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Rudraprasad D, Gandhi J, Naik P, N Naik M, Naidu C, Kumar Mishra D, Joseph J. A Novel and Low-cost Approach for Intravitreal Injection in an Experimental Model of Endophthalmitis. JOVR [Internet]. 2023Jul.23 [cited 2024May15];18(3):272–282. Available from: https://knepublishing.com/index.php/JOVR/article/view/13775

https://doi.org/10.18502/jovr.v18i3.13775

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authors

  • Dhanwini Rudraprasad

    Dhanwini Rudraprasad

    Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Jaishree Gandhi

    Jaishree Gandhi

    Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Poonam Naik

    Poonam Naik

    Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Milind N Naik

    Milind N Naik

    Ophthalmic Plastic Surgery & Facial Aesthetics, LV Prasad Eye Institute, Hyderabad, India
  • Chenchu Naidu

    Chenchu Naidu

    Ophthalmic pathology Laboratory, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Dilip Kumar Mishra

    Dilip Kumar Mishra

    Ophthalmic pathology Laboratory, LV Prasad Eye Institute, Hyderabad, Telangana, India
  • Joveeta Joseph

    Joveeta Joseph

    Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India

Published Date

  • Jul 23, 2023
This journal is publishing all the articles under Creative Commons Attribution License CC BY 4.0.

A Novel and Low-cost Approach for Intravitreal Injection in an Experimental Model of Endophthalmitis

Journal of Ophthalmic and Vision Research (JOVR) / July–September 2023, Volume 18, Issue 3 / Pages 272–282

Abstract

Purpose: Animal models are necessary in understanding the pathogenesis of endophthalmitis and are also necessary to assist the development of new therapeutics for this sight-threatening ocular inflammation. Hamilton syringes are usually preferred to inject pathogens when performing experiments on test subjects, however, this method has technical and financial disadvantages. In this study, we report the findings and assess the related benefits of applying a novel low-cost intravitreal injection technique to initiate endophthalmitis in a mouse model while using the Eppendorf tip and a 26G needle.


Methods: The 18-hr culture of clinical isolates of bacteria (Staphylococcus aureus and Pseudomonas aeruginosa) and fungus (Aspergillus flavus and Candida albicans) were resuspended to a final concentration of 10,000 colony forming units (CFU)/1 µL which were separately injected intravitreally into C57BL/6 mice (6–8 weeks) using a 0.1–2.5µL pipette attached to the modified Eppendorf tip with a 26G needle. The contralateral eye served as vehicle/uninjected control. Disease progression was determined by assessing the corneal haze, opacity, bacterial burden, and retinal histology of the eyes used in the model. Following euthanization, bacteria-infected mice were enucleated after 24 hr of the initial injection, and fungus-infected mice after 72 hr.


Results: Of the 50 mice injected, the modified technique was successful in 48 mice. Two mice were excluded due to cataract formed by accidental injury to the lens. The experimental endophthalmitis mice model successfully mimicked the natural clinical course. Clinical assessment and histopathology confirmed the influx of inflammatory cells into the posterior segment of the eye along with dissolution of retinal architecture.


Conclusion: Our novel method of injection using a modified Eppendorf tip and 26G needle yielded a cost-effective mouse model of clinical endophthalmitis, resulting in reproducible infection for understanding various aspects of its pathobiology.

Keywords:

Endophthalmitis; Histopathology; Infection; Mice Model; Pathobiology

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