Abstract:

AIM： To evaluate the effect of cryopreserved amniotic membrane in 100% glycerine and RPMI 1640 medium combined with limbal stem cells allograft transplantation in ocular surface reconstruction. METHODS： ①Totally 34 eyes of 29 patients with severe ocular surface burn were recruited by the Ophthalmic Center in Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA from 2000 to 2006. The experiment was approved by the hospital ethics committee. With the informed consents of pregnant women following cesarean delivery, the placenta was offered by the Department of Gynecology and Obstetrics in the same hospital. Fresh cornea within 6 hours ex vivo was provided by the Red Cross Society of China Fujian Branch.②Amniotic membrane was separated from chorionic vesicle by a blunt dissection. Facing upward, the epithelium of amniotic membrane was put on the thin-film for ophthalmologic operation by hautchen technique, and then placed in the aseptic bottle containing 5 mL 100% glycerine and 5 mL RPMI 1640 medium for preservation at -80 ℃, in a size of 3 cm × 4 cm. Toroid demarcation was cleared on transparent cornea 1 mm ahead and tunica dura 2 mm behind limbus of cornea to expose the epithelial with superficial matrix. For severe burnt patients, vascularized scars and damaged matrix tissues were eliminated, then amniotic membrane was upwards hatched on ocular surface and fixed on the superficial tunica dura in accordance with the defect area. Amniotic membrane was force to stick cornea tightly, with the apposition suture between amniotic membrane and tunica conjunctive bulbi. Afterwards the cryopreserved amniotic membrane was combined with limbal stem cells allograft for transplantation. Additionally four eyes with hypophasis underwent anchyloblepharon suturation at the same time. RESULTS： Thirty eyes （88.24%） of 29 patients with severe ocular surface burn were averagely followed up for 22.8 months. At 7-14 days after operation, the corneal edema was extinct. At 14-21 days, the corneas reepithelialized without infection. At 3-6 months, the corneal new vessels distinctively reduced, their clarity was improved, and the visual acuity was elevated 3-5 lines. As for 4 eyes （11.76%） occurred acute rejection reaction, deep layer of corneal transplant was conducted following anchyloblepharon suturation, and recovered two months later. All the 34 eyes resulted in the restoration of a normal and stable ocular surface. CONCLUSION： Cryopreserved amniotic membrane in 100% glycerine and RPMI 1640 medium combined with limbal stem cell allograft transplantation is a useful technique in promoting the rapid and stable reepithelialization of ocular surface following severe ocular burn. It not only utilizes the bionomics of the amniotic membrane, but also provides the limbal stem cells. Thus it is a feasible and effective way for therapy of severe ocular surface.[著者文摘]