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Biotechnology and Bioengineering Jul 2022In the past decades, bone tissue engineering developed and exploited many typologies of bioreactors, which, besides providing proper culture conditions, aimed at...
In the past decades, bone tissue engineering developed and exploited many typologies of bioreactors, which, besides providing proper culture conditions, aimed at integrating those bio-physical stimulations that cells experience in vivo, to promote osteogenic differentiation. Nevertheless, the highly challenging combination and deployment of many stimulation systems into a single bioreactor led to the generation of several unimodal bioreactors, investigating one or at mostly two of the required biophysical stimuli. These systems miss the physiological mimicry of bone cells environment, and often produced contrasting results, thus making the knowledge of bone mechanotransduction fragmented and often inconsistent. To overcome this issue, in this study we developed a perfusion and electroactive-vibrational reconfigurable stimulation bioreactor to investigate the differentiation of SaOS-2 bone-derived cells, hosting a piezoelectric nanocomposite membrane as cell culture substrate. This multimodal perfusion bioreactor is designed based on a numerical (finite element) model aimed at assessing the possibility to induce membrane nano-scaled vibrations (with ~12 nm amplitude at a frequency of 939 kHz) during perfusion (featuring 1.46 dyn cm wall shear stress), large enough for inducing a physiologically-relevant electric output (in the order of 10 mV on average) on the membrane surface. This study explored the effects of different stimuli individually, enabling to switch on one stimulation at a time, and then to combine them to induce a faster bone matrix deposition rate. Biological results demonstrate that the multimodal configuration is the most effective in inducing SaOS-2 cell differentiation, leading to 20-fold higher collagen deposition compared to static cultures, and to 1.6- and 1.2-fold higher deposition than the perfused- or vibrated-only cultures. These promising results can provide tissue engineering scientists with a comprehensive and biomimetic stimulation platform for a better understanding of mechanotransduction phenomena beyond cells differentiation.
Topics: Bioreactors; Bone and Bones; Cell Differentiation; Cells, Cultured; Mechanotransduction, Cellular; Osteogenesis; Tissue Engineering; Tissue Scaffolds
PubMed: 35383894
DOI: 10.1002/bit.28100 -
Cells, Tissues, Organs 2023Peristalsis is a nuanced mechanical stimulus comprised of multi-axial strain (radial and axial strain) and shear stress. Forces associated with peristalsis regulate...
Peristalsis is a nuanced mechanical stimulus comprised of multi-axial strain (radial and axial strain) and shear stress. Forces associated with peristalsis regulate diverse biological functions including digestion, reproductive function, and urine dynamics. Given the central role peristalsis plays in physiology and pathophysiology, we were motivated to design a bioreactor capable of holistically mimicking peristalsis. We engineered a novel rotating screw-drive based design combined with a peristaltic pump, in order to deliver multi-axial strain and concurrent shear stress to a biocompatible polydimethylsiloxane (PDMS) membrane "wall." Radial indentation and rotation of the screw drive against the wall demonstrated multi-axial strain evaluated via finite element modeling. Experimental measurements of strain using piezoelectric strain resistors were in close alignment with model-predicted values (15.9 ± 4.2% vs. 15.2% predicted). Modeling of shear stress on the "wall" indicated a uniform velocity profile and a moderate shear stress of 0.4 Pa. Human mesenchymal stem cells (hMSCs) seeded on the PDMS "wall" and stimulated with peristalsis demonstrated dramatic changes in actin filament alignment, proliferation, and nuclear morphology compared to static controls, perfusion, or strain, indicating that hMSCs sensed and responded to peristalsis uniquely. Lastly, significant differences were observed in gene expression patterns of calponin, caldesmon, smooth muscle actin, and transgelin, corroborating the propensity of hMSCs toward myogenic differentiation in response to peristalsis. Collectively, our data suggest that the peristalsis bioreactor is capable of generating concurrent multi-axial strain and shear stress on a "wall." hMSCs experience peristalsis differently than perfusion or strain, resulting in changes in proliferation, actin fiber organization, smooth muscle actin expression, and genetic markers of differentiation. The peristalsis bioreactor device has broad utility in the study of development and disease in several organ systems.
