Article Text

other Versions

PDF
Long-term preservation of donor corneas in glycerol for keratoplasty: exploring new protocols
  1. Himi Tripathi1,
  2. Manzoor Ul Mehdi1,
  3. Deepika Gupta2,
  4. Seema Sen3,
  5. Seema Kashyap3,
  6. Tapas Chandra Nag4,
  7. Manisha Purwar5,
  8. Manjeet Jassal2,
  9. Ashwini K Agrawal2,
  10. Sujata Mohanty6,
  11. Radhika Tandon1
  1. 1Department of Ophthalmology, Dr Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India
  2. 2SMITA Research Labs, Department of Textile Technology, Indian Institute of Technology, New Delhi, India
  3. 3Department of Ocular Pathology, Dr Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India
  4. 4Department of Anatomy, All India Institute of Medical Sciences, New Delhi, India
  5. 5Department of Pathology, All India Institute of Medical Sciences, New Delhi, India
  6. 6Stem Cell Facility, All India Institute of Medical Sciences, New Delhi, India
  1. Correspondence to Radhika Tandon, Cornea, Cataract, Refractive Surgery Services and Co-Chairperson National Eye Bank, Dr Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi 110029, India; radhika_tan{at}yahoo.com

Abstract

Aim To evaluate the role of temperature and adjunctive dehydration in better long-term preservation of human corneas when preserved and stored in glycerol.

Methods Different preservation temperatures and effects of adding silica gel in glycerol-preserved corneal tissues were evaluated. Human corneal tissues not suitable for optical keratoplasty initially preserved in McCarey–Kaufman medium were transferred to glycerol and stored at four different temperatures for 3 months as follows: tissues in anhydrous glycerol with and without silica gel at −80°C, −20°C, 4°C and at room temperature (RT). Parameters evaluated included microbial sterility, thickness (Digimatic micrometer), transparency (slit lamp examination, UV-Vis spectrophotometer), mechanical strength (Instron 5848 Microtester), tissue integrity (H&E staining), antigenicity (immunohistochemistry) and ultrastructure of collagen (transmission electron microscopy, TEM).

Results Microbial test after 3 months of glycerol preservation confirmed sterility of the tissues. The thickness increased in corneas preserved at RT with and without silica gel (p<0.001). RT corneas had the lowest transparency and tensile strength. Tissues in anhydrous glycerol stored with and without silica gel at −80°C were the most transparent (p<0.001) and had the highest tensile strength (p<0.001). Tissue integrity was maintained and expression of Human Leukocyte Antigen D related (HLA-DR) was less in glycerol-preserved corneas at −80°C. TEM studies indicated that parallel alignment of stromal collagen was disrupted at RT-preserved corneas.

Conclusions Corneal tissue preserved at −80°C was the best method for preservation as it maintained the sterility, thickness, optical transparency, mechanical strength and ultrastructural features.

  • Cornea
  • Eye (Tissue) Banking

Statistics from Altmetric.com

Request permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.

Linked Articles

  • At a glance
    Keith Barton James Chodosh Jost Jonas