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

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.

• 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

Východiska: Primární lidské B buňky chronické lymfocytární leukemie (CLL) podléhají při kultivaci in vitro buněčné smrti, nicméně jejich přežití lze signifikantně prodloužit kontaktem se stromálními buňkami nebo přítomností specifických solubilních faktorů. Pro účely výzkumu chování CLL buněk jsme vytvořili 3D in vitro model, ve kterém bylo simulováno vhodné mikroprostředí pro CLL buňky umožňující studium mechanizmu jejich přežívání v dlouhodobé kultivaci. Materiál a metody: Naším cílem bylo, aby struktura scaffoldu byla geometricky podobná 3D morfologii kostní dřeně, která vyplňuje trabekulární kost, aby měl 3D scaffold dostatečně velký povrch pro zachycení buněk a zároveň velkou pórovitost pro buněčnou migraci a transport živin. Dalším požadavkem byla také alespoň částečná transparentnost potřebná pro pozorování buněčného modelu pomocí optických metod. Připravili jsme 3D scaffoldy z porózního hydrogelu poly (2-hydroxyetyl metakrylát) (pHEMA), poly (2-hydroxyetyl metakrylát-co-2-aminoetyl metakrylát) p (HEMA-co-AEMA) a p (HEMA-co-AEMA) modifikovaný s často používaným adhezním peptidem Arg-Gly-Asp (RGD). Všechny hydrogelové scaffoldy byly vyrobeny ve čtyřech velikostech pórů (125, 200, 300 a 350–450 μm). Scaffoldy byly testovány pomocí HS-5 buněčné linie odvozené z lidských stromálních buněk kostní dřeně a HEK293 buněčné linie odvozené z lidských embryonálních buněk ledvin. Výsledky: Hydrogelový scaffold p (HEMA-co-AEMA) modifikovaný adhezním peptidem Arg-Gly-Asp (RGD) s velikostí pórů 350–450 μm prokázal, že je vhodným systémem pro 3D kultivace buněk, neboť podporuje interakce mezi buňkami navzájem a také mezi buňkami a materiálem. Tento scaffold byl použit pro nasazení kultivace složené z HS-5 buněk a CLL buněk, které byly stimulovány pomocí ligandu CD40 a cytokinu IL-4. Viabilita CLL buněk byla vyšší v přítomnosti obou stimulátorů zároveň než v případě každého zvlášť. Závěr: Ukázali jsme, že technologie 3D scaffoldů je velmi dobře využitelná pro modelování mikrosystémů, kde se nádorové buňky chovají jako ve svém přirozeném mikroprostředí. Klíčová slova: hematoonkologie – leukemie – hydrogel – stromální buňky

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. 2017 Accepted: 26. 3. 2017

• MeSH
• biokompatibilní materiály MeSH
• biologické modely MeSH
• chronická lymfatická leukemie patologie   MeSH
• hydrogely * MeSH
• kultivační techniky metody   MeSH
• mezenchymální kmenové buňky MeSH
• nádorové buňky kultivované * MeSH
• nádorové mikroprostředí MeSH
• techniky in vitro MeSH
• tkáňové podpůrné struktury * MeSH
• Publikační typ
• práce podpořená grantem 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.

• 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

INTRODUCTION: For fractures of the posterior part of the pelvis and disjunction of the sacroiliac joint, iliosacral screws are used, which transfix the sacral process of the hip bone with the sacrum in the region of the S1 or S2 segment. The correct placement of these screws in the sacrum is important in order to avoid injury to the sacral nerve fibres or soft tissue injury ventrally and dorsally to the sacrum. 3D navigation provides the possibility of more precise control of drilling and screw insertion. It works on the basis of a 3D scan taken directly in the operating room using a C-arm with the possibility of 3D scanning. The aim of the work is to determine the operating times, the dose of intraoperative X-ray radiation and also the occurrence of postoperative complications during the introduction of iliosacral screws into the posterior segment of the pelvis under the control of 3D navigation. MATERIAL AND METHODS: In the years 2014–2020, we performed 13 osteosyntheses of the posterior segment of the pelvis using iliosacral screws under the control of 3D navigation at the Department of Surgery and Trauma and Emergency Surgery. We performed osteosynthesis using one or two cannulated screws with a diameter of 7.0 mm. The Ziehm Flat panel 3D fluoroscopy device and Vector Vision computer navigation were used during the surgery. For individual surgeries, we monitored the operating time in minutes, the dose of intraoperative X-ray radiation in cGY per cm2, as well as the time of intraoperative fluoroscopy in seconds. Postoperatively, we performed an X-ray check of the location of the iliosacral screws in four projections, in case of uncertainty, we clarified the position of the screws by CT examination. We monitored the incidence of postoperative complications within 90 days after surgery. One year after the surgery, we performed a control X-ray of the pelvis in four projections with the aim of detecting possible migration of osteosynthetic material or dislocation of the posterior segment of the pelvis after osteosynthesis. RESULTS: The group of 13 patients consisted of eight men and five women. All fractures were type C according to the AO-OTA classification. Eight patients (74 %) underwent osteosynthesis with one screw and five patients (26 %) with two screws. The median age of all patients in the cohort is 51 years (range: 33–73 years). The median weight of all patients in the cohort is 83 kg (range: 65–107 kg). The median intraoperative X-ray time of all patients is 1.14 min (range: 0.50–2.35 min). The median X-ray radiation dose is 853 cGy/cm2 (range: 591–1369 cGy/cm2). The median total operative time is 42 minutes (range: 28–62 min). CONCLUSION: Osteosynthesis of the posterior segment of the pelvis using 3D navigation is more accurate than fluoroscopic control of this surgical procedure and also 2D navigation because it allows drilling to be monitored in individual sections of the sacral bone. It is possible to assess the intraosseous position throughout the drilling of the channel for the iliosacral screw. In a small group of patients, we did not observe post-operative complications in the sense of injury to neurovascular structures and also malposition of screws.

