scholarly journals Amorphous–Crystalline Calcium Phosphate Coating Promotes In Vitro Growth of Tumor-Derived Jurkat T Cells Activated by Anti-CD2/CD3/CD28 Antibodies

Materials ◽  
2021 ◽  
Vol 14 (13) ◽  
pp. 3693
Author(s):  
Yurii P. Sharkeev ◽  
Ekaterina G. Komarova ◽  
Valentina V. Chebodaeva ◽  
Mariya B. Sedelnikova ◽  
Aleksandr M. Zakharenko ◽  
...  
Keyword(s):  
T Cells ◽  
Calcium Phosphate ◽  
Biological Effects ◽  
Electrical Potential ◽  
Biological Properties ◽  
Modern Trend ◽  
Calcium Phosphate Coating ◽  
Jurkat T Cells ◽  
Micro Arc Oxidation

A modern trend in traumatology, orthopedics, and implantology is the development of materials and coatings with an amorphous–crystalline structure that exhibits excellent biocopatibility. The structure and physico–chemical and biological properties of calcium phosphate (CaP) coatings deposited on Ti plates using the micro-arc oxidation (MAO) method under different voltages (200, 250, and 300 V) were studied. Amorphous, nanocrystalline, and microcrystalline statesof CaHPO4 and β-Ca2P2O7were observed in the coatings using TEM and XRD. The increase in MAO voltage resulted in augmentation of the surface roughness Ra from 2.5 to 6.5 µm, mass from 10 to 25 mg, thickness from 50 to 105 µm, and Ca/P ratio from 0.3 to 0.6. The electrical potential (EP) of the CaP coatings changed from −456 to −535 mV, while the zeta potential (ZP) decreased from −53 to −40 mV following an increase in the values of the MAO voltage. Numerous correlations of physical and chemical indices of CaP coatings were estimated. A decrease in the ZP magnitudes of CaP coatings deposited at 200–250 V was strongly associated with elevated hTERT expression in tumor-derived Jurkat T cells preliminarily activated with anti-CD2/CD3/CD28 antibodies and then contacted in vitro with CaP-coated samples for 14 days. In turn, in vitro survival of CD4+ subsets was enhanced, with proinflammatory cytokine secretion of activated Jurkat T cells. Thus, the applied MAO voltage allowed the regulation of the physicochemical properties of amorphous–crystalline CaP-coatings on Ti substrates to a certain extent. This method may be used as a technological mechanism to trigger the behavior of cells through contact with micro-arc CaP coatings. The possible role of negative ZP and Ca2+ as effectors of the biological effects of amorphous–crystalline CaP coatings is discussed. Micro-arc CaP coatings should be carefully tested to determine their suitability for use in patients with chronic lymphoid malignancies.

scholarly journals Calcium Phosphate Coating Prepared by Microarc Oxidation Affects hTERT Expression, Molecular Presentation, and Cytokine Secretion in Tumor-Derived Jurkat T Cells

Materials ◽  
2020 ◽  
Vol 13 (19) ◽  
pp. 4307
Author(s):  
Larisa S. Litvinova ◽  
Olga G. Khaziakhmatova ◽  
Valeria V. Shupletsova ◽  
Kristina A. Yurova ◽  
Vladimir V. Malashchenko ◽  
...  
Keyword(s):  
T Cells ◽  
Calcium Phosphate ◽  
Tumor Cells ◽  
Surgical Stress ◽  
Microarc Oxidation ◽  
Calcium Phosphate Coating ◽  
Htert Expression ◽  
Cd4 Cells ◽  
Jurkat T Cells ◽  
X Ray

