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E-cadherin is the main component of epithelial adherens junctions (AJs), which play a crucial role in the maintenance of stable cell–cell adhesion and overall tissue integrity. Down-regulation of E-cadherin expression has been found in many carcinomas, and loss of E-cadherin is generally associated with poor prognosis in patients. During the last decade, however, numerous studies have shown that E-cadherin is essential for several aspects of cancer cell biology that contribute to cancer progression, most importantly, active cell migration. In this review, we summarize the available data about the input of E-cadherin in cancer progression, focusing on the latest advances in the research of the various roles E-cadherin-based AJs play in cancer cell dissemination. The review also touches upon the “cadherin switching” in cancer cells where N- or P-cadherin replace or are co-expressed with E-cadherin and its influence on the migratory properties of cancer cells.

This work deals with the deposition of bioactive TiCaPCON films onto biodegradable polycaprolactone (PCL) nanofibers using magnetron sputtering for bone tissue engineering. Low melting point of PCL nanofibers requires a careful tuning of processing parameters in order to achieve high film quality and avoid substrate degradation. SEM and XPS analyses showed that, with the magnetron current of 2 A, the TiCaPCON films deposited onto PCL nanofibers and Si wafers exhibit similar chemical states hereby indicating that the nanofiber structure is not affected by the deposition process. In vitro biological tests indicated that the osteoblasts adhesion and proliferation as well the ALP activity can be significantly improved by depositing the TiCaPCON film onto PCL. The reported findings suggest a new strategy for the preparation of biodegradable polymer-based biomaterials for bone tissue engineering.

The fabrication of biologically safe materials with enhanced antibacterial characteristics is of great significance. Here we propose B-based therapeutic strategy which offers a good alternative to the widely used approach based on the introduction of most famous bactericidal elements, such as Ag, Cu, and Zn. B-doped TiCaPCON films with 8, 11, and 15 at.% of B were obtained by simultaneous magnetron sputtering of two composite targets (TiC-CaO-Ti3POx and TiB2) in a gaseous mixture of Ar+15%N2. The B content in the films was controlled by changing magnetron current of the TiB2 target. The B-doped films were thoroughly studied by means of scanning and transmission electron microscopy, high-resolution TEM, X-ray diffraction, Raman spectroscopy, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, and glow discharge optical emission spectroscopy. The films were also characterized with respect to surface wettability, boron ion release, and electrochemical characteristics. To test biocompatibility and bioactivity of the films in vitro, spreading, actin cytoskeleton organization, proliferation and osteogenic differentiation of MC3T3-E1 osteoblastic cells grown on the film surface were studied. The antibacterial characteristics of the B-doped TiCaPCON samples against antibiotic-resistant Escherichia coli (E. coli) 261 strains were compared with two-layer BOx/TiCaPCON–B and TiOx/TiCaPCON–B films obtained by three different methods: (i) deposition of a B2O3 top layer, (ii) annealing and (iii) electrochemical oxidation of the as-deposited B-doped films. The obtained results clearly indicated that all types of B-containing films are not toxic for cells regardless of their surface composition. Antibacterial activity tests revealed that only the sample with B2O3 top layer completely inactivated E. coli cells and prevented biofilm formation. Thus the surface of two-layer B2O3/TiCaPCON–11%B film was simultaneously bactericidal towards E. coli strains and not-toxic for osteoblastic cells.

