Periodic Reporting for period 3 - EMAPS-Cardio (ElectroMechanoActive Polymer-based Scaffolds for Heart-on-Chip)
Período documentado: 2024-03-01 hasta 2025-08-31
EMAPS-Cardio is a response to the demand for more accurate in vitro models. Our vision is to increase the efficiency of drug development and safety screening. EMAPS-Cardio aims to develop models that can predict the success of a drug at a very early stage of drug development – prior to animal testing and clinical trials. This can significantly reduce the overall need for animal testing, failed clinical trials and drug withdrawals, and increase the rate of drug development and new drugs that are brought to market. Devices based on EMAPS can be used to generate organotypic in vitro models that require advanced stimulation, i.e. mechanical, electrical and biochemical, to achieve adult cell maturity. EMAPS-Cardio strives to develop a versatile platform that can be used to grow a variety of tissues, e.g. heart, lung, skin, muscle, etc.
As a first step towards this goal, the EMAPS-Cardio research focused on cardiac models to advance the current technological state-of-the-art by developing clinically relevant, accurate cardiac models for early-stage drug screening that facilitate the faster development of highly efficient cardiovascular drugs and limit the growing socioeconomic burden of cardiovascular diseases. The main objective was to provide in vitro cardiac models that are sufficiently accurate to detect drug potency in both healthy and diseased tissues.
More specifically, EMAPS-Cardio aimed to:
- Develop two devices that can be used for growing any healthy or diseased tissues that require electrical and mechanical stimulation, e.g. heart, lung, skin, muscle, bladder, etc. and validating them for cardiac models
- Deliver accurate healthy and diseased cardiac models that can be further used for understanding the disease progression, testing the impacts of various stimuli (nutrients, nanomaterials, mechanical stress, etc.) on onset and development of the cardiac diseases, testing the efficacy of the drugs, and their cardiotoxicity
- Develop microenvironment that provides all needed stimuli during the differentiation of hiPSC and maturation of tissues, i.e. electroactive, mechanoactive, bioactive scaffolds
- Develop hiPSC differentiation protocols optimized to produce matured myocardium
- Develop sensors for simultaneous and continuous sensing of cardiac health markers (contractility strength and frequency characterization)
- Develop advanced algorithms for detection of the drug efficacy on diseased in vitro myocardium to minimize false-negatives
- Commercialize mechanoactive scaffolds for 3D cell culture, trans-well inserts for heart-on-chip, miniaturized bioreactor for electromechanoactive cell cultures, and improved maturation quality hiPSC-CMs
As a first step towards this goal, the EMAPS-Cardio research focused on cardiac models to advance the current technological state-of-the-art by developing clinically relevant, accurate cardiac models for early-stage drug screening that facilitate the faster development of highly efficient cardiovascular drugs and limit the growing socioeconomic burden of cardiovascular diseases. The main objective was to provide in vitro cardiac models that are sufficiently accurate to detect drug potency in both healthy and diseased tissues.
More specifically, EMAPS-Cardio aimed to:
- Develop two devices that can be used for growing any healthy or diseased tissues that require electrical and mechanical stimulation, e.g. heart, lung, skin, muscle, bladder, etc. and validating them for cardiac models
- Deliver accurate healthy and diseased cardiac models that can be further used for understanding the disease progression, testing the impacts of various stimuli (nutrients, nanomaterials, mechanical stress, etc.) on onset and development of the cardiac diseases, testing the efficacy of the drugs, and their cardiotoxicity
- Develop microenvironment that provides all needed stimuli during the differentiation of hiPSC and maturation of tissues, i.e. electroactive, mechanoactive, bioactive scaffolds
- Develop hiPSC differentiation protocols optimized to produce matured myocardium
- Develop sensors for simultaneous and continuous sensing of cardiac health markers (contractility strength and frequency characterization)
- Develop advanced algorithms for detection of the drug efficacy on diseased in vitro myocardium to minimize false-negatives
- Commercialize mechanoactive scaffolds for 3D cell culture, trans-well inserts for heart-on-chip, miniaturized bioreactor for electromechanoactive cell cultures, and improved maturation quality hiPSC-CMs
EMAPS-Cardio project achieved impact across four main areas:
1. Strengthening the competitiveness and attractiveness of the European bio-medical and healthcare sector
2. Reducing the reliance on animal and clinical testing
3. Increasing awareness, knowledge, and acceptance of organ-on-chip tech-nologies
4. Enhancing understanding of medical regulatory frameworks, particularly among academia and SMEs
Main achievements:
- hiPSC-Cardiomyocytes cell culture bank established
- Standard characterization protocols for hiPSC-CMs established
- Biocompatibility tests of elastic and mechanoactive scaffolds performed
- Bioreactor for maturation studies with all the needed stimuli (biochemical, electrical, and mechanical) developed
- Multi-lens array-based contractility sensor developed and integrated by CSEM into readout module for continuous characterization of cardiac tissues. This sensor is fully compatible with the incubation system provided.
- Three different data analysis pipelines have successfully been implemented. Two are based on the use of biomarkers and the other one in calcium transient. The resulting models were validated by evaluating the effects of cardiovascular drugs with known effects.
- Electromechanical actuation on EMAPS scaffolds with a single 3D-printed insert validated before the full integration of the sensors and transwell inserts
- hiPSC-CMs seeding and culturing conditions optimized on Gel/Glu/PPy scaffolds. Cells are viable after long-term cultivation (at least 7 weeks).
- The detection of differences in early vs. advanced maturation level CM has been obtained by electrophysiology (patch-clamp techniques).
- The optical readout module is fully adapted to parallel reading of inserts in 24-well plates, even for small displacements (< 10 µm).
