Cell Culture Laboratory

Al Jīzah, Egypt

Cell Culture Laboratory

Al Jīzah, Egypt
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Ferri M.,University of Bologna | Graen-Heedfeld J.,Fraunhofer Institute for Environmental, Safety and Energy Technology | Bretz K.,Fraunhofer Institute for Environmental, Safety and Energy Technology | Guillon F.,Cell Culture Laboratory | And 7 more authors.
PLoS ONE | Year: 2017

Recently, the isolation of new health-related bioactive molecules derived from agro-food industrial by-products by means of environment-friendly extraction processes has become of particular interest. In the present study, a protein by-product from the rice starch industry was hydrolysed with five commercial proteolytic enzymes, avoiding the use of solvents or chemicals. The digestion processes were optimised, and the digestates were separated in fractions with four different molecular weight ranges by using a cross-flow membrane filtration technique. Total hydrolysates and fractions were tested in vitro for a wide range of biological activities. For the first time rice-derived peptides were assayed for anti-tyrosinase, anti-inflammatory, cytotoxicity and irritation capacities. Antioxidant and anti-hypertensive activities were also evaluated. Protamex, Alcalase and Neutrase treatments produced peptide fractions with valuable bioactivities without resulting cytotoxic or irritant. Highest levels of bioactivity were detected in Protamex-derived samples, followed by samples treated with Alcalase. Based on the present results, a future direct exploitation of isolated peptide fractions in the nutraceutical, functional food and cosmetic industrial fields may be foreseen. © 2017 Ferri et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Ferraz E.P.,Cell Culture Laboratory | Oliveira F.S.,Cell Culture Laboratory | de Oliveira P.T.,Cell Culture Laboratory | Crovace M.C.,Federal University of São Carlos | And 3 more authors.
Journal of Biomedical Materials Research - Part A | Year: 2016

The ability of Biosilicate® with two crystalline phases (BioS-2P) to drive osteoblast differentiation encourages the investigation of the cellular mechanisms involved in this process. Then, the aim of our study was to analyze the large-scale gene expression of osteoblasts grown on BioS-2P compared with Bioglass®45S5 (45S5). Osteoblasts differentiated from rat bone marrow mesenchymal stem cells were cultured under osteogenic conditions on BioS-2P, 45S5 and polystyrene (control). After 10 days, the expression of 23,794 genes was analyzed using mRNA Sequencing and the data were validated by real-time PCR. The BioS-2P exhibited 5 genes upregulated and 3 downregulated compared with 45S5. Compared with control, BioS-2P upregulated 15 and downregulated 11 genes, while 45S5 upregulated 25 and downregulated 21 genes. Eight genes were commonly upregulated and 4 downregulated by both bioactive glasses. In conclusion, our results demonstrated that bioactive glasses affect the gene expression profiling of osteoblasts. Most of the regulated genes by both BioS-2P and 45S5 are associated with the process of mineralization highlighting their osteostimulation property that is, at least in part, derived from the ability to modulate the intracellular machinery to promote osteoblast genotype expression. © 2016 Wiley Periodicals, Inc.

Abdelhady H.G.,Taibah University | Abdelhady H.G.,National Research Center of Egypt | Abdel-Salam H.A.,Taibah University | Abdel-Salam H.A.,Zagazig University | And 6 more authors.
Nanomedicine: Nanotechnology, Biology, and Medicine | Year: 2016

Suicide gene delivery is significant in cancer therapy but has not been fully investigated on a cellular scale. Here, Peak Force Quantitative Nanomechanical atomic force microscopy (PFQNM-AFM) was applied to visualize the effect of herpes simplex virus thymidine kinase dendriplexes (G4AcFaHSTK) on the morphological and nanomechanical properties of individual live and dividing HeLa cells. Cells were then exposed to G4AcFaHSTK, followed by ganciclovir, and directly imaged by real-time PFQNM-AFM. Cell membrane liquefaction, cytoplasmic shrinkage, and cytoskeleton structure loss were observed during cell division. The average Young's modulus of the nuclear region increased with time as the cell continued from metaphase (6.29 kPa) to telophase (13.6 kPa) and then decreased (2.25 kPa) upon apoptosis. In contrast, cells exposed to either ganciclovir or G4AcFaHSTK alone have no changes. Thus, understanding the real-time effects of suicide dendriplexes on the cytoskeletal and nanomechanical behaviors of cancer cells may provide new methods for cancer treatment. © 2016 Elsevier Inc.

