Skip to main content
NCBI home page
The British Journal of Ophthalmology logoLink to The British Journal of Ophthalmology
. 2001 Oct;85(10):1188–1197. doi: 10.1136/bjo.85.10.1188

Autologous serum eyedrops for dry eyes and epithelial defects: clinical and in vitro toxicity studies

A Poon 1, G Geerling 1, J Dart 1, G Fraenkel 1, J Daniels 1
PMCID: PMC1723716  PMID: 11567963

Abstract

BACKGROUND/AIMS—Autologous serum drops have been reported to be beneficial in keratoconjunctivitis sicca (KCS) and persistent epithelial defects (PED). A clinical pilot study was carried out to examine these potential uses and in vitro toxicity testing on corneal epithelial cell cultures was performed to compare the effect of serum drops with unpreserved hypromellose (hydroxypropylmethylcellulose 0.3%).
METHODS—Patients with KCS and PED, unresponsive to conventional treatment were recruited. Patients were examined before treatment, at 1 and 2 weeks after initiation, and then 2 weekly until treatment ceased. Symptoms were assessed at each visit. Clinical examination included Schirmer's test without anaesthesia, rose bengal staining, and fluorescein staining. Epithelial defects were measured with the slit beam. In the laboratory, cultured human corneal epithelial cells were exposed to serum drops and hypromellose, and their viability evaluated with fluorescent viability staining (Calcein AM ethidium homodimer) and an ATP assay.
RESULTS—Autologous serum was used in 15 eyes of 13 patients with PED and 11 eyes of nine patients with KCS. In two patients serum drops were started after penetrating keratoplasty (PK). The PKs were performed for perforations secondary to PEDs. Of the 15 eyes with PED, nine healed at a mean of 29 days and six failed. The mean duration of PED before the use of serum drops was 48.2 days. Of the 11 eyes with KCS, six had improved subjective scores and fluorescein scores, and five had improved rose bengal scores after the use of serum drops. For the two patients who used serum eyedrops post-PK, there was a stable and intact epithelium at 1 week. Cessation of serum drops during the postoperative period led to deterioration in the subjective and objective scores in both patients. One developed a PED that responded to reinstitution of serum drops. The morphology and ATP levels of cultured epithelial cells exposed to serum were better maintained than those exposed to hypromellose.
CONCLUSION—Autologous serum drops are useful for PED and KCS. This effect may be related to a number of active factors in serum including growth factors, fibronectin, vitamin A, and anti-proteases. In vitro toxicity testing demonstrated that serum drops have reduced toxicity compared with unpreserved hypromellose. Currently regulatory restrictions in the UK have prevented the establishment of a prospective randomised controlled trial examining the efficacy of autologous serum drops for the management of this group of ocular surface disorders.



Full Text

The Full Text of this article is available as a PDF (260.6 KB).

Figure 1  .

Figure 1  

Open in a new tab

This is a 74 year old man with KCS secondary to rheumatoid arthritis. History includes left infective corneal melt 3 years ago requiring a tectonic PK. This was followed by recurrent epithelial breakdown unresponsive to intensive lubrication and SRCL wear. The cornea perforated again requiring a repeat PK 3 months before the start of serum drops. The PED recurred and amniotic membrane transplantation was tried but failed to heal it. Serum drops were started subsequently. (A, B) Show a PED with and without fluorescein and cobalt blue illumination before the start of serum. (C) Shows that healing occurred in 2 months (epithelial bullae at site of PED). Serum was stopped after 2 months, and the PED recurred (D). It failed to respond to any therapy apart from the reinstitution of serum. (E) and (F) show healed defect with and without cobalt blue illumination. The PED recurred with further attempts at weaning and healed with reinstitution of serum. At the last review serum was weaned for the fourth time.    

Figure 2  .

Figure 2  

Open in a new tab

A 52 year old female with OCP, bilateral KCS, and left PK. Bilateral epitheliopathy was secondary to KCS. (A) and (B) show right and left eyes respectively. Serum drops were started and (C) and (D) were taken at 2 week review. Despite the absence of fluorescein in (C) and (D), there are obviously fewer surface irregularities and episcleral injection in the second set of photographs. Both objective and subjective scores had improved.    

Figure 3  .

