Park Jeong Su, Moon Dalnim, Kim Jin-Seok, Lee Jin Seok
J Biomed Nanotechnol. 2016 Mar;12(3):575-80. doi: 10.1166/jbn.2016.2192.
Nanotopological cues are popular tools for in vivo investigation of the extracellular matrix (ECM) and cellular microenvironments. The ECM is composed of multiple components and generates a complex microenvironment. The development of accurate in vivo methods for the investigation of ECM are important for disease diagnosis and therapy, as well as for studies on cell behavior. Here, we fabricated anodized aluminum oxide (AAO) membranes using sulfuric and oxalic acid under controlled voltage and temperature. The membranes were designed to possess three different pore and interpore sizes, AAO-1, AAO-2, and AAO-3 membranes, respectively. These membranes were used as tools to investigate nanotopology-signal induced cell behavior. Cancerous cells, specifically, the OVCAR-8 cell-line, were cultured on porous AAO membranes and the effects of these membranes on cell shape, proliferation, and viability were studied. AAO-1 membranes bearing small sized pores were found to maintain the spreading shape of the cultured cells. Cells cultured on AAO-2 and AAO-3 membranes, bearing large pore-sized AAO membranes, changed shape from spreading to rounding. Furthermore, cellular area decreased when cells were cultured on all three AAO membranes that confirmed decreased levels of focal adhesion kinase (FAK). Additionally, OVCAR-8 cells exhibited increased proliferation on AAO membranes possessing various pore sizes, indicating the importance of the nanosurface structure in regulating cell behaviors, such as cell proliferation. Our results suggest that porous-AAO membranes induced nanosurface regulated cell behavior as focal adhesion altered the intracellular organization of the cytoskeleton. Our results may find potential applications as tools in in vivo cancer research studies.
纳米拓扑线索是用于体内细胞外基质(ECM)和细胞微环境研究的常用工具。细胞外基质由多种成分组成,并产生复杂的微环境。开发用于研究细胞外基质的准确体内方法对于疾病诊断和治疗以及细胞行为研究都很重要。在这里,我们在控制电压和温度的条件下,使用硫酸和草酸制备了阳极氧化铝(AAO)膜。这些膜被设计为分别具有三种不同的孔径和孔间距尺寸,即AAO-1、AAO-2和AAO-3膜。这些膜被用作研究纳米拓扑信号诱导细胞行为的工具。具体而言,癌细胞系OVCAR-8在多孔AAO膜上培养,并研究了这些膜对细胞形状、增殖和活力的影响。发现带有小尺寸孔的AAO-1膜能维持培养细胞的铺展形状。在带有大孔径AAO膜的AAO-2和AAO-3膜上培养的细胞,形状从铺展变为圆形。此外,当细胞在所有三种AAO膜上培养时,细胞面积减小,这证实了粘着斑激酶(FAK)水平降低。此外,OVCAR-8细胞在具有各种孔径的AAO膜上增殖增加,表明纳米表面结构在调节细胞行为(如细胞增殖)中的重要性。我们的结果表明,多孔AAO膜诱导纳米表面调节细胞行为,因为粘着斑改变了细胞骨架的细胞内组织。我们的结果可能在体内癌症研究中作为工具找到潜在应用。