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3D Stem Cell Culture and Differentiation Using Alginate Hydrogel Scaffolds Incorporated with Silicon-Based Nanoparticles

Hassani, Masoud | 2019

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  1. Type of Document: Ph.D. Dissertation
  2. Language: Farsi
  3. Document No: 52374 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Yaghmaei, Soheila; Arpanaei, Ayyoob; Dolatshahi Pirouz, Alireza
  7. Abstract:
  8. Bone is one of the most vital parts of the body for almost any functional movement. Due to its many involved diseases, traumas and injuries, bone maintenance and regeneration are of high importance. Engineering and design of complex tissues, with impeccable mimicry of the native ones, are always accounted as the solution to dysfunction of body organs due to aging, various diseases, etc. However, the inability to complete mimicry of the native tissue architecture and cell microenvironment are the main barriers to functional tissue regeneration. In this project, in order to design a suitable scaffold for bone regeneration, hydrogel nanocomposites with polysaccharide basis were developed by incorporation of 2D nanomaterials including laponite, montmorillonite, and sumecton with a thickness of 1-2 nm and diameter to thickness ratio of 25, 140, and 260 nm, respectively. To this end, a plethora of experiment was accomplished using high-throughput systems. It was demonstrated that the incorporation of 2D nanosilicates in the hydrogel comprising alginate, hyaluronic acid, and polyethylene glycol improved mechanical properties and biocompatibility. The scaffold biocompatibility was measured using fluorescence microscopy and colorimetric cell cytotoxicity assay kit. In next steps, the hydrogels capability to induce osteogenesis was evaluated by measuring alkaline phosphatase activity and calcium deposition, FTIR technique, X-ray powder diffraction analysis, scanning electron microscopy, and push-out test. Among all, the samples incorporated with montmorillonite nanoparticles have significantly supplemented bone differentiation even in the absence of growth factors. In another part of this study, spherical mesoporous silica nanoparticles (MSN, d≈88 nm) with different surface chemistries (amine, carboxyl, and polymeric amine) were synthesized and investigated in terms of biocompatibility and their impact on the morphology of bone marrow-derived mesenchymal stem cells both in 2D and 3D culture systems. The results showed good biocompatibility for both bare MSNs and amine-functionalized MSNs in 2D and 3D culture systems. For these samples, the viability of about 80% was acquired after 2 weeks of 3D culture. Finally, amine-functionalized MSNs were employed as carriers for controlled release of dexamethasone and ascorbic acid in 2D and 3D culture systems. Alginate-gelatin beads were used for 3D culture. Measurement of alkaline phosphatase activity demonstrated that the smart shipping of these factors via nanoparticles can significantly induce osteogenesis in both culture systems when compared to common method of employing free factors in media
  9. Keywords:
  10. Tissue Engineering ; Hydrogel Nanocomposite ; Alginate ; Silica Nanoparticles ; Mesoporous Silica Nanoparticles ; Bone Regeneration

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