الفهرس 
Acknowledgment
Anatomy of the rabbit corneaOnly 14 pages are availabe for public view
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Abstract
The present study was carried out to evaluate the effect of spanlastic formulations of Hesperidin with and without olive oil and oxytetracycline on the induced alkali burn corneal ulcer in rabbits.
This study was done at the Department of Surgery Anesthesiology and Radiology, Faculty of Veterinary Medicine, Assiut university on thirty-eight clinically healthy New Zealand albino rabbits of both sexes (23 males and 15 non-pregnant non-lactating females). Their age varied from 7 to 8 months and their body weight ranged from 1700 to 2500 grams. The animals were divided into six groups, each of 5 except the control had 10 rabbits. Hesperidin spanlastics (SPL) were drawn up utilizing the ethanol injection method. Alkali burn’s corneal ulcer was created by using a sterile small cotton swab which was soaked in 1% NaOH and applied on the central cornea for 60 seconds. The right eye of each rabbit in all groups was used for ulcer induction. The groups of animals were; group 1: the control group left untreated (placebo; using 0.9% saline either topical or subconjunctival), group 2: sub-conjunctival injection of hesperidin, group 3: sub-conjunctival injection of hesperidin/ olive oil , group 4: topical application of hesperidin drops, group 5: topical application of hesperidin/olive oil drops, group 6: Sub-conjunctival injection of oxytetracycline 5%. Naked-eye examinations of each rabbit’s eye were performed on days 8, 15, 22, 29, and 36 by two observers. The treated eyes were inspected for the presence of lacrimation, pus formation, neovascularization, the diameter of the ulcer, corneal opacity, and perforation.. At the end of the study, the animals were euthanized and tissue samples from the cornea were harvested and fixed in 10 % neutral buffered formalin and sent for histopathological examination. Other specimens from the cornea were collected for electron microscopic examination. They were washed with normal saline. They were, subsequently, fixed in a mixture of 2.5% paraformaldehyde and 5% glutaraldehyde in 0.1 M. sodium phosphate buffer with a pH of 7.3 at 4℃ for 24 h.
The clinical findings of the topical use of Hesperidin (HD) and Hesperidin/olive oil combination (HOD) showed the corneal opacity increased gradually in the control group till the 3rd week, then persisted till the end of the experiment. It decreased in the other groups and reached a minimum at the end of the study. The corneal neovascularization increased sharply in the control group till the 3rd week, then persisted till the end of the experiment. It gradually increased, but slightly in comparison with the control one in the HOD group. It was not detected in the HD group. Lacrimation increased sharply in the control group till the 3rd week and then persisted till the end of the experiment. It decreased significantly in the HOD group relative to the control group, especially from the 3rd week. It was not detected in the HD group. There were significant decreases in the size of ulcers between the 1st and 5th week in the HOD and HD groups. There were significant decreases in pus formation in the HD and HOD groups relative to the control group starting from the 3rd week. The corneal perforation was detected only in the control group, but there was no significant difference between the treated groups and the control one.
The subconjunctival use of Hesperidin (HSC), Hesperidin/olive oil combination (HOSC), and Oxytetracycline 5% (T) displayed that there were no significant changes in the corneal opacity between groups except at the 1st week. In the HOSC group, there was a significant decrease in the corneal opacity score between the 1st and 5th week. There was a fluctuation in the corneal neovascularization in the HSC, and HOSC groups. In the T and control groups, there was a gradual increase in neovascularization till the 4th week. There was a substantial decrease in the lacrimation in the HSC and HOSC groups relative to the control group starting from the 3rd week till the end of the study. Also, there is a significant decrease in the level of lacrimation in the HOSC group relative to the HSC group in the 2nd week only. There was no difference between the T and control groups. There were significant decreases in the size of the corneal ulcer in the 5th week relative to the 1st week in both HSC and HOSC groups. In the 3rd and 4th weeks, there was an obvious decrease in the diameter of the corneal ulcer in the HSC relative to the control one. The presence of pus decreased significantly in both HSC and T groups in the 4th week compared to the control one. This refers to the antibacterial effect of the Hesperidin and oxytetracycline HCl. The perforation was not detected in the HOSC group. It was detected in other groups, especially the control and T groups, starting from the 3rd week.
The fluorescein staining of the corneal ulcer gradually decreased week by week in the HOD and HD groups compared to other groups (HSC, HOSC, T). The corneal ulcer in control groups did not take the staining at the end of the study due to the corneal perforation.
The histopathological and electron microscopic examination revealed that the healing of the corneal ulcer was better in the HOD group, followed by HD group. At the end of the study appeared slightly normal corneal epithelium with normal stroma and Descemet’s membrane in the HOD group. However, Hesperidin-treated group showed degenerated epithelium with no ulcers and normal stroma. The group of rabbits treated with hesperidin (HSC) showed necrosed epithelium and ulceration. The stroma revealed proliferated keratocytes and inflammatory cell infiltration as lymphocytes. Hesperidin olive oil treated group (HOSC) revealed necrosed epithelium with a small area of ulcer with normal stroma. In the Oxytetracycline treated group, there was a necrosis of the corneal epithelium with ulceration, edema of stroma, widening of its collagenous fibers, and damage to the Descemet’s membrane. In the control positive group, there were necrosed epithelium, desquamation, and multiple ulcerated areas covered with colonies of bacteria. The stroma swelled due to edema and was infiltrated with inflammatory cells, and there was damage in the Descemet’s membrane.