Development of air assisted lamellar keratectomy for corneal haze model and deep anterior lamellar keratoplasty in dogs
개에서 각막흐림모델과 심부표층각막이식을 위한 공기주입 각막절제술의 개발

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dc.description학위논문 (박사)-- 서울대학교 대학원 : 수의과대학 수의학과, 2016. 2. 서강문.-
dc.description.abstractThe purpose of the present study was development of air assisted lamellar keratectomy (AK) technique for the standardized corneal haze model and deep anterior lamellar keratoplasty in dogs.
The AK was performed to achieve a constant ablation depth and size and to generate the efficient corneal haze. The ex vivo porcine corneas were categorized into four groups depending on the following trephined depths: 250, 375, 500 and 750 µm. After 4 ml air was injected to the stroma at the base of the trephination groove, the fuzzy region of the white opaque cornea was removed. No significant differences were observed between the trephined corneal depths for resection and the ablated corneal thickness at depths deeper than 375 µm. AK and conventional keratectomy (CK) were applied to six beagles and development of corneal haze was evaluated weekly until postoperative day 28. The occurrence of corneal haze and α-smooth muscle actin (SMA) expression following AK were significantly higher than those following CK. Therefore, AK was used to achieve the desired corneal thickness after resection and produce a sufficient corneal haze.
The effect of onion extract ointment on corneal haze suppression was evaluated to verify the experimental utility of AK technique for the evaluation of haze development. After the AK was performed, an artificial tear (group C), prednisolone acetate (group P), onion extract ointment (group O), and transforming growth factor (TGF)-β1 (group T) were applied to each group. The haze was significantly suppressed in the group P and O compared with the group C from day-14 after the surgery. Also, the total green intensity for α-SMA was significantly less expressed in the group P and O than in the group C. Thus, the onion extract ointment was demonstrated to have the suppression effects for corneal haze development and AK technique was sufficient to evaluate these effects.
The feasibility of deep anterior lamellar keratoplasty (DALK) using big-bubble technique (BBT), upon which the AK technique was based, was evaluated in three dogs. The eyes were examined until 150 days after the operation of DALK. The central portion of the transplanted cornea remained transparent while corneal haze developed around the transplanted margin. The marginal haze was rarely observed between the donor and recipient corneas at 150 days after the operation. A spotted haze developed in the central part of the deep stroma near the Descemets membrane (DM). Alpha-SMA positive cells were detected at the transplanted margin in which the corneal haze appeared clinically.
Based on the results of the present studies, AK technique could be useful for the evaluation of corneal haze and contribute to improving the standardized corneal haze model.
dc.description.tableofcontentsGENERAL INTRODUCTION 1

CHAPTER I. Air assisted lamellar keratectomy for the corneal haze model 5
Abstract 6
Introduction 7
Materials and Methods 9
1. Experimental design 9
2. Animals 10
3. Ex vivo experiments on porcine eyes 11
4. In vivo experiments on canine eyes 14
5. Clinical grading of corneal haze 15
6. Quantitative corneal haze grading 16
7. Histopathological and immunofluorescence analyses 17
8. Quantification of α-SMA positive cells 18
9. Statistical analyses 19
Result 20
Discussion 30
Conclusions 34

CHAPTER II. Effect of onion extract on corneal haze suppression after air assisted lamellar keratectomy 35
Abstract 36
Introduction 37
Materials and Methods 40
1. Corneal fibroblast culture and cell viability test for onion extract 40
2. A process of manufacture for the onion extract ointment 42
3. Animals 43
4. Corneal haze generation by the air assisted lamellar keratectomy 44
5. Corneal haze grading 47
6. Immunofluorescence analyses 48
7. Quantification of α-SMA positive cells 49
8. Statistical analyses 50
Results 51
Discussion 59
Conclusions 63

CHAPTER III. Deep anterior lamellar keratoplasty of dog eyes using the big-bubble technique 64
Abstract 65
Introduction 66
Materials and Methods 68
1. Animals 68
2. Surgical technique 69
3. Post-operative care and evaluation 72
4. Histopathological evaluation 74
5. Immunofluorescence detection of α-SMA 75
Results 76
Discussion 84
Conclusions 88



dc.format.extent5344460 bytes-
dc.publisher서울대학교 대학원-
dc.subjectair assisted lamellar keratectomy-
dc.subjectbig-bubble technique-
dc.subjectcorneal haze-
dc.subjectdeep anterior lamellar keratopalsty-
dc.subjectonion extract ointment-
dc.titleDevelopment of air assisted lamellar keratectomy for corneal haze model and deep anterior lamellar keratoplasty in dogs-
dc.title.alternative개에서 각막흐림모델과 심부표층각막이식을 위한 공기주입 각막절제술의 개발-
dc.contributor.affiliation수의과대학 수의학과-
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College of Veterinary Medicine (수의과대학)Dept. of Veterinary Medicine (수의학과)Theses (Ph.D. / Sc.D._수의학과)
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