Topics: Humans; Peristalsis; Biomimetics; Actins; Cell Differentiation; Bioreactors
PubMed: 35008089
DOI: 10.1159/000521752 -
PeerJ 2023The myrtle () plant naturally grows in the temperate Mediterranean and subtropical regions and is used for various purposes; thus, it is among the promising species of...
The myrtle () plant naturally grows in the temperate Mediterranean and subtropical regions and is used for various purposes; thus, it is among the promising species of horticultural crops. This study aimed to evaluate and compare the performance of different propagation systems, including rooting, solid media propagation, rooting, and with the Plantform bioreactor system, in achieving healthy and rapid growth of four myrtle genotypes with diverse genetic origins and well-regional adaptation. The selection of myrtle genotypes with distinct genetic backgrounds and proven adaptability to specific regions allowed for a comprehensive assessment of the propagation systems under investigation. Present findings proved that the Plantform system, the new-generation tissue culture system, was quite successful in micropropagation and rooting myrtle genotypes. We succeeded micropropagation and rooting of diverse wild myrtle genotypes, enabling year-round propagation without reliance on specific seasons or environmental conditions. The process involved initiating cultures from explants and multiplying them through shoot proliferation in a controlled environment. This contributes to sustainable plant propagation, preserving and utilizing genetic resources for conservation and agriculture.
Topics: Myrtus; Agriculture; Bioreactors; Crops, Agricultural; Environment, Controlled
PubMed: 37744226
DOI: 10.7717/peerj.16061 -
Biotechnology Advances 2016The novel concept of reverse membrane bioreactors (rMBR) introduced in this review is a new membrane-assisted cell retention technique benefiting from the advantageous... (Review)
Review
The novel concept of reverse membrane bioreactors (rMBR) introduced in this review is a new membrane-assisted cell retention technique benefiting from the advantageous properties of both conventional MBRs and cell encapsulation techniques to tackle issues in bioconversion and fermentation of complex feeds. The rMBR applies high local cell density and membrane separation of cell/feed to the conventional immersed membrane bioreactor (iMBR) set up. Moreover, this new membrane configuration functions on basis of concentration-driven diffusion rather than pressure-driven convection previously used in conventional MBRs. These new features bring along the exceptional ability of rMBRs in aiding complex bioconversion and fermentation feeds containing high concentrations of inhibitory compounds, a variety of sugar sources and high suspended solid content. In the current review, the similarities and differences between the rMBR and conventional MBRs and cell encapsulation regarding advantages, disadvantages, principles and applications for biofuel production are presented and compared. Moreover, the potential of rMBRs in bioconversion of specific complex substrates of interest such as lignocellulosic hydrolysate is thoroughly studied.
Topics: Biofilms; Biofuels; Bioreactors; Diffusion; Membranes, Artificial
PubMed: 27238291
DOI: 10.1016/j.biotechadv.2016.05.009 -
Advanced Healthcare Materials Dec 2022Combining the sustainable culture of billions of human cells and the bioprinting of wholly cellular bioinks offers a pathway toward organ-scale tissue engineering....
Combining the sustainable culture of billions of human cells and the bioprinting of wholly cellular bioinks offers a pathway toward organ-scale tissue engineering. Traditional 2D culture methods are not inherently scalable due to cost, space, and handling constraints. Here, the suspension culture of human induced pluripotent stem cell-derived aggregates (hAs) is optimized using an automated 250 mL stirred tank bioreactor system. Cell yield, aggregate morphology, and pluripotency marker expression are maintained over three serial passages in two distinct cell lines. Furthermore, it is demonstrated that the same optimized parameters can be scaled to an automated 1 L stirred tank bioreactor system. This 4-day culture results in a 16.6- to 20.4-fold expansion of cells, generating approximately 4 billion cells per vessel, while maintaining >94% expression of pluripotency markers. The pluripotent aggregates can be subsequently differentiated into derivatives of the three germ layers, including cardiac aggregates, and vascular, cortical and intestinal organoids. Finally, the aggregates are compacted into a wholly cellular bioink for rheological characterization and 3D bioprinting. The printed hAs are subsequently differentiated into neuronal and vascular tissue. This work demonstrates an optimized suspension culture-to-3D bioprinting pipeline that enables a sustainable approach to billion cell-scale organ engineering.