Antibacterial coatings have become a rapidly developing field of research, strongly stimulated by the increasing urgency of identifying alternatives to the traditional administration of antibiotics. Such coatings can be deposited onto implants and other medical devices and prevent the inflammations caused by hospital-acquired infections. Nevertheless, the design of antibacterial yet biocompatible and bioactive surfaces is a challenge that biological community has faced for many years but the “materials of dream” have not yet been developed. In this work, the biocompatible yet antibacterial multi-layered films were prepared by a combination of magnetron sputtering (TiCaPCON film), ion implantation (Ag-doped TiCaPCON film), plasma polymerization (COOH layer), and the final immobilization of gentamicin (GM) and heparin (Hepa) molecules. The layer chemistry was thoroughly investigated by means of FTIR and X-ray photoelectron spectroscopies. It was found that the immobilization of therapeutic components occurs throughout the entire thickness of the plasma-deposited COOH layer. The influence of each type of bactericide (Ag+ ions, GM, and Hepa) on antibacterial activity and cell proliferation was analyzed. Our films were cytocompatible and demonstrated superior bactericidal efficiency toward antibiotic-resistant bacterial E. coli K261 strain. Increased toxicity while using the combination of Ag nanoparticles and COOH plasma polymer is discussed.

The microscopic environment inside a metazoan organism is highly crowded. Whether individual cells can tailor their behavior to the limited space remains unclear. In this study, we found that cells measure the degree of spatial confinement by using their largest and stiffest organelle, the nucleus. Cell confinement below a resting nucleus size deforms the nucleus, which expands and stretches its envelope. This activates signaling to the actomyosin cortex via nuclear envelope stretch-sensitive proteins, up-regulating cell contractility. We established that the tailored contractile response constitutes a nuclear ruler–based signaling pathway involved in migratory cell behaviors. Cells rely on the nuclear ruler to modulate the motive force that enables their passage through restrictive pores in complex three-dimensional environments, a process relevant to cancer cell invasion, immune responses, and embryonic development.

Drug-loaded nanocarriers have a great potential for tumor therapy. Such systems must have high drug-loading efficacy in an alkaline medium and effectively release therapeutic agent in an acidic med...

Implant-related bacterial infections remain a serious problem that is not solved yet. Herein we combined several antibacterial agents to achieve synergistic effects and broader protection of widely used metallic implants. Titanium samples with microcontainers for drug, produced by selective laser sintering, were coated with Ag-doped biocompatible and bioactive TiCaPCON film and loaded with an antibiotic (gentamicin or a mixture of gentamicin and amphotericin B). Bactericide release tests demonstrated that the release rate of one bactericide agent (Ag+ ions or gentamicin) depended on the presence of the other antibacterial component. The antibacterial activity of the biocide-doped samples was evaluated against clinically isolated Escherichia coli O78 (E. coli), Staphylococcus aureus (S. aureus) bacteria, and Neurospora crassa wt-987 (N. crassa) spores. It was found that samples loaded with low gentamicin concentration (0.2 and 0.02 mg/cm2), i.e. 10 and 100 times less than the standard gentamicin concentration (2 mg/cm2), demonstrated a superb antibacterial activity against E. coli bacteria. We showed that a crosslinking reaction between gentamicin and TiCaPCON film proceeded either through the formation of amide bonds or via the electrostatic interaction between amine groups of gentamicin and COOH groups of TiCaPCON and led to the formation of relatively stable drug/film conjugates that prevented a rapid dissolution of gentamicin and ensured its long-term (for 72 h) antibacterial protection. Leaching of silver ions provided an effective antibacterial protection after the depletion of the drug reservoirs. The obtained results clearly show a synergistic antibacterial action of Ag+ ions and gentamicin against S. aureus bacteria. In addition, in the presence of Ag+ ions, the antifungal activity of samples loaded with a mixture of gentamicin and amphotericin B against N. crassa fungus was observed to increase. Thus, it is demonstrated that silver can be successfully coupled with different types of antibiotics to provide innovative hybrid metal-ceramic bioconstructions that are able to deliver precise doses of bactericide agents within a certain period of time and are equally effective against Gram-negative E. coli bacteria, Gram-positive S. aureus, and N. crassa fungus.

