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Identifier 000433970
Title Laser microstructured scaffolds for tissue engineering under dynamic cell culture conditions
Alternative Title Κατασκεή ικριωμάτων με χρήση λέιζερ για ιστοτεχνολογία υπό συνθήκες δυναμικής κυτταροκαλλιέργειας
Author Μπαμπαλιάρη, Ελευθερία
Thesis advisor Μητράκη, Άννα
Reviewer Βαμβακάκη, Μαρία
Στρατάκης, Εμμανουήλ
Ρανέλλα, Ανθή
Παπάζογλου, Δημήτρης
Βελώνια, Καλλιόπη
Γκιζελή, Ηλέκτρα
Abstract Although the peripheral nervous system exhibits a higher rate of regeneration than that of the central nervous system through a spontaneous regeneration after injury, the functional recovery is fairly infrequent and misdirected. Thus, the development of successful methods to guide neuronal outgrowth, in vitro, is of high significance. Neural tissue engineering has emerged as a promising alternative field for the development of new nerve graft substitutes to overcome the limitations of the current grafts. The ultimate goal of a tissue-engineered construct is to sufficiently mimic the topographic features of the extracellular matrix and the surrounding environment of cells, such as shear stress and soluble factors, so that cells will function in the artificial environment as they would in vivo. Several approaches have been developed to create cellular substrates at the micro- and/or nano-scales that aim to reconstruct the extracellular matrix architecture in vitro. Among them, ultrafast laser structuring has proved to be important for engineering surface topography in various materials as a simple, fast, and effective method to fabricate micro- and nano-structures with controlled geometry and pattern regularity. Indeed, it has been well-reported that laser-induced topography significantly affects neuronal growth, orientation, and differentiation. Apart from topography, it has recently become increasingly evident that neurogenesis may be also driven by mechanical factors. Mechanical stress is a significant component of the host environment, as it influences the cellular signal transduction and the behavior of various cells. However, the combined effect of flow-induced shear stress and surface topography on neuronal outgrowth has been rarely reported. In the present thesis, a precise flow controlled microfluidic system with specific custom-designed chambers, incorporating the laser-microstructured substrates, was developed to investigate for the first time the combined effect of shear stress and topography on the growth, orientation, and elongation of Schwann (SW10) cells, as well as on the growth and differentiation of neuronal [Neuro-2a (N2a)] cells. Moreover, the effect of topography on the co-culture of SW10 and N2a cells was investigated. For this purpose, polymeric [polyethylene terephthalate (PET)] microstructured substrates with different geometries, microgrooves, chess-like, net-like, were fabricated by ultrafast laser structuring. The cell culture results on the combined effect of shear stress and topography were combined with computational flow simulations to precisely calculate the shear stress values. Our results demonstrate the ability to guide the outgrowth of Schwann cells via flow-induced shear stress and surface topography, which is crucial for neural tissue regeneration. Besides this, our findings revealed that shear stress may affect the differentiation of N2a cells. Thus, the microfluidic system presented here could be potentially used as a new model system to study the role of shear stress (induction and/or inhibition) on cell differentiation. Finally, our preliminary results on a co-culture system, comprising Schwann-SW10 glial cells and neuronal-N2a cells, showed that N2a cells preferentially grew and differentiated on top of the SW10 cells on the flat substrates. Further optimization of this co-culture patterned system will enable closer simulation of the in vivo microenvironment and hence will be useful in neural tissue-engineered dynamic microfluidic systems.
Language English
Subject Neural tissue engineering
Διαφοροποίηση νευρικών κυττάρων
Επιμήκυνση νευρικών κυττάρων
Κατεργασία με υπερβραχείς παλμούς λέιζερ
Μηχανική νευρικών ιστών
Προσανατολισμός νευρικών κυττάρων
Issue date 2021-03-26
Collection   School/Department--School of Sciences and Engineering--Department of Materials Science and Technology--Doctoral theses
  Type of Work--Doctoral theses
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