In situ IGF-1 gene delivery to cells emerging from the injured anterior cruciate ligament.
In situ IGF-1 gene delivery to cells emerging from the injured anterior cruciate ligament.
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DOI：
10.1016/j.biomaterials.2007.10.054
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发表时间：
2008-03
期刊：
影响因子：
14
通讯作者：
A. Steinert;Meike Weber;M. Kunz;G. Palmer;U. Nöth;C. Evans;M. Murray
中科院分区：
文献类型：
--
作者：
A. Steinert;Meike Weber;M. Kunz;G. Palmer;U. Nöth;C. Evans;M. Murray
前交叉韧带（ACL）断裂是常见的膝关节损伤，即使手术修复也无法愈合。我们的研究旨在开发新的生物方法，使缝合修复这一韧带。一种有前景的策略涉及在ACL的切断端之间插入胶原水凝胶。细胞从受损的韧带迁移到水凝胶中并产生修复组织。在这里，我们已经调查了潜在的增加这一进程的胰岛素样生长因子（IGF）1的cDNA转移到修复细胞使用腺病毒载体。目标是通过在插入缺损之前用载体加载水凝胶来实现直接的原位基因递送。在逐步评估这一过程的方法中，我们证实了单层ACL成纤维细胞被腺病毒载体有效转导，并在随后掺入水凝胶中时继续表达转基因;事实上，转基因表达在胶原凝胶中持续的时间比单层培养中更长。IGF-1 cDNA的转移增加了凝胶的细胞性，并导致I型和III型胶原蛋白、弹性蛋白、腱生蛋白和波形蛋白的合成和沉积量增加。即使在高病毒载量下，细胞仍保持活力。当胶原蛋白水凝胶在插入体外实验ACL损伤之前预先加载腺病毒时，获得了类似的结果。这些数据证实了使用载体负载的水凝胶原位递送基因到受损韧带内的细胞的前景，并提出了ACL生物修复的新可能性。
Ruptures of the anterior cruciate ligament (ACL) are common knee injuries that do not heal, even with surgical repair. Our research is directed towards developing novel, biological approaches that enable suture repair of this ligament. One promising strategy involves the insertion of a collagen hydrogel between the severed ends of the ACL. Cells migrate from the damaged ligament into the hydrogel and produce repair tissue. Here we have investigated the potential for augmenting this process by the transfer of insulin like growth factor (IGF) 1 cDNA to the repair cells using an adenovirus vector. The goal is to achieve direct, in situ gene delivery by loading the hydrogel with vector prior to its insertion into the defect. In a step-wise approach towards evaluating this process, we confirmed that monolayers of ACL fibroblasts were efficiently transduced by adenovirus vectors and continued to express transgenes when subsequently incorporated into the hydrogel; indeed, transgene expression persisted longer within collagen gels than in monolayer culture. Transfer of IGF-1 cDNA increased the cellularity of the gels and led to the synthesis and deposition of increased amounts of types I and III collagen, elastin, tenascin, and vimentin. The cells remained viable, even when subjected to high viral loads. Similar results were obtained when collagen hydrogels were preloaded with adenovirus prior to insertion into an experimental ACL lesion in vitro. These data confirm the promise of using vector-laden hydrogels for the in situ delivery of genes to cells within damaged ligaments and suggest novel possibilities for the biological repair of the ACL.