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Year : 1982  |  Volume : 30  |  Issue : 4  |  Page : 341-346

Cornel preservation in an indigenous medium­ reconstituted honey

SMS Medical College, Jaipur, India

Correspondence Address:
Indu Arora
SMS Medical College, Jaipur (Rajasthan)
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Source of Support: None, Conflict of Interest: None

PMID: 6762352

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How to cite this article:
Arora I, Kulshrestha O P, Ujjwaj N S. Cornel preservation in an indigenous medium­ reconstituted honey. Indian J Ophthalmol 1982;30:341-6

How to cite this URL:
Arora I, Kulshrestha O P, Ujjwaj N S. Cornel preservation in an indigenous medium­ reconstituted honey. Indian J Ophthalmol [serial online] 1982 [cited 2023 Feb 1];30:341-6. Available from: https://www.ijo.in/text.asp?1982/30/4/341/29467

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Table 1

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Table 1

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Having come across reports of the use of honey by Khanna et al[1] for preservation of Aortic valves and by Malti Gupta[2] for success­fully preserving skin graft in honey, we decid­ed to try out honey as a corneal preservative medium.

  Materials and methods Top

A total of 86 goat corneas were studied.

Honey from two best available sources was obtained and sent to the Public Health Labora­tory for detection of adulteration. Both samples were found unadulterated and were found to be of standard quality. Hence, one sample at random was selected.

20m1 of pure honey was poured into 6 sterile beakers, which were covered with alumi­nium foil. For preservation corneosclearal buttons were prepared from enucleated goat eyes and one cornea-scleral button was placed in each beaker and kept at 40° C. At the end of the 2nd day, Ist week, Ist month, 3rd month

and 4th month, one cornea was removed and the following tests performed :­

Corneal clarity by naked eye examination[3],[4]

Nitro-blue-tetrazolium staining.

Trypan Blue staining.

Bacterial and fungal cultures of the media. In the next phase of the study, honey was

diluted to solutions of 90%, 70%, 50% & 30% by adding distilled water. To the solutions was added 5% glycerine and 100mg/ml of Strepto Penicillin and the solutions were autoclaved.

A total of 40 corneo-scleral buttons, 10 for each dilution of honey were preserved as above. Two C-S buttons were removed from each dilution at the end of 1st, 2nd, 3rd, 4th & 8th days of preservation and the same para­meters as above were studied.

Next, 30 C-S buttons were preserved and studied as above, after preserving in 75%, 70% and 65% dilution of honey, with its pH adjust­ed to 7.4 by the addition of Sodium Carbonate and Sodium Bicarbonate buffer.

Lastly, 10 eyes were preserved by the con­ventional eye banking technique and the same parameters studied.

  Observations Top

The observations were shown in [Figure - 1] [Table - 1][Table - 2][Table - 3][Table - 4] and Graphs I, II, III & IV.

  Discussion Top

Purified honey is hygroscopic, has antisep­tic properties and contains 70 to 80% metabo­lisable glucose.

It is now an accepted fact that the success of a penetrating keratoplasty lies in a donor cornea with a viable endothelium. An estim­ated 70% viability of endothelial cells is the minimum required[5].

Nitro-Blue-Tetrazolium and Trypan Blue staining techniques were used as tests of endo­thelial viability on the basis of our previous


As observed from endothelial viability studies, it became apparent that the endothe­lium of corneas preserved in pure honey was not viable after two days of preservation though the corneas were optically clear up to the 4th month of preservation. Even though these corneas are unsuitable for penetrating keratoplasty because of the non-viable endoth­lium, they are suitable for Lamellar Keratop­lasty after rehydration.

All the samples of pure honey were found to be sterile after the requisite number of days of preservation, except the one kept for 4 months, which was found to grow Aspergillus.

To be on the safe side, in all subsequent studies the reconstituted honey was autoclaved.

Attributing the quick death of corneal cells to the extreme dehydration on being subjected to pure honey, the C-S buttons were preserved in randomly selected dilutions i.e. 70%, 50% & 30% so that the ideally suited dilution could be arrived at. It became apparent from the endothelial viability tests [Figure - 1] [Table - 1][Table - 2][Table - 3][Table - 4] and graphs I, II, 111 & IV) that the endothelium was best preserved in 70% dilution of honey, it being viable for 3 days.

In order to prolong the storage time, other parameters were looked into. It is known that honey is acidic and has a pH of 4 whereas the pH of Aqueous is 7.4. Three dilutions of honey closest to 70% i.e. 75%, 70% & 65% were prepared and the C-S buttons were preserved in these dilutions with the pH adjust­ed to 7.4.

It was observed that the endothelium could be preserved up to 4 days in the 75% dilution of pH adjusted honey.

Gorneas preserved by the standard eye banking technique were found to have viable endothelium for 2 days.

  Summary Top

The corneo scleral button being optically clear after 4 months of preservation in pure honey can be used for lamellar keratoplasty after rehydration.

By preserving the C-S button for 4 days in 75% dilution of pH adjusted honey, as against the conventional preservation of cornea up to 2 days, we present a very inexpensive, easily available and preparable medium which allows enough time for contacting and admitting even outstation patients for penerating keratoplasty. This would also allow elective scheduling of surgery, thereby providing better patient prepa­ration, improved hospital bed utilisation, and a rested surgeon operating with experienced operating room personnel.

Donor corneas can also be transported from any remote place to a centralised institute, where penetrating keratoplasty is routinely performed thus increasing availability of donor corneas.

Removal of only the corneo-scleral button from the cadaver without enucleating the eye may be looked upon with favour by the rela­tives of the deceased & thus may further increase the availability of donor corneas.

  Acknowledgements Top

This study was supported by a financial grant from the Indian Council of Medical Research, whose help is being generously acknowledged We are also thankful to Dr. M. Rai, Lecturer in Pathololy, S.M.S. Medical College, Jaipur for his valuable assistance.

  References Top

Khanna, S.K: ; Arora, H.L. ; Jain, T.C. & Gupta, M. : 1971 Ann, Ind. Med. Sci., 7 : 3.  Back to cited text no. 1
Malti Gupta 1977 : Ind. J. Surgery, 39 : 591.  Back to cited text no. 2
Arora Indu and Kulshrestha, O.P. ; 1979, VIIth Rajasthan Ophthalmol. Conference Jaipur in Press.  Back to cited text no. 3
Arora Indu and Kulshrestha, O.P.: 1980 Proc. 39th All. Ind Ophthalmol Conf, Manipal.  Back to cited text no. 4
Stocker. F.W. ; King, E.H. ; Lucas, D.O. and Giorgiade, N., 1970, Arch. Ophthalmol. 84 : 2.  Back to cited text no. 5


  [Figure - 1], [Figure - 2], [Figure - 3], [Figure - 4], [Figure - 5], [Figure - 6], [Figure - 7], [Figure - 8]

  [Table - 1], [Table - 2], [Table - 3], [Table - 4]


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