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Evaluation of histamine-induced conjunctival oedema in guinea pigs by means of image analysis
  1. A Takahashi1,
  2. T Sumi1,
  3. K Tada1,
  4. H Mibu2,
  5. D Shii2,
  6. A Kayasuga3,
  7. A Fukushima1
  1. 1Department of Ophthalmology and Visual Science, Kochi Medical School, Nankoku, Japan
  2. 2Santen Pharmaceutical Company, Osaka, Japan
  3. 3Cine-Science Laboratory Co, Ltd, Tokyo, Japan
  1. Correspondence to Dr Atsuki Fukushima, Department of Ophthalmology and Visual Science, Kochi Medical School, Kohasu, Oko-cho, Nankoku 783-8505, Japan; fukusima{at}kochi-u.ac.jp

Abstract

Background Conjunctival oedema is commonly observed in patients with allergic conjunctivitis and can be induced by histamine. In animal models of allergic conjunctivitis, conjunctival oedema is generally evaluated by measuring the extravasation of Evans blue dye into the conjunctiva. A limitation of this method is that it only allows evaluation at a single time point. The aim of the present study was to investigate kinetic changes in histamine-induced bulbar oedema.

Methods Evans blue dye was injected intravenously into male guinea pigs. Histamine eye-drops were administered 30 min later. One group of animals received levocabastine (an antihistamine) eye-drops 10 min before histamine challenge. A digital camera was used to obtain images of the bulbar conjunctiva at 1 min intervals until 30 min after histamine challenge. The conjunctivas were then harvested, and the concentration of Evans blue was measured. The ImageJ software was used to analyse the images by counting the number of absolute pixel values.

Results The degree of conjunctival oedema increased progressively until 20 min after histamine challenge and then stabilised. Correspondingly, the number of absolute pixel values increased significantly until 5 min after histamine challenge, then increased gradually until the 20 min time point and finally plateaued. Pixel values were significantly lower in animals treated with levocabastine than in control animals. A significant correlation was observed between the pixel values of the conjunctival images and the concentration of Evans blue in the conjunctiva.

Conclusions This is the first study to have quantitatively evaluated kinetic changes in histamine-induced bulbar oedema by means of image analysis.

  • Conjunctiva
  • hyperaemia
  • histamine
  • image analysis

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Footnotes

  • Competing interests Declared. All authors have completed the Unified Competing Interest form at http://www.icmje.org/coi_disclosure.pdf (available on request from the corresponding author) and declare financial support for the submitted work from Santen Pharmaceutical Company. All authors also declare that AF has received research grants and honorariums from Santen Pharmaceutical Company. No spouses, partners or children have relationships with commercial entities that might have an interest in the submitted work. There are no non-financial interests that may be relevant to the submitted work.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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