Periodic Reporting for period 2 - RISEUP (Regeneration of Injured Spinal cord by Electro pUlsed bio-hybrid imPlant)
Berichtszeitraum: 2022-05-01 bis 2023-04-30
To achieve RISEUP main goal of developing an electro-pulsed bio-hybrid (EPB) to attain functional regeneration after SCI, six specific objective were listed in the DoA (i.e. three technological and three biological) and are briefly summarized below:
1. Development of the EPB.
2. Implementation of a wireless system for energy supply and treatment control.
3. Implementation of numerical tools for electromagnetic and neuro-functionalized models to predict neuro-stimulation efficacy.
4. cell fate determination into neuronal phenotypes, cell survival, cell migration and paracrine activity control of iNSCs and MSCSs through a program of specific electrostimulations obtained with a combination of μsPEFs and DC.
5. neuronal activity and networking reorganization (synaptogenesis) in mature neuronal cells and organotypic cultures of spinal cord controlled by calcium oscillations.
6. Functional regeneration after acute SCI in rats by implanting the EPB.
The activity of this second year of the RISEUP project allowed to move steps towards the achievement of the above listed objectives. From the technological point of view, intense work was carried out towards the full development of the EPB (technological objective 1) with the creation of a completely flexible electrode, adaptable to the spinal cord anatomy of rats.
For what concerns the implementation of the wireless system (technological objective 2), the activity carried out lead to an ad-hoc design of a harvesting energy system based on an external monochromatic light source and a subdermal solar cell.
Finally, towards achieving the technological objective 3, the realization of 3D realistic models of iNSCss and mesenchymal stem cells (MSCSs) have been finalized. Such models have been employed in microdosimetric simulations to numerically investigate the electrical quantities induced on cellular and subcellular structures when exposed to the electrical stimulus generated by EPB. In parallel with the technological objectives of the project, the biological activities were implemented during this second year with the setting up of stimulation protocols using µs pulses and DC towards the achievement of the three biological objectives.
1. Development of the EPB.
2. Implementation of a wireless system for energy supply and treatment control.
3. Implementation of numerical tools for electromagnetic and neuro-functionalized models to predict neuro-stimulation efficacy.
4. cell fate determination into neuronal phenotypes, cell survival, cell migration and paracrine activity control of iNSCs and MSCSs through a program of specific electrostimulations obtained with a combination of μsPEFs and DC.
5. neuronal activity and networking reorganization (synaptogenesis) in mature neuronal cells and organotypic cultures of spinal cord controlled by calcium oscillations.
6. Functional regeneration after acute SCI in rats by implanting the EPB.
The activity of this second year of the RISEUP project allowed to move steps towards the achievement of the above listed objectives. From the technological point of view, intense work was carried out towards the full development of the EPB (technological objective 1) with the creation of a completely flexible electrode, adaptable to the spinal cord anatomy of rats.
For what concerns the implementation of the wireless system (technological objective 2), the activity carried out lead to an ad-hoc design of a harvesting energy system based on an external monochromatic light source and a subdermal solar cell.
Finally, towards achieving the technological objective 3, the realization of 3D realistic models of iNSCss and mesenchymal stem cells (MSCSs) have been finalized. Such models have been employed in microdosimetric simulations to numerically investigate the electrical quantities induced on cellular and subcellular structures when exposed to the electrical stimulus generated by EPB. In parallel with the technological objectives of the project, the biological activities were implemented during this second year with the setting up of stimulation protocols using µs pulses and DC towards the achievement of the three biological objectives.
After the setup of the Electro Pulsed Bio-hybrid (EPB) geometry, and the investigation of the biological outcomes to be pursued to induce the cell proliferation and differentiation with electrical stimulation, which were the main activities carried out in the first year of the project, the second year was dedicated to:
• The assessment of the technological procedure to manufacture the flexible and conformable electrode.
• The setting of electric stimulation protocols aimed at inducing the cells proliferation and differentiation.
