LSCD: TOWARDS A TISSUE ENGINEERED HEMICORNEA

TITLE: INNOVATIVE METHOD FOR THE TREATMENT OF TOTAL LIMBAL STEM CELLS DEFICIENCY (LSCD) THROUGH A TISSUE ENGINEERED HEMICORNEA
SPONSOR:

Ministry of Health

CO-SPONSOR:
TIMEFRAME:

November 2012 – November 2015

THEME: Limbal Stem Cell Deficiency
STATUS: ONGOING
GRANT AMOUNT:

275.700 €

FBOV ROLE: Leader
PARTNERS INVOLVED: University of Padova, IRCCS Fondazione G. Bietti Onlus, ULSS16, Villa Igea Hospital
P.I. : Enzo Di Iorio

 

SUMMARY: The principal goal of this project is to evaluate an innovative and single-stage approach for the treatment of visual impairment due to corneal opacities in patients with LSCD. The aim is (i) to reconstruct an innovative human corneal tissue-engineered (hemicornea) in vitro, containing autologous limbal stem cells, directly cultivated onto a corneal stroma obtained by human donor keratoplasty lenticules (HKLs) and (ii) to test the efficacy of this new approach of tissue engineering in patients suffering from LSCD through a single-stage surgical procedure termed keratolimbal autograft combined with anterior lamellar keratoplasty.

 

OBJECTIVES/SCOPE : The improvements outlined in this research project will increase the therapeutic rate of success of this cell therapy treatment. This will benefit at large for those patients who have long been waiting for treatments able to cure diseases that current corneal transplantation-based techniques could not solve. Research scientists of FBOV in collaboration with three Ophthalmological units, have agreed to evaluate the critical steps associated with the clinical application of corneal stem cells in order to reduce the percentage of unsuccessfully treated patients and increase the quality and therapeutic rate of success of corneal stem cell therapy treatment. The critical steps identified and that this project aims to improve are the following:

(1) the development of an innovative and single-stage surgical approach for the treatment of visual impairment due to corneal opacities in patients with LSCD;

(2) the development of new scaffolds/carriers alternative to fibrin-glue to generate hemicorneas by means autologous stem cells;

(3) the development of animal-free culture systems (no murine feeder layers or culturing media with animal-derived proteins/growth factors);

(4) maintenance of the stemness and proliferative potential of epithelial stem cells cultured in vitro and in the graft;

(5) evaluation of LSCD by means of impression cytology analysis;

(6) comparison between modern femtosecond laser-assisted and manual microkeratome technique effectiveness. This Project can represent the beginning of future cornea engineering studies, providing a new way for vision rehabilitation in patients suffering LSCD.

 

SUMMARY OF METHODS: To achieve this goal the Veneto Eye Bank Foundation (FBOV) will be collaborating with the IRCCS G.B. Bietti Foundation of Rome, and with two eye surgeons (Dr. Galan of S. Antonio Hospital, and Prof. Busin of Villa Serena Hospital) with long term experience in PK and limbal stem cell transplantation. The research activities of this grant application are divided into 2 phases for a full duration of 36 months.

1st Phase (21 months):

The first phase of the project will be focused on to the evaluation of HKLs as natural and reliable scaffolds compatible with the growth of corneal epithelial stem cells and stromal keratocytes. The biochemical, biomechanical and optical characterization of the HKLs will be performed by researchers at FBOV and Bietti locations.

2nd Phase (15 months):

patients recruited during the 1st phase (6 in total) will be admitted to the Clinical Sites for stem cell biopsy first and grafting 3 weeks later. Follow-up examinations will be carried out at defined time-intervals and rate of grafting success determined. During the second phase of the project, patients recruited during the 1st phase will be admitted to S. Antonio Hospital and Villa Serena Hospital for surgical treatments, i.e., limbus biopsy first and limbal stem cell grafting 3 weeks later. Follow-up examinations will be carried out at defined time-intervals and the rate of grafting success determined.

 

EXPECTED RESULTS: This new approach might open up new perspectives for the surgical treatment of LSCD patients. Being based on a single step, it would, in fact, (i) allow a rapid recovery of vision thus reducing waiting times and surgical complications usually caused by the multi-staged approach described earlier, and (ii) increase efficiency and success of corneal stem cell therapy. 

 

PUBLICATIONS:  

 

ABSTRACTS/POSTERS TO CONFERENCES:

 

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