Torres-Giner S.,Novel Materials and Nanotechnology Group | Gimeno-Alcaniz J.V.,Cell Culture Laboratory | Ocio M.J.,Novel Materials and Nanotechnology Group | Ocio M.J.,University of Valencia | Lagaron J.M.,Novel Materials and Nanotechnology Group
Journal of Applied Polymer Science | Year: 2011

Bone tissue interfacial scaffolds, which encourage cell growth, are critical determinants for clinical success after implant surgery. Over the years, a number of resorbable configurations have emerged for bone cell support and growth, but only a few have demonstrated clinical efficacy. Polymer coatings produced by electrospinning are regarded as very promising bone interfaces because of the ultrathin-scaled dimensions of its physical structure. In this study, the morphology, composition, thermal properties, and cell growth viability of a number of polylactide-based systems containing different binary and ternary formulations of this biomaterial with collagen and commercial hydroxyapatite nanoparticles were characterized. The best performance in terms of biocompatibility was obtained for the tricomponent system in which the submicron fibers were further subjected to uniaxial orientation process during formation. The in vitro proliferation of the cells, which harbored on these ultrathin-structured mats, was examined by means of a metabolic activity indicator and ensured by means of scanning electron microscopy, and cell anchorage was checked by fluorescent optical microscopy. Finally, the optimum tricomponent material was successfully sterilized for the first time by gamma radiation without noticeable losses in cell-seeding capacity. © 2011 Wiley Periodicals, Inc.

Torres-Giner S.,Novel Materials and Nanotechnology Group | Martinez-Abad A.,Novel Materials and Nanotechnology Group | Gimeno-Alcaniz J.V.,Cell Culture Laboratory | Ocio M.J.,Novel Materials and Nanotechnology Group | And 2 more authors.
Advanced Engineering Materials | Year: 2012

The purpose of this study was to generate ultrathin fibers based on polylactide (PLA) biopolyester with antimicrobial controlled release capacity to treat bacterial infections. To achieve this objective, gentamicin antibiotic was encapsulated into pure PLA fibers, a blend of PLA-collagen and coaxial fibers containing a skin of PLA and a core of collagen using the electrospinning technique. The morphology of the gentamicin-loaded fibers and the antibiotic distribution within the fibers were examined by SEM and TEM. The drug delivery profile of the different electrospun fibers was analyzed using a spectrophotometric method. The performance for treating common possible post-surgical infections was investigated against Staphylococcus epidermis, Pseudomonas aeruginosa and Escherichia coli. The morphology of the electrospun fibers as well as the hydrophilicity of the polymer blends ultimately determined the antibiotic release characteristics. The results indicated that such drug-loaded fibers can serve as advanced delivery platforms with strong and timing controllable antibacterial properties. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Perotti N.,Cell Culture Laboratory | Etcheverrigaray M.,Cell Culture Laboratory | Kratje R.,Cell Culture Laboratory | Oggero M.,Cell Culture Laboratory
Protein Expression and Purification | Year: 2013

A 7-mer hGM-CSF-derived linear epitope (APARSPS) is herein described as a novel and small tag taking into account its particular binding affinity in native conditions that could be easily modified by increasing or lowering the ionic strength. Thus, a 3.4 or 3.8-fold binding increment was observed in high NaCl concentration when the tag was fused to IFN-α2b or when the peptide was in its native environment, respectively. The high salt concentration increased the affinity of the binding interaction and improved the APARSPS epitope binding to the paratope allowing one to design an immunoaffinity chromatography purification step in which the high ionic strength was useful to adsorb the fusion protein to the immunoaffinity matrix and the low ionic strength at pH 9 was valuable to desorb it (a 470-fold purification with a 94%-purity was attained in only one step). Also, this short tag did not affect the functionality of the fusion protein and it was able to be detected both in the natural molecule (hGM-CSF) as in the tagged one with the same high detection limit: 273 pg of each protein. © 2013 Elsevier Inc. All rights reserved.

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