Project description:Titanium is a common implant material. However, in some patients titanium implants fail. Macrophages are key cells involved in foreign body response. To identify macrophage response to titainum, primary human macrophages were cultured on porous titanium discs for 6 days We used microarrays to determine the global expression pattern induced by porous titanium in macrophages and identify potential genes involved in implant failure.
Project description:Titanium is a common implant material. However, in some patients titanium implants fail. Macrophages are key cells involved in foreign body response. To identify macrophage response to titainum, primary human macrophages were cultured on polished titanium discs for 6 days We used microarrays to determine the global expression pattern induced by polished titanium in macrophages and identify potential genes involved in implant failure.
Project description:To evaluate mRNA expression profile (transcriptome), we identified mRNAs of osteoblastic cells from human alveolar crest culture in contact with different titanium surfaces: control (polished), nanotextured (polished titanium discs, etching at 25oC with 50% H2SO4conc and 50% H2O2aq) , nano+submicrotextured (polished titanium discs, etching at 50oC with 50% H2SO4conc and 50% H2O2aq) and rough microtexture (polished titanium discs, etching at 50oC with 100% H2O2aq). The osteoblastic cells were cultured in the alpha-minimum essential medium, supplemented with fetal bovine serum, gentamicin, fungizone, dexamethasone, ascorbic acid and β-glycerophosphate. The mRNA expression profiling was analyzed through hybridizations with whole-human genome Agilent microarray (4x44K format).
Project description:To evaluate miRNA expression profile (miRNOME), we identified microRNAs of osteoblastic cells from human alveolar crest culture in contact with different titanium surfaces: control (polished), nanotextured (polished titanium discs, etching at 25°C with 50% H2SO4conc and 50% H2O2aq) , nano+submicrotextured (polished titanium discs, etching at 50°C with 50% H2SO4conc and 50% H2O2aq) and rough microtexture (polished titanium discs, etching at 50°C with 100% H2O2aq). The osteoblastic cells were cultured in the alpha-minimum essential medium, supplemented with fetal bovine serum, gentamicin, fungizone, dexamethasone, ascorbic acid and β-glycerophosphate. The microRNA expression profiling was analyzed through hybridizations with Agilent miRNA-microarray (8x15K format).
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
Project description:Considering that PRF is clinically used in combination with dental implants and collagen membranes, we exposed titanium discs and collagen membranes to PRF lysates. We showed here that PRF-derived TGF-β activity adsorbs to titanium implants and collagen membranes indicated by the changes in gene expression and immunoassay analysis. Our study points towards TGF-β as major target of PRF and suggest that TGF-β activity released by PRF adsorbs to titanium surface and collagen membranes.
Project description:Kynureninase is a member of a large family of catalytically diverse but structurally homologous pyridoxal 5'-phosphate (PLP) dependent enzymes known as the aspartate aminotransferase superfamily or alpha-family. The Homo sapiens and other eukaryotic constitutive kynureninases preferentially catalyze the hydrolytic cleavage of 3-hydroxy-l-kynurenine to produce 3-hydroxyanthranilate and l-alanine, while l-kynurenine is the substrate of many prokaryotic inducible kynureninases. The human enzyme was cloned with an N-terminal hexahistidine tag, expressed, and purified from a bacterial expression system using Ni metal ion affinity chromatography. Kinetic characterization of the recombinant enzyme reveals classic Michaelis-Menten behavior, with a Km of 28.3 +/- 1.9 microM and a specific activity of 1.75 micromol min-1 mg-1 for 3-hydroxy-dl-kynurenine. Crystals of recombinant kynureninase that diffracted to 2.0 A were obtained, and the atomic structure of the PLP-bound holoenzyme was determined by molecular replacement using the Pseudomonas fluorescens kynureninase structure (PDB entry 1qz9) as the phasing model. A structural superposition with the P. fluorescens kynureninase revealed that these two structures resemble the "open" and "closed" conformations of aspartate aminotransferase. The comparison illustrates the dynamic nature of these proteins' small domains and reveals a role for Arg-434 similar to its role in other AAT alpha-family members. Docking of 3-hydroxy-l-kynurenine into the human kynureninase active site suggests that Asn-333 and His-102 are involved in substrate binding and molecular discrimination between inducible and constitutive kynureninase substrates.