Kino Rikako, Ikoma Toshiyuki, Yunoki Shunji, Nagai Nobuhiro, Tanaka Junzo, Asakura Tetsuo, Munekata Masanobu
Faculty and Graduate School of Engineering, Hokkaido University, Kita 13, Nishi 9, Kita-ku, Sapporo, Hokkaido 060-8628, Japan.
J Biosci Bioeng. 2007 Jun;103(6):514-20. doi: 10.1263/jbb.103.514.
We prepared multilayered films consisting of silk fibroin (SF) and hydroxyapatite (HAp) by alternating lamination using untreated SF and HAp-deposited SF films. Untreated SF films were prepared from a regenerated SF solution by air drying. HAp-deposited SF films were prepared by soaking methanol-treated SF films containing >5 wt% CaCl2 in a simulated body fluid with the ion concentration 1.5-fold higher than that of the standard one. The multilayered HAp/SF films had HAp layers with approximate thicknesses of 3-5 microm and SF layers with thicknesses of 40-70 microm. The bonding strength between the SF and HAp layers was significantly affected by temperature and compression time under the lamination method. The optimal conditions for achieving the maximum T-peel strength and beta-sheet contents were determined to be 130 degrees C for 4 min. The Young's modulus of the multilayered films (133.4 MPa) was higher than that of the films consisting of SF alone (92.5 MPa) under swollen conditions. The biocompatibility of the HAp-deposited SF films was analyzed by culturing of osteoblasts (MC3T3-E1) on a film. The results indicate that HAp-deposited SF films and SF films show similar degrees of cell adhesion and alkaline phosphatase activities.
我们通过交替层压使用未处理的丝素蛋白（SF）和羟基磷灰石（HAp）沉积的SF膜制备了由丝素蛋白（SF）和羟基磷灰石（HAp）组成的多层膜。未处理的SF膜由再生的SF溶液通过空气干燥制备。HAp沉积的SF膜通过将含有>5 wt% CaCl2的甲醇处理的SF膜浸泡在离子浓度比标准溶液高1.5倍的模拟体液中制备。多层HAp/SF膜具有厚度约为3-5微米的HAp层和厚度为40-70微米的SF层。在层压方法下，SF层和HAp层之间的结合强度受温度和压缩时间的显著影响。实现最大T型剥离强度和β-折叠含量的最佳条件确定为130℃保持4分钟。在肿胀条件下，多层膜的杨氏模量（133.4 MPa）高于仅由SF组成的膜（92.5 MPa）。通过在膜上培养成骨细胞（MC3T3-E1）分析了HAp沉积的SF膜的生物相容性。结果表明，HAp沉积的SF膜和SF膜显示出相似程度的细胞粘附和碱性磷酸酶活性。