Calcium phosphate (CaP) materials are among the best bone graft substitutes, but their use in the repair of damaged bone in tumor patients is still unclear. The human Jurkat T lymphoblast leukemia-derived cell line (Jurkat T cells) was exposed in vitro to a titanium (Ti) substrate (10 × 10 × 1 mm3) with a bilateral rough (average roughness index (Ra) = 2–5 μm) CaP coating applied via the microarc oxidation (MAO) technique, and the morphofunctional response of the cells was studied. Scanning electron microscopy (SEM), X-ray diffraction (XRD), and energy dispersive X-ray spectroscope (EDX) analyses showed voltage-dependent (150–300 V) growth of structural (Ra index, mass, and thickness) and morphological surface and volume elements, a low Ca/PaT ratio (0.3–0.6), and the appearance of crystalline phases of CaHPO4 (monetite) and β-Ca2P2O7 (calcium pyrophosphate). Cell and molecular reactions in 2-day and 14-day cultures differed strongly and correlated with the Ra values. There was significant upregulation of hTERT expression (1.7-fold), IL-17 secretion, the presentation of the activation antigens CD25 (by 2.7%) and CD95 (by 5.15%) on CD4+ cells, and 1.5–2-fold increased cell apoptosis and necrosis after two days of culture. Hyperactivation-dependent death of CD4+ cells triggered by the surface roughness of the CaP coating was proposed. Conversely, a 3.2-fold downregulation in hTERT expression increased the percentages of CD4+ cells and their CD95+ subset (by 15.5% and 22.9%, respectively) and inhibited the secretion of 17 of 27 test cytokines/chemokines without a reduction in Jurkat T cell survival after 14 days of coculture. Thereafter, cell hypoergy and the selection of an hTERT-independent viable CD4+ subset of tumor cells were proposed. The possible role of negative zeta potentials and Ca2+ as effectors of CaP roughness was discussed. The continuous (2–14 days) 1.5–6-fold reductions in the secretion of vascular endothelial growth factor (VEGF) by tumor cells correlated with the Ra values of microarc CaP-coated Ti substrates seems to limit surgical stress-induced metastasis of lymphoid malignancies.

Growth of Calcium Phosphate Coating on Ti-7.5Mo Alloy after Anodic Oxidation

2013 ◽  
Vol 334-335 ◽  
pp. 297-302 ◽  
Author(s):  
A.L.A. Escada ◽  
João Paulo Barros Machado ◽  
Roberto Zenhei Nakazato ◽  
Ana Paula Rosifini Alves Claro
Keyword(s):  
Calcium Phosphate ◽  
Simulated Body Fluid ◽  
Body Fluid ◽  
Calcium Phosphates ◽  
Sodium Titanate ◽  
Ammonium Fluoride ◽  
Biological Properties ◽  
Calcium Phosphate Coating ◽  
Phosphate Coating

Titanium and its alloys are widely used as biomaterials due to their mechanical, chemical and biological properties. To enhance the biocompatibility of titanium alloys, various surface treatments have been proposed. In particular, the formation of titanium oxide nanotubes layers has been extensively examined. Among the various materials for implants, calcium phosphates and hydroxyapatite are widely used clinically. In this work, titanium nanotubes were fabricated on the surface of Ti-7.5Mo alloy by anodization. The samples were anodized for 20 V in an electrolyte containing glycerol in combination with ammonium fluoride (NH4F, 0.25%), and the anodization time was 24 h. After being anodized, specimens were heat treated at 450 °C and 600°C for 1 h to crystallize the amorphous TiO2 nanotubes and then treated with NaOH solution to make them bioactive, to induce growth of calcium phosphate in a simulated body fluid. Surface morphology and coating chemistry were obtained respectively using, field-emission scanning electron microscopy (FEG-SEM), AFM and X-ray diffraction (XRD). It was shown that the presence of titanium nanotubes induces the growth of a sodium titanate nanolayer. During the subsequent in-vitro immersion in a simulated body fluid, the sodium titanate nanolayer induced the nucleation and growth of nanodimensioned calcium phosphate. It was possible to observe the formation of TiO2 nanotubes on the surface of Ti-7.5Mo. Calcium phosphate coating was greater in the samples with larger nanotube diameter. These findings represent a simple surface treatment for Ti-7.5Mo alloy that has high potential for biomedical applications.

