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Effects of riboflavin/UVA corneal cross-linking on keratocytes and collagen fibres in human cornea
- Source :
- Clinical & Experimental Ophthalmology. 38:49-56
- Publication Year :
- 2010
- Publisher :
- Wiley, 2010.
-
Abstract
- Purpose: To evaluate the effects of corneal cross-linking on keratocytes and collagen fibres in human corneas. Methods: Fifteen corneal buttons were examined. Ten were from patients with keratoconus submitted to penetrating keratoplasty and five of them were treated with cross-linking 6 months before penetrating keratoplasty. Five normal corneal buttons from healthy donors were used as controls. All samples were prepared for TUNEL assay and Western blot analysis for the detection of keratocyte apoptosis and immunohistochemical analysis for the morphological evaluation of keratocytes and collagen fibre diameter. Results: Normal corneas exhibited no TUNEL-positive keratocytes and keratoconic and cross-linked corneas showed moderate apoptotic cells mainly in the anterior part of the stroma. This apoptotic trend was confirmed by the cleavage of poly (ADP-ribose) polymerase assessed using Western blot. The Ki-67 staining showed a significant increase in the keratocyte proliferation in cross-linked corneas compared with normal and keratoconus. In cross-linked corneas CD34-positive keratocytes were regularly distributed throughout the whole corneal stroma as in the control, and keratoconus was associated with patchy loss of immunoreactivity. The immunohistochemical analysis of collagen type I showed a significant increase in fibre diameter of cross-linked corneas compared with control and keratoconus. Conclusion: Corneal cross-linking leads to keratocyte damage; after 6 months a repopulation by proliferating cells, a distribution of CD34-positive keratocytes as in control and an increase in collagen fibre diameter were observed. These modifications are the morphological correlate of the process leading to an increase in biomechanical stability.
- Subjects :
- Adult
Keratoconus
medicine.medical_specialty
genetic structures
Ultraviolet Rays
Corneal Stroma
Riboflavin
Blotting, Western
Antigens, CD34
Apoptosis
Immunoenzyme Techniques
Western blot
Stroma
Ophthalmology
Cornea
In Situ Nick-End Labeling
medicine
Humans
Cell Proliferation
Photosensitizing Agents
TUNEL assay
medicine.diagnostic_test
business.industry
Anatomy
Fibroblasts
medicine.disease
eye diseases
Staining
Blot
Ki-67 Antigen
medicine.anatomical_structure
Photochemotherapy
Immunohistochemistry
Collagen
sense organs
business
Keratoplasty, Penetrating
Subjects
Details
- ISSN :
- 14429071 and 14426404
- Volume :
- 38
- Database :
- OpenAIRE
- Journal :
- Clinical & Experimental Ophthalmology
- Accession number :
- edsair.doi.dedup.....958c97b2e60a61c8a5e63241f93838c3