Preparation and Characterization of Electrospun PCL/Silk Fibroin Scaffolds

Natural polymer-based scaffolds are generally considered as favourable matrices for the adhesion and growth of cells in tissue repair. One of the most popular materials in this respect is silk fibroin, known for its wide usage in biomedical applications. This work focuses on the development of electrospun scaffolds based on poly(ε-caprolactone) (PCL) and silk fibroin (SF) evaluated regarding the SF effect on their morphology, surface wetting ability, thermal properties, and HaCaT model cell line biocompatibility. The study revealed that the lowest PCL/SF concentration resulted in highest bead-like morphology formation, relatively thick fibers with the presence of random beads in the case of PCL, while uniform and thinner fibers in the case of increasing PCL/SF content scaffolds. The addition of SF reduced the degree of crystallinity in the PCL due to the less organized crystal structure, and decreased its thermal stability. Both SEM and MTT analyses showed cell presence on all scaffolds three days after cell seeding. Although SF improved PCL hydrophilicity, as shown quantitatively by the MTT assay for improved cytocompatibility properties, more structured electrospun PCL/SF scaffold strategies are required. This work is licensed under a Creative Commons Attribution 4.0 International License

The avian cell line AGE1.CR.pIX characterized by metabolic flux analysis

Lohr V, Haedicke O, Genzel Y, et al. The avian cell line AGE1.CR.pIX characterized by metabolic flux analysis. BMC Biotechnology. 2014;14(1): 72.Background: In human vaccine manufacturing some pathogens such as Modified Vaccinia Virus Ankara, measles, mumps virus as well as influenza viruses are still produced on primary material derived from embryonated chicken eggs. Processes depending on primary cell culture, however, are difficult to adapt to modern vaccine production. Therefore, we derived previously a continuous suspension cell line, AGE1.CR.pIX, from muscovy duck and established chemically-defined media for virus propagation. Results: To better understand vaccine production processes, we developed a stoichiometric model of the central metabolism of AGE1.CR.pIX cells and applied flux variability and metabolic flux analysis. Results were compared to literature dealing with mammalian and insect cell culture metabolism focusing on the question whether cultured avian cells differ in metabolism. Qualitatively, the observed flux distribution of this avian cell line was similar to distributions found for mammalian cell lines (e.g. CHO, MDCK cells). In particular, glucose was catabolized inefficiently and glycolysis and TCA cycle seem to be only weakly connected. Conclusions: A distinguishing feature of the avian cell line is that glutaminolysis plays only a minor role in energy generation and production of precursors, resulting in low extracellular ammonia concentrations. This metabolic flux study is the first for a continuous avian cell line. It provides a basis for further metabolic analyses to exploit the biotechnological potential of avian and vertebrate cell lines and to develop specific optimized cell culture processes, e.g. vaccine production processes

Green technology meets ecotoxicology

By applying concept and principles of green chemistry into different technological processes, green technologies are developed. The environmental and economic benefits of “green” approach is achieved through several directions, such as the use of renewable raw materials, creation of economic efficiency, the use of alternative reaction conditions, as well as the application of non-conventional solvents. From the point view of green chemistry, alternative solvents, in order to be a “green“ substitution to hazardous organic solvents, should be: non-volatile, non-flammable, stabile, synthesized by an environmentally friendly procedure, nontoxic and biodegradable. The toxic impact of all newly synthesized chemicals, such as alternative solvents, could be determined by methods and techniques of ecotoxicology. Ecotoxicology, an interdisciplinary scientific field, can serve as a way of monitoring the greenness of the processes. In vivo and in vitro experiments are used to study the effects of chemicals on different levels of organizations, from molecules to communities and ecosystem. The usage of in vitro methods is encouraged by a scientific community and regulatory agencies as an alternative to in vivo studies in order to reduce the number of laboratory animals used in the toxicological studies. Therefore, in this paper we gave a brief overview on the usage of animal cell cultures within the field of green chemistry and technology

Stable expression of chimeric heavy chain antibodies in CHO cells

Camelid single domain antibodies fused to noncamelid Fc regions, also called chimeric heavy chain antibodies (cHCAb), offer great potential as therapeutic and diagnostic candidates due to their relatively small size (80 kDa) and intact Fc. In this chapter, we describe two approaches, limiting dilution and minipools, for generating nonamplified Chinese hamster ovary cell lines stably expressing cHCAb in suspension and serum-free cultures using a stringent antibiotic selection. Neither of the protocols necessitates the acquisition or implementation of expensive automated infrastructures and thus could be applied in any lab with minimal cell culture setup. The given protocol allows the isolation of stable clones capable of generating up to 100 mg/L of antibody in batch mode performed in shaker flasks. \ua9 2012 Springer Science+Business Media, LLC.Peer reviewed: YesNRC publication: Ye

Subcritical water extraction as an environmentally-friendly technique to recover bioactive compounds from traditional Serbian medicinal plants

© 2017 Elsevier B.V. Subcritical water extraction (SWE) has become a popular green extraction technique for the isolation of different classes of compounds from natural matrices. Low price, safety and green character of water, good yields of target compounds and reduced energy consumption, make this technique favorable for potential industrial applications. The purpose of this study was to evaluate antioxidant, antimicrobial and cytotoxic activity of four medicinal plants traditionally used in folk medicine of Serbia. Black mulberry (Morus nigra L.), wall germander (Teucrium chamaedrys L.), wild geranium (Geranium macrorrhizum L.) and comfrey (Symphytum officinale L.) were extracted by subcritical water at different temperatures. Antioxidant activity of the extracts was defined by conventional spectrophotometric methods, such as the total phenolic content (TPC), DPPH-radical scavenging activity (DPPH-RSA), ferric reducing antioxidant power (FRAP) and total antioxidant capacity (TAC) assessed by a DNA-based sensor. Additionally, the main phenolic compounds contributing to the antioxidant activity of the produced extracts were also identified and quantified by high performance liquid chromatography with diode array detection (HPLC-DAD). Antimicrobial properties of extracts were evaluated against eight microbial strains. Furthermore, the cytotoxic activity was observed for two human cancer cell lines and a cell line derived from murine fibroblast