Calcium Phosphate Coating on Blast-Treated Titanium Implants by RF Magnetron Sputtering

2009 ◽  
Vol 631-632 ◽  
pp. 211-216 ◽  
Author(s):  
Kyosuke Ueda ◽  
Takayuki Narushima ◽  
Takashi Goto ◽  
T. Katsube ◽  
Hironobu Nakagawa ◽  
...  
Keyword(s):  
Calcium Phosphate ◽  
Magnetron Sputtering ◽  
Bonding Strength ◽  
Rf Magnetron Sputtering ◽  
Titanium Implants ◽  
Calcium Phosphate Coating ◽  
Phosphate Coating ◽  
Coating Films

Calcium phosphate coating films were fabricated on Ti-6Al-4V plates and screw-type implants with a blast-treated surface using radiofrequency (RF) magnetron sputtering and were evaluated in vitro and in vivo. Amorphous calcium phosphate (ACP) and oxyapatite (OAp) films obtained in this study could cover the blast-treated substrate very efficiently, maintaining the surface roughness. For the in vitro evaluations of the calcium phosphate coating films, bonding strength and alkaline phosphatase (ALP) activity were examined. The bonding strength of the coating films to a blast-treated substrate exceeded 60 MPa, independent of film phases except for the film after post-heat-treatment in silica ampoule. When compared with an uncoated substrate, the increase in the ALP activity of osteoblastic SaOS-2 cells on a calcium phosphate coated substrate was confirmed by a cell culture test. The removal torque of screw-type Ti-6Al-4V implants with a blast-treated surface from the femur of Japanese white rabbit increased with the duration of implantation and it was statistically improved by coating an ACP film 2 weeks after implantation. The in vitro and in vivo studies suggested that the application of the sputtered ACP film as a coating on titanium implants was effective in improving their biocompatibility with bones.

scholarly journals A Review on the Enhancement of Calcium Phosphate Cement with Biological Materials in Bone Defect Healing

Polymers ◽  
2021 ◽  
Vol 13 (18) ◽  
pp. 3075
Author(s):  
Sok Kuan Wong ◽  
Yew Hoong Wong ◽  
Kok-Yong Chin ◽  
Soelaiman Ima-Nirwana
Keyword(s):  
Calcium Phosphate ◽  
Calcium Phosphate Cement ◽  
Research Effort ◽  
Biological Materials ◽  
Biological Properties ◽  
Comprehensive Overview ◽  
Bone Defect Healing ◽  
Living Organisms

Calcium phosphate cement (CPC) is a promising material used in the treatment of bone defects due to its profitable features of self-setting capability, osteoconductivity, injectability, mouldability, and biocompatibility. However, the major limitations of CPC, such as the brittleness, lack of osteogenic property, and poor washout resistance, remain to be resolved. Thus, significant research effort has been committed to modify and reinforce CPC. The mixture of CPC with various biological materials, defined as the materials produced by living organisms, have been fabricated by researchers and their characteristics have been investigated in vitro and in vivo. This present review aimed to provide a comprehensive overview enabling the readers to compare the physical, mechanical, and biological properties of CPC upon the incorporation of different biological materials. By mixing the bone-related transcription factors, proteins, and/or polysaccharides with CPC, researchers have demonstrated that these combinations not only resolved the lack of mechanical strength and osteogenic effects of CPC but also further improve its own functional properties. However, exceptions were seen in CPC incorporated with certain proteins (such as elastin-like polypeptide and calcitonin gene-related peptide) as well as blood components. In conclusion, the addition of biological materials potentially improves CPC features, which vary depending on the types of materials embedded into it. The significant enhancement of CPC seen in vitro and in vivo requires further verification in human trials for its clinical application.