• The detailed numerical modeling of the EPB and of realistically shaped stem cells, including the representation of neuronal behavior and calcium oscillations.
• The in vitro and in vivo evaluation of the biocompatibility of the materials composing the EPB.
• The first evaluation of immune cells response (microglia and macrophages cells) to the same electric stimulation protocol applied on the stem cells.
As already occurred during the first year, due to tight interconnection between the biological and engineering activities, the beneficiaries worked in a synergistic way to reach the proposed goals.
• The assessment of the technological procedure to manufacture the flexible and conformable electrode.
• The setting of electric stimulation protocols aimed at inducing the cells proliferation and differentiation.
• The detailed numerical modeling of the EPB and of realistically shaped stem cells, including the representation of neuronal behavior and calcium oscillations.
• The in vitro and in vivo evaluation of the biocompatibility of the materials composing the EPB.
• The first evaluation of immune cells response (microglia and macrophages cells) to the same electric stimulation protocol applied on the stem cells.
As already occurred during the first year, due to tight interconnection between the biological and engineering activities, the beneficiaries worked in a synergistic way to reach the proposed goals.
Two years after the starting of the project, the consortium still sees the same impacts as at the beginning.
From a scientific and technological point of view the possibility of determining the fate of stem cells, using electrical stimulation, allows an improvement in the knowledge of the molecular response induced by this exposure, leading to an optimization and expansion of their use in biomedical applications.
A miniaturized electrode, able to exert sequentially different stimulation protocols, is going through the manufacturing process, applying a new technology. The use of porous gold, in place of more rigid solid materials, allows to have a completely flexible structure, that can conform to the shape of the spinal cord and any other curve of the human body, making it potentially suitable for the treatment of all those pathologies where electric stimuli are therapeutic, such as nerve regeneration, or inflammatory diseases. This aspect could impact the current market of flexible electrodes. Indeed, those currently available in the market cannot maintain the conductive characteristics after deep torsions or bending. Whereas the one developed in RISEUP is able to bend even at 180° without structural damage, owed to its intrinsic porous nature.
Furthermore, the technological development to produce an electrify biocompatible scaffolds opens the possibility of exploring new fields of application of these bio-hybrid device. All these aspects will expand the physical agents application in bio-medicine, giving a significant boost to electroceuticals in general.
The advance in the knowledge of stem cells response to electric stimuli will improve their use in tissue regeneration.
From a general point of view, RISEUP represents a great opportunity for personal and professional growth for all the young scientists involved. They can be considered the central core of all the experimental activities, allowing the progression of the project. A mid and long-term impact is the training of these young researchers to develop their own career.
From a scientific and technological point of view the possibility of determining the fate of stem cells, using electrical stimulation, allows an improvement in the knowledge of the molecular response induced by this exposure, leading to an optimization and expansion of their use in biomedical applications.
A miniaturized electrode, able to exert sequentially different stimulation protocols, is going through the manufacturing process, applying a new technology. The use of porous gold, in place of more rigid solid materials, allows to have a completely flexible structure, that can conform to the shape of the spinal cord and any other curve of the human body, making it potentially suitable for the treatment of all those pathologies where electric stimuli are therapeutic, such as nerve regeneration, or inflammatory diseases. This aspect could impact the current market of flexible electrodes. Indeed, those currently available in the market cannot maintain the conductive characteristics after deep torsions or bending. Whereas the one developed in RISEUP is able to bend even at 180° without structural damage, owed to its intrinsic porous nature.
Furthermore, the technological development to produce an electrify biocompatible scaffolds opens the possibility of exploring new fields of application of these bio-hybrid device. All these aspects will expand the physical agents application in bio-medicine, giving a significant boost to electroceuticals in general.
The advance in the knowledge of stem cells response to electric stimuli will improve their use in tissue regeneration.
From a general point of view, RISEUP represents a great opportunity for personal and professional growth for all the young scientists involved. They can be considered the central core of all the experimental activities, allowing the progression of the project. A mid and long-term impact is the training of these young researchers to develop their own career.