Topics: Humans; Induced Pluripotent Stem Cells; Cell Culture Techniques; Cell Proliferation; Cell Line; Bioreactors
PubMed: 36314397
DOI: 10.1002/adhm.202201138 -
Sheng Wu Gong Cheng Xue Bao = Chinese... Oct 2019Industrial fermentation focuses on realizing the uniform of high titer, high yield, and high productivity. Multi-scale analysis and regulation, including molecule level,... (Review)
Review
Industrial fermentation focuses on realizing the uniform of high titer, high yield, and high productivity. Multi-scale analysis and regulation, including molecule level, cell level, and bioreactor level, facilitate global optimization and dynamic balance of fermentation process, which determine high efficiency of biosynthesis, targeted directionality of bioconversion, process robustness, and well-organized system. In this review, we summariz and discuss advances in multi-scale analysis and regulation for fermentation process focusing on the following four aspects: 1) kinetic modeling of metabolic pathways, 2) characteristic of cell metabolism, 3) co-coupling fermentation and purification, and 4) bioreactor design. Integrating multi-scale analysis of fermentation process and integrating multi-scale regulation are expected as an important strategy for realizing highly efficient fermentation by industrial microorganisms.
Topics: Bioreactors; Fermentation; Industrial Microbiology; Kinetics; Metabolic Networks and Pathways
PubMed: 31668044
DOI: 10.13345/j.cjb.190244 -
Tissue Engineering. Part B, Reviews Apr 2013Tendon and ligament injury is a worldwide health problem, but the treatment options remain limited. Tendon and ligament engineering might provide an alternative tissue... (Review)
Review
Tendon and ligament injury is a worldwide health problem, but the treatment options remain limited. Tendon and ligament engineering might provide an alternative tissue source for the surgical replacement of injured tendon. A bioreactor provides a controllable environment enabling the systematic study of specific biological, biochemical, and biomechanical requirements to design and manufacture engineered tendon/ligament tissue. Furthermore, the tendon/ligament bioreactor system can provide a suitable culture environment, which mimics the dynamics of the in vivo environment for tendon/ligament maturation. For clinical settings, bioreactors also have the advantages of less-contamination risk, high reproducibility of cell propagation by minimizing manual operation, and a consistent end product. In this review, we identify the key components, design preferences, and criteria that are required for the development of an ideal bioreactor for engineering tendons and ligaments.
Topics: Animals; Bioreactors; Equipment Design; Humans; Ligaments; Regeneration; Tendons; Tissue Engineering
PubMed: 23072472
DOI: 10.1089/ten.TEB.2012.0295 -
International Orthopaedics May 2014Our aim was to explore the effect of varying in vitro culture conditions on general chondrogenesis of minced cartilage (MC) fragments.
PURPOSE
Our aim was to explore the effect of varying in vitro culture conditions on general chondrogenesis of minced cartilage (MC) fragments.
METHODS
Minced, fibrin-associated, bovine articular cartilage fragments were cultured in vitro within polyurethane scaffold rings. Constructs were maintained either free swelling for two or four weeks (control), underwent direct mechanical knee-joint-specific bioreactor-induced dynamic compression and shear, or they were maintained free swelling for two weeks followed by two weeks of bioreactor stimulation. Samples were collected for glycosaminoglycan (GAG)/DNA quantification; collagen type I, collagen type II, aggrecan, cartilage oligomeric matrix protein (COMP), proteoglycan-4 (PRG-4) messenger RNA (mRNA) analysis; histology and immunohistochemistry.