• Klíčová slova
• Vector Vision,
• MeSH
• chirurgie s pomocí počítače metody   MeSH
• kostní šrouby MeSH
• lidé MeSH
• sakroiliakální kloub chirurgie   zranění   MeSH
• sakrokokcygeální krajina chirurgie   zranění   MeSH
• vnitřní fixace fraktury * metody   MeSH
• výsledek terapie MeSH
• zobrazování trojrozměrné MeSH
• Check Tag
• lidé 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

It is currently challenging to adequately model the growth and migration of glioblastoma using two-dimensional (2D) in vitro culture systems as they quickly lose the original, patient-specific identity and heterogeneity. However, with the advent of three-dimensional (3D) cell cultures and human-induced pluripotent stem cell (iPSC)-derived cerebral organoids (COs), studies demonstrate that the glioblastoma-CO (GLICO) coculture model helps to preserve the phenotype of the patient-specific tissue. Here, we aimed to set up such a model using mature COs and develop a pipeline for subsequent analysis of cocultured glioblastoma. Our data demonstrate that the growth and migration of the glioblastoma cell line within the mature COs are significantly increased in the presence of extracellular matrix proteins, shortening the time needed for glioblastoma to initiate migration. We also describe in detail the method for the visualization and quantification of these migrating cells within the GLICO model. Lastly, we show that this coculture model (and the human brain-like microenvironment) can significantly transform the gene expression profile of the established U87 glioblastoma cell line into proneural and classical glioblastoma cell types.

• MeSH
• buněčné kultury metody   MeSH
• buněčné linie MeSH
• glioblastom * genetika   metabolismus   MeSH
• lidé MeSH
• mozek MeSH
• nádorové mikroprostředí MeSH
• organoidy metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Cell infiltration is a critical parameter for the successful development of 3D matrices for tissue engineering. Application of electrospun nanofibers in tissue engineering has recently attracted much attention. Notwithstanding several of their advantages, small pore size and small thickness of the electrospun layer limit their application for development of 3D scaffolds. Several methods for the pore size and/or electrospun layer thickness increase have been recently developed. Nevertheless, tissue engineering still needs emerging of either novel nanofiber-enriched composites or new techniques for 3D nanofiber fabrication. Forcespinning(®) seems to be a promising alternative. The potential of the Forcespinning(®) method is illustrated in preliminary experiment with mesenchymal stem cells.

• Klíčová slova
• odstředivé zvlákňování, elektrostatické zvlákňování,
• MeSH
• mezenchymální kmenové buňky MeSH
• nanovlákna MeSH
• polyestery MeSH
• tkáňové inženýrství MeSH
• tkáňové podpůrné struktury MeSH

AIM: This study aimed to investigate the phytochemical composition of Psychotria montana extract (PME) and evaluate its inhibitory effects on MCF7 breast cancer cells. METHODS: The chemical composition of PME was analyzed using UPLC-QToF-MS. The effects of PME on cell proliferation were evaluated using the MTT assay. Flow cytometry was used for cell cycle and apoptosis analysis. The effects of PME on the transcription of cell cycle control genes were assessed using real-time PCR. RESULTS: UPLC-QToF-MS analysis revealed major compounds of PME, including terpenoids and flavonoids, with the potential to inhibit proliferation, migration, and induce apoptosis in MCF7 cancer cells. PME effectively suppressed MCF7 cell proliferation under 2D culture, with a low IC50 value of 34.7 μg/ml. PME also hindered cell migration (p < 0.01) and reduced spheroid number (p < 0.001) and size (p < 0.001) in serum-free 3D culture. Apoptosis analysis via nuclear staining with DAPI and flow cytometry revealed an increase in the number of apoptotic cells after PME treatment (p < 0.001). Additionally, the PME induced cell cycle arrest at the G0/G1 phase (p < 0.05). PME altered the expression of cell cycle control genes (cyclins and CDKs) as well as cancer suppressor genes including p16, p27, and p53 at the transcriptional level (mRNA). The results of molecular docking suggest that the compounds present in PME exhibit a high binding affinity for CDK3, CDK4, CDK6, and CDK8 proteins, which are essential regulators of the cell cycle. CONCLUSION: Psychotria montana has the potential to inhibit cancer cells by inducing apoptosis and halting the cell cycle of MCF7 breast cancer cells.

• MeSH
• apoptóza * účinky léků   MeSH
• buněčný cyklus účinky léků   MeSH
• fytogenní protinádorové látky farmakologie   chemie   MeSH
• lidé MeSH
• MFC-7 buňky MeSH
• nádory prsu * farmakoterapie   patologie   genetika   metabolismus   MeSH
• počítačová simulace MeSH
• pohyb buněk účinky léků   MeSH
• proliferace buněk * účinky léků   MeSH
• Psychotria * chemie   MeSH
• rostlinné extrakty * farmakologie   chemie   MeSH
• simulace molekulového dockingu MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

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