Ludwig Boltzmann Institute for Experimental and Clinical Traumatology/AUVA Research Centre, Linz/Vienna, Austria.
J Tissue Eng Regen Med. 2009 Dec;3(8):651-4. doi: 10.1002/term.207.
Preserved amniotic membrane (AM) has been used in the field of ophthalmology and wound care due to its bacteriostatic, antiphlogistic, protease-inhibiting, re-epithelialization, wound-protecting and scar formation-reducing properties. Typically, AM is applied after banking in a glycerol-preserved or freeze-dried state. Cell viabilities in different forms of preparation vary substantially, which in consequence may also be reflected in the amount and type of growth factors released from the preserved material. Therefore, we characterized the angiogenic factor (AF) profile released from different AM preparations. For this, medium was conditioned with non-preserved, glycerol- and cryo-preserved AM for 48 h, which was screened for AFs using a protein array system. In parallel, the preparations were tested for cell viability. Non-preserved as well as cryo-preserved AM maintained viabilities at 106.5 +/- 23.9% and 21.9 +/- 23.3%, respectively, whereas glycerol-preserved AM was found to be non-viable. Of the 20 investigated factors, high levels of angiogenin, GRO, IL-6/8, TIMP-1/2 and MCP-1 and low levels of EGF, IFNgamma, IGF-1, leptin, RANTES, TGFbeta1 and thrombopoietin were identified to be secreted from non-preserved AM. Cryo-preserved AM secreted high levels of IL-8, intermediate levels of GRO and TIMP-1/2 but only low levels of angiogenin, IFNgamma, IL-6 and MCP-1 and no detectable EGF, IGF-1, leptin, RANTES, TGFbeta1 and thrombopoietin. After banking in glycerol, AM releases only minute amounts of TIMP-1/2. Along with viability, the AF profile of amniotic membrane largely depends on the preparation method applied for banking. This should be considered for selection of an AM product for a specific clinical application.
保存的羊膜（AM）由于其具有抑菌、抗炎、抑制蛋白酶、再上皮化、保护伤口和减少疤痕形成的特性，已在眼科和伤口护理领域得到应用。通常，AM 在甘油保存或冻干状态下保存后再使用。不同形式的制备物中的细胞活力有很大差异，这可能也反映了从保存材料中释放的生长因子的数量和类型。因此，我们对不同 AM 制剂释放的血管生成因子（AF）谱进行了表征。为此，将非保存、甘油保存和冷冻保存的 AM 用培养基孵育 48 小时，然后使用蛋白质阵列系统筛选 AF。同时，对制剂进行了细胞活力测试。非保存和冷冻保存的 AM 的活力分别保持在 106.5%+/-23.9%和 21.9%+/-23.3%，而甘油保存的 AM 则没有活力。在所研究的 20 种因子中，发现大量分泌的因子有血管生成素、GRO、IL-6/8、TIMP-1/2 和 MCP-1，而 EGF、IFNgamma、IGF-1、瘦素、RANTES、TGFbeta1 和血小板生成素的水平则较低，这些因子均从非保存的 AM 中分泌。冷冻保存的 AM 分泌高水平的 IL-8、中水平的 GRO 和 TIMP-1/2，但仅低水平的血管生成素、IFNgamma、IL-6 和 MCP-1，且无法检测到 EGF、IGF-1、瘦素、RANTES、TGFbeta1 和血小板生成素。甘油保存后，AM 仅释放微量的 TIMP-1/2。与活力一样，羊膜的 AF 谱在很大程度上取决于用于保存的制备方法。在为特定的临床应用选择 AM 产品时，应考虑这一点。