Due to their good mechanical stability compared to gelatin, collagen or polyethylene glycol nanofibers and slow degradation rate, biodegradable poly-&epsilon, caprolactone (PCL) nanofibers are promising material as scaffolds for bone and soft-tissue engineering. Here, PCL nanofibers were prepared by the electrospinning method and then subjected to surface functionalization aimed at improving their biocompatibility and bioactivity. For surface modification, two approaches were used: (i) COOH-containing polymer was deposited on the PCL surface using atmospheric pressure plasma copolymerization of CO2 and C2H4, and (ii) PCL nanofibers were coated with multifunctional bioactive nanostructured TiCaPCON film by magnetron sputtering of TiC&ndash, CaO&ndash, Ti3POx target. To evaluate bone regeneration ability in vitro, the surface-modified PCL nanofibers were immersed in simulated body fluid (SBF, 1&times, ) for 21 days. The results obtained indicate different osteoblastic and epithelial cell response depending on the modification method. The TiCaPCON-coated PCL nanofibers exhibited enhanced adhesion and proliferation of MC3T3-E1 cells, promoted the formation of Ca-based mineralized layer in SBF and, therefore, can be considered as promising material for bone tissue regeneration. The PCL&ndash, COOH nanofibers demonstrated improved adhesion and proliferation of IAR-2 cells, which shows their high potential for skin reparation and wound dressing.

The microscopic environment inside a metazoan organism is highly crowded. Whether individual cells can tailor their behavior to the limited space remains unclear. Here, we found that cells measure the degree of spatial confinement using their largest and stiffest organelle, the nucleus. Cell confinement below a resting nucleus size deforms the nucleus, which expands and stretches its envelope. This activates signaling to the actomyosin cortexvianuclear envelope stretch-sensitive proteins, upregulating cell contractility. We established that the tailored contractile response constitutes a nuclear ruler-based signaling pathway involved in migratory cell behaviors. Cells rely on the nuclear ruler to modulate the motive force enabling their passage through restrictive pores in complex three-dimensional (3D) environments, a process relevant to cancer cell invasion, immune responses and embryonic development.One Sentence SummaryNuclear envelope expansion above a threshold triggers a contractile cell response and thus acts as a ruler for the degree of cell deformation.

The aim of this work is to obtain antibacterial yet biocompatible coatings using pulsed electrospark deposition (PED). For this purpose new composite electrodes were fabricated from reaction mixtures Ti–C–20%Fe-10%Ca 3 (PO 4 ) 2 –3.4%Mg–X%Ag with different amount of antibacterial component (X = 0, 0.5, 1.0, 1.5 and 2.0 at% of Ag) using self-propagating high-temperature synthesis method. The electrodes consisted of TiC grains surrounded by TiFe 2 and TiFeP intermetallic matrix, CaO and MgO inclusions, and Ag-based phase. The influence of Ag content on the electrode mass transfer kinetics was studied by comparing the total substrate weight gain and electrode mass loss during PED. The structure, elemental composition, and surface roughness of coatings were studied by means of X-ray diffraction, scanning electron microscopy, and optical profilometry. The coatings were characterized in terms of Ag + ion release, mechanical and electrochemical properties, as well as biocompatibility. The antibacterial characteristics of Ag-doped PED coatings were compared with those obtained by PED using Ag-free electrode and then implanted with Ag + ions. The results indicated that an increase in the Ag content in electrode leads to a decrease in electrode erosion and substrate weight gain, but the efficiency of the PED process increases. Doping with a small amount of Ag (≤ 1 at%) resulted in 100% antibacterial effect against both gram-positive S. aureus and gram-negative E. сoli bacteria. In addition, the dynamics of МС3Т3-Е1 cell proliferation on the surface of PED coatings with 0.6–0.7 at% of Ag was similar to that in control samples, hereby indicating their biocompatibility. The coating biological characteristics were discussed based on the results of Ag + ion release and electrochemical tests.

