Graduate Program in Health Science, School of Medicine, Pontifícia Universidade Católica do Paraná, Curitiba, Paraná, Brazil.
Department of Mechanical Engineering, Colorado State University, Fort Collins, CO, USA.
Mater Sci Eng C Mater Biol Appl. 2019 Oct;103:109850. doi: 10.1016/j.msec.2019.109850. Epub 2019 Jun 3.
The aim of this work was to evaluate the cellular response to titanium nanotube arrays with variable crystalline structure. Cytotoxicity, viability and the ability of the titania nanotube arrays to stimulate adhesion and proliferation of adipose derived stem cells (ADSCs) was evaluated. Titania nanotube arrays were fabricated by electrochemical anodization of titanium in diethyleneglycol/hydrofluoric acid electrolyte at 60 V for 6 h, then annealed at 300, 530 and 630 °C for 5 h. The nanotube arrays were characterized using scanning electron microscopy (SEM), contact angle goniometry, x-ray diffraction (XRD) and protein adsorption. ADSCs were cultured on titania nanotube arrays at a density of 1 × 10 cells/ml. The cells were allowed to adhere and to proliferate for 1, 4 and 7 days. Cell viability was characterized by the CellTiter-Blue® Cell Viability Assay; and cell morphology was characterized by SEM. Cell adhesion, proliferation and morphology were characterized using fluorescence microscopy by staining the cells with DAPI and rhodamine/phalloidin. The results from this study showed that the annealing at 300 and 530 °C formed anatase phase, and annealing at 630 °C formed anatase/rutile phase. The results indicated that the modification of the crystalline structure (i.e. anatase/rutile phase) of titania nanotube arrays influenced the ADSC adhesion and proliferation. Future studies are now directed towards evaluating differentiation of this cellular model in osteoblasts.
本工作旨在评估具有不同结晶结构的钛纳米管阵列的细胞反应。评估了钛纳米管阵列的细胞毒性、存活率以及刺激脂肪来源干细胞 (ADSCs) 黏附和增殖的能力。钛纳米管阵列通过在二甘醇/氢氟酸电解质中以 60V 电化学阳极氧化钛 6 小时，然后在 300、530 和 630°C 下退火 5 小时来制备。使用扫描电子显微镜 (SEM)、接触角测角法、X 射线衍射 (XRD) 和蛋白质吸附对纳米管阵列进行了表征。将 ADSCs 以 1×10 个细胞/ml 的密度培养在钛纳米管阵列上。使细胞附着并增殖 1、4 和 7 天。通过 CellTiter-Blue®细胞活力测定法来表征细胞活力；并通过 SEM 来表征细胞形态。通过用 DAPI 和罗丹明/鬼笔环肽对细胞进行染色，通过荧光显微镜来表征细胞黏附、增殖和形态。本研究的结果表明，在 300 和 530°C 下退火形成锐钛矿相，在 630°C 下退火形成锐钛矿/金红石相。结果表明，钛纳米管阵列结晶结构（即锐钛矿/金红石相）的修饰影响 ADSC 的黏附和增殖。目前的研究方向是评估该细胞模型在成骨细胞中的分化。
