Amniotic membrane for ocular surface reconstruction:: Donor variations and the effect of handling on TGF-ß content
Amniotic membrane for ocular surface reconstruction:: Donor variations and the effect of handling on TGF-ß content
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DOI：
10.1167/iovs.05-1415
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发表时间：
2006-10-01
影响因子：
4.4
通讯作者：
Dua, Harminder S.
中科院分区：
文献类型：
--
作者：
Hopkinson, Andrew;McIntosh, Richard S.;Dua, Harminder S.
目的.羊膜移植是眼外科公认的手术方法。然而，很少有人知道的供体间和供体内的变异性内的膜。此外，处理、储存和术前准备方法对膜的影响尚未完全阐明。本研究的目的是使用TGF-β作为一个例子，以调查供体间和供体内的变异性，并确定"处理"对新鲜，加工和储存，移植准备AM（TRAM）内TGF-β 1的影响。通过实时聚合酶链反应、免疫组织化学、SDS-PAGE和Western印迹法分析了17例新鲜和处理的人AM的TGF-β 1。TGF-β 1是所有样品中TGF-β的最高标准化表达同种型，但它在不同供体的膜之间以及在同一膜内的不同位点处变化。海绵层中的浓度最高。去除海绵层成功地从TRAM中去除了大量TGF-β 1。潜伏相关蛋白（LTP）和潜伏性TGF-β结合蛋白（LTBP）也被检测到。TGF-β 1以各种调节形式存在于AM中。膜间和膜内变化的程度通过处理来改变。除非采用标准化方案，提供具有一致成分的膜，否则临床结果可能会有所不同，比较可能无效。
PURPOSE. Amniotic membrane (AM) transplantation is an accepted procedure in ocular surgery. However, little is known of the interdonor and intradonor variability within the membrane. In addition, the effects of the methods of processing, storage, and preoperative preparation on the membrane are not fully elucidated. The purpose of this study was to use TGF-beta as an example to investigate interdonor and intradonor variability and to determine the effect of "handling" on TGF-beta 1 within fresh, processed and stored, and transplantation-ready AM (TRAM).METHODS. Seventeen human AMs, both fresh and handled, were analyzed for TGF-beta 1 by real-time polymerase chain reaction, immunohistochemistry, SDS-PAGE, and Western blotting.RESULTS. TGF-beta 1 was the highest normalized expressed isoform of TGF-beta in all samples, but it varied between membranes of different donors and at different sites within the same membrane. The highest concentration was noted in the spongy layer. Removal of the spongy layer successfully removed the bulk of TGF-beta 1 from TRAM. Latency-associated protein (LAP) and a latent TGF-beta-binding protein (LTBP) were also detected.CONCLUSION. TGF-beta 1 is present in various regulatory forms in the AM. A degree of intermembrane and intramembrane variation is modified by handling. Unless a standardized protocol is adopted that delivers a membrane with consistent constituents, clinical outcomes may vary and comparisons may be invalid.