Epithelial-mesenchymal transition (EMT) is a fundamental process of morphogenesis whereby epithelial cells acquire the mesenchymal phenotype. Multiple data suggest a critical role of EMT in tumor progression. In carcinomas, EMT can be initiated and promoted by many oncogenic signaling pathways, hypoxia, and signals of tumor microenvironment resulting in epithelial cells losing their cell polarity and cell-cell adhesion and gaining the migratory and invasive properties. Downregulation of expression of the cell adhesion protein E-cadherin is considered a poor prognostic factor in cancer. Many tumors are characterized by incomplete EMT, where tumor cells acquire mesenchymal characteristics but retain their epithelial markers, in particular, E-cadherin. In cells with the hybrid epithelial-mesenchymal phenotype, E-cadherin is accumulated in adherens junctions which are less stable than adherens junctions in normal epithelial cells. E-cadherin-based adherens junctions are essential for efficient collective migration and invasion of carcinoma cells, and their survival in metastasis. The plasticity of the hybrid epithelial-mesenchymal phenotype improves adaptive capabilities of cancer cells. By undergoing EMT, carcinoma cells become resistant to chemotherapy and acquire the ability to suppress immune response. Emergence of cancer stem cells after EMT activation has been observed in many types of carcinoma.

There is ample evidence that, instead of a binary switch, epithelial-mesenchymal transition (EMT) in cancer results in a flexible array of phenotypes, each one uniquely suited to a stage in the invasion-metastasis cascade. The phenotypic plasticity of epithelium-derived cancer cells gives them an edge in surviving and thriving in alien environments. This review describes in detail the actin cytoskeleton and E-cadherin-based adherens junction rearrangements that cancer cells need to implement in order to achieve the advantageous epithelial/mesenchymal phenotype and plasticity of migratory phenotypes that can arise from partial EMT.

Thick (up to 30 μm) multicomponent biocompatible coatings with high surface roughness (R a > 6 μm) and hardness up to 11 GPa have been obtained by pulsed electrospark deposition (PED). The surface of Ti substrates (BT1-0 brand mark) was treated using TiC-CaO-Ti 3 PO (x) and TiC-CaO-Ti 3 PO (x) -Ag 2 Ca electrodes obtained by self-propagating high-temperature synthesis from the reaction mixtures of 90%(Ti + 0.5C) + 10%Ca 3 (PO 4 ) 2 and 81%(Ti + 0.5C) + 9% Ca 3 (PO 4 ) 2 + 10% Ag. The Ag was doped into the electrode composition in order to provide the antibacterial activity of the PED coatings. The different contents of O and N in the coatings were achieved through the treatment in various environments: Ar atmosphere, air, or distilled water. The PED was performed at single-discharge energies of 3 and 30 mJ. Depending on the PED mode and processing environments, the coatings with different phase compositions were obtained. Cell proliferation tests showed good biocompatibility of Ag-free and Ag-doped PED coatings with low Ag content. The present paper gives a detailed description of PED process and structural transformations at the tip of electrodes during the PED that affect final coating structure.

Epithelial-mesenchymal transition (EMT) plays an important role in development and also in initiation of metastasis during cancer. Disruption of cell-cell contacts during EMT allowing cells to detach from and migrate away from their neighbors remains poorly understood. Using immunofluorescent staining and live-cell imaging, we analyzed early events during EMT induced by epidermal growth factor (EGF) in IAR-20 normal epithelial cells. Control cells demonstrated stable adherens junctions (AJs) and robust contact paralysis, whereas addition of EGF caused rapid dynamic changes at the cell-cell boundaries: fragmentation of the circumferential actin bundle, assembly of actin network in lamellipodia, and retrograde flow. Simultaneously, an actin-binding protein EPLIN was phosphorylated, which may have decreased the stability of the circumferential actin bundle. Addition of EGF caused gradual replacement of linear E-cadherin&ndash, based AJs with dynamic and unstable punctate AJs, which, unlike linear AJs, colocalized with the mechanosensitive protein zyxin, confirming generation of centripetal force at the sites of cell-cell contacts during EMT. Our data show that early EMT promotes heightened dynamics at the cell-cell boundaries&mdash, replacement of stable AJs and actin structures with dynamic ones&mdash, which results in overall weakening of cell-cell adhesion, thus priming the cells for front-rear polarization and eventual migration.

