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가시아메바각막염 마우스 모델의 개발
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 신호준 | - |
| dc.contributor.author | 강희경 | - |
| dc.date.accessioned | 2019-04-01T16:42:26Z | - |
| dc.date.available | 2019-04-01T16:42:26Z | - |
| dc.date.issued | 2019-02 | - |
| dc.identifier.other | 28837 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/15219 | - |
| dc.description | 학위논문(박사)--아주대학교 일반대학원 :의생명과학과,2019. 2 | - |
| dc.description.tableofcontents | INTRODUCTION MATERIALS AND METHODS A. Cultivation of Acanthamoeba castellanii B. Encystation of A. castellanii trophozoites C. Experimental acanthamoebic keratitis development in mouse (1) Inoculation of A. castellanii into mouse eye (2) Observation of experimental mouse keratitis symptom D. Confirmation of experimental mouse keratitis (1) Genomic DNA extraction from keratitis-induced mouse eyeball (2) PCR with A. castellanii-specific primers (3) 18S-rDNA sequencing and homology analysis E. Optimal numbers of A. castellanii for the development of mouse keratitis F. Effect of commercial lens solutions on AK development in mouse (1) Pre-treatment of commercial lens solutions on A. castellanii (2) Effect of commercial lens solutions on AK mouse model G. Ethics statement III. RESULTS A. Morphological observation of A. castellanii cyst formation B. The occurrence of acanthamoebic keratitis in mouse model C. A. castellanii DNA amplification from mouse eye tissue developed acanthamoebic keratitis D. Optimal numbers of A. castellanii for keratitis development in mouse E. Establishment of acanthamoebic keratitis mouse model F. Effect of commercial contact lens solution on the AK development in mice (1) Morphology of A. castellanii pre-treatment with solution A or B (2) Change of acanthamoebic keratitis development in mice IV. DISCUSSION V. CONCLUSION REFERENCES 국문요약 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | 가시아메바각막염 마우스 모델의 개발 | - |
| dc.title.alternative | Heekyoung Kang | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Heekyoung Kang | - |
| dc.contributor.department | 일반대학원 의생명과학과 | - |
| dc.date.awarded | 2019. 2 | - |
| dc.description.degree | Doctoral | - |
| dc.identifier.localId | 905171 | - |
| dc.identifier.uci | I804:41038-000000028837 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/common/orgView/000000028837 | - |
| dc.description.alternativeAbstract | 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. | - |
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- Graduate School of Ajou University > Department of Biomedical Sciences > 4. Theses(Ph.D)
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