Figure 3  

Open in a new tab

Human corneal epithelial cells after 2 hours' incubation with (A) hypromellose 0.3% and (B) 50% serum drops. The green fluorescent calcein demonstrates viable intracellular esterase activity. The red fluorescence results from ethidium-homodimer bound to nucleic acids as a consequence of a pathological increase in cell membrane permeability.    

graphic file with name bjo-csoonm.f1.jpg

Open in a new tab

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Avisar R., Creter D., Levinsky H., Savir H. Comparative study of tear substitutes and their immediate effect on the precorneal tear film. Isr J Med Sci. 1997 Mar;33(3):194–197. [PubMed] [Google Scholar]
  2. Berman Collagenase inhibitors: rationale for their use in treating corneal ulceration. Int Ophthalmol Clin. 1975 Winter;15(4):49–66. doi: 10.1097/00004397-197501540-00006. [DOI] [PubMed] [Google Scholar]
  3. Berry M., Gurung A., Easty D. L. Toxicity of antibiotics and antifungals on cultured human corneal cells: effect of mixing, exposure and concentration. Eye (Lond) 1995;9(Pt 1):110–115. doi: 10.1038/eye.1995.17. [DOI] [PubMed] [Google Scholar]
  4. Burstein N. L. The effects of topical drugs and preservatives on the tears and corneal epithelium in dry eye. Trans Ophthalmol Soc U K. 1985;104(Pt 4):402–409. [PubMed] [Google Scholar]
  5. Collins M. K., Perkins G. R., Rodriguez-Tarduchy G., Nieto M. A., López-Rivas A. Growth factors as survival factors: regulation of apoptosis. Bioessays. 1994 Feb;16(2):133–138. doi: 10.1002/bies.950160210. [DOI] [PubMed] [Google Scholar]
  6. Daniels J. T., Harris I. R., Kearney J. N., Ingham E. Calcium: a crucial consideration in serum-free keratinocyte culture. Exp Dermatol. 1995 Aug;4(4 Pt 1):183–191. doi: 10.1111/j.1600-0625.1995.tb00243.x. [DOI] [PubMed] [Google Scholar]
  7. Fox R. I., Robinson C. A., Curd J. G., Kozin F., Howell F. V. Sjögren's syndrome. Proposed criteria for classification. Arthritis Rheum. 1986 May;29(5):577–585. doi: 10.1002/art.1780290501. [DOI] [PubMed] [Google Scholar]
  8. Fredj-Reygrobellet D., Plouet J., Delayre T., Baudouin C., Bourret F., Lapalus P. Effects of aFGF and bFGF on wound healing in rabbit corneas. Curr Eye Res. 1987 Oct;6(10):1205–1209. doi: 10.3109/02713688709025230. [DOI] [PubMed] [Google Scholar]
  9. Gordon J. F., Johnson P., Musch D. C. Topical fibronectin ophthalmic solution in the treatment of persistent defects of the corneal epithelium. Chiron Vision Fibronectin Study Group. Am J Ophthalmol. 1995 Mar;119(3):281–287. doi: 10.1016/s0002-9394(14)71168-7. [DOI] [PubMed] [Google Scholar]
  10. Göbbels M., Spitznas M. Corneal epithelial permeability of dry eyes before and after treatment with artificial tears. Ophthalmology. 1992 Jun;99(6):873–878. doi: 10.1016/s0161-6420(92)31879-2. [DOI] [PubMed] [Google Scholar]
  11. Kirschner S. E. Persistent corneal ulcers. What to do when ulcers won't heal. Vet Clin North Am Small Anim Pract. 1990 May;20(3):627–642. doi: 10.1016/s0195-5616(90)50054-6. [DOI] [PubMed] [Google Scholar]
  12. Lemp M. A. Report of the National Eye Institute/Industry workshop on Clinical Trials in Dry Eyes. CLAO J. 1995 Oct;21(4):221–232. [PubMed] [Google Scholar]
  13. McCluskey P., Wakefield D., York L. Topical fibronectin therapy in persistent corneal ulceration. Aust N Z J Ophthalmol. 1987 Nov;15(4):257–262. doi: 10.1111/j.1442-9071.1987.tb00081.x. [DOI] [PubMed] [Google Scholar]
  14. McDonnell P. J., Schanzlin D. J., Rao N. A. Immunoglobulin deposition in the cornea after application of autologous serum. Arch Ophthalmol. 1988 Oct;106(10):1423–1425. doi: 10.1001/archopht.1988.01060140587028. [DOI] [PubMed] [Google Scholar]
  15. Nakamura M., Nishida T., Mishima H., Otori T. Effects of antimicrobials on corneal epithelial migration. Curr Eye Res. 1993 Aug;12(8):733–740. doi: 10.3109/02713689308995769. [DOI] [PubMed] [Google Scholar]
  16. Nelson J. D., Drake M. M., Brewer J. T., Jr, Tuley M. Evaluation of a physiological tear substitute in patients with keratoconjunctivitis sicca. Adv Exp Med Biol. 1994;350:453–457. doi: 10.1007/978-1-4615-2417-5_78. [DOI] [PubMed] [Google Scholar]