The epithelial-mesenchymal transition (EMT) plays an important role in development and cancer progression. Upon EMT, epithelial cells lose stable cell-cell adhesions and reorganize their cytoskeleton to acquire migratory activity. Recent data demonstrated that EMT drives cancer cells from the epithelial state to a hybrid epithelial/mesenchymal phenotype with retention of some epithelial markers (in particular, E-cadherin), which is important for cancer cell dissemination. In vitro studies of the effect of growth factors (in particular, epidermal growth factor (EGF)) on cultured cells can be highly advantageous for understanding the details of the early stages of EMT. The methods described in this chapter are intended for studying intermediate phenotypes of EMT. Time-lapse DIC microscopy is used for visualization of changes in morphology and motility of the cells stimulated with EGF. The transwell migration assay allows the evaluation of the migratory activity of the cells. Studying of dynamics of a fluorescently labeled actin-binding protein F-tractin-tdTomato using confocal microscopy allows detection of EGF-induced changes in the organization of the actin cytoskeleton. Live-cell imaging of cells stably expressing GFP-E-cadherin visualizes reorganization of stable tangential E-cadherin-based adherens junctions (AJs) into unstable radial AJs during the early stages of EMT.

International audience; It is very important to prevent bacterial colonization at the early postoperative stages. There are four major strategies and their corresponding types of antibacterial surfaces specifically designed to fight infection bactericide release, anti-adhesion, pH-sensitive, and contact-killing. Herein, we aimed at determining the antibacterial efficiency of different types of bactericidal ions and revealing the possible contribution of surface microgalvanic effects arising from a potential difference on heterogeneous surfaces. We considered five types of TiCaPCON films, with Ag, Zn, Pt, Ag + Zn, and Pt + Zn nanoparticles (NPs) on their surface. The Ag-modified film demonstrated a pronounced antibacterial effect at a very low Ag ion concentration of 0.11 ppb in physiological solution that was achieved already after 3 h of immersion in Escherichia coli (E. coli) bacterial culture. The Zn-containing sample also showed a noticeable antibacterial effect against E. coli and Staphylococcus aureus (S. aureus) strains, wherein the concentration of Zn ions was 2 orders of magnitude higher (15 ppb) compared with the Ag ions. The presence of Ag NPs accelerated the leaching of Zn ion out of the TiCaPCON-Ag-Zn film, but no synergistic effect of the simultaneous presence of the two bactericidal components was observed. After the incubation of the samples with Ag, Zn, and Ag + Zn NPs in E. coli and S. aureus suspensions for 24 and 8 h, respectively, all bacterial cells were completely inactivated. The Pt-containing film showed a very low Pt ion release, and therefore the contribution of this type of ions to the total bactericidal effect could be neglected. The results of the electrochemical studies and Kelvin probe force microscopy indicated that microgalvanic couples were formed between the Pt NPs and the TiCaPCON film, but no noticeable antibacterial effect against either E. coli or S. aureus strains was observed. All ion-modified samples provided good osteoblastic cell attachment, spreading, and proliferation and therefore were concluded to be nontoxic for cells. In addition, the TiCaPCON films with Ag, Pt, and Zn NPs on their surface demonstrated good osteoconductive characteristics.

International audience; Nanoparticles (NPs) have a great potential as nanosized drug-delivery carriers. Such systems must safely deliver the drug to the site of the tumor without drug leakage, effectively penetrate inside cancer cells, and provide intracellular drug release. Herein we developed an original and simple method aimed at the fabrication of spherical boron nitride NPs (BNNPs), 100-200 nm in diameter, with peculiar petal-like surfaces via chemical vapor deposition. Such structures were found to be able to absorb a large amount of antitumor drug-killing tumor cells. They revealed low cytotoxicity and rapid cellular uptake. BNNPs were saturated with doxorubicin (DOX) and then dispersed. The BNNPs loaded with DOX (BNNPs-DOX) were stable at neutral pH but effectively released DOX at pH 4.5-5.5. MTT assay and cell growth testing showed that the BNNPs-DOX nanocarriers had been toxic for IAR-6-1 cells. BNNPs loaded with DOX penetrated into the neoplastic IAR-6-1 cells using endocytic pathways, and then DOX released into the cytoplasm and cell nuclei and resulted in cell death. © 2015 American Chemical Society.

