bioreactor dynamics
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Cardiovascular diseases are the most important cause of morbidity and mortality in the civilized world. Stenosis or occlusion of blood vessels leads not only to events that are directly life-threatening, such as myocardial infarction or stroke, but also to a significant reduction in quality of life, for example in lower limb ischemia as a consequence of metabolic diseases. The first synthetic polymeric vascular replacements were used clinically in the early 1950s. However, they proved to be suitable only for larger-diameter vessels, where the blood flow prevents the attachment of platelets, pro-inflammatory cells and smooth muscle cells on their inner surface, whereas in smaller-diameter grafts (6 mm or less), these phenomena lead to stenosis and failure of the graft. Moreover, these polymeric vascular replacements, like biological grafts (decellularized or devitalized), are cell-free, i.e. there are no reconstructed physiological layers of the blood vessel wall, i.e. an inner layer of endothelial cells to prevent thrombosis, a middle layer of smooth muscle cells to perform the contractile function, and an outer layer to provide innervation and vascularization of the vessel wall. Vascular substitutes with these cellular components can be constructed by tissue engineering methods. However, it has to be admitted that even about 70 years after the first polymeric vascular prostheses were implanted into human patients, there are still no functional small-diameter vascular grafts on the market. The damage to small-diameter blood vessels has to be addressed by endovascular approaches or by autologous vascular substitutes, which leads to some skepticism about the potential of tissue engineering. However, new possibilities of this approach lie in the use of modern technologies such as 3D bioprinting and/or electrospinning in combination with stem cells and pre-vascularization of tissue-engineered vascular grafts. In this endeavor, sex-related differences in the removal of degradable biomaterials by the cells and in the behavior of stem cells and pre-differentiated vascular cells need to be taken into account. Key words: Blood vessel prosthesis, Regenerative medicine, Stem cells, Footprint-free iPSCs, sr-RNA, Dynamic bioreactor, Sex-related differences.
Natural products have many healing effects on the skin with minimal or no adverse effects. In this study, we analyzed the regenerative properties of a waste product (hydrolate) derived from Helichrysum italicum (HH) on scratch-tested skin cell populations seeded on a fluidic culture system. Helichrysum italicum has always been recognized in the traditional medicine of Mediterranean countries for its wide pharmacological activities. We recreated skin physiology with a bioreactor that mimics skin stem cell (SSCs) and fibroblast (HFF1) communication as in vivo skin layers. Dynamic culture models represent an essential instrument for recreating and preserving the complex multicellular organization and interactions of the cellular microenvironment. Both cell types were exposed to two different concentrations of HH after the scratch assay and were compared to untreated control cells. Collagen is the constituent of many wound care products that act directly on the damaged wound environment. We analyzed the role played by HH in stimulating collagen production during tissue repair, both in static and dynamic culture conditions, by a confocal microscopic analysis. In addition, we performed a gene expression analysis that revealed the activation of a molecular program of stemness in treated skin stem cells. Altogether, our results indicate a future translational application of this natural extract to support skin regeneration and define a new protocol to recreate a dynamic process of healing.
- MeSH
- fibroblasty metabolismus účinky léků MeSH
- Helichrysum * chemie MeSH
- hojení ran * účinky léků MeSH
- kmenové buňky metabolismus účinky léků cytologie MeSH
- kolagen * metabolismus MeSH
- kultivované buňky MeSH
- kůže * metabolismus účinky léků MeSH
- lidé MeSH
- regenerace * účinky léků MeSH
- rostlinné extrakty * farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Although phosphine is ubiquitously present in anaerobic environments, little is known regarding the microbial community dynamics and metabolic pathways associated with phosphine formation in an anaerobic digestion system. This study investigated the production of phosphine in anaerobic digestion, with results indicating that phosphine production mainly occurred during logarithmic microbial growth. Dehydrogenase and hydrogen promoted the production of phosphine, with a maximum phosphine concentration of 300 mg/m3. The abundance of Ruminococcaceae and Escherichia was observed to promote phosphine generation. The analysis of metabolic pathways based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) and the MetaCyc pathway database revealed the highest relative abundance of replication and repair in genetic information processing; further, the cofactor, prosthetic group, electron carrier, and vitamin biosynthesis were observed to be closely related to phosphine formation. A phylogenetic tree was reconstructed based on the neighbor-joining method. The results indicated the clear evolutionary position of the isolated Pseudescherichia sp. SFM4 strain, adjacent to Escherichia, with a stable phosphate-reducing ability for a maximum phosphine concentration of 26 mg/m3. The response surface experiment indicated that the initial optimal conditions for phosphine production by SFM4 could be achieved with nitrogen, carbon, and phosphorus loads of 6.17, 300, and 10 mg/L, respectively, at pH 7.47. These results provide comprehensive insights into the dynamic changes in the microbial structure, isolated single bacterial strain, and metabolic pathways associated with phosphine formation. They also provide information on the molecular biology associated with phosphorus recycling.