  17. Nishida T., Nakamura M., Ofuji K., Reid T. W., Mannis M. J., Murphy C. J. Synergistic effects of substance P with insulin-like growth factor-1 on epithelial migration of the cornea. J Cell Physiol. 1996 Oct;169(1):159–166. doi: 10.1002/(SICI)1097-4652(199610)169:1<159::AID-JCP16>3.0.CO;2-8. [DOI] [PubMed] [Google Scholar]
  18. Nishida T., Ohashi Y., Awata T., Manabe R. Fibronectin. A new therapy for corneal trophic ulcer. Arch Ophthalmol. 1983 Jul;101(7):1046–1048. doi: 10.1001/archopht.1983.01040020048007. [DOI] [PubMed] [Google Scholar]
  19. Pasternak A. S., Miller W. M. First-order toxicity assays for eye irritation using cell lines: parameters that affect in vitro evaluation. Fundam Appl Toxicol. 1995 May;25(2):253–263. doi: 10.1006/faat.1995.1061. [DOI] [PubMed] [Google Scholar]
  20. Pastor J. C., Calonge M. Epidermal growth factor and corneal wound healing. A multicenter study. Cornea. 1992 Jul;11(4):311–314. doi: 10.1097/00003226-199207000-00007. [DOI] [PubMed] [Google Scholar]
  21. Phan T. M., Foster C. S., Boruchoff S. A., Zagachin L. M., Colvin R. B. Topical fibronectin in the treatment of persistent corneal epithelial defects and trophic ulcers. Am J Ophthalmol. 1987 Nov 15;104(5):494–501. doi: 10.1016/s0002-9394(14)74107-8. [DOI] [PubMed] [Google Scholar]
  22. Reim M., Kehrer T., Lund M. Clinical application of epiderm growth factor in patients with most severe eye burns. Ophthalmologica. 1988;197(4):179–184. doi: 10.1159/000309941. [DOI] [PubMed] [Google Scholar]
  23. Rodeck U., Jost M., Kari C., Shih D. T., Lavker R. M., Ewert D. L., Jensen P. J. EGF-R dependent regulation of keratinocyte survival. J Cell Sci. 1997 Jan;110(Pt 2):113–121. doi: 10.1242/jcs.110.2.113. [DOI] [PubMed] [Google Scholar]
  24. Scardovi C., De Felice G. P., Gazzaniga A. Epidermal growth factor in the topical treatment of traumatic corneal ulcers. Ophthalmologica. 1993;206(3):119–124. doi: 10.1159/000310376. [DOI] [PubMed] [Google Scholar]
  25. Schultz G. S., Davis J. B., Eiferman R. A. Growth factors and corneal epithelium. Cornea. 1988;7(2):96–101. [PubMed] [Google Scholar]
  26. Singh G., Foster C. S. Epidermal growth factor in alkali-burned corneal epithelial wound healing. Am J Ophthalmol. 1987 Jun 15;103(6):802–807. doi: 10.1016/s0002-9394(14)74397-1. [DOI] [PubMed] [Google Scholar]
  27. Tseng S. C., Tsubota K. Important concepts for treating ocular surface and tear disorders. Am J Ophthalmol. 1997 Dec;124(6):825–835. doi: 10.1016/s0002-9394(14)71700-3. [DOI] [PubMed] [Google Scholar]
  28. Tsubota K., Goto E., Fujita H., Ono M., Inoue H., Saito I., Shimmura S. Treatment of dry eye by autologous serum application in Sjögren's syndrome. Br J Ophthalmol. 1999 Apr;83(4):390–395. doi: 10.1136/bjo.83.4.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tsubota K., Goto E., Shimmura S., Shimazaki J. Treatment of persistent corneal epithelial defect by autologous serum application. Ophthalmology. 1999 Oct;106(10):1984–1989. doi: 10.1016/S0161-6420(99)90412-8. [DOI] [PubMed] [Google Scholar]
  30. Tsubota K., Satake Y., Ohyama M., Toda I., Takano Y., Ono M., Shinozaki N., Shimazaki J. Surgical reconstruction of the ocular surface in advanced ocular cicatricial pemphigoid and Stevens-Johnson syndrome. Am J Ophthalmol. 1996 Jul;122(1):38–52. doi: 10.1016/s0002-9394(14)71962-2. [DOI] [PubMed] [Google Scholar]
  31. Ubels J. L., Edelhauser H. F., Austin K. H. Healing of experimental corneal wounds treated with topically applied retinoids. Am J Ophthalmol. 1983 Mar;95(3):353–358. doi: 10.1016/s0002-9394(14)78305-9. [DOI] [PubMed] [Google Scholar]
  32. Ubels J. L., Williams K. K., Lopez Bernal D., Edelhauser H. F. Evaluation of effects of a physiologic artificial tear on the corneal epithelial barrier: electrical resistance and carboxyfluorescein permeability. Adv Exp Med Biol. 1994;350:441–452. doi: 10.1007/978-1-4615-2417-5_77. [DOI] [PubMed] [Google Scholar]
  33. Wang X. M., Terasaki P. I., Rankin G. W., Jr, Chia D., Zhong H. P., Hardy S. A new microcellular cytotoxicity test based on calcein AM release. Hum Immunol. 1993 Aug;37(4):264–270. doi: 10.1016/0198-8859(93)90510-8. [DOI] [PubMed] [Google Scholar]

Articles from The British Journal of Ophthalmology are provided here courtesy of BMJ Publishing Group

RESOURCES