Two approaches (surface doping with bactericidal element and loading of antibiotic into specially formed surface microcontainers) to the fabrication of antibacterial yet biocompatible and bioactive surfaces are described. A network structure with square-shaped blind pores of 2.6 ± 0.6 × 10 −3 mm 3 for drug loading was obtained by selective laser sintering (SLS). The SLS-fabricated samples were loaded with 0.03, 0.3, 2.4, and 4 mg/cm 2 of co-amoxiclav (amoxicillin and clavulanic acid). Ag-doped TiCaPCON films with 0.4, 1.2, and 4.0 at.% of Ag were obtained by co-sputtering of composite TiC 0.5 -Ca 3 (PO 4 ) 2 and metallic Ag targets. The surface structure of SLS-prepared samples and cross-sectional morphology of TiCaPCON-Ag films were studied by scanning electron microscopy. The through-thickness of Ag distribution in the TiCaPCON-Ag films was obtained by glow discharge optical emission spectroscopy. The kinetics of Ag ion release in normal saline solution was studied using inductively coupled plasma mass spectrometry. Bacterial activity of the samples was evaluated against S. epidermidis , S. aureus , and K. pneum. ozaenae using the agar diffusion test and photometric method by controlling the variation of optical density of the bacterial suspension over time. Cytocompatibility of the Ag-doped TiCaPCON films was observed in vitro using chondrocytic and MC3T3-E1 osteoblastic cells. The viability and proliferation of chondrocytic cells were determined using the MTS assay and PicoGreen assay tests, respectively. The alkaline phosphatase (ALP) activity of the SLS-fabricated samples loaded with co-amoxiclav was also studied. The obtained results showed that the moderate bacteriostatic effect of the Ag-doped TiCaPCON films is mainly manifested in the change of bacterial colony morphology and optical densities of bacteria suspensions. In contrast, the SLS-prepared samples showed a very rapid initial drug release resulting in strong bactericidal effect just from the start of the test and for as long as several days. Cytocompatibility and ALP activity tests demonstrated that it is possible to achieve a pronounced antibacterial effect compatible or even higher than that in the control sample (standard disk loaded with the amoxicillin/clavulanic acid mixture (30 μg)) without compromising the material biocompatibility and bioactivity.

Herein we study the effect of doxorubicin-loaded BN nanoparticles (DOX-BNNPs) on cell lines that differ in the multidrug resistance (MDR), namely KB-3-1 and MDR KB-8-5 cervical carcinoma lines, and K562 and MDR i-S9 leukemia lines. We aim at revealing the possible differences in the cytotoxic effect of free DOX and DOX-BNNP nanoconjugates on these types of cells. The spectrophotometric measurements have demonstrated that the maximum amount of DOX in the DOX-BNNPs is obtained after saturation in alkaline solution (pH 8.4), indicating the high efficiency of BNNPs saturation with DOX. DOX release from DOX-BNNPs is a pH-dependent and DOX is more effectively released in acid medium (pH 4.0–5.0). Confocal laser scanning microscopy has shown that the DOX-BNNPs are internalized by neoplastic cells using endocytic pathway and distributed in cell cytoplasm near the nucleus. The cytotoxic studies have demonstrated a higher sensitivity of the leukemia lines to DOX-BNNPs compared with the carcinoma lines: IC50(DOX-BNN...

