Periodic Reporting for period 3 - IMBIBE (Innovative technology solutions to explore effects of the microbiome on intestine and brain pathophysiology)
Período documentado: 2020-10-01 hasta 2022-03-31
The human gut is host to over 100 trillion bacteria that are known to be essential for human health. Intestinal microbes can affect the function of the gastrointestinal (GI) tract, via immunity, nutrient absorption, energy metabolism and intestinal barrier function. Alterations in the microbiome have been linked with many disease phenotypes including colorectal cancer, Crohn’s disease, obesity, diabetes as well as neuropathologies such as autism spectrum disorder (ASD), stress and anxiety. Animal studies remain one of the sole means of assessing the importance of microbiota on development and well-being, however the use of animals to study human systems is increasingly questioned due to ethics, cost and relevance concerns. In vitro models have developed at an accelerated pace in the past decade, benefitting from advances in cell culture (in particular 3D cell culture and use of human cell types), increasing the viability of these systems as alternatives to traditional cell culture methods. This in turn will allow refinement and replacement of animal use. In particular in basic science, or high throughput approaches where animal models are under significant pressure to be replaced, in vitro human models can be singularly appropriate. The development of in vitro models with microbiota has not yet been demonstrated even though the transformative role of the microbiota appears unquestionable. The IMBIBE project is focussed on using engineering and materials science approaches to develop complete (i.e. human and microbe) in vitro models to truly capture the human situation. IMBIBE benefits from cutting edge organic electronic technology which will allow real-time monitoring thus enabling iterative improvements in the models employed. The result from this project will be a platform to study host-microbiome interactions and consequences for pathophysiology, in particular, of the GI tract and brain.
So far we have focused on a model of the human gut which we have now integrated into the platform using human-derived cells representative of those found in the body. We have demonstrated that cells grow readily and form tissue-like architectures within the conducting polymer scaffold device. The process of tissue formation inside the conducting polymer channel gradually modulates the transistor characteristics. We have demonstrated long-term operation (1 month) of non-invasive, in-line electrical monitoring of tissue formation. The next phase of this work will involve optimising the fluidics and connection of multiple modules to allow for gut-brain connection to the brain module which is currently undergoing testing, expected to transition to the tubistor platform shortly. Initial studies have also begun on integration of bacteria into our human intestinal model, with promising results showing that we can maintain bacterial and human cells in culture together.
Although our ultimate goal is the connected model of gut-brain and microbiome, the individual modules are already garnering interest as test models for a variety of laboratories worldwide, in Korea, Finland, Wales and Ireland so far. Many groups are keen to test bacterial products or mimic diseases such as Crohn’s disease to determine how they might use microbes or their derivatives to protect against or even reverse deleterious effects on the gut (and brain). In the short term, this project has the potential to dramatically change how such things are studied in vitro. In the long term, our work will contribute towards the understanding of diseases related to microbial imbalances in the human body.
Although our ultimate goal is the connected model of gut-brain and microbiome, the individual modules are already garnering interest as test models for a variety of laboratories worldwide, in Korea, Finland, Wales and Ireland so far. Many groups are keen to test bacterial products or mimic diseases such as Crohn’s disease to determine how they might use microbes or their derivatives to protect against or even reverse deleterious effects on the gut (and brain). In the short term, this project has the potential to dramatically change how such things are studied in vitro. In the long term, our work will contribute towards the understanding of diseases related to microbial imbalances in the human body.
Our tubistor platform is widely considered to be both novel and highly unconventional compared to existing organ on chip type platforms. Although a growing number of organ-on-chip applications have begun to integrate electrodes to allow for real-time monitoring of barrier integrity very few of these are adapted to 3D cultures. Not only do we provide a highly biomimetic tissue environment, we show evidence of real-time tissue monitoring.
The tunable properties of PEDOT:PSS and the in situ fabrication process we adopted allowed us to fine-tune the electrical, mechanical and biochemical properties of the scaffolds. This allows really interesting tissue engineering applications, where cell growth and fate can be determined by changing the combination of these stimuli in vitro.
Our new generation Tubistor platforms allows us to mimic luminal architecture of the human gastrointestinal tract. The new version of the device supports maintenance and monitoring of a biological model of the human gut epithelium for 26 days. Our approach to gradually build the tri-culture intestinal cell model by firstly culturing fibroblasts in the porous bulk compartment of the hollow scaffolds and later injecting intestinal epithelial cells to line the lumen lining and form the epithelial layer anchoring on the basal lamina was proven successful for the reconstruction of the desired tissue microenvironment. The configuration/design of our device enables real-time, non-invasive monitoring of cell activity and tissue formation on the scaffolds, as evidenced by modulation of their electrical properties. In addition, the configuration of our bioelectronic platform enables bi-modal operation of the device – both as electrode and as a transistor – thereby providing us with more electrical readouts, analysis of which reveals valuable information for the biological model in real time, cross-validated with optical analysis. The unique integration of in-line sensing components in a 3D intestinal system achieved with our system highlights the potential of this technology for building more advanced experimental models of the human gut as tools for studying disease pathology, host-pathogen and host-microbiome interactions, as well as for identifying novel therapeutic targets.
The tunable properties of PEDOT:PSS and the in situ fabrication process we adopted allowed us to fine-tune the electrical, mechanical and biochemical properties of the scaffolds. This allows really interesting tissue engineering applications, where cell growth and fate can be determined by changing the combination of these stimuli in vitro.
Our new generation Tubistor platforms allows us to mimic luminal architecture of the human gastrointestinal tract. The new version of the device supports maintenance and monitoring of a biological model of the human gut epithelium for 26 days. Our approach to gradually build the tri-culture intestinal cell model by firstly culturing fibroblasts in the porous bulk compartment of the hollow scaffolds and later injecting intestinal epithelial cells to line the lumen lining and form the epithelial layer anchoring on the basal lamina was proven successful for the reconstruction of the desired tissue microenvironment. The configuration/design of our device enables real-time, non-invasive monitoring of cell activity and tissue formation on the scaffolds, as evidenced by modulation of their electrical properties. In addition, the configuration of our bioelectronic platform enables bi-modal operation of the device – both as electrode and as a transistor – thereby providing us with more electrical readouts, analysis of which reveals valuable information for the biological model in real time, cross-validated with optical analysis. The unique integration of in-line sensing components in a 3D intestinal system achieved with our system highlights the potential of this technology for building more advanced experimental models of the human gut as tools for studying disease pathology, host-pathogen and host-microbiome interactions, as well as for identifying novel therapeutic targets.