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Development and Characterization of Novel Bioink by Using Decellularized Extracellular Matrix for Bone Tissue Engineering Applications
(01. Izmir Institute of Technology, 2023) Kara Özenler, Aylin; Tıhmınlıoğlu, Funda; Havıtçıoğlu, Hasan
Bone tissue engineering has focused on the development of functional scaffolds that can organize bone regeneration with appropriate structures and properties. Three-dimensional (3D) printing technology enables the development of personalized scaffolds. In addition, biological scaffolds obtained by decellularization have various advantages for developing natural-based scaffolds. The development of printable, patient-specific bioinks derived from decellularized extracellular matrix could provide 3D fabrication of tissues and organs with high potential to mimic native tissues. The presented thesis study demonstrates the development of various bioink compositions for bone tissue engineering applications. In this regard, bone tissues were decellularized with a novel method and then characterized in order to verify the removal of whole cellular components for eliminating immunological reactions. After the pulverization of tissues, decellularized bone (DB) particles were used as an additive within various ink combinations (alginate-, gelatin- and alginate-gelatin-based). Thus, various bioink formulations were developed containing DB particles, biopolymers and mesenchymal stem cells (MSC). All prepared bioinks were bioprinted, then the viability, proliferation and differentiation capacity of the cells inside the structures as well as the physical, rheological, and printability properties of the inks were assessed. The results revealed that all bioink combinations were suitable for bioprinting and the addition of DB particles improved cell proliferation and osteogenic differentiation in all bioink formulations. Alginate-based bioinks exhibited the greatest printability and shape fidelity, gelatin-based bioinks showed the highest cell proliferation and attachment, also, gelatin incorporation into alginate-based bioinks improved the biological activity of cells. In conclusion, cytocompatible, functional composite bioinks developed in this thesis study are of value for bone tissue engineering research in future to explore their functions in the living system and show complete bone regeneration while maintaining their stability for a long time.
Molecular and Cellular Level Adaptations of Bone Marrow Mesenchymal Progenitor Cells To Chemical and Physical Signals
(Izmir Institute of Technology, 2020) Baskan Erbilgiç, Öznur; Özçivici, Engin; Atabey, Safiye Neşe
Mechanical forces are the integral determinants in cell and tissue homeostasis and regeneration, and they can affect numerous biological process from proliferation to fate determination. Mechanical forces that possess low magnitude and high frequency characteristics are also known as low intensity vibrations (LIVs). These signals were studied widely on many cell types for regenerative purposes, however most of these studies select components of LIV signals (e.g. magnitude, frequency, duration, etc.) arbitrarily. Here, we addressed the effect of LIV applied frequency, LIV daily exposure time and fate induction on the viability of preadipocyte 3T3-L1 cells. For this, we performed a frequency sweep that was ranging from 30 to 120 Hz with 15 Hz increments applied for 5, 10 or 20 minutes during quiescent growth or adipogenesis for up to 10 days. Results suggest that the applied frequency and fate induction was an important determinant of cell viability, lipid droplet physiology, triglyceride concentration, cell density and adipogenic-specific gene expression while daily exposure time had no effect. These findings contribute to the effort of optimizing a relevant mechanical stimulus that can inhibit adipogenesis. On the other hand, random and aligned PAN/PPy nanofibers were investigated as a scaffold material for osteogenic differentiation of D1 ORL UVA mouse bone marrow mesenchymal stem cells. Cells were able to attach and grow on nanofibers confirmed by cell viability results. Stem cells that were cultured with osteogenic induction were able to mineralize on electrospun nanofibers based on alizarin red and Von Kossa dye staining. For aligned PPy nanofibers, mineralization occurred in the fiber alignment direction. Consequently, PAN/PPy nanofibrous mats in both random and aligned forms would be potential candidates for bone tissue engineering.

Phd Degree / Doktora

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