Technique for Processing and Preservation of Human Amniotic Membrane for Ocular Surface Reconstruction

Human amniotic membrane (HAM) is a useful biological material for the reconstruction of damaged ocular surface. The processing and preservation of HAM is critical to prevent the patients undergoing amniotic membrane transplant (AMT) from cross infections. For HAM preparation human placenta is obtained after an elective cesarean delivery. Before collection, the donor is screened for seronegativity of HCV, Hbs Ag, HIV and Syphilis. After collection, placenta is washed in balanced salt solution (BSS) in sterile environment. Amniotic membrane is then separated from the placenta as well as chorion while keeping the preparation in BSS. Scrapping of HAM is then carried out manually until all the debris is removed and clear transparent membrane is acquired. Nitrocellulose membrane filters are then placed on the stromal side of HAM, cut around the edges with little membrane folded towards other side making it easy to separate during surgery. HAM is finally stored in solution of glycerine and Dulbecco-s Modified Eagle Medium (DMEM) in 1:1 ratio containing antibiotics. The capped borosil vials containing HAM are kept at -80°C until use. This vial is thawed to room temperature and opened under sterile operation theatre conditions at the time of surgery.

Anticoagulatory Role of an Ergot Mesylate: Hydergine

Thrombosis can be life threatening, necessitating therefore its instant treatment. Hydergine, a nootropic agent is used as a cognition enhancer in stroke patients but relatively little is known about its anti-thrombolytic effect. To investigate this aspect, in vivo and ex vivo experiments were designed and conducted. Three groups of rats were injected 1.5mg, 3.0mg and 4.5mg hydergine intraperitonealy with and without prior exposure to fresh plasma. Positive and negative controls were run in parallel. Animals were sacrificed after 1.5hrs and BT, CT, PT, INR, APTT, plasma calcium levels were estimated. For ex vivo analyses, each 1ml blood aspirated was exposed to 0.1mg, 0.2mg, 0.3mg dose of hydergine with parallel controls. Parameters analyzed were as above. Statistical analysis was through one-way ANOVA. Dunken-s and Tukey-s tests provided intra-group variance. BT, CT, PT, INR and APTT increased while calcium levels dropped significantly (P