Jakub Jaroszewicz

3D bioprinting of BM-MSCs-loaded ECM biomimetic hydrogels for in vitro neocartilage formation.

In this work we demonstrate how to print 3D biomimetic hydrogel scaffolds for cartilage tissue engineering with high cell density (>10(7) cells ml(-1)), high cell viability (85 ÷ 90%) and high printing resolution (≈100 μm) through a two coaxial-needles system. The scaffolds were composed of modified biopolymers present in the extracellular matrix (ECM) of cartilage, namely gelatin methacrylamide (GelMA), chondroitin sulfate amino ethyl methacrylate (CS-AEMA) and hyaluronic acid methacrylate (HAMA). The polymers were used to prepare three photocurable bioinks with increasing degree of biomimicry: (i) GelMA, (ii) GelMA + CS-AEMA and (iii) GelMA + CS-AEMA + HAMA. Alginate was added to the bioinks as templating agent to form stable fibers during 3D printing. In all cases, bioink solutions were loaded with bone marrow-derived human mesenchymal stem cells (BM-MSCs). After printing, the samples were cultured in expansion (negative control) and chondrogenic media to evaluate the possible differentiating effect exerted by the biomimetic matrix or the synergistic effect of the matrix and chondrogenic supplements. After 7, 14, and 21 days, gene expression of the chondrogenic markers (COL2A1 and aggrecan), marker of osteogenesis (COL1A1) and marker of hypertrophy (COL10A1) were evaluated qualitatively by means of fluorescence immunocytochemistry and quantitatively by means of RT-qPCR. The observed enhanced viability and chondrogenic differentiation of BM-MSCs, as well as high robustness and accuracy of the employed deposition method, make the presented approach a valid candidate for advanced engineering of cartilage tissue.

Highly ordered and tunable polyHIPEs by using microfluidics

We demonstrate how to generate highly ordered porous matrices from dextran-methacrylate (DEX-MA) using microfluidics. We use a flow focusing device to inject an aqueous solution of DEX-MA and surfactant to break the flow of an organic solvent (cyclohexane) into monodisperse droplets at a high volume fraction (above 74% v/v) to form an ordered high internal phase emulsion (HIPE). We collect the crystalline HIPE structure and freeze it by gelling. The resulting polyHIPEs are characterized by an interconnected and ordered morphology. The size of pores and interconnects ranges between hundreds and tens of micrometers, respectively. The technique that we describe allows for precise tuning of all the structural parameters of the matrices, including their porosity, the size of the pores and the lumen of interconnects between the pores. The resulting ordered and precisely tailored HIPE gels represent a new class of scaffolds for applications in tissue engineering.

Microfluidic Foaming: A Powerful Tool for Tailoring the Morphological and Permeability Properties of Sponge-like Biopolymeric Scaffolds

Ordered porous polymeric materials can be engineered to present highly ordered pore arrays and uniform and tunable pore size. These features prompted a number of applications in tissue engineering, generation of meta materials, and separation and purification of biomolecules and cells. Designing new and efficient vistas for the generation of ordered porous materials is an active area of research. Here we investigate the potential of microfluidic foaming within a flow-focusing (FF) geometry in producing 3D regular sponge-like polymeric matrices with tailored morphological and permeability properties. The challenge in using microfluidic systems for the generation of polymeric foams is in the high viscosity of the continuous phase. We demonstrate that as the viscosity of the aqueous solution increases, the accessible range of foam bubble fraction (Φb) and bubble diameter (Db) inside the microfluidic chip tend to narrow progressively. This effect limits the accessible range of geometric properties of the resulting materials. We further show that this problem can be rationally tackled by appropriate choice of the concentration of the polymer. We demonstrate that via such optimization, the microfluidic assisted synthesis of porous materials becomes a facile and versatile tool for generation of porous materials with a wide range of pore size and pore volume. Moreover, we demonstrate that the size of interconnects among pores-for a given value of the gas fraction-can be tailored through the variation of surfactant concentration. This, in turn, affects the permeability of the materials, a factor of key importance in flow-through applications and in tissue engineering.

Correlation between porous texture and cell seeding efficiency of gas foaming and microfluidic foaming scaffolds

In the design of scaffolds for tissue engineering applications, morphological parameters such as pore size, shape, and interconnectivity, as well as transport properties, should always be tailored in view of their clinical application. In this work, we demonstrate that a regular and ordered porous texture is fundamental to achieve an even cell distribution within the scaffold under perfusion seeding. To prove our hypothesis, two sets of alginate scaffolds were fabricated using two different technological approaches of the same method: gas-in-liquid foam templating. In the first one, foam was obtained by insufflating argon in a solution of alginate and a surfactant under stirring. In the second one, foam was generated inside a flow-focusing microfluidic device under highly controlled and reproducible conditions. As a result, in the former case the derived scaffold (GF) was characterized by polydispersed pores and interconnects, while in the latter (μFL), the porous structure was highly regular both with respect to the spatial arrangement of pores and interconnects and their monodispersity. Cell seeding within perfusion bioreactors of the two scaffolds revealed that cell population inside μFL scaffolds was quantitatively higher than in GF. Furthermore, seeding efficiency data for μFL samples were characterized by a lower standard deviation, indicating higher reproducibility among replicates. Finally, these results were validated by simulation of local flow velocity (CFD) inside the scaffolds proving that μFL was around one order of magnitude more permeable than GF.

