新型自组装水凝胶NBD/RADA16可促进前成骨细胞的分化

背景：RADA16是较成熟的自组装纳米短肽材料，在亲水面往复形成互补离子键，可组装为纳米纤维，并且能够促MC3T3 E1细胞的黏附、伸展和增殖。目的：观察新型自组装多肽水凝胶NBD/RADA16对小鼠前成骨细胞MC3T3 E1成骨分化能力的影响。方法：将MC3T3 E1细胞分别接种于自组装多肽水凝胶NBD/RADA16与RADA16水凝胶中，进行成骨诱导培养，以单纯成骨诱导培养的细胞为对照。诱导培养1，3，6 d检测细胞碱性磷酸酶活性；诱导培养7 d后， Western Blot检测细胞骨形态发生蛋白2的表达；诱导培养21 d后，茜素红染色观察细胞钙化结节。结果与结论：MC3T3 E1细胞在NBD/RADA16多肽水凝胶上生长状态良好，优于在RADA16上生长的细胞。自组装多肽水凝胶NBD/RADA16上MC3T3 E1细胞的碱性磷酸酶活性高于RADA16水凝胶上及单纯成骨诱导培养的细胞（P〈0.01）。自组装多肽水凝胶NBD/RADA16上MC3T3 E1细胞的矿化基质沉积、骨形态发生蛋白2表达高于RADA16水凝胶上的细胞（P〈0.01）。结果提示NBD/RADA16自组装多肽水凝胶较RADA16水凝胶更能促进MC3T3 E1细胞的成骨分化。

Preosteoblastic differentiation of MC3T3 E1 cells on the new-type self-assembling peptide hydrogel NBD/RADA16

BACKGROUND：RADA16 is a mature amphiphilic self-assembling peptide, which can be assembled into nanofibers, and promote MC3T3 E1 cellattachment, spreading and proliferation.
OBJECTIVE：To observe the effects of the new-type self-assembling peptide hydrogel NBD/RADA16 on osteogenic differentiation of mouse preosteoblasts MC3T3 E1.
METHODS：The MC3T3 E1 cells were inoculated in NBD/RADA16 self-assembling peptide hydrogel and RADA16 hydrogel for osteogenic induction. cells undergoing simple osteogenic induction served as controls. After culture for 1, 3, 6 days, the activity of alkaline phosphatase was detected. After 7 days, western blot assay was used to determine the expression of bone morphogenetic protein-2. After 21 days, alizarin red staining was used to observe calcified nodules.
RESULTS AND CONCLUSION：MC3T3 E1 cells grew wel on the NBD/RADA16 peptide hydrogel, which were superior to those on the RADA16 hydrogel. The activity of alkaline phoshpatase was higher in the NBD/RADA16 group than the RADA16 and control groups （P〈0.01）. Compared with the RADA16 hydrogel, mineralized matrix deposition and expression of bone morphogenetic protein-2 were higher on the NBD/RADA16 peptide hydrogel （P〈0.01）. These findings indicate that the NBD/RADA16 peptide hydrogel is superior to the RADA16 hydrogel for promoting the osteogenic differentiation of MC3T3 E1 cells.