Indian Journal of Ophthalmology

: 1985  |  Volume : 33  |  Issue : 4  |  Page : 243--244

In-vitro adherence of candida albicans to conjunctival epithelial cells of patients using topical steroids

Gabu Bhardwaj, Shashi Vashist, Imtiaz A Khan 
 Department of Microbiology and Ophthalmology Sher-i-Kashmir Institute of Medical Sciences, Srinagar, India

Correspondence Address:
Gabu Bhardwaj
Department of Microbiology, Sher-i-Kashmir Institute of Medical Sciences, Soura- 190011, Srinagar

How to cite this article:
Bhardwaj G, Vashist S, Khan IA. In-vitro adherence of candida albicans to conjunctival epithelial cells of patients using topical steroids.Indian J Ophthalmol 1985;33:243-244

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Bhardwaj G, Vashist S, Khan IA. In-vitro adherence of candida albicans to conjunctival epithelial cells of patients using topical steroids. Indian J Ophthalmol [serial online] 1985 [cited 2021 Jun 20 ];33:243-244
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Full Text

Steroids are used topically in a number of eye ailments. Increase in opportunistic fungal infections of the eye has coincid­ed in time with the general use of corticosteroids in Ophthalmology. It is believed that steroids abrogate natural defences of eye, leading thereby to fungal colonization. There is significant fungal flora in the normal eye[1] and even greater flora in the diseased eye. Adherence plays a major role in the pathogenesis of many bacterial infections' and perhaps in case of Candida albicans too, as in mucocutaneous Candidia­sis. We wanted to see if long term local administration of steroids in various eye diseases leads to any difference in in-vitro adherence of C. albicans to conjunctival epithelial cells compared to conjunctional cells from a person having normal healthy eyes, who served as a control.


Three patients of chronic spring catarrh were included in the study who were on the topical steroid drops (dexamethasone, beta­methasone) for durations given in the table. Patients were using drops for approximately 8-10 months continuously in a year. Both the eyes were involved.

Human conjunctival epithelial cells : Eyes were first anaesthetized by instillation of 2% xylocaine drops every 5 minutes for three times. Lower palpebral conjunctiva of both eyes was gently scraped with a sterile swab and the swab was swirled in phosphate buffe­red saline (PBS) pH 7.2. Cells were concen­trated by centrifugation at 1000 rpm, 10 minutes, washed thrice with PBS and adjust­ed to concentration of 10 5 cells/ml of PBS.

Yeasts : Candida albicans, a clinical iso­late was grown on Sabouraud dextrose agar (SDA), slants for 48 hours at 37° C, harvested washed thrice with PBS pH 7.2 and suspend­ed to a concentration of 10 6 cells/ml.

Adherence test : 0.5ml samples of yeast and epithelial cells (ratio of yeast to epithe­lial cells, 10 ; 1) were taken into tubes and incubated in a rotatory water both for 1 hour at 37°C. Unattached yeasts were removed by differential centrifugation, instead of 12 um pore size polycarborate filters which are commonly employed for such purposes. We did not have such filters. With centrifuga­tion at 500 rpm for 10 minutes, majority of epithelial cells with attached yeasts got set­tled down while unattached yeasts remained at the top. Supernatant was decanted and deposit was gently washed twice with PBS. One drop of final pellet was placed on a glass slide; air dried, heat fixed and Gram stained. Number of yeasts attached to 100 epithelial cells was determined by light microscopy at x400. All experiments were performed in triplicate and repeated again.


[Table 1] shows the mean number of yeasts adhering to 100 epithelial cells in the three cases studied as well as in the control. All the three cases showed enhanced adherence of Candida albicans to conjunctival epithelial cells but this increase was not significant (p>0.05). There was no relation between duration of steroid usage and number of adhering yeasts. There was no previous colo­nization of the conjunctival sac by Candida albicans as shown by culture done before commencement of the study. Control too did not have any resident mycotic flora.


C.albicans has been shown to adhere in vitro to human epithelial cells[3],[4],[5]. It is an opportunistic pathogen and is believed to colonize eyes compromised by use of steroids. Another possibility could be steroid's direct action on the yeast. In fact there are reports wherein an increase in the pathogenicity of C.albicans and Aspergillus niger has been shown by pre-treatment of the fungi with steroids[6], as shown by production of corneal ulcer. We have found that pre-treatment of C.albicans with dexamethasone does not significantly affect its adherence to conjunc­tival epithelial cells. In the present study we wanted to see if prolonged use of local ste­roids affects the conjunctival epithelial cells in a way so that there is increased adherence of yeasts. We found that this was not so.


The present study shows that prolonged use of topical steroid drops in the eyes does not lead to any greater adherence of Candida albicans to conjunctival epithelial cells.


1Hammeke J. C. and Ellis P. P., 1960, Amer. J., Ophthalmol, 49 : 1174.
2Gibbons R. J. and Van Haute J., 1975, Ann. Rev. Microbiol., 29: 19.
3Kimura L. H. and Pearsall N. N„ 1978. Infect. Immun., 21 : 6
4King R. D., Lee J. C. and Morris A. L., 1980, Infect Immun, 27 : 667.
5Sandin R. L.. Rogars A. L., Patterson R. J. and Beneke E. S., 1982, Infect Immun., 25: 79.
6Hasany S. N., Basu P. K. and Kozdan J. J., 1973, Canad. J., Ophtbalmol, 8 ; 119.