Application of cross-linked salmon atelocollagen to the scaffold of human periodontal ligament cells
Application of cross-linked salmon atelocollagen to the scaffold of human periodontal ligament cells
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DOI：
10.1016/s1389-1723(04)70224-8
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发表时间：
2004-06-01
影响因子：
2.8
通讯作者：
Munekata, M
中科院分区：
文献类型：
--
作者：
Nagai, N;Yunoki, S;Munekata, M
本研究的目的是研究鲑鱼去端胶原（SAC）的应用，以支架。SAC具有低变性温度，并且在用作支架之前需要交联。本文采用1-乙基-3-（3-二甲氨基丙基）碳二亚胺盐酸盐（EDC）或去水热处理（DHT）对SAC进行交联。评价了由EDC（EDC-SAC）和DHT（DHT-SAC）交联的SAC支架的材料性质（交联度和在磷酸盐缓冲盐水中的溶解度）。结果表明，EDC-SAC与DT-SAC相比，具有较高的交联度和较高的稳定性。体外培养人牙周膜（HPDL）细胞2周，比较EDC-SAC和DHT-SAC与EDC交联的牛去端胶原（BAC）（EDC-BAC）和DHT交联的牛去端胶原（DHT-BAC）中HPDL细胞的增殖率和碱性磷酸酶（ALP）活性。EDC-SAC培养的HPDL细胞增殖率与EDC-BAC相当，且EDC-SAC中ALP活性显著高于EDC-BAC。在DHT交联中，DT-SAC的细胞增殖率和ALP活性均低于DT-BAC。DHT似乎提供了不足的交联，并且发现DHT-SAC是可断裂的和可收缩的，导致细胞活性降低。相反，EDC-SAC和EDC-BAC之间的热稳定性、多孔结构和细胞增殖率没有差异。此外，EDC-SAC的胶原螺旋被发现部分变性，这种结构导致HPDL细胞的ALP活性比EDC-BAC的增强。总之，我们的结果表明EDC-SAC可以用作体外培养的支架。
The purpose of this study was to investigate the application of salmon atelocollagen (SAC) to a scaffold. SAC has a low denaturation temperature and needs to be cross-linked before being used as a scaffold. In the present study, SAC was cross-linked by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) or dehydrothermal treatment (DHT). The material properties (degree of cross-linking and solubility in phosphate-buffered saline) of the SAC scaffolds crosslinked by EDC (EDC-SAC) and DHT (DHT-SAC) were evaluated. It was found that EDC-SAC had a high degree of cross-linking and high stability compared with DHT-SAC. Human periodontal ligament (HPDL) cells were cultured in the scaffolds for 2 weeks in vitro, and the activities (proliferation rate and alkaline phosphatase [ALP] activity) of HPDL cells cultured in EDC-SAC and DHT-SAC were compared with those cultured in bovine atelocollagen (BAC) scaffolds crosslinked by EDC (EDC-BAC) and DHT (DHT-BAC), respectively. The proliferation rate of HPDL cells cultured in EDC-SAC was equivalent to that in EDC-BAC, and the ALP activity in EDC-SAC was found to be significantly higher than that in EDC-BAC. In the cross-linking by DHT, the cell proliferation rate and the ALP activity in DHT-SAC were lower than those in DHT-BAC. DHT seemed to provide insufficient cross-linking, and DHT-SAC was found to be breakable and contractile, resulting in less cell activity. In contrast, there was no difference in the thermal stability, porous structure, and cell proliferation rate between EDC-SAC and EDC-BAC. In addition, the collagen helix of EDC-SAC was found to be partially denatured, and this structure resulted in the enhancement of ALP activity of HPDL cells compared with that using EDC-BAC. In conclusion, our results indicate that EDC-SAC could be used as a scaffold for in vitro culture.