A Tissue Engineering product development pathway

Tissue engineering is a field of inquiry and research that uses engineering techniques and principles of biological sciences to develop functional substitutes for reconstruction of damaged organs. Commercial translation of tissue engineering products is currently in progress all over the world. Many companies are moving their interest towards this market segment that grows by 6% per year. Aim of this thesis is to probe the possibility of developing tissue engineering products in the most cost-effective way, minimizing the industrial risk and developing a specific fund raising model. Tissue engineering is based on three main features: cells, scaffolds and bioreactors. Cells are seeded on a scaffold and cultured in a bioreactor in order to obtain a tissue engineering product. Nevertheless, developing cell carrying products is hampered by certification claims ("advanced therapies" certification rules) that unbearably increase R&D and certification costs and can be faced by either big companies or start-ups of big companies and spin-offs of complex aggregates of research centers involved in advanced cell research. On the other hand, scaffolds (certification class IIb) and bioreactors for tissue engineering (certification class I) can be developed with a lower economic effort, being the competition based on innovation, since their market is in the "growth phase" for scaffolds and in the "introduction phase" for bioreactors in the Levitt's product life cycle theory. Purpose of this thesis is to basically study scaffold and bioreactor features, then to preliminarily design some models of bioreactors and, eventually, to set a business model, based on private and public fund raising, aimed to the development of scaffolds for dental implantology and of bioreactors for cardiovascular and bone tissue engineering. Finally, a business plan of a company being spin-off of Politecnico di Torino and industrial start-up has been elaborate

Computational fluid dynamic analysis of bioprinted self-supporting perfused tissue models

Natural tissues are incorporated with vasculature, which is further integrated with a cardiovascular system responsible for driving perfusion of nutrient‐rich oxygenated blood through the vasculature to support cell metabolism within most cell‐dense tissues. Since scaffold‐free biofabricated tissues being developed into clinical implants, research models, and pharmaceutical testing platforms should similarly exhibit perfused tissue‐like structures, we generated a generalizable biofabrication method resulting in self‐supporting perfused (SSuPer) tissue constructs incorporated with perfusible microchannels and integrated with the modular FABRICA perfusion bioreactor. As proof of concept, we perfused an MLO‐A5 osteoblast‐based SSuPer tissue in the FABRICA. Although our resulting SSuPer tissue replicated vascularization and perfusion observed in situ, supported its own weight, and stained positively for mineral using Von Kossa staining, our in vitro results indicated that computational fluid dynamics (CFD) should be used to drive future construct design and flow application before further tissue biofabrication and perfusion. We built a CFD model of the SSuPer tissue integrated in the FABRICA and analyzed flow characteristics (net force, pressure distribution, shear stress, and oxygen distribution) through five SSuPer tissue microchannel patterns in two flow directions and at increasing flow rates. Important flow parameters include flow direction, fully developed flow, and tissue microchannel diameters matched and aligned with bioreactor flow channels. We observed that the SSuPer tissue platform is capable of providing direct perfusion to tissue constructs and proper culture conditions (oxygenation, with controllable shear and flow rates), indicating that our approach can be used to biofabricate tissue representing primary tissues and that we can model the system in silico

A dual flow bioreactor with controlled mechanical stimulation for cartilage tissue engineering

In cartilage tissue engineering bioreactors can create a controlled environment to study chondrocyte behavior under mechanical stimulation or produce chondrogenic grafts of clinically relevant size. Here we present a novel bioreactor, which combines mechanical stimulation with a two compartment system through which nutrients can be supplied solely by diffusion from opposite sides of a tissue engineered construct. This design is based on the hypothesis that creating gradients of nutrients, growth factors and growth factor antagonists can aid in the generation of zonal tissue engineered cartilage. Computational modeling predicted that the design facilitates the creation of a biologically relevant glucose gradient. This was confirmed by quantitative glucose measurements in cartilage explants. In this system it is not only possible to create gradients of nutrients, but also of anabolic or catabolic factors. Therefore, the bioreactor design allows control over nutrient supply and mechanical stimulation useful for in vitro generation of cartilage constructs that can be used for the resurfacing of articulated joints or as a model for studying OA disease progression

