No abstract

Tungsten chemical-mechanical polished integrated circuits were used to study the alignment and immobilization of mammalian (Vero) cells. These devices consist of blanket silicon oxide thin films embedded with micro- and nano-meter scale tungsten metal line structures on the surface. The final surfaces are extremely flat and smooth across the entire substrate, with a roughness in the order of nanometers. Vero cells were deposited on the surface and allowed to adhere. Microscopy examinations revealed that cells have a strong preference to adhere to tungsten over silicon oxide surfaces with up to 99% of cells adhering to the tungsten portion of the surface. Cells self-aligned and elongated into long threads to maximize contact with isolated tungsten lines as thin as 180 nm. The orientation of the Vero cells showed sensitivity to the tungsten line geometric parameters, such as line width and spacing. Up to 93% of cells on 10 μm wide comb structures were aligned within ± 20° of the metal line axis. In contrast, only ~22% of cells incubated on 0.18 μm comb patterned tungsten lines were oriented within the same angular interval. This phenomenon is explained using a simple model describing cellular geometry as a function of pattern width and spacing, which showed that cells will rearrange their morphology to maximize their contact to the embedded tungsten. Finally, it was discovered that the materials could be reused after cleaning the surfaces, while maintaining cell alignment capability.

The primary goal of this work was to investigate the resulting morphology of a mammalian cell deposited on three-dimensional nanocomposites constructed of tantalum and silicon oxide. Vero cells were used as a model. The nanocomposite materials contained comb structures with equal-width trenches and lines. High-resolution scanning electron microscopy and fluorescence microscopy were used to image the alignment and elongation of cells. Cells were sensitive to the trench widths, and their observed behavior could be separated into three different regimes corresponding to different spreading mechanism. Cells on fine structures (trench widths of 0.21 to 0.5 μm) formed bridges across trench openings. On larger trenches (from 1 to 10 μm), cells formed a conformal layer matching the surface topographical features. When the trenches were larger than 10 μm, the majority of cells spread like those on blanket tantalum films; however, a significant proportion adhered to the trench sidewalls or bottom corner junctions. Pseudopodia extending from the bulk of the cell were readily observed in this work and a minimum effective diameter of ~50 nm was determined for stable adhesion to a tantalum surface. This sized structure is consistent with the ability of pseudopodia to accommodate ~4–6 integrin molecules.

Tantalum is one of the most important biomaterials used for surgical implant devices. However, little knowledge exists about how nanoscale-textured tantalum surfaces affect cell morphology. Mammalian (Vero) cell morphology on tantalum-coated comb structures was studied using high-resolution scanning electron microscopy and fluorescence microscopy. These structures contained parallel lines and trenches with equal widths in the range of 0.18 to 100 μm. Results showed that as much as 77% of adherent cell nuclei oriented within 10° of the line axes when deposited on comb structures with widths smaller than 10 μm. However, less than 20% of cells exhibited the same alignment performance on blanket tantalum films or structures with line widths larger than 50 μm. Two types of line-width-dependent cell morphology were observed. When line widths were smaller than 0.5 μm, nanometer-scale pseudopodia bridged across trench gaps without contacting the bottom surfaces. In contrast, pseudopodia structures covered the entire trench sidewalls and the trench bottom surfaces of comb structures with line-widths larger than 0.5 μm. Furthermore, results showed that when a single cell simultaneously adhered to multiple surface structures, the portion of the cell contacting each surface reflected the type of morphology observed for cells individually contacting the surfaces.

Nano-and micro-scale engineered surface structures are often used to control cell morphology and mimic the extracellular matrix in tissue engineering. However, there is little understanding of how toxins produced by common bacteria might affect cell adhesion to these structures. In this study, human dermal fibroblast (GM5565) cells were incubated on patterned tungsten/silicon oxide nanocomposite in media in the presence or absence of Antimycin A. This composite consists of parallel tungsten and silicon oxide lines with identical widths in the range of 0.18 and 50 lm. The morphology of the cells and of their mitochondria was characterized by using high-resolution scanning electron microscopy and fluorescence confocal microscopy. Results show that cells preferentially align along the line axes in a patterndependent manner, with a maximum population of cells oriented within 10 of the line axes on the structures containing 10 lm wide lines. Cells treated with Antimycin A, however, show a smaller proportion of cells oriented in this direction as compared to cells cultured in Antimycin A-free media (34.4% vs 53.0%). The majority of mitochondria in cells growing in Antimycin A-free media are tubular in shape and are preferentially positioned on the tungsten lines, whereas these organelles exhibit a circular geometry and are less attracted to the metal lines in the presence of Antimycin A.