Mietz Holger, Welsandt Gerhard, Hueber Arno, Esser Christina, Krieglstein Günter K
Department of Ophthalmology, University of Cologne, Cologne, Germany.
Graefes Arch Clin Exp Ophthalmol. 2007 Sep;245(9):1367-75. doi: 10.1007/s00417-007-0547-z. Epub 2007 Feb 21.
Highly toxic antimetabolites have gained access to routine clinical use to modulate and reduce the amount of postoperative scarring following glaucomatous filtering procedures. It could be speculated that by combining two different antiproliferative substances with different mechanisms of action total amounts of the substances could be decreased and side effects reduced.
Twenty-two substances were tested that had antiproliferative effects by acting cytotoxically, inhibiting growth factors, or inducing apoptosis. With combinations of each two substances, cell culture experiments using 3T3 and human Tenon's capsule fibroblasts were performed evaluating cell toxicity, proliferation and migration, the extent of free radicals, and the amount of apoptosis (TUNEL, electron microscopy). The five most potent combinations were used in an animal experiment with rabbits performing filtering procedures. The extent of episcleral scarring was evaluated by histopathology.
The results of the various assays revealed consistently strong effects in 5 of the 462 combinations. Of these five combinations, two were highly effective in the rabbit model. Substances with strong effects when applied in combination included staurosporine, mitomycin, and CD95L.
We found synergistic effects in assays that evaluated different aspects of cell function. The amount of scarring in an animal experiment was inhibited to a level comparable with a high single dose of mitomycin. Combination therapy of two antiproliferative acting substances may be a promising concept.
高毒性抗代谢物已用于常规临床，以调节和减少青光眼滤过术后的瘢痕形成量。可以推测，通过将两种具有不同作用机制的抗增殖物质联合使用，可减少物质的总量并降低副作用。
测试了22种通过细胞毒性作用、抑制生长因子或诱导凋亡发挥抗增殖作用的物质。将每两种物质组合，使用3T3细胞和人Tenon囊成纤维细胞进行细胞培养实验，评估细胞毒性、增殖和迁移、自由基程度以及凋亡量（TUNEL法、电子显微镜检查）。将五种最有效的组合用于对兔子进行滤过手术的动物实验。通过组织病理学评估巩膜上瘢痕形成的程度。
各种检测结果显示，在462种组合中的5种组合中始终呈现出强烈的效果。在这五种组合中，有两种在兔子模型中非常有效。联合应用时效果显著的物质包括星形孢菌素、丝裂霉素和CD95L。
我们在评估细胞功能不同方面的检测中发现了协同效应。动物实验中的瘢痕形成量被抑制到与高剂量单剂量丝裂霉素相当的水平。两种抗增殖作用物质的联合治疗可能是一个有前景的概念。