Insights into the macroporosity of freeze-cast hierarchical geopolymers

Geopolymer monoliths with controlled lamellar macroporosity and total porosity ranging from 60% to 70% were prepared by ice-templating a partially geopolymerized slurry. Both the maturation treatment of the starting mixture and the water specifically added for freeze-casting were tailored to modify both the geopolymerization and viscosity of the slurry, and, consequently, its freezing behavior, in order to optimize the final lamellar architecture. Following a room temperature maturation treatment, a 50% water content added for freezing developed thick lamellae and wide pores. A lower water content (30%) and curing at 80 °C after maturation at room temperature (for both 50% and 30% H2O) was conducive to a narrow lamellar pore width distribution in the 30–130 μm range. However, the consumption of water due to geopolymerization in samples cured at 80 °C led to a decreased length and thickness of the lamellae. Lastly, the interparticle meso- and macropores (0.003 to 1 μm) within the geopolymer lamellae were only slightly modified by the maturation treatment.

Nanocrystalline Cu-Al2O3 Composites Sintered by the Pulse Plasma Technique

The paper presents the results of examination of the structure and properties of nanocrystalline Cu-Al2O3 composites with the two different Al2O3 contents: 10 and 20 vol.%. The composites were produced using a mixture of copper and Al2O3 powders with an average crystallite size of about 60nm for Cu and about 40nm for Al2O3. The powders were consolidated by pulse plasma sintering (PPS) for 5 minutes at a temperature of 650oC under a load of 60 MPa. Irrespective of the volumetric content of Al2O3, the relative density of the composites was about 92%, and the average Cu crystallite size was about 80nm. The hardness of the composites varied with the volumetric content of Al2O3, and was equal to 270 HV0.1 for 20 and to 240 HV0.1 for 10% of Al2O3. The Cu-20%Al2O3 composite had a resistivity of 0.386 while that with 10% of Al2O3 was 0.149 56m.

Pulse Plasma Sintering of Nano-Crystalline Cu Powder

Nanocrystalline copper powders, produced by the reduction of the CuO with hydrogen, were consolidated using the pulse plasma sintering (PPS) method. The sintering process was carried out at temperatures between 500 and 900 oC under a load of 60 MPa for 5 min. The average crystallite size of the sintered component obtained at 500 oC was about 80nm and at 900 oC 1880 nm. The components produced at 500 oC had a relative density of 90 %, and those sintered at 900 oC 92 %; their hardness was 215 and 140 HV0.1, respectively.

Characterization of Single-Crystal Dendrite Structure and Porosity in Nickel-Based Superalloys Using X-Ray Micro-Computed Tomography

X-ray micro-computed tomography system has been used for visualization in two- (2-D) and three dimensions (3-D) of the dendrite structure and pores in single-crystals fabricated by Bridgman investment casting technique. The system described in the paper reconstructs 3-D geometry from a set of 2-D images obtained by multiple slicing of an X-ray radiography image. The results obtained in this study demonstrate the effect of withdrawal rate on the primary dendrite arm spacing and porosity in single-crystal made of CMSX-4 alloy.

Osteogenesis around CaP-coated titanium implants visualized using 3D histology and micro-computed tomography

Calcium phosphate (CaP) coatings can enhance the performance of bone implants in compromised conditions, such as osteoporosis. Therefore, this study compared non-coated vs. CaP-coated (n = 8) titanium implants in osteoporotic ovariectomized (OVX) rats. Bone volume (BV) was assessed using micro-computer tomography (micro-CT) and three-dimensional (3D) histology, in three zones from the implant surface. Bone remodeling was assessed using fluorochrome labels and osteoclast staining. Micro-CT and 3D histology showed a BV reduction in OVX animals, of respectively 22.4 and 10.5%. BV was significantly increased inside all zones around CaP coatings, especially in the inner zone of the OVX animals. Fluorochrome labels were predominantly seen when the coating was applied. Osteoclasts were mainly found in the area remote from the surface of non-coated implants in control animals. For the coated implants, osteoclasts were distributed evenly, and present in direct vicinity of the surface. In conclusion, 3D histology is a suitable technique to obtain data and insight into bone architecture around implants at relatively high resolution. Bone formation was significantly reduced in osteoporotic animals. CaP coatings resulted in a higher BV directly around implants installed in osteoporotic animals, enhanced turnover, and a shift of remodeling activity toward the implant surface.

Skeletal Ontogeny in Basal Scleractinian Micrabaciid Corals

The skeletal ontogeny of the Micrabaciidae, one of two modern basal scleractinian lineages, is herein reconstructed based on serial micro‐computed tomography sections and scanning electron micrographs. Similar to other scleractinians, skeletal growth of micrabaciids starts from the simultaneous formation of six primary septa. New septa of consecutive cycles arise between septa of the preceding cycles from unique wedge‐shaped invaginations of the wall. The invagination of wall and formation of septa are accompanied by development of costae alternating in position with septa. During corallite growth, deepening invagination of the wall results in elevation of septa above the level of a horizontal base. The corallite wall is regularly perforated thus invaginated regions consist of pillars inclined downwardly and outwardly from the lower septal margins. Shortly after formation of septa (S2 and higher cycles) their upper margins bend and fuse with the neighboring members of a previous cycle, resulting in a unique septal pattern, formerly misinterpreted as “septal bifurcation.” Septa as in other Scleractinia are hexamerally arranged in cycles. However, starting from the quaternaries, septa within single cycles do not appear simultaneously but are inserted in pairs and successively flank the members of a preceding cycle, invariably starting from those in the outermost parts of the septal system. In each pair, the septum adjacent to older septa arises first (e.g., the quinaries between S2 and S4 before quinaries between S3 and S4). Unique features of micrabaciid skeletal ontogeny are congruent with their basal position in scleractinian phylogeny, which was previously supported by microstructural and molecular data. J. Morphol., 2013.