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C.J. Connon, T. Nakamura, A.J. Quantock, S. Kinoshita; The Persistence of Transplanted Amniotic Membrane in Corneal Stroma . Invest. Ophthalmol. Vis. Sci. 2005;46(13):4988.
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Purpose: In recent years the amniotic membrane (AM) has been used as a treatment substrate for reconstruction of the ocular surface in patients with limbal stem cell deficiency or a persistent epithelia defect. However, the AM’s interaction with corneal stroma has yet to be properly characterized. A study was therefore undertaken to investigate any histological changes to the stroma or AM following its insertion in to the cornea for up to 10 months. Methods: Rabbit AM was collected from 7 foetuses from a heavily pregnant rabbit, washed in PBS and cut into 5mmx5mm squares. These squares were incubated in 0.5% dichlorotriazinyl aminofluorescein (DTAF) dye in 0.2mol/l sodium bicarbonate for 1 min and stored in PBS until used. Four young rabbits were anaesthetised and stromal pockets measuring 6–7mm in diameter with a 2mm opening were cut into the central stroma of each eye. To the right eye DTAF treated AM squares were carefully inserted. Antibiotics (Tarvid Ophthalmic ointment) were added to each eye twice a day for 5 days. The eyes were photographed, enucleated and then examined by transmission electron microscopy and fluorescent microscopy at 1, 3, 7 and 10 months following surgery. Results: From photographs, visibility of the inserted AM decreased overtime. However florescent microscopy clearly showed the presence of DTAF treated AM after 10 months and that the inserted AM did not elicit an immune response. Furthermore, transmission electron microscopy revealed little or no change to the AM ultrastructure or invasion by stromal keratocytes. Conclusions: AM allografts persist intact within the stroma for many months post–op and are not broken down or dissolved. The possibility of long–term effects on corneal health and transparency from the continued presence of AM within the eye should be considered before using AM as a treatment substrate for reconstruction of the ocular surface.
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