- MeSH
- anaerobióza MeSH
- bioreaktory mikrobiologie MeSH
- Clostridiales genetika metabolismus MeSH
- dusík metabolismus MeSH
- Escherichia genetika metabolismus MeSH
- fosfáty metabolismus MeSH
- fosfiny analýza metabolismus MeSH
- fosfor metabolismus MeSH
- fylogeneze MeSH
- metabolické sítě a dráhy * genetika MeSH
- mikrobiota * MeSH
- odpadní vody mikrobiologie MeSH
- vodík metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Purpose: Nanomaterials for antimicrobial applications have gained interest in recent years due to the increasing bacteria resistance to conventional antibiotics. Wound sterilization, water treatment and surface decontamination all avail from multifunctional materials that also possess excellent antibacterial properties, eg zinc oxide (ZnO). Here, we assess and compare the effects of synthesized hedgehog-like ZnO structures and commercial ZnO particles with and without mixing on the inactivation of bacteria on surfaces and in liquid environments. Methods: Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria in microbial culture medium were added to reverse spin bioreactors that contained different concentrations of each ZnO type to enable dynamic mixing of the bacteria-ZnO suspensions. Optical density of the bacteria-ZnO suspensions was measured in real-time and the number of viable bacteria after 24 h exposure was determined using standard microbiological techniques. The concentration of zinc ion generated from ZnO dissolution in different liquid types was estimated from the dynamic interaction exposure. Static antibacterial tests without agitation in liquid media and on agar surface were performed for comparison. Results: A correlation between increasing ZnO particle concentration and reduction in viable bacteria was not monotonous. The lowest concentration tested (10 µg/mL) even stimulated bacteria growth. The hedgehog ZnO was significantly more antibacterial than commercial ZnO particles at higher concentrations (up to 1000 µg/mL tested), more against E. coli than S. aureus. Minimum inhibitory concentration in microwell plates was correlated with those results. No inhibition was detected for any ZnO type deposited on agar surface. Zinc ion release was greatly suppressed in cultivation media. Scanning electron microscopy images revealed that ZnO needles can pierce membrane of bacteria whereas the commercial ZnO nanoparticles rather agglomerate on the cell surface. Conclusion: The inhibition effects are thus mainly controlled by the interaction dynamics between bacteria and ZnO, where mixing greatly enhances antibacterial efficacy of all ZnO particles. The efficacy is modulated also by ZnO particle shapes, where hedgehog ZnO has superior effect, in particular at lower concentrations. However, at too low concentrations, ZnO can stimulate bacteria growth and must be thus used with caution.
- MeSH
- antibakteriální látky chemie farmakologie MeSH
- biomimetické materiály chemie farmakologie MeSH
- Escherichia coli účinky léků růst a vývoj MeSH
- ježkovití * MeSH
- mikrobiální testy citlivosti MeSH
- oxid zinečnatý chemie farmakologie MeSH
- Staphylococcus aureus účinky léků růst a vývoj MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Sufficient mixing is crucial for the proper performance of anaerobic digestion (AD), creating a homogeneous distribution of soluble substrates, biomass, pH, and temperature. The opaqueness of the sludge and mode of operation make it challenging to study AD mixing experimentally. Therefore, hydrodynamics modelling employing computational fluid dynamics (CFD) is often used to investigate this mixing. However, CFD models mostly do not include biochemical reactions and, hence, ignore the effect of diffusion-induced transport on AD heterogeneity. The novelty of this work is the partial integration of Anaerobic Digestion Model no. 1 (ADM1) into the CFD model. The aim is to better understand the effect of advection-diffusion transport on the homogenization of soluble substrates and biomass. Furthermore, AD homogeneity analysis in terms of concentration distribution is proposed rather than the traditional velocity distributions. The computed results indicate that including diffusion-induced transport affects the homogeneity of AD.
