Experiment of amnion epithelial cell suspension liquid used for acute rabbit corneal alkali burn / 国际眼科杂志(Guoji Yanke Zazhi)

ABSTRACT

AIM:To investigate the effects of amnion epithelial cell ( AEC) suspension liquid on the biological behavior of the rabbit's corneal epithelium, combined with the in vitro and in vivo experiments. · METHODS: The rabbit's corneal epithelium were cultured in the lower chamber of transwell, and AEC suspension liquid was dropwised in the upper chamber. There was only culture medium in the upper chamber of the control group. The proliferation of rabbit's corneal epithelium was observed with CCK-8 automated colorimetry and the expression of PCNA was detected by immunocytochemistry. We used the scratch wound assay to detect the migration of corneal epithelial cell ( CEC ) . The in vivo models were established by placing a 10mm diameter corneal trephine in the center of the cornea, within 1mol/L NaOH for 1min. We divided those into three groups treatment group of AEC suspension liquid eye drop, AEC suspension liquid subconjunctival injection and the control group without any treatment. Using the slit- lamp biomicroscope and fluorescence staining to observe the cornea per week. After 28d we took the eyeballs with the HE staining. The expression of VEGF was detected by immunohistochemistry. ·RESULTS: The activity of CEC with AEC treatment was much higher than the control group ( P< 0. 05 ). The expression of PCNA increased in AEC group (P<0. 05). And the migration of CEC in the AEC group was faster than the control one. In vivo, the inflammation of the corneal and the CNV of the AEC group were all significantly reduced compared with the control group (P<0. 05 ). There were less invasive cells and more ordered organization arrangement in ACE group observed by the HE staining. The expression of VEGF and mcp-1 in these two AEC treatment groups all significantly decreased compared with the control group (P<0. 05). ·CONCLUSION: AEC suspension liquid can promote the proliferation and migration of the rabbit's corneal epithelium. The potential of AEC suspension liquid as a therapy for acute corneal alkali burn.