Dehydrated form of plasmid expressing basic fibroblast growth factor-polyethylenimine complex is a novel and accurate method for gene transfer to the cornea
Dehydrated form of plasmid expressing basic fibroblast growth factor-polyethylenimine complex is a novel and accurate method for gene transfer to the cornea
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DOI：
10.1080/02713680500330512
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发表时间：
2005-11-01
影响因子：
2
通讯作者：
Tai, MH
中科院分区：
文献类型：
--
作者：
Kuo, CN;Yang, LC;Tai, MH
目的：我们描述了一种新的载体系统的非病毒基因转移到角膜使用脱水形式的质粒表达碱性成纤维细胞生长因子-聚乙烯亚胺（p-bFGF-PEI）复合物诱导血管生成。研究方法：在植入含有1 μ g绿色荧光蛋白的脱水形式的PEI（p-GFP- PEI;对照组）或通过在环境温度下旋转真空引入的10 μ g、1 μ g或0.1 μ g的p-bFGF-PEI后，在Sprague-Dawley大鼠的48只眼睛中评价角膜新血管形成。从第1天至第45天观察并定量新生血管形成。Western blot和免疫组化检测18 kDa bFGF蛋白表达。结果：p-bFGF-PEI组角膜缘血管在第3天开始出芽。脱水形式的p-bFGF-PEI复合物诱导剂量依赖性角膜新生血管形成，其在10 μ g bFGF组中在第24-30天达到最大值，在1 μ g bFGF组中在第18-24天达到最大值，在0.1 μ g bFGF组中在第15-21天达到最大值，然后逐渐消退。在GFP组中没有观察到新血管形成。结论：p-bFGF-PEI复合物的脱水形式是一种用于控制剂量、定位试剂和避免在转染到角膜组织中期间损失液体形式的新颖且精确的方法。
Purpose: We describe a novel vector system of nonviral gene transfer into the cornea using a dehydrated form of a plasmid expressing basic fibroblast growth factor - polyethylenimine (p-bFGF-PEI) complex to induce angiogenesis. Methods: Corneal neovascularization was evaluated in 48 eyes of Sprague-Dawley rats after implantation of a dehydrated form of PEI containing 1 mu g green fluorescent protein (p-GFP- PEI; control group), or 10 mu g, 1 mu g, or 0.1 mu g of p-bFGF-PEI introduced by spin vacuum at ambient temperature. Neovascularization was observed and quantified from day 1 to day 45. Eighteen kDa bFGF protein expression was analyzed by Western blot and immunohistochemistry. Results: Limbal vessels began to sprout on day 3 in the p-bFGF-PEI groups. The dehydrated form of the p-bFGF-PEI complex induced dose-dependent corneal neovascularization, which reached a maximum on days 24-30 in the 10 mu g bFGF group, days 18-24 in the 1 mu g bFGF group, and days 15-21 in 0.1 mu g bFGF group, and then regressed progressively. No neovascularization was observed in the GFP group. Conclusions: The dehydrated form of the p-bFGF-PEI complex is a novel and precise method for controlling the dose, localizing the reagents, and avoiding loss of liquid form during transfection into corneal tissue.