The aim of this project was to analyse the cellular and biomechanical changes after collagen cross-linking (CXL) treatment on postmortem eye-banked human corneas using different UVA intensities and repeated treatments, and to explore the effects of standard collagen cross-linking on keratoconic corneal buttons, in-vitro. Preliminary studies were conducted to assess the feasibility of using eye-banked corneas to assess the effects of collagen cross-linking, and the possibility of applying scanning acoustic microscopy (SAM) to measure the speed of sound/elasticity of corneal tissue. Eye-banked human corneas were successfully cross-linked allowing the effects of CXL to be studied in-vitro and SAM was used effectively to determine the mechanical properties of corneal tissue at different depths. The results of two experiments comparing UVA intensity suggested that no statistically significant difference was found in the histological changes or in the induced stiffness after applying low and high intensity cross-linking on normal human corneas. However, the number of apoptotic cells was found to be significantly less but deeper into the posterior stroma in the high intensity cross-linked corneas. Collectively, these results confirmed the safety and efficacy of both techniques with the advantage of reducing the treatment time using the higher-intensity treatment. In another in-vitro study, keratoconic corneal tissue was used. Different histological and biomechanical outcomes were found between the cross-linked and control keratoconic tissue. The effects of cross-linking were found to penetrate deeper in the keratoconic tissue compared to in the normal corneal tissue found in previous studies. This could be due to the altered collagens and extracellular matrix of the keratoconic corneas, as they were taken from patients in advanced stages of the disease. This study confirmed the importance of having corneal thickness of at least 400μm after epithelial debriding to maintain the endothelial cell density and integrity. Finally, further cross-links were induced when collagen cross-linking treatment was repeated. However, repeating cross-linking three times a deeper cell death close to the endothelium was noticed which suggests that multiple treatments could be unsafe. Additionally, lower speed of sound than the cross-linking twice. This could be due to elimination of the induced cross-links by longer exposure to UVA irradiation. In conclusion, eye-banked human corneas were successfully used to evaluate the effects of cross-linking treatment and repeated treatment. Additionally, keratoconic corneal buttons were used to study the effects of collagen cross-linking in-vitro. This model of using eye-banked human corneas and keratoconic corneal tissue enabled us to study the effects of cross-linking treatment using different protocols and the effects of repeated treatment, and it could ultimately be used to compare the results with in-vivo studies.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:632199 |
| Date | January 2013 |
| Creators | Beshtawi, Ithar |
| Contributors | O'Donnell, Clare; Hillarby, Mary; Radhakrishnan, Hema |
| Publisher | University of Manchester |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | https://www.research.manchester.ac.uk/portal/en/theses/the-structural-and-functional-effects-of-corneal-collagen-crosslinking-on-human-corneal-tissue(12f210fe-82b6-4855-a3ea-c5abd828642d).html |
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