Migration of parasitic worms through the host tissues, which may occasionally result in fatal damage to the internal organs, represents one of the major risks associated with helminthoses. In order to track the parasites, traditionally used 2D imaging techniques such as histology or squash preparation do not always provide sufficient data to describe worm location/behavior in the host. On the other hand, 3D imaging methods are widely used in cell biology, medical radiology, osteology or cancer research, but their use in parasitological research is currently occasional. Thus, we aimed at the evaluation of suitability of selected 3D methods to monitor migration of the neuropathogenic avian schistosome Trichobilharzia regenti in extracted spinal cord of experimental vertebrate hosts. All investigated methods, two of them based on tracking of fluorescently stained larvae with or without previous chemical clearing of tissue and one based on X-ray micro-CT, exhibit certain limits for in vivo observation. Nevertheless, our study shows that the tested methods as ultramicroscopy (used for the first time in parasitology) and micro-CT represent promising tool for precise analyzing of parasite larvae in the CNS. Synthesis of these 3D imaging techniques can provide more comprehensive look at the course of infection, host immune response and pathology caused by migrating parasites within entire tissue samples, which would not be possible with traditional approaches.

• Klíčová slova
• 3D imaging, Light sheet microscopy, Neuroinfection, Parasite migration, Trichobilharzia regenti, X-ray micro-CT,
• MeSH
• infekce červy třídy Trematoda veterinární   MeSH
• larva MeSH
• nemoci zvířat diagnóza   parazitologie   MeSH
• obratlovci MeSH
• protozoární infekce centrálního nervového systému veterinární   MeSH
• Schistosomatidae * MeSH
• zobrazování trojrozměrné metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The authors report on three cases in which a custom-made 3D printed titanium acetabular component of total hip arthroplasty was used to manage an advanced acetabular bone defect with pelvic discontinuity. The implant surface structure impeded long-term bone integration. Nonetheless, the stable bridging of the acetabular defect resulted in full integration of impacted bone allografts at the base of the implant. The pelvic continuity was restored within 12 months after surgery, and thus the acetabulum was prepared for potential further implantation of a standard revision acetabular component. Only one of the three female patients underwent a revision surgery at 18 months after surgery, the other two female patients were satisfied to such a degree with the clinical outcome at 6 years and 5 years, respectively, after surgery that they refused to undertake the revision surgery, despite X-ray images showing signs of loosening of the custom-made titanium acetabular component. The authors concluded that the implantation technique of three-point fixed custom-made 3D printed acetabular component made of titanium combined with impaction grafting of the acetabular base is a good alternative in managing the advanced bone defects of acetabulum with pelvic discontinuity after the failure of total hip arthroplasty. Even though inadequate surface porosity of the thus produced component did not allow its permanent osteointegration, the assembly was stable enough to allow the bone allografts to rebuild and restore continuity of the pelvis and facilitated future implantation of the standard revision acetabular component. Key words: 3D printing, individual acetabular component, titanium, total hip prosthesis, revision hip arthroplasty, acetabular reconstruction, custom-made implants.

• MeSH
• 3D tisk MeSH
• acetabulum diagnostické zobrazování   chirurgie   MeSH
• kyčelní protézy * MeSH
• lidé MeSH
• náhrada kyčelního kloubu * MeSH
• následné studie MeSH
• protézy - design MeSH
• reoperace MeSH
• selhání protézy MeSH
• titan MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• titan MeSH

Additive manufacturing, also called 3D printing, is an effective method for preparing scaffolds with defined structure and porosity. The disadvantage of the technique is the excessive smoothness of the printed fibers, which does not support cell adhesion. In the present study, a 3D printed scaffold was combined with electrospun classic or structured nanofibers to promote cell adhesion. Structured nanofibers were used to improve the infiltration of cells into the scaffold. Electrospun layers were connected to 3D printed fibers by gluing, thus enabling the fabrication of scaffolds with unlimited thickness. The composite 3D printed/nanofibrous scaffolds were seeded with primary chondrocytes and tested in vitro for cell adhesion, proliferation and differentiation. The experiment showed excellent cell infiltration, viability, and good cell proliferation. On the other hand, partial chondrocyte dedifferentiation was shown. Other materials supporting chondrogenic differentiation will be investigated in future studies.