scholarly journals In-vitro Assessment of Biological Properties of Lipids Isolated from Gonads of Stomopneustes variolaris

2020 ◽  
Vol 2 (1) ◽  
pp. 66
Keyword(s):  
Biological Effects ◽  
Lipid Fraction ◽  
Radical Scavenging ◽  
Biological Properties ◽  
Total Phenolic ◽  
Klebsiella Pneumonia ◽  
Spectroscopic Techniques ◽  
Lipid Fractions ◽  
Anti Fungal

Lipid fractions of gonads present in sea urchins serves as a source of bioactive agents with potent pharmaceutical properties. The present study reports the in-vitro biological effects of lipids isolated from gonads of sea urchin: Stomopneustes variolaris collected from the East coast of India. The extracted lipids were characterized by spectroscopic techniques such as GCMS and FTIR and tested for in-vitro biological effects. GCMS analysis of the lipid extract detected high levels of hexa triacontane (17.023 %), tetratetracontane (15.913%), and octacosane (15.628%) and low concentrations of oleic acid (2.206%) and sulfurous acid, pentadecy 2-propyl ester (1.744%). FTIR analysis identified rich composition of functional groups present in the lipids such as 3418.93 cm-1 (hydroxyl), 2921.08cm-1 and 2854.81 cm-1 (alkane), 2660.69 cm-1 (carboxylic acid), 1596.11 cm-1 (amine), 1291.76 cm-1 (aromatic amine). The lipid fraction evaluated by agar diffusion assay measured in terms of zone of inhibition showed bactericidal effects against gram-positive bacteria: Streptococcus aureus (30 mm); Pseudomonas aeruginosa (28.5 mm) and gram-negative bacteria: Escherichia coli (29.5 mm); Klebsiella pneumonia (27.5 mm) and Vibrio cholera (28 mm) respectively. The lipid fraction also showed an effective anti-fungal effect against C.albicans (25 mm). Further, the lipid fractions showed good radical scavenging effect against total phenolic, flavonoid content (15.12 mg GAE/g and 32.72 mg QE/g), and hydrogen peroxide radicals (IC50- 48.28mg/ml) confirming its anti-oxidant potential. Based on the observed results, it was identified that the lipid fraction of gonads of Stomopneustes variolaris demonstrated various biological effects such as bactericidal, anti-fungal and radical scavenging activities which could have a great scope in the formulation of biopharmaceutical agents.

In-vitro assessment of Jurkat T-cells response to 1966 MHz electromagnetic fields in a GTEM cell

2015 ◽  
Author(s):  
Nektarios Moraitis ◽  
Maria Christopoulou ◽  
Konstantina S. Nikita ◽  
Georgia-Persephoni Voulgaridou ◽  
Ioannis Anestopoulos ◽  
...  
Keyword(s):  
T Cells ◽  
Electromagnetic Fields ◽  
Jurkat T Cells ◽  
In Vitro Assessment ◽  
Gtem Cell

scholarly journals Bisphenol A modulates the metabolic regulator oestrogen-related receptor-α in T-cells

Reproduction ◽  
2014 ◽  
Vol 147 (4) ◽  
pp. 419-426 ◽  
Author(s):  
Riccardo Cipelli ◽  
Lorna Harries ◽  
Katsuhiro Okuda ◽  
Shin'ichi Yoshihara ◽  
David Melzer ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Bisphenol A ◽  
Oxidative Metabolism ◽  
Biological Effects ◽  
Control Point ◽  
White Blood Cells ◽  
Fold Increase ◽  
Metabolic Effects

