The Viability Control of Human Endothelial Cells Before Keratoplasty (V-CHECK) Study (V-CHECK)

Preoperative Surgeon Evaluation of Corneal Endothelial Status: the Viability Control of Human Endothelial Cells Before Keratoplasty (V-CHECK) Study

The purpose of this international cohort study is to validate a new method for preoperative assessment of endothelial viability in donor corneal tissues for transplantation, and to correlate endothelial health as assessed by the surgical team to functional and structural long-term outcomes in the cohort of patients receiving them.

Study Overview

• Status: Recruiting
• Conditions: Corneal Endothelial Cell Loss; Corneal Transplant Failure
• Intervention / Treatment: Diagnostic test: V-Check

Detailed Description

The corneal endothelium is responsible for preserving corneal transparency by maintaining a dynamic equilibrium of corneal stromal hydration. Corneal Endothelial Cells (CECs) health is approximated by evaluating the Endothelial Cell Density (ECD) of corneal tissues though specular biomicroscopy. ECD is a key determinant of long-term tissue survival after corneal transplantation, as corneal graft function is directly dependent on the number of viable CECs. However, ECD represents only an estimation of total corneal endothelial health and is a suboptimal proxy for future endothelial failure.

In the current chain of supply of donor corneal tissues, no independent evaluation of endothelial cell viability by the surgical team tasked with transplanting the donor corneal tissue is anticipated. The investigators have previously shown a poor correlation between ECD declared by the eye bank providing the tissue and tissue quality as assessed independently by a cornea surgeon performing the transplantation (unpublished data).

In this study, the investigators describe a simple method to independently evaluate the overall endothelial viability of donor corneal tissue which can be adopted both by eye banks before shipment and by the corneal surgeons before transplantation in the operating theatre. This method uses basic image analysis to assess the extent of endothelial cell death and/or denuded endothelial areas with the use of Trypan Blue, a vital dye commonly employed during corneal transplantation. Endothelial cell loss is visualized and quantified as Trypan Blue Positive Areas (TBPA).

The primary objective of this study is to quantify the extent of TBPA in the operating theatre before corneal transplantation and correlate it to overall donor mortality declared by the eye bank. Secondary objectives include the assessment of the correlation of TBPA with declared ECD and CECs mortality, on folds, as well as the longitudinal analysis of endothelial cell loss as a function of time from surgery and initial TBPA over a follow-up of 12 months, final ECD, incidence of graft failure, and risk of graft failure attributable to preoperative TBPA.

• Study Type: Observational
• Enrollment (Estimated): 135

Contacts and Locations

• Study Contact: Name: Matteo Airaldi, MD; Phone Number: +390303995847; Email: matteo.airaldi@unibs.it

Study Locations

• Belgium
  • Edegem, Belgium, 2650
    • Not yet recruiting
    • Antwerp University Hospital
    • Contact: Sorcha Ni Dhubhghaill, MD
    • Principal Investigator: Sorcha Ni Dhubhghaill, MD
• Germany
  • Cologne, Germany, 50937
    • Not yet recruiting
    • Department of Ophthalmology, University of Cologne
    • Contact: Björn Bachmann, PhD
    • Principal Investigator: Björn Bachmann, PhD
  • Münster, Germany, 48149
    • Not yet recruiting
    • Münster University Hospital
    • Contact: Lamis Baydoun, MD
    • Principal Investigator: Lamis Baydoun, MD
• Italy
  • Bologna, Italy, 40138
    • Not yet recruiting
    • IRCCS Azienda Ospedaliero-Universitaria di Bologna
    • Contact: Luigi Fontana, MD
    • Principal Investigator: Luigi Fontana, MD
    • Sub-Investigator: Antonello Moramarco, MD
  • Catanzaro, Italy, 88100
    • Not yet recruiting
    • Department of Medical and Surgical Sciences, University Magna Græcia
    • Contact: Vincenzo Scorcia, MD
    • Principal Investigator: Vincenzo Scorcia, MD
  • Roma, Italy, 00133
    • Not yet recruiting
    • Department of Experimental Medicine, Tor Vergata School of Medicine and Surgery
    • Contact: Francesco Aiello, MD
    • Principal Investigator: Francesco Aiello, MD
  • Venice, Italy, 30174
    • Not yet recruiting
    • Fondazione Banca degli Occhi del Veneto
    • Contact: Diego Ponzin, MD
    • Principal Investigator: Diego Ponzin, MD
    • Sub-Investigator: Stefano Ferrari, PhD
    • Sub-Investigator: Alessandro Ruzza, MSc
    • Sub-Investigator: Mohit Parekh, PhD
  • Vicenza, Italy, 36100
    • Not yet recruiting
    • San Bortolo Hospital
    • Contact: Pietro Viola, MD
    • Principal Investigator: Pietro Viola, MD
  • BS
    • Brescia, BS, Italy, 25123
      • Recruiting
      • Azienda Socio Sanitaria Territoriale degli Spedali Civili di Brescia
      • Contact: Matteo Airaldi, MD; Phone Number: +390303995847; Email: matteo.airaldi@unibs.it
      • Principal Investigator: Matteo Airaldi, MD
      • Principal Investigator: Francesco Semeraro, MD
• Netherlands
  • Maastricht, Netherlands, 6229
    • Not yet recruiting
    • Maastricht University Medical Center
    • Contact: Mor Dickman, MD
    • Principal Investigator: Mor Dickman, MD
• Spain
  • Málaga, Spain, 29010
    • Not yet recruiting
    • Hospital Regional Universitario de Malaga
    • Contact: Marina Rodriguez Calvo de Mora, MD
    • Principal Investigator: Marina Rodriguez Calvo de Mora, MD
    • Sub-Investigator: Carlos Rocha de Lossada
• United Kingdom
  • Liverpool, United Kingdom, L7 8YE
    • Not yet recruiting
    • Royal Liverpool University Hospital
    • Contact: Stephen B Kaye, MD
    • Principal Investigator: Stephen B Kaye, MD
    • Sub-Investigator: Kunal A Gadhvi, MD
    • Sub-Investigator: Hannah J Levis, PhD
    • Sub-Investigator: Yalin Zheng, PhD