• Klíčová slova
• 3D printing, cell infiltration, chondrocytes, electrospinning, nanofibres,
• MeSH
• 3D tisk * MeSH
• buněčná adheze fyziologie   MeSH
• buněčná diferenciace fyziologie   MeSH
• chondrocyty cytologie   MeSH
• kultivované buňky fyziologie   MeSH
• lidé MeSH
• nanovlákna * chemie   MeSH
• proliferace buněk fyziologie   MeSH
• tkáňové inženýrství metody   MeSH
• tkáňové podpůrné struktury * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Primary human B cells chronic lymphocytic leukemia undergoes apoptosis, from which they can be rescued by contact with stromal cells or by the addition of specific soluble factor, when cultured in vitro. For research purposes of the behavior of CLL cells we created 3D in vitro model in which we simulated appropriate microenvironment for CLL cells to allow study the mechanism of survival of these cells in long-term cultivation. MATERIAL AND METHODS: Our aim was the scaffold structure to be geometrically similar to the 3D morphology of supporting bone marrow tissue in a trabecular bone; the 3D scaffold was also designed to conform to biocompatibility, sufficiently large surface area for cell attachment, high porosity for cell migration, proliferation and transport of nutrients. Another requirement was a partial transparency for inspection of cell model with optical techniques. We prepared 3D scaffolds from porous hydrogel poly (2-hydroxyethyl methacrylate) (pHEMA), poly (2-hydroxyethyl methacrylate-co-2-aminoethyl methacrylate) p (HEMA-co-AEMA) and p (HEMA-co-AEMA) modified with frequently used cell adhesion peptide Arg-Gly-Asp (RGD). All hydrogel scaffolds were manufactured in four pore diameters (125, 200, 300 and 350-450 μm). Scaffolds were tested with human bone marrow stromal cell line HS-5 and human embryonic kidney cell line HEK293. RESULTS: Hydrogel scaffold p (HEMA-co-AEMA) modified with adhesion peptide Arg-Gly-Asp (RGD) with pore diameter of 350-450 μm demonstrated that it is a convenient system for 3D cell cultivation, since it promotes interaction between the cells and also between the cells and the material. This scaffold was used for seeding of co-cultivation system of HS-5 cells with CLL-cells, which were stimulated through the CD40L signaling pathway as well as via the IL-4 pathway. Viability of B-CLL cells was higher in the presence of both stimulators than with each alone. CONCLUSIONS: We have shown that 3D scaffold technology is very useful for modeling of microsystems where the cancer cells behave like in their natural microenvironment.Key words: hematooncology - leukemia - hydrogel - stromal cells This work was supported by grant COST CZ LD15144 "Cellular and acellular grounds for regeneration of bones and teeth" awarded by the Ministry of Education, Youth and Sport of the Czech Republic. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 6. 3. 2017Accepted: 26. 3. 2017.

• MeSH
• B-lymfocyty patologie   MeSH
• chronická lymfatická leukemie patologie   MeSH
• HEK293 buňky MeSH
• hydrogely chemie   MeSH
• kokultivační techniky MeSH
• lidé MeSH
• proliferace buněk MeSH
• tkáňové podpůrné struktury * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• hydrogely MeSH

Total hip arthroplasty (THA) significantly improves the quality of life in majority of patients with severe osteoarthritis. However, long-term outcomes of THAs are compromised by aseptic loosening and periprosthetic osteolysis which needs revision surgery. Both of these are causally linked to a prosthetic wear deliberated from the prosthetic articulating surfaces. As a result, there is a need to measure the mode and magnitude of wear. The paper evaluates three optical methods proposed for construction of a device for the non-contact prosthetic wear measurement. Of them, the scanning profilometry achieved promising combination of accuracy and repeatability. Simultaneously, it is time efficient to enable the development of a sensor for wear measurement.

• MeSH
• algoritmy MeSH
• artróza kyčelních kloubů chirurgie   MeSH
• Fourierova analýza MeSH
• kyčelní protézy * MeSH
• lidé MeSH
• mikroskopie metody   MeSH
• náhrada kyčelního kloubu přístrojové vybavení   MeSH
• osteolýza MeSH
• protézy - design MeSH
• reoperace MeSH
• reprodukovatelnost výsledků MeSH
• selhání protézy * MeSH
• software MeSH
• teoretické modely MeSH
• zobrazování trojrozměrné metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