RESULTS
Cellular outgrowth and neomatrix formation was successfully accomplished among all groups. GAG/DNA and collagen type I mRNA were not different between groups; chondrogenic genes collagen type II, aggrecan and COMP revealed a significant downregulation among free-swelling constructs over time (week two through week four). Mechanical loading was able to maintain chondrogenic expression with significantly stronger expression at long-term time points (four weeks) in comparison with four-week control. Histology and immunohistochemistry revealed that bioreactor culture induced stronger cellular outgrowth than free-swelling constructs. However, weaker collagen type II and aggrecan expression with an increased collagen type I expression was noted among this outgrowth neotissue.
CONCLUSIONS
The method of MC culture is feasible under in vitro free-swelling and dynamic loading conditions, simulating in vivo posttransplantation. Mechanical stimulation significantly provokes cellular outgrowth and long-term chondrogenic maturation at the mRNA level, whereas histology depicts immature neotissue where typical cartilage matrix is expected.
Topics: Animals; Bioreactors; Cartilage; Cattle; Chondrogenesis; Pressure; Stress, Mechanical; Tissue Culture Techniques
PubMed: 24287980
DOI: 10.1007/s00264-013-2194-9 -
The Science of the Total Environment Oct 2021This study consists of a review on the removal efficiencies of a wide spectrum of micropollutants (MPs) in biological treatment (mainly membrane bioreactor) coupled with... (Review)
Review
This study consists of a review on the removal efficiencies of a wide spectrum of micropollutants (MPs) in biological treatment (mainly membrane bioreactor) coupled with activated carbon (AC) (AC added in the bioreactor or followed by an AC unit, acting as a post treatment). It focuses on how the presence of AC may promote the removal of MPs and the effects of dissolved organic matter (DOM) in wastewater. Removal data collected of MPs are analysed versus AC dose if powdered AC is added in the bioreactor, and as a function of the empty bed contact time in the case of a granular activated carbon (GAC) column acting as a post treatment. Moreover, the enhancement in macropollutant (organic matter, nitrogen and phosphorus compounds) removal is analysed as well as the AC mitigation effect towards membrane fouling and, finally, how sludge properties may change in the presence of AC. To sum up, it was found that AC improves the removal of most MPs, favouring their sorption on the AC surface, promoted by the presence of different functional groups and then enhancing their degradation processes. DOM is a strong competitor in sorption on the AC surface, but it may promote the transformation of GAC in a biologically activated carbon thus enhancing all the degradation processes. Finally, AC in the bioreactor increases sludge floc strength and improves its settling characteristics and sorption potential.
Topics: Adsorption; Bioreactors; Charcoal; Wastewater; Water Pollutants, Chemical; Water Purification
PubMed: 34091341
DOI: 10.1016/j.scitotenv.2021.148050 -
Journal of Environmental Management May 2024This study focused on the economic feasibility of two potential industrial-scale bioleaching technologies for metal recovery from specific metallurgical by-products,...
This study focused on the economic feasibility of two potential industrial-scale bioleaching technologies for metal recovery from specific metallurgical by-products, mainly basic oxygen steelmaking dust (BOS-D) and goethite. The investigation compared two bioleaching scaling technology configurations, including an aerated bioreactor and an aerated and stirred bioreactor across different scenarios. Results indicated that bioleaching using Acidithiobacillus ferrooxidans proved financially viable for copper extraction from goethite, particularly when 5% and 10% pulp densities were used in the aerated bioreactor, and when 10% pulp density was used in the aerated and stirred bioreactor. Notably, a net present value (NPV) of $1,275,499k and an internal rate of return (IRR) of 65% for Cu recovery from goethite were achieved over 20-years after project started using the aerated and stirred bioreactor plant with a capital expenditure (CAPEX) of $119,816,550 and an operational expenditure (OPEX) of $5,896,580/year. It is expected that plant will start to make profit after one year of operation. Aerated and stirred bioreactor plant appeared more reliable alternative compared to the aerated bioreactor plant as the plant consists of 12 reactors which can allow better management and operation in small volume with multiple reactors. Despite the limitations, this techno-economic assessment emphasized the significance of selective metal recovery and plant design, and underscored the major expenses associated with the process.
Topics: Bioreactors; Metallurgy; Acidithiobacillus; Copper; Minerals; Iron Compounds
PubMed: 38643624
DOI: 10.1016/j.jenvman.2024.120904