Model of murine ventricular cardiac tissue for in vitro kinematic-dynamic studies of electromagnetic and beta2-adrenergic stimulation

In a model of murine ventricular cardiac tissue in vitro, we have studied the inotropic effects of electromagnetic stimulation (frequency, 75 Hz), isoproterenol administration (10 μM), and their combination. In particular, we have performed an image processing analysis to evaluate the kinematics and the dynamics of beating cardiac syncytia starting from the video registration of their contraction movement. We have found that the electromagnetic stimulation is able to counteract the β-adrenergic effect of isoproterenol and to elicit an antihypertrophic response

Training-to-Beat bioreactor for investigating Engineered Cardiac Tissues: design, development & validation

In last the decades, advances relevant to the generation of 3D Engineered Cardiac Tissues (ECTs) have been made. In reason of this, ECTs are now considered a great promise for in vitro studies of cardiac development, disease and, eventually, for strategies for the repair of the structure and function of the injured myocardium. Among the several physical stimuli which have been exploited to improve the functionality and maturation of ECTs, a preeminent role has been ascribed to mechanical stimulation. Appropriate mechanical stimulation can be recreated and maintained within bioreactors, which are devices/platforms devoted to mimic the physiological milieu in a monitored/controlled culture environment, where the engineered constructs can be properly stimulated. One main limitation of the bioreactor-based strategy for cardiac tissue engineering applications is that the devices which are currently used are meant to passively apply to ECTs a stimulus predefined by the user, regardless of their level of maturation along the duration of the in vitro culture. In this scenario, and trying to overcome current limitations, a novel bioreactor design has been conceived for the investigation of 3D ECTs with a biomimetic approach. Technically, the here proposed bioreactor is capable (1) to apply native-like or pathologic mechanical stimuli (cyclic strain) by means of a reliable linear actuator operating in a wide range of strains and frequencies, and (2) to monitor in real-time both chemo-physical parameters (e.g. oxygen tension, pH) of the milieu and the mechanical stiffness of ECTs by means of dedicated sensors, eventually adapting the stimulation to the actual stage of maturation of the constructs. As a proof of concept, a first experimental campaign has been carried out with a double aim: (1) to verify the bioreactor feasibility in delivering mechanical cyclical stimulation to 3D fibrin-based, ring shaped Engineered Cardiac Tissues (ECTs); (2) to assess the effect of cyclic strain on tissue maturation, contractility and modification on its mechanical properties. In detail, the bioreactor platform has been preliminarily tested to verify protocols for hold on, sterilization, and control of the delivered mechanical stimuli. Firstly, the suitability of the bioreactor platform in culturing ad-hoc designed constructs, in terms of ease of use and capability in setting the stimulation parameters, has been tested. Then, the observed maturation of ring shaped ECTs subject to sinusoidal cyclic strain within the bioreactor has confirmed the potency of the proposed approach and the instrumental role of mechanical stimulation in ECTs maturation and in the development of an adult-like cardiac phenotype responsive to electrical excitation. Even if further validation steps are required before the implementation of culture strategy fully adaptive in terms of mechanical stimuli applied to the engineered cardiac constructs, the developed bioreactor represents a valuable proof of concept for, in its most advanced operational mode, biomimetic culturing of engineered cardiac constructs

Cells in Space

Discussions and presentations addressed three aspects of cell research in space: the suitability of the cell as a subject in microgravity experiments, the requirements for generic flight hardware to support cell research, and the potential for collaboration between academia, industry, and government to develop these studies in space. Synopses are given for the presentations and follow-on discussions at the conference and papers are presented from which the presentations were based. An Executive Summary outlines the recommendations and conclusions generated at the conference

IV.3. Bioreactors in tissue engineering.

IV.3. Bioreactors in tissue engineering