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Technique for Processing and Preservation of Human Amniotic Membrane for Ocular Surface Reconstruction
Abstract: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.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1082431Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 3181
 M. Muraine, G. Descargues, O. Franck, F. Villeroy, D. Toubeau, E. Menguy, J. Martin, G. Brasseur. "Amniotic membrane graft in ocular surface disease. Prospective study with 31 cases." J Fr Ophtalmol., vol. 24, 2001, pp.798-812.
 N. J. Koizumi, T. J. Inatomi, C. J. Sotozono. "Growth factor mRNA and protein in preserved human amniotic membrane." Curr Eye Res. Vol. 20, 2000, pp. 173-177.
 Q. Zhang, K. Shimya, A. Moriyama. "Production of secretory leukocyte protease inhibitor by human amniotic membranes and regulation of its concentration in amniotic fluid." Mol Hum Reprod. vol. 7, 2001, pp. 573-579.
 Hao Y, Ma DH, Hwang DG. "Identification of antiangiogenic and antiinflammatory proteins in human amniotic membrane." Cornea vol. 19, 2000, pp. 348-352.
 H. J. Chen, R. T. Pires, S. C. Tseng. "Amniotic membrane transplantation for severe neurotrophic corneal ulcers." Br J Ophthalmol. vol. 84, 2000, pp. 826-833.
 Linnala A , Balza E, Zardi L, Virtanen I . "Human amnion epithelial cells assemble tenascins and three fibronectin isoforms in the extracellular matrix." FEBS Lett. vol. 317, 1993, pp. 74-8.
 Shimmura S, Shimazaki J, Ohashi Y, Tsubota K. "Anti inflammatory effects of amniotic membrane transplantation in ocular surface disorders." Cornea. vol. 20, 2001, pp. 408-413.
 O. Franck, G. Descargues, E. Menguy. "Technique of harvesting and preparation of amniotic membranes." J Fr Ophthalmol. vol. 23, 2000, pp.729-734.
 A. DeRoth. "Plastic repair of conjunctival defects with fetal membrane." Arch Ophthalmol. vol. 23, 1940, pp. 522-25.
 P. J. Adds, C. Hunt, S. "Hartley. Bacterial contamination of amniotic membrane." Br J Ophthalmol. vol. 85, 2001, pp. 228-30.
 A. Blanco, C. T. Pillai, H. S. Dua. "Amniotic membrane transplantation for ocular surface reconstruction." Br J Ophthalmol. vol. 83, 1999, pp.399-402.
 D. Meller, S. C. Tseng. "Conjunctival epithelial cell differentiation on amniotic membrane." Invest Ophthalmol Vis Sci. vol. 40, 1999, pp 878-886.
 S. C. Tseng, , D. Q. Li, , X. Ma. "Suppression of transforming growth factor-beta isoforms, TGF-beta receptor type II, and myofibroblast differentiation in cultured human corneal and limbal fibroblasts by amniotic membrane matrix." J Cell Physiol. Vol. 179, 1999, pp. 325- 335.
 J. C. I. Kim, S. C. G. Tseng. "Transplantation of preserved human amniotic membrane for surface reconstruction in severely damaged rabbit corneas." Cornea. Vol. 14, 1995, pp. 473-484.
 F. E. Kruse, K. Rohrschneider, H. E. Voelcker. "Multilayer amniotic membrane transplantation for reconstruction of deep corneal ulcers." Ophthalmology. vol. 106, 1999, pp. 1504-1511.
 M. A. Ganatra, S Bhura. "Management of skin graft donor site by irradiated amniotic membrane." Pakistan J Surg. vol. 19, 2003, pp. 82- 85.
 H. S. Dua, J. A. Gomes, A. J. King, V. S. Maharajan. "The amniotic membrane in ophthalmology." Surv Ophthalmol. vol. 49, 2004, pp. 51- 77.
 K. Martin, M. Horst, B. J├╝rgen. "Reconstruction of the contracted ocular socket with free full-thickness mucosa graft." Intl J Oral Maxillofacial Surg. vol. 29, 2000, pp. 96-98.
 K. C. Sippel, J. J. K. Ma, C. S. Foster. "Amniotic membrane surgery." Curr Opin Ophthalmol. vol. 12, 2001, pp. 269-281.
 F. E. Kruse, A. M. Joussen, K. Rohrschneider. "Cryopreserved human amniotic membrane for ocular surface reconstruction." Graefes Arch Clin Exp Ophthalmol. vol. 238, 2000, pp. 68-75.
 M. Kubo, Y. Sonoda, R. Muramatsu, M. Usui. "Immunogenicity of human amniotic membrane in experimental xenotransplantation." Invest Ophthalmol Vis Sci. vol. 42, 2001, pp. 1539-1546.
 P. J. Adds, C. Hunt, S. Hartley. "Bacterial contamination of amniotic membrane." Br J Ophthalmol. vol. 85, 2001, pp. 228-230.
 A. Chandra, O. P. Maurya, B. Reddy, G. Kumar, K. Pandey, G. Bhaduri. "Amniotic membrane transplantation in ocular surface disorders." J Indian Med Assoc. vol. 103, 2005, pp. 364-368.
 H. N. Madhavan, K. Priya, J. Malathi, P. R. Joseph. "Preparation of amniotic membrane for ocular surface reconstruction." Indian J Ophthalmol. vol. 50, 2002, pp. 227-31.