Mechanical stress and patterned surface of the scaffolds has been recognized as a crucial factor in determining cell func- tionality and tissue development, which in turn can direct the cell responses. In this study, fibroblasts M-3T3 in three-dimensional （3D） honeycomb patterning Chitosan/Poly（L-Lactic Acid） （CS/PLLA） composites was stimulated by a 15% sinusoidal （1 Hz） strain applied by a biodynamic test instrument. The effects of mechanical stimulus on the cell proliferation and basic Fibroblast Growth Factor （bFGF） secretion were studied in comparison to the non-strain groups and blank control. Results show that fibroblasts are able to sense the mechanical stimulation and respond, resulting in a time dependent increase of bFGF secretion and promoting cell proliferation. Moreover, the cells seeded in the scaffolds showed a higher cell proliferation and bFGF secretion. These findings support the hypothesis that suitable mechanical stimulus has positive effect on fibroblasts, and such a 3D honeycomb patterned scaffold may play a positive role in regulating cell behaviors in vitro.
通过对壳聚糖(CS)进行改性,成功制备了壳聚糖季铵盐(HACC)。经过FT-IR、1 H NMR表征,证实了其分子结构。采用在仿生溶液中添加CS或HACC的方法,在微弧氧化前后的镁合金样品表面修饰得到4种不同的仿生涂层。利用SEM和XRD对涂层分析表明,仿生矿化后的镁合金表面为含有CaP/CS或CaP/HACC的复合涂层。降解实验表明,微弧氧化后仿生矿化的样品失重率小于微弧氧化前仿生矿化的样品;利用ICP测定释放的镁、铝离子浓度,发现镁离子浓度先升高后降低,铝离子浓度越来越小。抑菌实验结果表明,含有CaP/HACC比含有CaP/CS的复合涂层样品对金黄色葡萄球菌具有更明显的抑菌作用;对于仿生溶液中添加同样浓度的CS或HACC,微弧氧化后比微弧氧化前矿化的样品抑菌率高。比浊法测定显示,4种涂层在4d内均具有不同程度的抑菌效果,微弧氧化后的镁合金样品在添加有HACC的仿生溶液中矿化得到的涂层抑菌率最高。研究说明,改性后的HACC比CS具有更强的抑菌作用;微弧氧化后的样品具有更强的抗腐蚀能力和载药性能。
We prepared a cholesterol-conjugated chitosan（CHCS） material and evaluated its potential application as a bone tissue repair material by in vitro cell experiments. Cell proliferation, differentiation and morphology on CHCS membrane surfaces with different graft degrees were assessed in mouse pre-osteoblasts MC3T3-E1 cells. The results indicate that CHCS materials could promote the proliferation of MC3T3-E1 cells at low graft degrees, but the CHCS material with high graft degree inhibits the proliferation of cells in contrast to the pure chitosan membrane. However, the alkaline phosphatase（ALP） activity of MC3T3-E1 ceils on different CHCS membrane surface increased with in- creasing graft degrees of cholesterol. The area of cells stretched onto the surface of CHCS materials was larger than on the surface of CS materials, and more microfilaments and stress fibers in cells were observed on CHCS materials than on the pure chitosan material surface. After 7 d, the expression of related osteogenic marker genes, such as rum-related transcription factor 2（Runx2）, osterix（OSX）, osteocalcin（OCN）, osteopontin（OPN）, ALP and collagen I（COL-I） were all up-regulated in CHCS materials to different degrees compared to pure chitosan material, which in- dicated that the CHCS materials facilitated MC3T3-EI cell differentiation and maturation, Characterizing CHCS materials is useful in designing and developing strategies for bone tissue engineering.
LI ZhenzhenWEN JianhuaJIA WeijianDING ShanXIA XiaohuiZHOU ChangrenHUANG Yadong
