| Corneal wound is a common clinical ophthalmological disease in dogs which often accompanies with corneal turbidity, ulceration. Persistent wound may lead to corneal neovascularization or chromatosis, which can affect normal vision, even result in the permanent lose of vision. At present, treatment measures that could be taken for corneal wound diseases are limited.Amniotic membrane has many properties such as anti-inflammatory, antibacterial, antiviral, inhibiting the formation of blood vessels as well as promoting epithelial growth which make it valuable in research and application for clinical ophthalmology. However, there are few studies about the use of amniotic membrane in canine eye diseases.Thymosin beta4(Tβ4) can inhibit apoptosis, decrease the formation of CNV, reduce inflammatory, promote migration and adhesion of corneal epithelial cells. The effectiveness of amniotic membrane transplantation combined with Tβ4for corneal wound healing still unclear.Amniotic membrane transplantation combined with T04was used for the canine corneal injury treatment in this experiment. The object of this study was to provide clinical basis for seeking an effective treatment for corneal injury in dogs.In vitro, the corneal epithelial tissues were used to culture the primary canine corneal epithelial cells. The third generation of canine corneal epithelial cells was respectively incubated at a density of5×105/mL in a96-well plate and exposed to different concentrations Tβ4(1ng/mL,10ng/mL,50ng/mL,100ng/mL) mediums, and the effect of Tβ4is tested with the CCK-8kit after24h and48h and showed with the optical density (OD). In the corneal alkali burn model in vivo, the circular filter paper pieces were soaked with0.5mol/mL NaOH, and then applied into the center of the cornea for1minute in every dog’s right eye. Twenty-four dogs were divided on average into four groups randomly:the control group, the Tβ4group, the amniotic membrane transplantation group and the amniotic membrane transplantation combined with TP4group. The amniotic membrane transplantation surgery was performed immediately following the alkali burn in the amniotic membrane transplantation group and the amniotic membrane transplantation combined with Tβ4group. The Tβ4were used in the TP4group and the amniotic membrane transplantation combined with Tβ4group, at the same time PBS were dropped into the eyes of the control dogs and the amniotic membrane transplantation treated dogs. The ofloxacin were dropped into the eye of every dog. The corneal turbidity and neovascularization were valuated by slit lamp from day1to day10.In vitro, the effect of Tβ4to stimulate corneal epithelial cells proliferation was dose-dependent and reached the maximal effect at50ng/mL. The100ng/mL Tβ4accelerated the corneal epithelial cells proliferation more significantly than the control group. The effect of Tβ4promoted corneal epithelium proliferation at the48h point is better than24h point. In vivo, the use of amniotic membrane transplantation combined with Tβ4in the canine corneal alkali burn model could effectively promote the corneal wound healing, reduce the corneal opacity and inhibit CNV. Day1～10the scores of the corneal turbidity of the amniotic membrane transplantation combined with Tβ4group and the amniotic membrane transplantation group were significantly lower than the control group and the Tβ4group. The CNV appeared at the third day in every group. Day3-10the CNV scores of the amniotic membrane transplantation group were higher significantly than the Tβ4group. The result of the CNV scores had no significant difference between the Tβ4group and the amniotic membrane transplantation combined with Tβ4group.In conclusion, Tβ4can promote corneal epithelium proliferation and the maximal effective dose was50ng/mL. The amniotic membrane transplantation combined with Tβ4can significantly advance corneal wound healing, inhibit CNV, reduce the corneal opacity and may be an effective treatment for canine corneal injury. |
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