Bisphenol A (BPA) is a widely used plastics constituent that has been associated with endocrine, immune and metabolic effects. Evidence for how BPA exerts significant biological effects at chronic low levels of exposure has remained elusive. In adult men, exposure to BPA has been associated with higher expression of two nuclear receptors, oestrogen receptor-β (ERβ) and oestrogen-related-receptor-α (ERRα), in peripheral white blood cells in vivo. In this study, we explore the expression of ESR2 (ERβ) and ESRRA (ERRα) in human leukaemic T-cell lymphoblasts (Jurkat cells) exposed to BPA in vitro. We show that exposure to BPA led to enhanced expression of ESRRA within 6 h of exposure (mean±s.e.m.: 1.43±0.08-fold increase compared with the control, P<0.05). After 72 h, expression of ESRRA remained significantly enhanced at concentrations of BPA ≥1 nM. Oxidative metabolism of BPA by rat liver S9 fractions yields the potent oestrogenic metabolite, 4-methyl-2,4-bis(4-hydroxyphenyl)pent-1-ene (MBP). Exposure of cells to 1–100 nM MBP increased the expression of both ESRRA (significantly induced, P<0.05, at 1, 10, 100 nM) and ESR2 (1.32±0.07-fold increase at 100 nM exposure, P<0.01). ERRα is a major control point for oxidative metabolism in many cell types, including T-cells. Following exposure to both BPA and MBP, we found that cells showed a decrease in cell proliferation rate. Taken together, these results confirm the bioactivity of BPA against putative T-cell targets in vitro at concentrations relevant to general human exposure.

scholarly journals Bactericidal and Bioresorbable Calcium Phosphate Cements Fabricated by Silver-Containing Tricalcium Phosphate Microspheres

2020 ◽  
Vol 21 (11) ◽  
pp. 3745
Author(s):  
Michiyo Honda ◽  
Yusuke Kawanobe ◽  
Kohei Nagata ◽  
Ken Ishii ◽  
Morio Matsumoto ◽  
...  
Keyword(s):  
Calcium Phosphate ◽  
Tricalcium Phosphate ◽  
Bacterial Adhesion ◽  
Inflammatory Responses ◽  
Spray Pyrolysis Technique ◽  
Biological Properties ◽  
Histological Evaluation ◽  
Bone Apposition ◽  
Calcium Phosphate Cements

Bacterial adhesion to the calcium phosphate surface is a serious problem in surgery. To prevent bacterial infection, the development of calcium-phosphate cements (CPCs) with bactericidal properties is indispensable. The aim of this study was to fabricate antibacterial CPCs and evaluate their biological properties. Silver-containing tricalcium phosphate (Ag-TCP) microspheres consisting of α/β-TCP phases were synthesized by an ultrasonic spray-pyrolysis technique. The powders prepared were mixed with the setting liquid to fabricate the CPCs. The resulting cements consisting of β-TCP and hydroxyapatite had a porous structure and wash-out resistance. Additionally, silver and calcium ions could be released into the culture medium from Ag-TCP cements for a long time accompanied by the dissolution of TCP. These data showed the bioresorbability of the Ag-TCP cement. In vitro antibacterial evaluation demonstrated that both released and immobilized silver suppressed the growth of bacteria and prevented bacterial adhesion to the surface of CPCs. Furthermore, histological evaluation by implantation of Ag-TCP cements into rabbit tibiae exhibited abundant bone apposition on the cement without inflammatory responses. These results showed that Ag-TCP cement has a good antibacterial property and good biocompatibility. The present Ag-TCP cements are promising for bone tissue engineering and may be used as antibacterial biomaterials.

scholarly journals Tyrosine 192 within the SH2 domain of the Src-protein tyrosine kinase p56Lck regulates T-cell activation independently of Lck/CD45 interactions

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Matthias Kästle ◽  
Camilla Merten ◽  
Roland Hartig ◽  
Thilo Kaehne ◽  
Ardiyanto Liaunardy-Jopeace ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Tyrosine Kinase ◽  
Enzymatic Activity ◽  
T Cell Activation ◽  
Protein Tyrosine Kinase ◽  
Cell Activation ◽  
Sh2 Domain ◽  
Jurkat T Cells

