In vitro characterization of chitosan-gelatin scaffolds for tissue engineering
In vitro characterization of chitosan-gelatin scaffolds for tissue engineering
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DOI：
10.1016/j.biomaterials.2005.05.036
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发表时间：
2005-12-01
期刊：
影响因子：
14
通讯作者：
Madihally, SV
中科院分区：
文献类型：
--
作者：
Huang, Y;Onyeri, S;Madihally, SV
近年来，壳聚糖 - 明胶支架在各种组织工程应用中备受关注。然而，除了支架的降解和力学特性外，潜在的细胞 - 基质相互作用仍不清楚。在本研究中，我们评估了：（i）在溶菌酶浓度为10mg/L的条件下，壳聚糖和壳聚糖 - 明胶支架在两个月期间的降解动力学，包括尺寸稳定性、重量损失和pH值变化；（ii）在37℃湿润状态下薄膜和支架的拉伸和压缩性能；（iii）成纤维细胞和人脐静脉内皮细胞（HUVECs）在支架上的存活率；（iv）在静态以及4.5、8.5、13和18 dyn/cm²剪切应力条件下，HUVECs的细胞铺展特性、细胞骨架肌动蛋白分布、粘着斑激酶（FAK）分布和PECAM - 1表达的变化。降解结果表明，含明胶的壳聚糖支架比壳聚糖降解速度更快，材料损失更显著。明胶的加入对壳聚糖的力学性能有影响，尽管没有明显的趋势。三维壳聚糖和壳聚糖 - 明胶支架对成纤维细胞的存活率支持程度相同。然而，壳聚糖膜减少了HUVECs的细胞铺展面积，破坏了F - 肌动蛋白，并使FAK在细胞核内定位。重要的是，在壳聚糖上进行3小时最低测试剪切应力（4.5 dyn/cm²）会冲走细胞，表明细胞粘附性较弱。在混合物中，明胶的作用占主导；在静态培养中，肌动蛋白和FAK的分布与明胶相当。然而，在较高的剪切应力下，壳聚糖的存在抑制了剪切诱导的细胞铺展增加，并削弱了细胞的粘附强度。PECAM - 1表达未观察到显著差异。总之，这些结果表明明胶与壳聚糖混合对支架性能和细胞行为有显著影响。（c）2005 Elsevier Ltd.保留所有权利。
Recently, chitosan-gelatin scaffolds have gained much attention in various tissue engineering applications. However, the underlying cell-matrix interactions remain unclear in addition to the scaffold degradation and mechanical characteristics. In this study, we evaluated (i) the degradation kinetics of chitosan and chitosan-gelatin scaffolds in the presence of 10 mg/L of lysozyme for dimensional stability, weight loss, and pH changes for a period of 2 months, (ii) tensile and compressive properties of films and scaffolds in wet state at 37 degrees C, (iii) viability of fibroblasts and human umbilical vein endothelial cells (HUVECs) on scaffolds, and (iv) the alteration in cell spreading characteristics, cytoskeletal actin distribution, focal adhesion kinase (FAK) distribution and PECAM-1 expression of HUVECs under static and 4.5, 8.5, 13 and 18 dyn/cm(2) shear stress conditions. Degradation results showed that gelatin-containing chitosan scaffolds had faster degradation rate and significant loss of material than chitosan. Mechanical properties of chitosan are affected by the addition of gelatin although there was no clear trend. Three-dimensional chitosan and chitosan-gelatin scaffolds supported fibroblast viability equally. However, chitosan membranes decreased cell-spreading area, disrupted F-actin and localized FAK in the nucleus of HUVECs. Importantly, the lowest shear stress tested (4.5 dyn/cm(2)) for 3 h washed away cells on chitosan suggesting weak cell adhesion. In the blends, effect of gelatin was dominant; actin and FAK distribution were comparable to gelatin in static culture. However, at higher shear stresses, presence of chitosan inhibited shear-induced increase in cell spreading and weakened cell adhesive strength. No significant differences were observed in PECAM-1 expression. In summary, these results showed significant influence of blending gelatin with chitosan on scaffold properties and cellular behavior. (c) 2005 Elsevier Ltd. All rights reserved.