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: No

• Sampling Method: Probability Sample

Study Population

Consecutive patients undergoing keratoplasty at participating referral cornea clinics

Description

Inclusion Criteria:

• Age older than 18 years old
• Planned corneal transplantation: perforating keratoplasty (PK), Descemet stripping automated endothelial keratoplasty (DSAEK), ultra-thin DSAEK (UT-DSAEK) or Descemet membrane endothelial keratoplasty (DMEK)
• Indication for corneal transplantation (low-moderate risk of rejection): keratoconus, Fuchs endothelial dystrophy
• Ability to provide written informed consent.

Exclusion Criteria:

• Prior corneal transplantation
• Planned preloaded DSAEK or preloaded DMEK
• Planned keratoplasty not involving corneal endothelium transplantation (i.e., superficial/deep anterior lamellar keratoplasty)
• Evidence of concurrent microbial keratitis
• Evidence of non-infectious or autoimmune keratitis
• Impending or frank corneal perforation
• Uncontrolled, elevated intraocular pressure (IOP)- Previous glaucoma surgery
• Acute or chronic inflammatory/infectious anterior segment uveitis
• Presence of an anterior chamber intraocular lens (IOL) for phakic or aphakic purposes

Study Plan

How is the study designed?

Number of groups / cohorts

1

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Keratoplasty group; Patients undergoing keratoplasty and their respective donor tissue	Diagnostic test: V-Check; Preoperative analysis of endothelial tissue staining with vital dye (Trypan Blue 0.05%) and image analysis to identify the percentage of Trypan Blue Positive Areas, corresponding to dead endothelial cells and/or denuded areas of endothelium, in tissues destined to keratoplasty

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Trypan Blue Positive Areas (TBPA) percentage and Donor mortality	The comparison of the endothelial mortality of included donor corneas measured preoperatively (TBPA) and the overall endothelial mortality as declared by the eye bank providing the tissue	Preoperative

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Final ECD	Final ECD of transplanted corneas at 12 months	1 year after enrollment
Rate of endothelial cell loss	Rate of change of ECD over the course of the entire follow-up	1 year after enrollment
Incidence of endothelial graft failure	The occurrence of clinically definite corneal edema and/or need for endothelial keratoplasty due to endothelial failure	1 year after enrollment
Correlation of TBPA to declared endothelial cell density (dECD)	Spearman's rank correlation coefficient of TBPA and declared Endothelial Cell Density (dECD) as provided by the Eye Bank supplying the tissue	Preoperative
Correlation of TBPA to declared endothelial mortality on corneal folds	Spearman's rank correlation coefficient of TBPA and declared Endothelial Cell mortality on corneal folds as provided by the Eye Bank supplying the tissue	Preoperative
Relative risk of endothelial graft failure associated with TBPA	Calculation through a proportional hazard model of the relative risk of endothelial graft failure attributable to preoperatively measured TBPA	Preoperative

Collaborators and Investigators

• Sponsor: Azienda Socio Sanitaria Territoriale degli Spedali Civili di Brescia
• Investigators: Principal Investigator: Vito Romano, MD, Azienda Socio Sanitaria Territoriale degli Spedali Civili di Brescia

Publications and helpful links

General Publications

• Airaldi M, Zheng Y, Aiello F, Bachmann B, Baydoun L, Ni Dhubhghaill S, Dickman MM, Kaye SB, Fontana L, Gadhvi KA, Moramarco A, Rodriguez Calvo de Mora M, Rocha de Lossada C, Scorcia V, Viola P, Calza S, Levis HJ, Parekh M, Ruzza A, Ferrari S, Ponzin D, Semeraro F, Romano V. Preoperative surgeon evaluation of corneal endothelial status: the Viability Control of Human Endothelial Cells before Keratoplasty (V-CHECK) study protocol. BMJ Open Ophthalmol. 2023 Sep;8(1):e001361. doi: 10.1136/bmjophth-2023-001361.

Study record dates

Study Major Dates

• Study Start (Actual): December 1, 2023
• Primary Completion (Estimated): March 30, 2026
• Study Completion (Estimated): September 30, 2026

Study Registration Dates

• First Submitted: April 27, 2023
• First Submitted That Met QC Criteria: April 27, 2023
• First Posted (Actual): May 6, 2023

Study Record Updates

• Last Update Posted (Estimated): October 2, 2025
• Last Update Submitted That Met QC Criteria: September 29, 2025
• Last Verified: September 1, 2025

More Information

Terms related to this study

• Keywords: Corneal endothelium; Keratoplasty; Eye Banking
• Additional Relevant MeSH Terms: Postoperative Complications; Pathologic Processes; Eye Diseases; Corneal Diseases; Eye Manifestations; Pathological Conditions, Signs and Symptoms; Signs and Symptoms; Corneal Endothelial Cell Loss
• Other Study ID Numbers: VC1

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES
• IPD Plan Description: All collected IPD with appropriated anonymization of sensible data will be stored to a dedicated repository after completion of the study
• IPD Sharing Time Frame: After completion of study
• IPD Sharing Access Criteria: Upon approval of individual sharing request
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No