Abstract Background Upon engagement of the T-cell receptor (TCR), the Src-family protein tyrosine kinase p56Lck phosphorylates components of the TCR (e.g. the TCRζ chains), thereby initiating T-cell activation. The enzymatic activity of Lck is primarily regulated via reversible and dynamic phosphorylation of two tyrosine residues, Y394 and Y505. Lck possesses an additional highly conserved tyrosine Y192, located within the SH2 domain, whose role in T-cell activation is not fully understood. Methods Knock-in mice expressing a phospho-mimetic (Y192E) form of Lck were generated. Cellular and biochemical characterization was performed to elucidate the function of Y192 in primary T cells. HEK 293T and Jurkat T cells were used for in vitro studies. Results Co-immunoprecipitation studies and biochemical analyses using T cells from LckY192E knock-in mice revealed a diminished binding of LckY192E to CD45 and a concomitant hyperphosphorylation of Y505, thus corroborating previous data obtained in Jurkat T cells. Surprisingly however, in vitro kinase assays showed that LckY192E possesses a normal enzymatic activity in human and murine T cells. FLIM/FRET measurements employing an LckY192E biosensor further indicated that the steady state conformation of the LckY192E mutant is similar to Lckwt. These data suggest that Y192 might regulate Lck functions also independently from the Lck/CD45-association. Indeed, when LckY192E was expressed in CD45−/−/Csk−/− non-T cells (HEK 293T cells), phosphorylation of Y505 was similar to Lckwt, but LckY192E still failed to optimally phosphorylate and activate the Lck downstream substrate ZAP70. Furthermore, LckY19E was recruited less to CD3 after TCR stimulation. Conclusions Taken together, phosphorylation of Y192 regulates Lck functions in T cells at least twofold, by preventing Lck association to CD45 and by modulating ligand-induced recruitment of Lck to the TCR. Major findings Our data change the current view on the function of Y192 and suggest that Y192 also regulates Lck activity in a manner independent of Y505 phosphorylation.

scholarly journals Very late antigen 4-dependent adhesion and costimulation of resting human T cells by the bacterial beta 1 integrin ligand invasin.

1993 ◽  
Vol 177 (1) ◽  
pp. 207-212 ◽  
Author(s):  
E Ennis ◽  
R R Isberg ◽  
Y Shimizu
Keyword(s):  
T Cells ◽  
T Cell ◽  
Adhesion Molecules ◽  
Cd4 T Cells ◽  
Yersinia Pseudotuberculosis ◽  
Biological Properties ◽  
Host Cells ◽  
Late Antigen ◽  
Integrin Ligand

Bacteria and viruses often use the normal biological properties of host adhesion molecules to infect relevant host cells. The outer membrane bacterial protein invasin mediates the attachment of Yersinia pseudotuberculosis to human cells. In vitro studies have shown that four members of the very late antigen (VLA) integrin family of adhesion molecules, VLA-3, VLA-4, VLA-5, and VLA-6, can bind to invasin. Since CD4+ T cells express and use these integrins, we have investigated the interaction of CD4+ T cells with purified invasin. Although VLA integrin-mediated adhesion of T cells to other ligands such as fibronectin does not occur at high levels unless the T cells are activated, resting T cells bind strongly to purified invasin. The binding of resting T cells to invasin requires metabolic activity and an intact cytoskeleton. Although CD4+ T cells express VLA-3, VLA-4, VLA-5, and VLA-6, monoclonal antibody (mAb) blocking studies implicate only VLA-4 as a T cell invasin receptor. Like other integrin ligands, invasin can facilitate T cell proliferative responses induced by a CD3-specific mAb. These results suggest that the nature of the integrin ligand is a critical additional factor that regulates T cell integrin activity, and that direct interactions of T cells with bacterial pathogens such as Yersinia may be relevant to host immune responses to bacterial infection.

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