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가시아메바각막염 마우스 모델의 개발
- Alternative Title
- Heekyoung Kang
- Author(s)
- 강희경
- Alternative Author(s)
- Heekyoung Kang
- Advisor
- 신호준
- Department
- 일반대학원 의생명과학과
- Publisher
- The Graduate School, Ajou University
- Publication Year
- 2019-02
- Language
- eng
- Alternative Abstract
- Acanthamoeba castellanii invades the cornea and results in acanthamoebic keratitis (AK). It is mainly triggered by risk factors such as corneal injury, faulty eye surgery, and poor contact lens use. AK is typically chronic and progressive with inflammatory cells infiltrating the corneal stroma. Many in vitro studies of the pathogenesis of AK and development of therapeutic drugs need to be confirmed by the in vivo experiments. In this study, development of AK animal model was attempted. BALB/c mouse cornea was scratched using a syringe needle and ophthalmic surgical blade under anesthesia. A. castellanii (trophozoites equally mixed with cysts) cultures were serially diluted from 1 × 106 to 0.3 × 105 and placed on a 2-mm contact lens to be inserted into the mouse eye. To allow complete contact of A. castellanii with the damaged cornea, the eyelid was sutured. The keratitis symptoms in mouse eyes were grossly observed from day 1 to 7, post-inoculation. In addition, to confirm the AK development, the genomic DNA was extracted from the mouse ocular tissue with AK. For the amplification of acanthamoebic 18S-rRNA, PCR was performed with P-FLA primers. It was observed that the experimental mouse AK, characterized by typical hazy blurring and melting of the mouse cornea, developed on day 1 post-inoculation and was induced with at least 0.3 × 105 amoeba cells. With 0.5 × 105 amoeba inoculation, the experimental mouse AK was prolonged for 2 months (no more extended observation). On the contrary, the sham-operated mouse eye was clear during the observation periods. In addition, PCR products amplified from the extracted mouse eye DNA, using P-FLA primers confirmed the AK development during infection periods. Many commercial lens solutions are less effective against amoebicidal effects of Acanthamoeba. To confirm the amoebicidal effect of the commercial lens solutions, 0.5 × 105 amoebae were pre-treated with solutions A or B for 1, 6, 12 or 24 h, and inoculated into the eye of a previously established AK mouse model. AK occurrence by A. castellanii treated with lens solution A or B for 1 or 6 h was not grossly observed on day 1 and 2, but the infection progressed with a typical circulatory edema and keratitis, on days 3, 5, and 7. Upon pre-treatment for 12 or 24 h, AK development was not observed on days 1, 2, or 3 but progressed to corneal infection on days 5 and 7. Finally, the commercial contact lens solutions did not display a protective effect against AK development, although they affected the occurrence by delaying the AK. In conclusion, it is suggested that the present AK mouse model may serve as an important in vivo model for various future studies.
- Fulltext
- Appears in Collections:
- Graduate School of Ajou University > Department of Biomedical Sciences > 4. Theses(Ph.D)
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