Cardiac Tissue Engineering
1. In vitro 3D engineered cardiac models
Functional in vitro biomimetic cardiac models are a valid tool to investigate processes of myocardial repair/regeneration. We exploit our long-lasting expertise to generate functional engineered contractile 3D micro- and macro-scale cardiac tissues by recapitulating in vitro key biophysical stimuli. Direct perfusion of the culture medium through the cardiac constructs is employed to mimic the highly dense capillary network present in the myocardium to ensure the cardiomyocyte survival in vitro in several mm-thick engineered cardiac constructs.
Electrical and mechanical stimuli are the most investigated physiological stimuli, known to enhance the cardiomyocyte maturation and ultra-structural organization during 3D cell culture. We are currently working on the generation of a miniaturized 3D culture system for the integration of the mechanical and electrical stimulation, with the possibility to assess in real-time the contractile force and evaluate changes and functional improvements in small-scale 3D cardiac engineered tissues (e.g. stripes with a proximal size of 8 x 5 x 1 mm).
Recently, we exploited the organ-on-a-chip technology (in collaboration with Marco Rasponi at the Politecnico of Milano, Italy) to achieve a fine micro-environmental control over the mechanical stimulation to reproducibly engineer contractile 3D cardiac micro-tissues (Marsano et al., Lab on a chip, 2016).
Human inducible pluripotent stem cells (iPSC) are used for the generation of these functional cardiac 3D in vitro models.
Marsano A et al. Beating heart on a chip: a novel microfluidic platform to generate functional 3D cardiac microtissues. Lab Chip. 2016 Jan 26;16(3):599-610. Pubmed.
Cerino G et al. Three dimensional multi-cellular muscle-like tissue engineering in perfusion-based bioreactors. Biotechnol Bioeng. 2015 Jun 30. doi: 10.1002/bit.25688. Pubmed.
Tandon N and Marsano A et al. Surface-patterned electrode bioreactor for electrical stimulation. Lab Chip. 2010 Mar 21;10(6):692-700. Pubmed.
Marsano A et al. Scaffold stiffness affects the contractile function of three-dimensional engineered cardiac constructs. Biotechnol Prog, 2010. Pubmed.
Tandon N et al. Electrical stimulation systems for cardiac tissue engineering. Nat Protoc. 2009;4(2):155-73. Pubmed.
Radisic M et al. Cardiac tissue engineering using perfusion bioreactor systems. Nat Protoc. 2008; 3 (3). Pubmed.
2. Engineered tissues (patches) for the treatment of chronic cardiac ischemia: angiogenic and anti-apoptotic potential.
Chronic myocardial ischemia causes progressive deterioration of cardiac function often leading to end-stage heart failure. However, if blood flow is restored, the low-perfused tissue (hibernating myocardium) is capable of resuming full function. Our proposed treatment consists in using an engineered tissue (patch) generated by a heterogeneous cell population, namely adipose tissue-derived Stromal Vascular Fraction (SVF) (in collaboration with Arnaud Scherberich at the University of Basel). The SVF includes mesenchymal stromal cells, endothelial/mural mature and progenitor cells conferring it a high vasculogenic potential and a broad range of releasing factors (including anti-apoptotic and angiogenic factors). The patches are generated by culturing the human adipose tissue-derived SVF in a 3D perfusion-based bioreactor-based system. We recently observed that compared to static condition the perfusion culture could alone enhance the initial composition of freshly isolated SVF cells by enriching the heterogeneous population for endothelial/mural cells (in particular the pericytes). The engineered tissues (generated in perfusion-based bioreactors) function as angiogenic niches capable to remarkably accelerate the angiogenesis and support the formation of blood vessels by human origin grafted cells upon in vivo implantation.
Cerino G et al. Pericytes determine the development of an engineered pro-angiogenic niche. Scientific Reports 2017 Oct 27;7(1):14252. Pubmed.
Staubli S and Cerino G et al. Control of angiogenesis and host response by modulating the cell adhesion properties of an Elastin-Like Recombinamer-based hydrogel. Biomaterials 2017 Aug;135:30-41. Pubmed.
3. Genetically modified cell-based patches to induce therapeutic angiogenesis
This angiogenic approach aims at engineering cell-loaded patches as controlled Vascular Endothelial Growth Factor (VEGF)-releasing devices to restore the micro-vascularization in cardiac chronic ischemia (in collaboration with Andrea Banfi at the University of Basel). For this purpose, we combined cell-based gene therapy with tissue engineering: transduced human adipose tissue-derived mesenchymal stromal cells (ASC), purified to release a sustained, safe and efficient VEGF dose, were organized in three-dimensional engineered tissues (patches). The VEGF-releasing patches improved the cell survival and promoted a normal and efficient angiogenesis in the surrounding area. In this approach, mesenchymal cells act mainly as vehicle to deliver a safe VEGF dose in the microenvironment, no strong paracrine effects or direct cell contribution to blood vessel formation was indeed observed.
Melly L and Cerino G et al. Myocardial infarction stabilization by cell-based expression of controlled VEGF levels. *equally contributing authors. Journal of Cellular and Molecular Medicine, in press.
Gaudiello E, Melly L, Cerino G, Boccardo S, Jalili-Firoozinezhad S, Xu L, Eckstein F, Martin I, Kaufmann BA, Banfi A, Marsano A. Scaffold composition determines the angiogenic outcome of cell-based Vascular Endothelial Growth Factor expression by modulating its micro-environmental distribution. Advanced Healthcare Materials 2017 Oct 10. Pubmed.
Boccardo S and Gaudiello E et al. “Engineered mesenchymal cell-based patches as controlled VEGF delivery systems to induce extrinsic angiogenesis" Acta Biomater. 2016 Sep 15;42:127-35. Pubmed.
Marsano A et al. Controlled VEGF expression by transduced cells in a cardiac patch improves vascularization and cardiac function in a mouse model of myocardial infarction. Biomaterials, 2013 Jan;34(2):393-401. Pubmed.