Li Na, Chen Gang, Liu Jue, Xia Yang, Chen Hanbang, Tang Hui, Zhang Feimin, Gu Ning
Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University , Nanjing 210029, China.
ACS Appl Mater Interfaces. 2014 Oct 8;6(19):17134-43. doi: 10.1021/am5047944. Epub 2014 Sep 25.
The effects of bioactive properties and surface topography of biomaterials on the adhesion and spreading properties of mouse preosteoblast MC3T3-E1 cells was investigated by preparation of different surfaces. Poly lactic-co-glycolic acid (PLGA) electrospun fibers (ES) were produced as a porous rough surface. In our study, coverslips were used as a substrate for the immobilization of 3,4-dihydroxyphenylalanine (DOPA) and collagen type I (COL I) in the preparation of bioactive surfaces. In addition, COL I was immobilized onto porous electrospun fibers surfaces (E-COL) to investigate the combined effects of bioactive molecules and topography. Untreated coverslips were used as controls. Early adhesion and growth behavior of MC3T3-E1 cells cultured on the different surfaces were studied at 6, 12, and 24 h. Evaluation of cell adhesion and morphological changes showed that the all the surfaces were favorable for promoting the adhesion and spreading of cells. CCK-8 assays and flow cytometry revealed that both topography and bioactive properties were favorable for cell growth. Analysis of β1, α1, α2, α5, α10 and α11 integrin expression levels by immunofluorescence, real-time RT-PCR, and Western blot and indicated that surface topography plays an important role in the early stage of cell adhesion. However, the influence of topography and bioactive properties of surfaces on integrins is variable. Compared with any of the topographic or bioactive properties in isolation, the combined effect of both types of properties provided an advantage for the growth and spreading of MC3T3-E1 cells. This study provides a new insight into the functions and effects of topographic and bioactive modifications of surfaces at the interface between cells and biomaterials for tissue engineering.
通过制备不同表面，研究了生物材料的生物活性特性和表面形貌对小鼠前成骨细胞MC3T3-E1细胞黏附与铺展特性的影响。制备了具有多孔粗糙表面的聚乳酸-乙醇酸共聚物（PLGA）电纺纤维（ES）。在我们的研究中，在制备生物活性表面时，使用盖玻片作为固定3,4-二羟基苯丙氨酸（DOPA）和I型胶原蛋白（COL I）的底物。此外，将COL I固定在多孔电纺纤维表面（E-COL）上，以研究生物活性分子和形貌的联合作用。未处理的盖玻片用作对照。在6、12和24小时研究了在不同表面上培养的MC3T3-E1细胞的早期黏附与生长行为。细胞黏附及形态变化评估表明，所有表面均有利于促进细胞的黏附与铺展。CCK-8检测和流式细胞术显示，形貌和生物活性特性均有利于细胞生长。通过免疫荧光、实时RT-PCR和蛋白质印迹分析β1、α1、α2、α5、α10和α11整合素的表达水平，结果表明表面形貌在细胞黏附早期起着重要作用。然而，表面形貌和生物活性特性对整合素的影响是可变的。与单独的任何一种形貌或生物活性特性相比，两种特性的联合作用为MC3T3-E1细胞的生长和铺展提供了优势。本研究为组织工程中细胞与生物材料界面处表面形貌和生物活性修饰的功能及作用提供了新的见解。
