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Updated: Jul 17, 2026
A Simplified Technique for In situ Excision of Cornea and Evisceration of Retinal Tissue from Human Ocular Globe
Published on: June 12, 2012
The new future perspective in corneal tissue utilisation - methods of preparation and preservation
Martina Polachova1,2, Magdalena Netukova3,4, Oldrich Benada5
1Department of Ophthalmology, University Hospital Kralovske Vinohrady and 3rd Faculty of Medicine, Srobarova 1150/50, Prague 10, 100 34, Czech Republic. martina.polachova@fnkv.cz.
Purpose:
The goal of our study is to find an optimal approach to the preparation and preservation of corneal stromal tissue. We want to compare different methods of corneal stromal tissue creation and storage to optimize the efficacy of this process under the conditions of an eye bank. After we find the most suitable method to create a safe high quality product, we want to prove the possibility of using a single donor cornea for more than one patient. We would also like to verify the feasibility of making more corneal lenticules after the removal of a corneal endothelium for DMEK transplantation.
Methods:
We provided morphological (histology, scanning electron microscope) and microbiological analysis in order to compare different methods of corneal lenticule and corneal stromal lamellae preparation and preservation. We also tested the surgical handling of the tissue to secure a safe manipulation of the tissue for clinical use. We compared two methods of corneal lenticule preparation: microkeratome dissection and femtosecond laser. As methods of preservation, we tested hypothermia, cryopreservation at -80 degrees Celsius in DMSO (dimethyl sulfoxide) and storage at room temperature with glycerol. Some intrastromal lenticules and lamellae in each group were previously irradiated with gamma radiation of 25 kGy (KiloGray).
Results:
Corneal stromal lamellae prepared with a microkeratome have a smoother cut - side surface compared to lamellae prepared with a femtosecond laser. Femtosecond laser preparation caused more irregularities on the surface and we detected more conglomerates of the fibrils, while lamellae made with microkeratome had more sparse network. Using femtosecond laser, we were able to make more than five lenticules from a single donor cornea. Gamma irradiation led to damage of collagen fibrils in corneal stroma and a loss of their regular arrangement. Corneal tissue stored in glycerol showed collagen fibril aggregates and empty spaces between fibrils caused by dehydration. Cryopreserved tissue without previous gamma irradiation showed the most regular structure of the fibrils comparable to storage in hypothermia.
Conclusion:
Our results suggest that formation of a corneal lenticule lamellae by microkeratome results in smoother corneal lenticules, while being much cheaper than formation by femtosecond laser. Gamma irradiation of 25 kGy caused damage of the collagen fibres as well as their network arrangement, which correlated with loss of transparency and stiffer structure. These changes impair possible surgical utilisation of gamma irradiated corneas. Storage in glycerol at room temperature and cryopreservation had similar outcomes and we believe that both methods are appropriate and safe for further clinical use .
Insights
Microkeratome preparation yields smoother corneal lenticules for transplantation. Both glycerol storage and cryopreservation are suitable for preserving corneal stromal tissue for eye bank use.
Area of Science:
- Ophthalmology
- Biomaterials Science
- Tissue Engineering
Background:
- Corneal stromal tissue preparation and preservation are critical for eye banking.
- Optimizing these processes can enhance the quality and availability of corneal grafts.
- Current methods require evaluation for improved clinical outcomes.
Purpose of the Study:
- To determine the optimal method for preparing and preserving corneal stromal tissue for eye bank applications.
- To compare microkeratome dissection versus femtosecond laser for corneal lenticule creation.
- To evaluate different preservation techniques including hypothermia, cryopreservation, and glycerol storage.
Main Methods:
- Morphological analysis (histology, SEM) and microbiological testing were performed.
- Surgical handling properties were assessed for clinical safety.
- Corneal lenticules were prepared using microkeratome and femtosecond laser.
- Preservation methods included hypothermia, cryopreservation (-80°C in DMSO), and room temperature storage in glycerol.
- The impact of 25 kGy gamma irradiation was also investigated.
Main Results:
- Microkeratome preparation resulted in smoother corneal lenticules compared to femtosecond laser, which caused surface irregularities.
- Femtosecond laser enabled the creation of over five lenticules per cornea.
- Gamma irradiation at 25 kGy damaged collagen fibrils, leading to loss of transparency and structural integrity.
- Glycerol storage caused collagen aggregation and dehydration artifacts.
- Cryopreserved tissue (without irradiation) and hypothermic storage showed the most regular collagen fibril structure.
Conclusions:
- Microkeratome dissection is a cost-effective method for producing smoother corneal lenticules.
- Gamma irradiation at 25 kGy is detrimental to corneal stromal tissue structure and surgical utility.
- Both glycerol storage at room temperature and cryopreservation are viable and safe preservation methods for corneal tissue.
