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Establishing a Porcine Ex Vivo Cornea Model for Studying Drug Treatments against Bacterial Keratitis
Published on: May 12, 2020
Developing a Pseudomonas aeruginosa keratitis model in Swiss albino mice for clinical assessment
Lalan Kumar Arya1, Anita Kumari1, Rakhi Kusumesh1
1Regional Institute of Ophthalmology, Indira Gandhi Institute of Medical Sciences, Patna, Bihar, India.
Abstract:
Background & objectives Pseudomonas aeruginosa is a leading cause of bacterial keratitis, known for its rapid progression, severe corneal damage, and resistance to treatment. Existing animal models exist to study disease mechanisms and therapeutic options require specialised conditions or complex procedures. This study aimed to develop a simplified, cost-effective, and reproducible murine model of P. aeruginosa keratitis using Swiss albino mice for translational research applications. Methods Four female Swiss albino mice (6-8 wk old) were maintained under standard conditions. Baseline ocular evaluation was done using a handheld slit lamp. The left corneas were abraded with a sterile 26G needle and inoculated topically with 10 μL of P. aeruginosa (1.0 × 10⁶ CFU/mL, ATCC 19660). The right eyes served as uninfected controls. Clinical signs were assessed on days 1, 2, and 3 using a standardised 0-4 scoring scale. Infection was confirmed through culture, biochemical tests, and PCR targeting the exotoxin A gene. Statistical analysis was performed using one-way ANOVA with Tukey's post hoc test. Results All infected eyes developed progressive keratitis marked by lid swelling, corneal haze, and stromal involvement. Control eyes remained unaffected. Clinical scores increased significantly over time (P< 0.05). Culture and molecular analyses confirmed successful infection. Interpretation & conclusions This simplified Swiss albino mouse model effectively mimics human P. aeruginosa keratitis and enables cost-efficient study of pathogenesis and therapeutic testing. It can facilitate standardised ophthalmic infection research.
Insights
A new, simplified mouse model using Swiss albino mice effectively replicates Pseudomonas aeruginosa bacterial keratitis. This cost-effective model aids research into disease mechanisms and treatments for this severe eye infection.
Area of Science:
- Ophthalmology
- Microbiology
- Infectious Diseases
Background:
- Pseudomonas aeruginosa is a primary cause of bacterial keratitis, leading to rapid, severe corneal damage and treatment resistance.
- Current animal models for studying P. aeruginosa keratitis are often complex and require specialized conditions.
- There is a need for a simplified, reproducible, and cost-effective model for translational research.
Purpose of the Study:
- To develop and validate a simplified, cost-effective, and reproducible murine model of P. aeruginosa keratitis.
- To utilize Swiss albino mice for studying bacterial keratitis relevant to human disease.
- To facilitate translational research applications in ophthalmic infections.
Main Methods:
- Swiss albino mice underwent corneal abrasion followed by topical inoculation with P. aeruginosa.
- Clinical signs of keratitis were assessed using a standardized scoring scale over three days.
- Infection confirmation involved bacterial culture, biochemical tests, and PCR for the exotoxin A gene.
Main Results:
- All infected mouse eyes developed progressive keratitis, including lid swelling, corneal haze, and stromal involvement.
- Control eyes remained healthy, demonstrating the model's specificity.
- Clinical scores significantly increased over time (P<0.05), and infection was confirmed by culture and molecular methods.
Conclusions:
- This simplified Swiss albino mouse model accurately mimics human P. aeruginosa keratitis.
- The model provides a cost-effective platform for investigating bacterial keratitis pathogenesis.
- It enables efficient therapeutic testing and supports standardized ophthalmic infection research.