Adherens junctions (AJs) are molecular complexes that mediate cell-cell adhesive interactions and play pivotal roles in maintenance of tissue organization in adult organisms and at various stages of development. AJs consist of cadherin adhesion receptors, providing homophilic ligation with cadherins on adjacent cells, and members of the catenin protein family: p120, β- and α-catenin. α-catenin's linkage with the actin cytoskeleton defines the linear or punctate organization of AJs in different cell types. Myosin II-dependent tension drives vinculin recruitment by α-catenin and stabilizes the linkage of the cadherin/catenin complex to F-actin. Neoplastic transformation leads to prominent changes in the organization, regulation and stability of AJs. Epithelial-mesenchymal transition (EMT) whereby epithelial cells lose stable cell-cell adhesion, and reorganize their cytoskeleton to acquire migratory activity, plays the central role in cancer cell invasion and metastasis. Recent data demonstrated that a partial EMT resulting in a hybrid epithelial/mesenchymal phenotype with retention of E-cadherin is essential for cancer cell dissemination. E-cadherin and E-cadherin-based AJs are required for collective invasion and migration, survival in circulation, and metastatic outgrowth.

A key property of multicomponent bioactive nanostructured Ti(C,N)-based films doped with Ca, P, and O (TiCaPCON) that can be improved further is their antibacterial effect that should be achieved without compromising the implant bioactivity and biocompatibility. The present work is focused on the study of structure, chemical, mechanical, tribological, and biological properties of Ag- and Cu-doped TiCaPCON films. The films with Ag (0.4–4 at.%) and Cu (13 at.%) contents were obtained by simultaneous sputtering of a TiC0.5–Ca3(PO4)2 target and either an Ag or a Cu target. The film structure was studied using X-ray diffraction, transmission and scanning electron microscopy, energy dispersive X-ray spectroscopy, glow discharge optical emission spectroscopy, and Raman-shift and IR spectroscopy. The films were characterized in terms of their hardness, elastic modulus, dynamic impact resistance, friction coefficient and wear rate (both in air and normal saline), surface wettability, electrochemical behavior and Ag or Cu ion release in normal saline. Particular attention was paid to the influence of inorganic bactericides (Ag and Cu ions) on the bactericidal activity against unicellular yeast fungus Saccharomyces cerevisiae and gram-positive bacteria Lactobacillus acidophilus, as well as on the attachment, spreading, actin cytoskeleton organization, focal adhesions, and early stages of osteoblastic cell differentiation. The obtained results show that the Ag-doped films are more suitable for the protection of metallic surfaces against bacterial infection compared with their Cu-doped counterpart. In particular, an excellent combination of mechanical, tribological, and biological properties makes Ag-doped TiCaPCON film with 1.2 at.% of Ag very attractive material for bioengineering and modification of load-bearing metal implant surfaces.

The present work focuses on the surface modification of Ti alloys using a combination of various techniques such as cold spray (CS), selective laser sintering (SLS), pulsed electro-erosion treatment (PEET), and magnetron sputtering to control surface topography (roughness and blind porosity), surface chemistry, and wettability, i.e. the characteristics which affect osseointegration. The sample structure, elemental composition, surface topography, and wettability were studied using X-ray diffraction, optical and scanning electron microscopy, glow discharge optical emission spectroscopy, energy dispersive spectroscopy, and water contact angle measurements. The obtained results show that Ti coatings deposited by CS can be divided into three groups with a characteristic value of average roughness R a : (i) 4 μm (single particles and agglomerates on the surface), (ii) 22 μm (thin coatings), and (iii) 80 μm (thick coatings). PEET with pulse discharge energies of 0.025 and 0.38 J resulted in the average values of surface roughness of 3 and 8 μm, respectively. During SLS, Ti powder paths were sintered by a laser beam in mutually perpendicular directions to form surface network structures. By varying the distance between the tracks, samples with blind porosity 1.0–5.1 × 10 − 3 mm 3 were obtained. In order to modify the surface chemistry, multifunctional bioactive nanostructured TiCaPCON films, 1–2 μm thick, were deposited atop the CS, PEET, and SLS samples by sputtering a composite TiC 0.5 + Ca 3 (PO 4 ) 2 target. The wettability measurements showed that the CS and PEET modified surfaces exhibit high values of water contact angle. Ion etching in vacuum and TiCaPCON film deposition made the samples highly hydrophilic. The influence of the surface chemistry and surface topography on adhesion, proliferation, and early stages of osteoblasts differentiation was studied. The combination of high surface roughness and blind porosity with hydrophilicity and biocompatibility makes the fabricated metal-ceramic materials promising candidates for applications involving tissue regeneration.

