Lee Haram, Lim Sooseok, Birajdar Mallinath S, Lee Soo-Hong, Park Hansoo
School of Integrative Engineering, Chung-Ang University, Republic of Korea.
Department of Biomedical Science, College of Life Science, CHA University, Republic of Korea.
Int J Biol Macromol. 2016 Dec;93(Pt B):1559-1566. doi: 10.1016/j.ijbiomac.2016.07.041. Epub 2016 Jul 15.
Conjugated gelatin nanofibers were fabricated by electrospinning, followed by a simple glutaraldehyde cross-linking procedure and avidin conjugation. Then, biotinylated growth factors were immobilized onto the surface of the fibers through avidin-biotin covalent binding. The immobilization of growth factors was confirmed through immunostaining using fluorescence microscopy and microplate spectrophotometry. Adipose derived stem cells (ASCs) were cultured to examine the effect of immobilized growth factors on cell proliferation using the cell counting Kit-8 (CCK-8) assay. Gelatin nanofibers with no growth factors attached and growth factors in suspension within media were used as controls. Growth factors were successfully immobilized onto the surface, in amounts corresponding to the concentrations applied, and increased cell proliferation to a higher extend than growth factors in suspension. Our results suggest that this controllable scaffolding strategy provides an effective system for growth factors delivery in tissues, suitable for engineering applications.
通过静电纺丝制备共轭明胶纳米纤维，随后进行简单的戊二醛交联程序和抗生物素蛋白共轭。然后，通过抗生物素蛋白-生物素共价结合将生物素化生长因子固定在纤维表面。使用荧光显微镜和酶标仪通过免疫染色确认生长因子的固定化。培养脂肪干细胞（ASC），使用细胞计数试剂盒-8（CCK-8）测定法检查固定化生长因子对细胞增殖的影响。未附着生长因子的明胶纳米纤维和培养基中悬浮的生长因子用作对照。生长因子成功固定在表面，其数量与应用浓度相对应，并且比悬浮的生长因子更能促进细胞增殖。我们的结果表明，这种可控的支架策略为组织中的生长因子递送提供了一个有效的系统，适用于工程应用。
