PhD Report – Ludovica Cacopardo
PhD Program in Information Engineering – Cycle XXXIResearch activity
The aim of my PhD project was to contribute to the engineering of physiologically relevant in-vitro models with the design of systems able to mimic and monitor the dynamic environment of cellular constructs. These advanced models are emerging as a powerful solution to bridge the gap between basic and clinical research, currently hindered by the low predictivity and limited successful translation rate of animal models. In particular, a cellular impedance-meter was designed and interfaced with a dual-flow bioreactor to monitor the electrical properties of biological barriers. Indeed, epithelial and endothelial tissue allow the separation of different compartments in the human body and are normally exposed to shear stress on both sides (e.g. blood and interstitial fluid flow). The system was validated with an intestinal model, demonstrating that dynamic conditions contribute significantly to the tightness of the barrier. Possible applications of the system range from drug testing to the study of several pathologic conditions related with an alteration of barrier functions. Since it is well known that mechanical properties of tissues dynamically evolve during growth, ageing or diseases like fibrosis, different methods were investigated to modulate spatial and temporal viscoelastic properties of hydrogel-based scaffolds. In order to monitor mechanical properties of cellular constructs in real-time, the MechanoCultureTR (MCTR) bioreactor was re-engineered in collaboration with the Canadian company CellScale. Finally, a model of liver fibrosis was implemented mimicking in-vitro the ‘healthy’-fibrotic transition with an enzymatic stiffening method and monitoring the mechanical properties of hepatocyte laden gels in the MCTR. The results obtained represent a first step towards physiologically relevant models useful in the understanding of fibrosis mechanobiological mechanisms and progression or in the study of antifibrotic drugs.
In conclusion, during the PhD, advanced models with integrated sensing for the real-time monitoring of electrical and mechanical properties of cell construct were designed, tested and validated. The results obtained have relevant implications both in the medical field, contributing to the study of disease development, and in the pharmaceutic and cosmetic fields, providing reliable tools for testing chemical compounds and drugs.
Publications
International Journals
L. Cacopardo, N. Guazzelli, R. Nossa, G. Mattei and A. Ahluwalia: “Engineering hydrogel viscoelasticity’’, Journal of Mechanical Behaviour of Biomedical Materials (Accepted September 2018 - doi: 10.1016/j.jmbbm.2018.09.031, publication: January 2019, vol. 89, pp. 162-167)
O.Ramirez, L. Cacopardo, B. Leon, J. Costa “Design and development of a dual-flow bioreactor mimicking intestinal peristalsis and permeability in epithelial tissue barriers for drug transport assessment”, 2019, Biocell (Vol. 43.1, p. 29-35, doi:10.32604/biocell.2019.04790)
G. Mattei, L. Cacopardo, A. Ahluwalia: ‘’Micro-Mechanical Viscoelastic Properties of Crosslinked Hydrogels Using the Nano-Epsilon Dot Method. Materials’’, 2017, MDPI (Vol. 10, p. 889, doi:10.3390/ma10080889).
S. Giusti, D. Mazzei, L. Cacopardo, G. Mattei, C. Domenici, A. Ahluwalia. ‘’Environmental Control in Flow Bioreactors’’, 2017, Processes, MDPI (Vol. 5, p. 16).
International Conferences with peer review
L. Cacopardo, F. Burgio., F. Riccio, G. Mattei, A. Ahluwalia: ‘’New force-controlled testing method for real-time viscoelastic measurements in bioreactors’’. World Congress of Biomechanics, July 2018, Dublin (Ireland) – Oral presentation
L. Cacopardo, N. Guazzelli, R. Nossa, G. Mattei, A. Ahluwalia: ‘’Engineering Viscoelasticity in Biomaterials’’. World Congress of Biomechanics, July 2018, Dublin (Ireland) – Oral presentation
L. Cacopardo, N. Guazzelli, R. Nossa, G. Mattei, A. Ahluwalia: ‘’Engineering Viscoelasticity in Biomaterials’’. Nanoengineering for Mechanobiology, March 2018, Camogli (Italy) – Oral presentation L. Cacopardo, G. Mattei, A. Ahluwalia: ‘’Alginate gel with spatially tunable mechanical properties for
pathophysiological in-vitro models’’. European Society of Biomechanics Congress, July 2017, Seville (Spain) - Oral Presentation
G. Mattei, L. Cacopardo, Nossa R., Guazzelli N., A. Ahluwalia: ‘Effect of testing sample and geometry on biomaterial mechanical properties’’. European Society of Biomechanics Congress, July 2017, Seville (Spain) - Oral Presentation
L. Cacopardo, L. Buoncompagni, J. Costa, S. Giusti, S. Meucci, C. Domenici, A. Ahluwalia: ‘’Cellular Impedance Monitoring and Real Time Imaging in a Bioreactor for In-Vitro Models of Biological Barriers’’. Advance in Cell & Tissue Culture, May 2017, Manchester (UK) – Poster
L. Cacopardo, G. Mattei, A. Ahluwalia: ‘’Alginate gel with spatially tunable mechanical properties for pathophysiological in-vitro models’’. Nanoengineering for Mechanobiology, March 2017, Camogli (Italy) – Poster
L. Cacopardo, G. Mattei, A. Ahluwalia: ‘’Enzymatically Stiffenable Gelatin Hydrogels for Engineering Pathophysiological Organ Models’’. European Society of Biomechanics Congress, July 2016, Lyon (France) – Oral Presentation
L. Cacopardo, J. Costa, S. Giusti, C. Domenici, A. Ahluwalia. ‘’Cellular Impedance Monitoring and Real Time Imaging in a Bioreactor for In-Vitro Models of Biological Barriers’’. Advance in Cell & Tissue Culture, May 2016, Barcelona (Spain) – Poster
Formation Activities
English for Academic Purpose C1: 4 CFU PhD+: 9 CFU
Meccanica dei Continui: 4 CFU
Nanoscale Biohybrid Engineering: 5 CFU Rapid Prototyping for Engineers: 3 CFU English for Academic Purpose C1+: 4 CFU
STMicroelectronics - “Semiconductor trip: From a simple idea to a complex manufacturing”: 6CFU TOTAL CFU: 35 (ext 0 int 35)
Teaching Activities
Support for teaching activities (20 hours) the course of Biomimetic and Smart Material - Biomedical Engineering (Master), 2nd semester A.A 2018/19, University of Pisa
Support for teaching activities (20 hours) the course of Biomimetic and Smart Material - Biomedical Engineering (Master), 2nd semester A.A 2017/18, University of Pisa
Support for teaching activities (20 hours) for the course of Transport Phenomena - Biomedical Engineering (Bachelor), 1st semester A.A 2016/17, University of Pisa
