Comparison of corneal keratocytes before and after corneal collagen cross-linking in keratoconus patients

Abstract Keratoconus is a bilateral and asymmetric corneal ectasia. Cross-linking uses ultra-violate rays to enhance corneal tissue. The purpose of this study was to use in vivo confocal microscopy to quantitatively analyze microstructural changes over time, after corneal collagen cross-linking (CXL) in keratoconus patients. In this before-and-after study, a total of 45 keratoconic eyes were selected for cross-linking among patients referred to Al-Zahra ophthalmology clinic during 2013–2014. All patients underwent complete ophthalmologic examinations. Keratocytes and the present of activated keratocytes were calculated preoperatively and at 1, 3, 6, and 12 months postoperatively using confocal microscopy. One year after CXL, best corrected and best uncorrected visual acuity was increased significantly (p < 0.001). Spherical equivalent and spherical refractive errors reduced significantly (p < 0.001). The reduction in density of anterior and mid-stromal keratocytes was significant initially (p < 0.001). During follow-up, the density of keratocytes increased significantly in all layers reaching near normal values by 12 months. The percentage of activated keratocytes showed a significant increase 1 month after cross-linking (p < 0.001) albeit this percent reduced as the corneal healing proceeded by month 12. The endothelial cells showed no significant reduction during the follow-up. Collagen cross-linking-induced significant reduction in keratocyte density. The percent of activated keratocytes increased significantly after cross-linking which showed reduction with improvement of corneal healing. After collagen cross-linking, hyper-reflective structures were observed consistent with the stromal collagen structures. Further studies are needed to assess possible changes on corneal biomechanics. The consistency in corneal endothelium numbers proves the safety of this technique.

[1]  C. Mazzotta,et al.  Corneal healing after riboflavin ultraviolet-A collagen cross-linking determined by confocal laser scanning microscopy in vivo: early and late modifications. , 2008, American journal of ophthalmology.

[2]  J. Croxatto,et al.  Sequential in vivo confocal microscopy study of corneal wound healing after cross-linking in patients with keratoconus. , 2010, Journal of refractive surgery.

[3]  J. Holladay,et al.  Visual acuity measurements. , 2004, Journal of cataract and refractive surgery.

[4]  O. Yılmaz,et al.  Efficacy of corneal collagen cross-linking using a custom epithelial debridement technique in thin corneas: a confocal microscopy study. , 2011, Journal of refractive surgery.

[5]  G. Kymionis,et al.  Posterior linear stromal haze formation after simultaneous photorefractive keratectomy followed by corneal collagen cross-linking. , 2010, Investigative ophthalmology & visual science.

[6]  W. Chaidaroon CONFOCAL MICROSCOPY OF THE HUMAN CORNEA , 2007 .

[7]  S. Kaufman,et al.  Corneal Collagen Cross-Linking: A Confocal, Electron, and Light Microscopy Study of Eye Bank Corneas , 2009, Cornea.

[8]  T. Seiler,et al.  Scheimpflug Imaging of Corneas After Collagen Cross-Linking , 2009, Cornea.

[9]  E. Spoerl,et al.  Induction of cross-links in corneal tissue. , 1998, Experimental eye research.

[10]  R. Naranjo-tackman,et al.  Hallazgos tempranos por microscopia confocal en cross-linking , 2013 .

[11]  O. Stachs,et al.  Results of Confocal Microscopy Examinations after Collagen Cross-Linking with Riboflavin and UVA Light in Patients with Progressive Keratoconus , 2010, Ophthalmologica.

[12]  K. Zadnik,et al.  Collaborative Longitudinal Evaluation of Keratoconus (CLEK) Study: methods and findings to date. , 2007, Contact lens & anterior eye : the journal of the British Contact Lens Association.

[13]  D. Patel,et al.  In vivo confocal microscopy analyses of corneal microstructural changes in a prospective study of collagen cross-linking in keratoconus. , 2014, Ophthalmology.

[14]  M. Mannis,et al.  Riboflavin/Ultraviolet A Corneal Collagen Cross-linking for the Treatment of Keratoconus: Visual Outcomes and Scheimpflug Analysis , 2011, Cornea.

[15]  N. Efron,et al.  Corneal confocal microscopy following conventional, transepithelial, and accelerated corneal collagen cross-linking procedures for keratoconus. , 2012, Journal of refractive surgery.

[16]  Ecosse Lamoureux,et al.  A randomized controlled trial of corneal collagen cross-linking in progressive keratoconus: preliminary results. , 2008, Journal of refractive surgery.

[17]  Marcella Q. Salomão,et al.  Corneal wound healing after ultraviolet-A/riboflavin collagen cross-linking: a rabbit study. , 2011, Journal of refractive surgery.

[18]  L. Mastropasqua,et al.  Morphological Modification of the Cornea After Standard and Transepithelial Corneal Cross-linking as Imaged by Anterior Segment Optical Coherence Tomography and Laser Scanning In Vivo Confocal Microscopy , 2013, Cornea.

[19]  Eberhard Spoerl,et al.  Biomechanical evidence of the distribution of cross‐links in corneastreated with riboflavin and ultraviolet A light , 2006, Journal of cataract and refractive surgery.

[20]  W. Abdelghaffar,et al.  Corneal collagen cross-linking: promises and problems , 2010, British Journal of Ophthalmology.

[21]  T. Seiler,et al.  Riboflavin/ultraviolet-a-induced collagen crosslinking for the treatment of keratoconus. , 2003, American journal of ophthalmology.

[22]  Cristina Tommasi,et al.  Treatment of Progressive Keratoconus by Riboflavin-UVA-Induced Cross-Linking of Corneal Collagen: Ultrastructural Analysis by Heidelberg Retinal Tomograph II In Vivo Confocal Microscopy in Humans , 2007, Cornea.

[23]  S. Rabinowitz MAJOR REVIEW OF KERATOCONUS , 1998 .

[24]  Mirko R Jankov II,et al.  Corneal collagen cross linking: Crossing out the microbes , 2019, Journal of EuCornea.

[25]  E. Hernández-Quintela,et al.  Early confocal microscopy findings after cross-linking treatment , 2013 .

[26]  S. Wilson,et al.  Keratocyte apoptosis after corneal surgery. , 1998, Investigative ophthalmology & visual science.

[27]  Theo Seiler,et al.  Keratocyte Apoptosis After Corneal Collagen Cross-linking Using Riboflavin/UVA Treatment , 2004, Cornea.