1. Strengthening the competitiveness and attractiveness of the European bio-medical and healthcare sector
2. Reducing the reliance on animal and clinical testing
3. Increasing awareness, knowledge, and acceptance of organ-on-chip tech-nologies
4. Enhancing understanding of medical regulatory frameworks, particularly among academia and SMEs
Main achievements:
- hiPSC-Cardiomyocytes cell culture bank established
- Standard characterization protocols for hiPSC-CMs established
- Biocompatibility tests of elastic and mechanoactive scaffolds performed
- Bioreactor for maturation studies with all the needed stimuli (biochemical, electrical, and mechanical) developed
- Multi-lens array-based contractility sensor developed and integrated by CSEM into readout module for continuous characterization of cardiac tissues. This sensor is fully compatible with the incubation system provided.
- Three different data analysis pipelines have successfully been implemented. Two are based on the use of biomarkers and the other one in calcium transient. The resulting models were validated by evaluating the effects of cardiovascular drugs with known effects.
- Electromechanical actuation on EMAPS scaffolds with a single 3D-printed insert validated before the full integration of the sensors and transwell inserts
- hiPSC-CMs seeding and culturing conditions optimized on Gel/Glu/PPy scaffolds. Cells are viable after long-term cultivation (at least 7 weeks).
- The detection of differences in early vs. advanced maturation level CM has been obtained by electrophysiology (patch-clamp techniques).
- The optical readout module is fully adapted to parallel reading of inserts in 24-well plates, even for small displacements (< 10 µm).
- The EMAPS-Cardio project produced high quality human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), enabling the investigation of the state-of-the-art maturation processes with novel approaches. The multifactorial maturation perspectives are expected to achieve a new level of cardiomyocytes of human origin for in vitro drug screenings, toxicology and to reduce the number of laboratory animals.
- The initial results of the S-LCA analysis have confirmed that the technology under development does have the potential to create a wide range of social impacts, e.g. better understanding of the cardiovascular diseases, faster, cheaper and more efficient drug development, more personalized medicine in practice, the development of new products and new workplaces, rescued patients, saved hospitalization days, healthier society, greater competitiveness and attractiveness of European biomedical companies. The S-LCA is the guarantee that the development of the technology will be steered towards safe and socially acceptable production, and that awareness will be raised to the intended and expected benefits.
- Electromechanoactive elastic scaffolds have been produced by electrospinning and electrochemical deposition. The electromechanical actuation is efficient in biological medium and will help in increasing the maturation of cardiomyocytes through mechanical stimulation. This technology can also be used in the field of soft robotics, artificial muscles, flexible sensors, etc.
- The development of a novel multi-lens array, high resolution, parallel tracking contractility sensor system for 24 well plates has been achieved enabling the long-term, real-time tissue monitoring
- We created a model that processes cardiac electrical signals to characterize the drug profile of the emitting tissue. The novelty of this approach is the usage of ML models to transfer knowledge from in-vivo to in-vitro tissue. This will significantly reduce the cost of cardiac's drugs experimentation.
- Preparation of controlled drug delivery systems with fluorescence monitoring for both, in-vitro and in-vivo experiments has been achieved and the production of flexible nanofiber membranes for microfluidic detection has been done. All these results will permit to produce novel theranostic systems for early-stage diagnosis and treatment. It will also help in decreasing the medical costs for diagnostics.
- New insert systems with compliant pillars have been developed overcoming the state-of-art by several means. The bendability is regulated using combination of flexible SLA resin and design with fenestration allowing regulation of bendability in different parts of pillars printed in single step process. Moreover, the pillars have a non-published design enabling combination with metallic part for electrical stimulation. Especially the clip-based connectors is a new concept playing role in mechanical positioning and electrical connection.
- The initial results of the S-LCA analysis have confirmed that the technology under development does have the potential to create a wide range of social impacts, e.g. better understanding of the cardiovascular diseases, faster, cheaper and more efficient drug development, more personalized medicine in practice, the development of new products and new workplaces, rescued patients, saved hospitalization days, healthier society, greater competitiveness and attractiveness of European biomedical companies. The S-LCA is the guarantee that the development of the technology will be steered towards safe and socially acceptable production, and that awareness will be raised to the intended and expected benefits.
- Electromechanoactive elastic scaffolds have been produced by electrospinning and electrochemical deposition. The electromechanical actuation is efficient in biological medium and will help in increasing the maturation of cardiomyocytes through mechanical stimulation. This technology can also be used in the field of soft robotics, artificial muscles, flexible sensors, etc.
- The development of a novel multi-lens array, high resolution, parallel tracking contractility sensor system for 24 well plates has been achieved enabling the long-term, real-time tissue monitoring
- We created a model that processes cardiac electrical signals to characterize the drug profile of the emitting tissue. The novelty of this approach is the usage of ML models to transfer knowledge from in-vivo to in-vitro tissue. This will significantly reduce the cost of cardiac's drugs experimentation.
- Preparation of controlled drug delivery systems with fluorescence monitoring for both, in-vitro and in-vivo experiments has been achieved and the production of flexible nanofiber membranes for microfluidic detection has been done. All these results will permit to produce novel theranostic systems for early-stage diagnosis and treatment. It will also help in decreasing the medical costs for diagnostics.
- New insert systems with compliant pillars have been developed overcoming the state-of-art by several means. The bendability is regulated using combination of flexible SLA resin and design with fenestration allowing regulation of bendability in different parts of pillars printed in single step process. Moreover, the pillars have a non-published design enabling combination with metallic part for electrical stimulation. Especially the clip-based connectors is a new concept playing role in mechanical positioning and electrical connection.