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Browsing by Subject "bioreactor"

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  • Fermentation of organic waste for bioplastic production : A potential sustainable alternative to fossil-fuel plastic 
    Demeslay, Lise (2023)
    The plastic pollution has become a massive problem in the Arctic, affecting aquatic, and terrestrial ecosystems, the cryosphere, and the atmosphere. One of the solutions proposed by the Arctic Council is to improve waste management by using renewable and sustainable materials. This is where bioplastics reveal their importance. They can be bio-produced by microorganisms from organic waste, they are biodegradable and can be reused. Their production relies on a circular economy system making it sustainable. Here lies the relevance of developing the bioplastic bioproduction and technology. The present research focused on the development of a specific production of polyhydroxyalkanoates (PHAs) from organic waste, in collaboration with the start-up Dionymer (Bordeaux, France). First, the purpose of the study was to up scale the process from the fermentation of chemical volatile fatty acids in flasks (400 mL culture medium) to 2 L bioreactors (BR) by characterizing the main differences in the two processes. Secondly, the research consisted in implementing and testing different set-up for the BR to enhance and improve bioplastic and biomass yields, including aeration and agitation. The characterization of the culture parameters differences between BR and flask pointed out; a higher viscosity of the medium at the end of the process, a darker PHA product and a lower final optical density (OD) (8 versus 12) respectively. Secondly, the focus was on the increase of the OD in BR and finding the origin of the stress, to do so, the following parameters were tested; - three aerations strategies; pO2<10%, <20% and >20%; - two agitations blades; marine and Rushton with baffles; - two aerations spargers; circular and micro. The results revealed that; the pO2 needs to be higher to 20% and it may be linked with the reduction of stress induced to the cells; the marine blades increased the OD and reduced the medium viscosity; the impact of the micro sparger seemed to improve aeration and tent to be very sensitive to antifoam agent that reduced the aeration of the medium. So far, the optimum BR set-up seemed to include the use of marine blades and a pO2 above 20%. More experiments of optimization still need to be performed to unsure a stable and higher production performance.
  • Preparation of light-sensitive ICG-Doxorubicin liposomes and developing of Quasi-Vivo® -based two-cell model for drug efficacy and toxicity testing. 
    Haapalainen, Joonatan (2022)
    Traditional 2D cell cultivating vessels and experimental models cannot often simulate natural chemical and physical environment of different cell types. For example, availability of oxygen, chemical gradients, messaging molecules, fluid pressure, flow and surface topography are factors that may affect significantly in cell differentiation, growth, cellular structure, and metabolism. Modular bioreactors like Quasi-Vivo® -system can be used to simulate these factors. Liposomes are particles of phospholipid bilayer with aqueous space enclosed within. They can be modified in numerous ways, like loading them with hydrophobic and hydrophilic molecules, changing their transition temperature or coating them according to different needs. Doxorubicin is effective and widely used cytostatic agent, but when administered as a free drug it has often severe side-effects, like cardiotoxicity. Goal of this thesis is to determine appropriate manufacturing parameters and verify adequate shelf-life of ICG-Doxorubicin liposomes, that they are applicable for future in vitro experiments. Then survival of HepG2 cell line under flow in Quasi-Vivo®-equipment is determined, after which A549 and HepG2 will be then combined into one two-cell model. Finally, a simple illumination experiment in this cell model with previously made liposomes is conducted, and the effect in whole system is examined. Using protocol presented in this thesis it is possible to produce successfully and repeatedly liposomes with both ICG and doxorubicin encapsulation over 70%. Their shelf-life was at least 14 days when stored in 4°C protected from light. This was determined to be sufficient for in vitro testing. Cultivating A549 and HepG2 cell lines combined in the same system with shared media and fluid flow conditions was successful. Neither of the cell lines show significant difference in viability when compared to static control. When light-activating liposomes are administered to the system and then illuminated, from preliminary results we can see significant difference in drug effect. Both illuminated chambers and off-target chambers connected via Quasi-Vivo® show increased suppression, which shows promise that this in vitro model would be useful for future experiments.

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