Using confocal microscopy, we analyzed the behavior of IAR-6-1, IAR1170, and IAR1162 transformed epithelial cells seeded onto the confluent monolayer of normal IAR-2 epithelial cells. Live-cell imaging of neoplastic cells stably expressing EGFP and of normal epithelial cells stably expressing mKate2 showed that transformed cells retaining expression of E-cadherin were able to migrate over the IAR-2 epithelial monolayer and invade the monolayer. Transformed IAR cells invaded the IAR-2 monolayer at the boundaries between normal cells. Studying interactions of IAR-6-1 transformed cells stably expressing GFP-E-cadherin with the IAR-2 epithelial monolayer, we found that IAR-6-1 cells established E-cadherin-based adhesions with normal epithelial cells: dot-like dynamic E-cadherin-based adhesions in protrusions and large adherens junctions at the cell sides and rear. A comparative study of a panel of transformed IAR cells that differ by their ability to form E-cadherin-based AJs, either through loss of E-cadherin expression or through expression of a dominant negative E-cadherin mutant, demonstrated that E-cadherin-based AJs are key mediators of the interactions between neoplastic and normal epithelial cells. IAR-6-1DNE cells expressing a dominant-negative mutant form of E-cadherin with the mutation in the first extracellular domain practically lost the ability to adhere to IAR-2 cells and invade the IAR-2 epithelial monolayer. The ability of cancer cells to form E-cadherin-based AJs with the surrounding normal epithelial cells may play an important role in driving cancer cell dissemination in the body.

E-cadherin–mediated cell–cell adhesion, which is essential for the maintenance of the architecture and integrity of epithelial tissues, is often lost during carcinoma progression. To better understand the nature of alterations of cell–cell interactions at the early stages of neoplastic evolution of epithelial cells, we examined the line of nontransformed IAR-2 epithelial cells and their descendants, lines of IAR-6-1 epithelial cells transformed with dimethylnitrosamine and IAR1170 cells transformed with N-RasG12D. IAR-6-1 and IAR1170 cells retained E-cadherin, displayed discoid or polygonal morphology, and formed monolayers similar to IAR-2 monolayer. Fluorescence staining, however, showed that in IAR1170 and IAR-6-1 cells the marginal actin bundle, which is typical of nontransformed IAR-2 cells, disappeared, and the continuous adhesion belt (tangential adherens junctions (AJs)) was replaced by radially oriented E-cadherin–based AJs. Time-lapse imaging of IAR-6-1 cells stably transfected with GFP-E-cadherin revealed that AJs in transformed cells are very dynamic and unstable. The regulation of AJ assembly by Rho family small GTPases was different in nontransformed and in transformed IAR epithelial cells. As our experiments with the ROCK inhibitor Y-27632 and the myosin II inhibitor blebbistatin have shown, the formation and maintenance of radial AJs critically depend on myosin II-mediated contractility. Using the RNAi technique for the depletion of mDia1 and loading cells with N17Rac, we established that mDia1 and Rac are involved in the assembly of tangential AJs in nontransformed epithelial cells but not in radial AJs in transformed cells. Neoplastic transformation changed cell–cell interactions, preventing contact paralysis after the establishment of cell–cell contact and promoting dynamic cell–cell adhesion and motile behavior of cells. It is suggested that the disappearance of the marginal actin bundle and rearrangements of AJs may change the adhesive function of E-cadherin and play an active role in migratory activity of carcinoma cells.