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Scaffold Design and Fabrication for Retinal Tissue Engineering

Abdi, Romina | 2023

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 56359 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Mashayekhan, Shohreh
  7. Abstract:
  8. The retina, a photosensitive area in the central nervous system, is delicate and intricate. It is susceptible to degenerative disorders such as age-related macular degeneration (AMD), retinitis pigmentosa (RP), Stargardt disease (SD), and glaucoma. These diseases can lead to severe vision loss and ultimately irreversible blindness by causing destruction or dysfunction of different types of retinal cells. Unfortunately, there are no proven treatment strategies to cure or reverse these degenerative disorders. However, cell transplantation therapies may be an alternative to replace distorted cells and improve an individual's vision. Recent clinical outcomes show that transplanted cells in the retina have better cell survival when supported by a scaffold compared to cell suspensions. Therefore, retinal tissue scaffolds can increase cell viability and adhesion in the appropriate position within the retina. Among various synthetic and natural materials, retinal ECM-based hydrogel scaffolds are highly regarded due to their ability to mimic the body's conditions and microenvironment for cultured cells. In this project, we designed and fabricated a hydrogel scaffold based on retinal ECM. We added oxidized alginate with an oxidation degree of 5% to the mixture to improve mechanical properties and control biodegradability. Oxidized alginate can make ionic bonding with calcium ions and also form crosslinks with the amine groups of retinal ECM through covalent bonding to improve mechanical properties. We performed a DAPI assay to analyze the decellularization process and determine the appropriate protocol for retinal tissue decellularization. We also carried out FTIR and spectrophotometry assays to identify the characteristics of oxidized alginate. FTIR and FE-SEM analysis showed the formation of bonds and a porous network structure in the hydrogel. We optimized the mechanical properties of the scaffolds using compressive strength tests and rheological evaluations, which showed that the hydrogel with 3% alginate concentration is in the range of native retinal tissue compressive modulus. The hydrogel scaffolds also showed suitable swelling profiles and biodegradability. We performed MTT and cell adhesion assays using FE-SEM analysis to evaluate the biocompatibility of the scaffolds. Eventually, the hydrogel with 3% alginate showed the best cell survival rate and cell growth compared to other groups
  9. Keywords:
  10. Extracellular Matrix ; Retina ; Oxidized Alginate ; Tissue Engineering ; Retinal Degenerative Diseases ; Retinal Tissue Engineering

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