PURPOSE: To evaluate the inflammatory response following the insertion of corneal epithelium into rabbit corneal stroma. METHODS: Newzealand white rabbits were underwent corneal flap procedure using Hansatome and corneal epithelium was inserted. We divided the rabbits into three groups: Group A: flap only, Group B: flap with central epithelium insertion, Group C: flap with peripheral epithelium insertion. Eyes are enucleated, 4, 24, 48, 72 hours, 1 week, 1 month. Immunohistochemical stain fro CD4, CD8, and CD11b were used to test frozen section. RESULTS: Cd4 was expressed weakly at 24 hours in group A, B, C. around limbus and peripheral cornea. CD8 was expressed at 24 hours in group a, b, c especially strongly at group 3. In group B, C, CD8 was persistently expressed at 72 housr. CD11b was expressed at 24 hours in all groups. But in group C. CD11b was expressed strongly and persists at 72 hours and 1 week. CONCLUSION: Macrophage and Cytotoxic T cell maybe play an important role in corneal stromal wound healing after iatrogenic corneal epithelial insertion and further evaluation will be needed.
Cornea ; Corneal Stroma* ; Epithelium ; Epithelium, Corneal ; Frozen Sections ; Macrophages ; Rabbits ; Wound Healing

PURPOSE: To investigate that the effects of early wound healing stage after corneal epithelial scrape injury. METHODS: We studied the change of scraped corneal wound like corneal cells, corneal thickness, acelluar zone, and celluar morphology occurring at the time points of 1, 4, 12, 24, 48, 72 hours, and 7 days after corneal epithelial scrape injury by the confocal microscopy and EM findings in 4 each group rabbits. RESULTS: By normal confocal microscopy, the mean cell density was 891 cells/mm2 in the anterior stroma and decreased to 814 cells/mm2 in the middle stroma, 731 cells/mm2 in the posterior stroma, and the endothelial density was 3236 cells/mm2. The change in the morphology of the keratocyte nuclei from an elliptical shape anteriorly, to a more elongated shape posteriorly. Apoptosis revealed like as condensation or fragmentation of chromatin and nuclei, vesicle formation, apoptotic bodies after corneal scraped injury by EM findings. The mean thickness of normal cornea was as follow; 47 micrometer in the epithelium, 334 micrometer in the stroma, and 392 micrometer in total cornea. The thickness of postoperative cornea including stromal thickness and total thickness increased at the early wound healing stage, and then decreased to the postoperative 48 hours(P<0.001). Mean range of acellular zone in the stroma increased at the early wound healing, but significantly decreased at the postoperative 48 hours, 79 micrometer(P<0.001). CONCLUSIONS: Keratocyte cell density and corneal thickness at the three portions of cornea, the thickness of stromal acelluar zone, and the changes of cellular morphology were related with a kind of the early post-inflammatory reaction, especially 24 hours, of corneal scraped injury. It should be needed more studies concerned with control of early post-inflammatory reaction.
Apoptosis ; Cell Count ; Chromatin ; Cornea ; Epithelium ; Microscopy, Confocal ; Rabbits ; Wound Healing ; Wounds and Injuries

PURPOSE: This study was aimed to evaluate changes in the stromal keratocyte after ablation of 50 micrometer and 100 micrometer with use of photorefractive keratectomy(PRK). METHODS: At 4 hours, 24 hours, 72 hours, 7 days and 1 month after PRK, each group of rabbits including normal control group was treated with terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling(TUNEL) staining using ApopTag(R) kit in vivo, then apoptotic keratocytes were evaluated with light microscope. RESULTS: There was no response with TUNEL staining of the epithelial cells, stromal keratocyte, and endothelium in normal cornea. In the ablation group, however, regardless of the depth of photorefractive ablation, the TUNEL signal was maximal after 4 hours, and it decreased with time. The signal was more intense in 100 micrometer ablation group than 50 micrometer ablation group, although the signal was not observed at the endothelial cells in both groups. The number of apoptotic stromal keratocytes at each time point of 4 hr, 24 hr, 72 hr, and 1 week was 57+/-8.9, 49+/-7.5, 36+/-5.1, and 12+/-1.3 cells/field in 100 micrometer ablation group, and 31+/-4.4, 28+/-4.6, 21+/-3.9, and 5+/-1.1 cells/field in 50 micrometer ablation group, and the difference between the two groups was statistically significant(P<0.05). CONCLUSION: The more the amount of ablation with photorefractive keratectomy, the stronger the apoptotic response. The postoperative apoptotic response was observed especially within 1 week. These findings suggest that early suppression of postoperative apoptosis within 1 week will influence on the prognosis of visual quality after photorefractive keratectomy, and more studies will be needed in the future.
Apoptosis* ; Cornea ; Endothelial Cells ; Endothelium ; Epithelial Cells ; In Situ Nick-End Labeling ; Photorefractive Keratectomy* ; Prognosis ; Rabbits

The present study aimed at evaluation of prophylactic efficacy and possible mechanisms of asiaticoside (AS) based standardized extract of Centella asiatica (L.) Urban leaves (INDCA) in animal models of migraine. The effects of oral and intranasal (i.n.) pretreatment of INDCA (acute and 7-days subacute) were evaluated against nitroglycerine (NTG, 10 mg·kg(-1), i.p.) and bradykinin (BK, 10 μg, intra-arterial) induced hyperalgesia in rats. Tail flick latencies (from 0 to 240 min) post-NTG treatment and the number of vocalizations post-BK treatment were recorded as a measure of hyperalgesia. Separate groups of rats for negative (Normal) and positive (sumatriptan, 42 mg·kg(-1), s.c.) controls were included. The interaction of INDCA with selective 5-HT1A, 5-HT1B, and 5-HT1D receptor antagonists (NAN-190, Isamoltane hemifumarate, and BRL-15572 respectively) against NTG-induced hyperalgesia was also evaluated. Acute and sub-acute pre-treatment of INDCA [10 and 30 mg·kg(-1) (oral) and 100 μg/rat (i.n.) showed significant anti-nociception activity, and reversal of the NTG-induced hyperalgesia and brain 5-HT concentration decline. Oral pre-treatment with INDCA (30 mg·kg(-1), 7 d) showed significant reduction in the number of vocalization. The anti-nociceptive effects of INDCA were blocked by 5-HT1A and 5-HT1B but not 5-HT1D receptor antagonists. In conclusion, INDCA demonstrated promising anti-nociceptive effects in animal models of migraine, probably through 5-HT1A/1B medicated action.
Administration, Intranasal ; Administration, Oral ; Animals ; Bradykinin ; Female ; Hyperalgesia ; chemically induced ; prevention & control ; Male ; Migraine Disorders ; chemically induced ; prevention & control ; Models, Animal ; Nitroglycerin ; Nociception ; drug effects ; Plant Leaves ; chemistry ; Pre-Exposure Prophylaxis ; Rats ; Rats, Wistar ; Reaction Time ; Receptors, Serotonin, 5-HT1 ; drug effects ; Serotonin 5-HT1 Receptor Antagonists ; metabolism ; Tail ; physiology ; Triterpenes ; administration & dosage ; pharmacology