OBJECTIVES: We prepared 3D poly (ε-caprolactone) (PCL) nanofibre scaffolds and tested their use for seeding, proliferation, differentiation and migration of mesenchymal stem cell (MSCs). MATERIALS AND METHODS: 3D nanofibres were prepared using a special collector for common electrospinning; simultaneously, a 2D PCL nanofibre layer was prepared using a classic plain collector. Both scaffolds were seeded with MSCs and biologically tested. MSC adhesion, migration, proliferation and osteogenic differentiation were investigated. RESULTS: The 3D PCL scaffold was characterized by having better biomechanical properties, namely greater elasticity and resistance against stress and strain, thus this scaffold will be able to find broad applications in tissue engineering. Clearly, while nanofibre layers of the 2D scaffold prevented MSCs from migrating through the conformation, cells infiltrated freely through the 3D scaffold. MSC adhesion to the 3D nanofibre PCL layer was also statistically more common than to the 2D scaffold (P < 0.05), and proliferation and viability of MSCs 2 or 3 weeks post-seeding, were also greater on the 3D scaffold. In addition, the 3D PCL scaffold was also characterized by displaying enhanced MSC osteogenic differentiation. CONCLUSIONS: We draw the conclusion that all positive effects observed using the 3D PCL nanofibre scaffold are related to the larger fibre surface area available to the cells. Thus, the proposed 3D structure of the nanofibre layer will find a wide array of applications in tissue engineering and regenerative medicine.

• MeSH
• buněčná diferenciace * MeSH
• buněčné kultury přístrojové vybavení   metody   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• nanovlákna chemie   ultrastruktura   MeSH
• osteogeneze MeSH
• osteokalcin metabolismus   MeSH
• pohyb buněk MeSH
• polyestery chemie   MeSH
• povrchové vlastnosti MeSH
• proliferace buněk MeSH
• pružnost MeSH
• regenerativní lékařství MeSH
• sialoprotein vázající integrin metabolismus   MeSH
• tkáňové inženýrství MeSH
• tkáňové podpůrné struktury * MeSH
• viabilita buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• osteokalcin MeSH
• polycaprolactone MeSH Prohlížeč
• polyestery MeSH
• sialoprotein vázající integrin MeSH

The plasticity of cancer cell invasion represents substantial hindrance for effective anti-metastatic therapy. To better understand the cancer cells' plasticity, we performed complex transcriptomic and proteomic profiling of HT1080 fibrosarcoma cells undergoing mesenchymal-amoeboid transition (MAT). As amoeboid migratory phenotype can fully manifest only in 3D conditions, all experiments were performed with 3D collagen-based cultures. Two previously described approaches to induce MAT were used: doxycycline-inducible constitutively active RhoA expression and dasatinib treatment. RNA sequencing was performed with ribo-depleted total RNA. Protein samples were analysed with tandem mass tag (TMT)-based mass spectrometry. The data provide unprecedented insight into transcriptome and proteome changes accompanying MAT in true 3D conditions.

• MeSH
• fibrosarkom patologie   MeSH
• invazivní růst nádoru * MeSH
• kolagen chemie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• pohyb buněk * MeSH
• proteom * MeSH
• rhoA protein vázající GTP MeSH
• sekvenční analýza RNA MeSH
• tandemová hmotnostní spektrometrie MeSH
• transkriptom * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• dataset MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kolagen MeSH
• proteom * MeSH
• rhoA protein vázající GTP MeSH
• RHOA protein, human MeSH Prohlížeč

Observation and analysis of cancer cell behaviour in 3D environment is essential for full understanding of the mechanisms of cancer cell invasion. However, label-free imaging of live cells in 3D conditions is optically more challenging than in 2D. Quantitative phase imaging provided by coherence controlled holographic microscopy produces images with enhanced information compared to ordinary light microscopy and, due to inherent coherence gate effect, enables observation of live cancer cells' activity even in scattering milieu such as the 3D collagen matrix. Exploiting the dynamic phase differences method, we for the first time describe dynamics of differences in cell mass distribution in 3D migrating mesenchymal and amoeboid cancer cells, and also demonstrate that certain features are shared by both invasion modes. We found that amoeboid fibrosarcoma cells' membrane blebbing is enhanced upon constriction and is also occasionally present in mesenchymally invading cells around constricted nuclei. Further, we demonstrate that both leading protrusions and leading pseudopods of invading fibrosarcoma cells are defined by higher cell mass density. In addition, we directly document bundling of collagen fibres by protrusions of mesenchymal fibrosarcoma cells. Thus, such a non-invasive microscopy offers a novel insight into cellular events during 3D invasion.