- MeSH
- anaerobióza MeSH
- bioreaktory MeSH
- difuze MeSH
- hydrodynamika * MeSH
- odpadní vody MeSH
- teoretické modely * MeSH
- Publikační typ
- časopisecké články MeSH
Natural cosmetic products have recently re-emerged as a novel tool able to counteract skin aging and skin related damages. In addition, recently achieved progress in nanomedicine opens a novel approach yielding from combination of modern nanotechnology with traditional treatment for innovative pharmacotherapeutics. In the present study, we investigated the antiaging effect of a pretreatment with Myrtus communis natural extract combined with a polycaprolactone nanofibrous scaffold (NanoPCL-M) on skin cell populations exposed to UV. We set up a novel model of skin on a bioreactor mimicking a crosstalk between keratinocytes, stem cells and fibroblasts, as in skin. Beta-galactosidase assay, indicating the amount of senescent cells, and viability assay, revealed that fibroblasts and stem cells pretreated with NanoPCL-M and then exposed to UV are superimposable to control cells, untreated and unexposed to UV damage. On the other hand, cells only exposed to UV stress, without NanoPCL-M pretreatment, exhibited a significantly higher yield of senescent elements. Keratinocyte-based 3D structures appeared disjointed after UV-stress, as compared to NanoPCL-M pretreated samples. Gene expression analysis performed on different senescence associated genes, revealed the activation of a molecular program of rejuvenation in stem cells pretreated with NanoPCL-M and then exposed to UV. Altogether, our results highlight a future translational application of NanoPCL-M to prevent skin aging.
- MeSH
- exprese genu účinky léků MeSH
- fibroblasty účinky léků MeSH
- keratinocyty účinky léků MeSH
- kmenové buňky účinky léků MeSH
- kultivované buňky MeSH
- kůže účinky léků MeSH
- lidé MeSH
- Myrtus chemie MeSH
- nanovlákna chemie MeSH
- polyestery chemie MeSH
- proliferace buněk účinky léků MeSH
- rostlinné extrakty farmakologie MeSH
- stárnutí buněk účinky léků MeSH
- stárnutí kůže účinky léků MeSH
- ultrafialové záření škodlivé účinky MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Design and development of scale-down approaches, such as microbioreactor (μBR) technologies with integrated sensors, are an adequate solution for rapid, high-throughput and cost-effective screening of valuable reactions and/or production strains, with considerably reduced use of reagents and generation of waste. A significant challenge in the successful and widespread application of μBRs in biotechnology remains the lack of appropriate software and automated data interpretation of μBR experiments. Here, it is demonstrated how mathematical models can be usedas helpful tools, not only to exploit the capabilities of microfluidic platforms, but also to reveal the critical experimental conditions when monitoring cascade enzymatic reactions. A simplified mechanistic model was developed to describe the enzymatic reaction of glucose oxidase and glucose in the presence of catalase inside a commercial microfluidic platform with integrated oxygen sensor spots. The proposed model allowed an easy and rapid identification of the reaction mechanism, kinetics and limiting factors. The effect of fluid flow and enzyme adsorption inside the microfluidic chip on the optical sensor response and overall monitoring capabilities of the presented platform was evaluated via computational fluid dynamics (CFD) simulations. Remarkably, the model predictions were independently confirmed for μL- and mL- scale experiments. It is expected that the mechanistic models will significantly contribute to the further promotion of μBRs in biocatalysis research and that the overall study will create a framework for screening and evaluation of critical system parameters, including sensor response, operating conditions, experimental and microbioreactor designs.
