inner tube surface forming a compact cell layer. So, the fibrin appears helpful to achieve an immediate high cell seeding efficiency and an almost homogeneous cell distribution inside the tubes. (3) Although using the spheroid technique the scaffold internal surface was not homogeneously colonized with cells, after 7 d cell migration to the inner scaffold surface from the attaching spheroids could be observed. In longitudinal sections cells were elongated like typical ligament fibroblasts parallel to the longitudinal tube axis. Therefore, it can be affirmed that employment of tubular scaffolds produced by DIPS could be a promising approach of ligament TE. In the future, it would be interesting to evaluate the effectiveness of seeding by combining the spheroids and the fibrin gel

Current strategies for tracheal replacement: A review

Airway cancers have been increasing in recent years. Tracheal resection is commonly performed during surgery and is burdened from post-operative complications severely affecting quality of life. Tracheal resection is usually carried out in primary tracheal tumors or other neoplasms of the neck region. Regenerative medicine for tracheal replacement using bio-prosthesis is under current research. In recent years, attempts were made to replace and transplant human cadaver trachea. An effective vascular supply is fundamental for a successful tracheal transplantation. The use of biological scaffolds derived from decellularized tissues has the advantage of a three-dimensional structure based on the native extracellular matrix promoting the perfusion, vascularization, and differentiation of the seeded cell typologies. By appropriately modulating some experimental parameters, it is possible to change the characteristics of the surface. The obtained membranes could theoretically be affixed to a decellularized tissue, but, in practice, it needs to ensure adhesion to the biological substrate and/or glue adhesion with biocompatible glues. It is also known that many of the biocompatible glues can be toxic or poorly tolerated and induce inflammatory phenomena or rejection. In tissue and organ transplants, decellularized tissues must not produce adverse immunological reactions and lead to rejection phenomena; at the same time, the transplant tissue must retain the mechanical properties of the original tissue. This review describes the attempts so far developed and the current lines of research in the field of tracheal replacement