• MeSH
• buněčná membrána metabolismus   MeSH
• buněčné kultury metody   MeSH
• fibrosarkom diagnostické zobrazování   patologie   MeSH
• holografie přístrojové vybavení   metody   MeSH
• intravitální mikroskopie přístrojové vybavení   metody   MeSH
• invazivní růst nádoru diagnostické zobrazování   patologie   MeSH
• kolagen metabolismus   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• pohyb buněk * MeSH
• pseudopodia metabolismus   MeSH
• zobrazování trojrozměrné přístrojové vybavení   metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kolagen MeSH

Cranial neural crest cells populate the future facial region and produce ectomesenchyme-derived tissues, such as cartilage, bone, dermis, smooth muscle, adipocytes, and many others. However, the contribution of individual neural crest cells to certain facial locations and the general spatial clonal organization of the ectomesenchyme have not been determined. We investigated how neural crest cells give rise to clonally organized ectomesenchyme and how this early ectomesenchyme behaves during the developmental processes that shape the face. Using a combination of mouse and zebrafish models, we analyzed individual migration, cell crowd movement, oriented cell division, clonal spatial overlapping, and multilineage differentiation. The early face appears to be built from multiple spatially defined overlapping ectomesenchymal clones. During early face development, these clones remain oligopotent and generate various tissues in a given location. By combining clonal analysis, computer simulations, mouse mutants, and live imaging, we show that facial shaping results from an array of local cellular activities in the ectomesenchyme. These activities mostly involve oriented divisions and crowd movements of cells during morphogenetic events. Cellular behavior that can be recognized as individual cell migration is very limited and short-ranged and likely results from cellular mixing due to the proliferation activity of the tissue. These cellular mechanisms resemble the strategy behind limb bud morphogenesis, suggesting the possibility of common principles and deep homology between facial and limb outgrowth.

• Klíčová slova
• Early face development, clonal envelopes, embryonic development, migration, morphogenesis, neural crest cells,
• MeSH
• anatomické modely MeSH
• buněčná diferenciace * MeSH
• buněčné klony cytologie   MeSH
• crista neuralis cytologie   MeSH
• dánio pruhované MeSH
• ektoderm cytologie   embryologie   MeSH
• exprese genu MeSH
• fenotyp MeSH
• mezoderm cytologie   embryologie   MeSH
• morfogeneze * MeSH
• myši MeSH
• obličej embryologie   MeSH
• organogeneze * MeSH
• pohyb buněk MeSH
• reportérové geny MeSH
• zobrazování trojrozměrné MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

This paper investigates the effect of plasticizer structure on especially the printability and mechanical and thermal properties of poly(3-hydroxybutyrate)-poly(lactic acid)-plasticizer biodegradable blends. Three plasticizers, acetyl tris(2-ethylhexyl) citrate, tris(2-ethylhexyl) citrate, and poly(ethylene glycol)bis(2-ethylhexanoate), were first checked whether they were miscible with poly(3-hydroxybutyrate)-poly(lactic acid) (PHB-PLA) blends using a kneading machine. PHB-PLA-plasticizer blends of 60-25-15 (wt.%) were then prepared using a corotating meshing twin-screw extruder, and a single screw extruder was used for filament preparation for further three-dimensional (3D) fused deposition modeling (FDM) printing. These innovative eco-friendly PHB-PLA-plasticizer blends were created with a majority of PHB, and therefore, poor mechanical properties and thermal properties of neat PHB-PLA blends were improved by adding appropriate plasticizer. The plasticizer also influences the printability of blends, which was investigated, based on our new specific printability tests developed for the optimization of printing conditions (especially printing temperature). Three-dimensional printed test samples were used for heat deflection temperature measurements and Charpy and tensile-impact tests. Plasticizer migration was also investigated. The macrostructure of 3D printed samples was observed using an optical microscope to check the printing quality and printing conditions. Tensile tests of 3D printed samples (dogbones), as well as extruded filaments, showed that measured elongation at break raised, from 21% for non-plasticized PHB-PLA reference blends to 84% for some plasticized blends in the form of filaments and from 10% (reference) to 32% for plasticized blends in the form of printed dogbones. Measurements of thermal properties (using modulated differential scanning calorimetry and oscillation rheometry) also confirmed the plasticizing effect on blends. The thermal and mechanical properties of PHB-PLA blends were improved by the addition of appropriate plasticizer. In contrast, the printability of the PHB-PLA reference seems to be slightly better than the printability of the plasticized blends.

• Klíčová slova
• 3D printing, biodegradable polymeric blends, citrates, plasticizers, poly(ethylene glycol) PEG, poly(hydroxybutyrate)-poly(lactic), printability,
• Publikační typ
• časopisecké články MeSH