AIMS: Stress urinary incontinence and pelvic organ prolapse are very common conditions with a proportion of patients requiring implantation of synthetic materials for a durable repair. However increasing numbers of post-surgical complications have been reported related to the use of polypropylene meshes. One hypothesis for the adverse response is poor mechanical matching of the relatively stiff polypropylene mesh particularly as materials in the pelvic floor will need to cope with decades of distension as occurs with increase of intraabdominal pressure on coughing, laughing, or sneezing. METHODS: In this study we have undertaken a very simple fatigue testing regime to compare the mechanical abilities of six materials. Four commercial meshes in clinical use and two novel electrospun materials not yet evaluated in the clinic were assessed using a uniaxial tensile test. This was performed on six samples of each dry material and on another six samples of each material after just 3 days of fatigue conditions using a dynamic bioreactor. RESULTS: The four commercial materials showed permanent mechanical deformation after just 3 days of stretching these materials by 25% elongation on a regular dynamic cycle, whereas the two new materials presented more elastic properties without deformation. CONCLUSIONS: We suggest that a test as simple as this 3-day fatigue testing is sufficient to distinguish between materials which have already been found to cause complications clinically and newer materials yet to be tested clinically which will hopefully prove more mechanically appropriate for implantation in the pelvic floor.
- MeSH
- chirurgické síťky * MeSH
- lidé MeSH
- mechanické jevy MeSH
- pánevní dno chirurgie MeSH
- pevnost v tahu MeSH
- polypropyleny MeSH
- prolaps pánevních orgánů chirurgie MeSH
- stresová inkontinence moči chirurgie MeSH
- testování materiálů * MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Process fluctuation caused by temperature modification of anaerobic digestion is routinely monitored via operational parameters, such as pH and gas production, but these parameters are lagging on microbial community performance. In this study, 13C isotope fractionation in CH4 and CO2 of biogas together with microbial community dynamics were applied to evaluate process stability in response to temperature increment. Results showed that the weakening correlated links between Firmicutes affiliated families and Methanomicrobiaceae were found regarding temperature increase. In contrast, Methanosarcinaceae and Methanobacteriaceae strengthened their links with multiple bacterial groups. This suggests that the 13C isotope fractionation in CH4 can predict the collapse of certain microbial interconnections and process instability, the new reinforced microbial links directly reflect the microbial community redundancy for maintaining function of syntrophic populations.
- MeSH
- anaerobióza MeSH
- Bacteria MeSH
- bioreaktory * MeSH
- Euryarchaeota * MeSH
- methan MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
One of the main aims of bone tissue engineering, regenerative medicine and cell therapy is development of an optimal artificial environment (scaffold) that can trigger a favorable response within the host tissue, it is well colonized by resident cells of organism and ideally, it can be in vitro pre-colonized by cells of interest to intensify the process of tissue regeneration. The aim of this study was to develop an effective tool for regenerative medicine, which combines the optimal bone-like scaffold and colonization technique suitable for cell application. Accordingly, this study includes material (physical, chemical and structural) and in vitro biological evaluation of scaffolds prior to in vivo study. Thus, porosity, permeability or elasticity of two types of bone-like scaffolds differing in the ratio of collagen type I and natural calcium phosphate nanoparticles (bCaP) were determined, then analyzes of scaffold interaction with mesenchymal stem cells (MSCs) were performed. Simultaneously, dynamic seeding using a perfusion bioreactor followed by static cultivation was compared with standard static cultivation for the whole period of cultivation. In summary, cell colonization ability was estimated by determination of cell distribution within the scaffold (number, depth and homogeneity), matrix metalloproteinase activity and gene expression analysis of signaling molecules and differentiation markers. Results showed, the used dynamic colonization technique together with the newly-developed collagen-based scaffold with high content of bCaP to be an effective combined tool for producing bone grafts for bone implantology and regenerative medicine.
- MeSH
- buněčná diferenciace MeSH
- fosforečnany vápenaté metabolismus MeSH
- kolagen chemie MeSH
- kosti a kostní tkáň chemie MeSH
- kultivované buňky MeSH
- mezenchymální kmenové buňky metabolismus MeSH
- nanočástice MeSH
- osteogeneze účinky léků MeSH
- prasata MeSH
- regenerativní lékařství MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury chemie MeSH
- transplantace mezenchymálních kmenových buněk metody MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
