Photochemically cross-linked collagen gels as three-dimensional scaffolds for tissue engineering
Photochemically cross-linked collagen gels as three-dimensional scaffolds for tissue engineering
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DOI：
10.1089/ten.2006.0153
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发表时间：
2007-08-01
影响因子：
--
通讯作者：
Gill, Thomas J.
中科院分区：
文献类型：
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作者：
Ibusuki, Shinichi;Halbesma, Gerrit J.;Gill, Thomas J.
胶原蛋白凝胶对于组织工程具有许多有利的属性，但是当加入细胞时，由于在自发凝胶化期间形成的弱非共价键，凝胶经历急剧收缩。我们假设，光化学交联胶原凝胶将使合适的支架组织工程具有良好的细胞活力和最小的凝胶收缩。分别采用532和458 nm波长的光作为凝胶交联的候选光引发剂.软骨细胞活力测定后，初始凝胶化的几个浓度的引发剂。然后使用软骨细胞和成纤维细胞在7天的培养中测量细胞活力和凝胶收缩。在凝胶化所需浓度下使用的虎红导致细胞活力很少或没有。短期存活率结果显示，0.25-或0.5- mM浓度的核黄素和40秒的光照允许超过90%的细胞存活率.使用0.25或0.5mM的核黄素浓度，在第7天，长期软骨细胞活力分别为113.1 +/-11.6%和25.4 +/-2.7%。尽管到第7天，非交联的软骨细胞结构收缩至其原始直径的59.9 +/-11.8%，成纤维细胞收缩至其原始直径的24.9 +/-5.0%，但交联的结构分别保留了原始直径的88.8 +/-7.4%和85.5 +/-5.0%.总之，通过使用核黄素和可见光的光化学交联胶原凝胶，可以产生具有有利细胞存活的稳定凝胶支架。
Collagen gels have many favorable attributes for tissue engineering, but the gels undergo dramatic contraction when cells are added because of the weak noncovalent bonds that form during spontaneous gelation. We hypothesized that photochemically cross- linking collagen gels would make suitable scaffolds for tissue engineering with favorable cell viability and minimal gel contraction. Rose Bengal and riboflavin were chosen as candidate photo- initiators for gel cross- linking using 532- and 458- nm- light wavelengths, respectively. Chondrocyte viability was measured after initial gelation for several concentrations of initiators. Cell viability and gel contraction were then measured using chondrocytes and fibroblasts over 7 days of culture. Rose Bengal used at concentrations necessary for gelation resulted in little or no cell viability. Short- term viability results showed that 0.25- or 0.5- mM concentrations of riboflavin, and 40 s of illumination permitted more than 90% cell viability. Using riboflavin concentrations of 0.25 or 0.5mM, long- term chondrocyte viability was 113.1 +/- 11.6% and 25.4 +/- 2.7%, respectively, at day 7. Although non- cross- linked chondrocyte constructs contracted to 59.9 +/- 11.8% of their original diameter and fibroblasts contracted to 24.9 +/- 5.0% of their original diameter by day 7, the cross- linked constructs retained 88.8 +/- 7.4% and 85.5 +/- 5.0% of the original diameter, respectively. In conclusion, by photochemically cross- linking collagen gels using riboflavin and visible light, stable gel scaffolds with favorable cell survival can be produced.